CN1791431A - Micro-cluster compositions - Google Patents

Abstract

Micro-clustered liquids, methods of manufacture and use. Culture media and cultures comprising micro-clustered water; use of micro-clustered culture media and cultures for cell, tissue and organ maintenance and growth; use in microbial biotechnology. Micro-clustered water compositions of bio-affecting agents, body-treating agents, and adjuvants or carriers, pharmaceutical and diagnostic compositions thereof. Methods of using the compositions involving administering them ex vivo to cells, tissues or organs, or in vivo to living bodies; and methods of making the compositions. Methods of hydrating foods and food ingredients in food processing systems with micro-clustered water. Edible foods, ingredients, flavoring and sweetening compositions containing micro-clustered water.

Description

Micro-cluster compositions

Invention field

Generally speaking, the present invention relates to micelle grain (micro-cluster) liquid and manufacturing and the method for using them.The invention provides program and the using method thereof of making micelle grain liquid.

The application content

I. micelle grain liquid and manufacturing and the method for using them

II. culture medium and manufacturing and the method for using culture medium

III. medicine, biotic influence (bio-affecting) compositions and health are handled (body-treating) compositions

IV. food or Edible material and beverage; Method, compositions and product

I. micelle grain liquid and manufacturing and the method for using them

Background of invention

Water is by single H ₂The O molecular composition, they mutually combine by hydrogen and form the granule (cluster) with following five category features: disjunct molecule, in a similar tetrahedral arrangement by five (5) individual H ₂The molecule of the molecular four sides of O hydrogen bonded, and the molecule that links to each other by the surface that 1,2 or 3 hydrogen bonded is connected to granule, (United States Patent (USP) 5,711,950Lorenzen; Lee H.).These can form by the molecular big array of varying number micelle grain, to form array by one or more this power by faint remote Van der Waals (Van der Waals) gravitation; (1) dipole-dipole interaction promptly has the electrostatic attraction between two molecules of permanent dipole moment; (2) dipole induction dipolar interaction, promptly the dipole of a molecule makes adjacent molecular polarization; And (3) are by the dispersion force of instantaneous dipole generation faint in the atom.Under household condition, tetrahedron micelle grain less stable, the vibration that overcomes Van der Waals repulsive force by generation London Force (London forces) can form big array again.When these molecules period of the day from 11 p.m. to 1 a.m of close another minute,, thereby cause the single peplos of intratomic parton trade shape to be out of shape because the motion of relative position and two hydrones can produce dispersion force.Each molecule stops and causes ever-increasing counteracting force by continuous modification, until reaching the points of proximity that London induction force (London Inductive Force) comes into force.If the speed of these molecules is big to enough making their mutual close distances equal Van derWaals radius, then can be combined into hydrone.

At present, we need the processing method of a kind of advantageously fractionated (fractionate) liquid macromole array.And, also need to can be used for to consume, the more small water molecule (for example, micelle grain) of medicine and chemical field processing.

Summary of the invention

Inventors find, the liquid (for example, water) of the macromole array that forms by various static and van der Waal power can be cracked into fractionated or micelle grain molecule (for example, theoretical tetrahedron micelle grain water) by cavitation.Inventors have further found to utilize Vander Waals repulsive force to stablize the method for newly-built micelle grain water.This method is that the micelle grain is water-cooled to required density, and wherein micelle grain water can be oxidized then.When still cooling off, micelle grain water is bottled.In addition, when micelle grain oxidize water density big (being under the state of cooling), fill bottle and add a cover, by reducing the vibration that in the bottle that part is filled, may occur and dissolved gases is (for example in solution simultaneously, oxygen) provide dividing potential drop, London forces can be reduced, thereby when depositing under 40 to 70 the temperature conditions, the micelle grain can be stablized about 6 to 9 months.

The invention provides a kind of method for preparing micelle grain liquid (as water), comprise make the liquid cavitation so that in this liquid the dissolved gas of carrying secretly form a large amount of cavitation bubbles; And make the liquid that comprises a large amount of cavitation bubbles stand the decompression, in this process pressure reduce can cause big fluid molecule matrix to be broken into less fluid molecule matrix.In another embodiment, liquid is substantially free of mineral, can be the water that also is substantially free of mineral.This embodiment provides multiple processing method, reaches about 140 °F (about 60 ℃) until water.Cavitation can provide like this: liquid is applied first pressure, and fast decompression to the second pressure is to form cavitation bubble then.Can use pump to pressurize.In one embodiment, first pressure is about 55psig to more than the 120psig.In another embodiment, second pressure is about atmospheric pressure.Embodiment can be carried out like this, breaks or explosion so that pressure change causes a large amount of cavitation bubbles.Can implement pressure change, with dissociate local atom and reformulate the plasma of atom with difference in conjunction with angle and intensity of formation.In another embodiment, liquid cools is to about 4 ℃ to 15 ℃.Other embodiment comprises in micelle grain liquid and infeeds gas that gas is oxygen as described.In further embodiment, oxygen supply time is about 5 to 15 minutes.In further embodiment, the invention provides the method for making micelle grain liquid, comprise liquid is applied enough big pressure; Discharge fluid under pressure to generate successive liquid stream; Make continuous flow form the rotating multi eddy current, it has partial vacuum pressure, so that dissolving and the gas carried secretly generate a large amount of cavitation bubbles in the liquid; To containing the liquid decompression of a large amount of cavitation bubbles, cavitation bubbles a large amount of in this process can break or explosion then, and this can cause makes big fluid molecule matrix form the shock wave of less fluid molecule matrix.In further embodiment, liquid is substantially free of mineral, and in additional embodiment, liquid is water, preferably is substantially free of mineral.The invention provides multiple processing method, reach about 140 °F (about 60 ℃) until water.In another embodiment, cavitation can provide like this: liquid is applied first pressure, and fast decompression to the second pressure is to form cavitation bubble then.In addition, the pressurization among the present invention can use pump to provide.In further embodiment, first pressure is about 55psig to more than the 120psig, and in another embodiment, second pressure is about atmospheric pressure, comprises that second pressure is lower than the embodiment of 5psig.Embodiment can be carried out like this, breaks or explosion so that pressure change causes a large amount of cavitation bubbles.The present invention also provides micelle grain liquid, and wherein pressure change causes that a large amount of cavitation bubbles break or explosion.In another embodiment, pressure change forms and to dissociate local atom and to reformulate the plasma of atom with difference in conjunction with angle and intensity.The present invention also provides a kind of method, wherein with extremely about 4 ℃ to 15 ℃ of liquid cools.In another embodiment, the invention provides in micelle grain liquid and infeed gas.Preferred described gas is that oxygen, particularly oxygen supply time are about 5 to 15 minutes, and more preferably pressure is about 15 to 20psig.

The present invention also provides the compositions that contains the micelle grain water of making by above-mentioned steps.

And, another aspect of the present invention is the micelle grain water with any or all of following attribute: conductivity is about 3.0 to 4.0 μ mhos/cm, at the FTIR microphotohram that about 2650 wave numbers have main obvious characteristic, press the definite steam pressure of thermogravimetric analysis between 40 ℃ to 70 ℃ ¹⁷O NMR peak shift with respect to reverse osmosis water at least about+30 hertz, preferably at least about+40 hertz.

The present invention also further provides the purposes of micelle grain water of the present invention, is used for such as regulating cell performance and the purpose that reduces cell free radical level by cell is contacted with micelle grain water.

In addition, the present invention also provides the drug-supplying system that contains micelle grain water (for example, oxidation micelle grain water) and activating agent (for example, nutrient, medicine etc.).

And micelle grain water of the present invention also can be used for by fabric is contacted the spot of removing on the fabric with micelle grain water.

Accompanying drawing and following description detailed one or more embodiments of the present invention.Further feature of the present invention, purpose and advantage will be by description and accompanying drawings and apparent by claims.

All publications cited herein, patent and patent application are all introduced especially with reference to being used for all purposes.

Brief Description Of Drawings

Fig. 1 shows the net dipole moment of hydrone and initiation.

Fig. 2 shows the big flood molecular array.

Fig. 3 shows the micelle grain water with 5 hydrones that forms a tetrahedron shape.

Fig. 4 shows that liquid forms the utility unit example of cavitation phenomenon.This device is equipped with the liquid inlet, makes liquid form the rotating multi eddy current then, reaches the partial vacuum pressure of about 27 inches of mercury.Liquid is expelled to the cavity (for example, about atmospheric pressure) that is lower than the device internal pressure by accelerating tube from the A point that installs.

Fig. 5 shows the FTIR spectrum of RO water (Fig. 5 (a)) and treated micelle grain water (Fig. 5 (b)).

Fig. 6 shows the TGA figure of the micelle grain water of RO water and oxidation.

Fig. 7 shows RO water (Fig. 7 (a)), unoxidized micelle grain water (Fig. 7 (b)) and through the NMR spectrum of the micelle grain water (Fig. 7 (c)) of oxidation.

Fig. 8 shows Raman (Raman) spectroscopy schematic representation of apparatus.

Fig. 9 shows the influence of micelle granulocyte culture medium to the macrophage plasma membrane.

Figure 10 shows the influence of micelle granulocyte culture medium to pH in the born of the same parents.

Figure 11 shows the influence of micelle granulocyte culture medium to the 293T cell viability.

Figure 12 a and 12b show that micelle grain water is to two types of human cells' the growth and the influence of transfection.

Figure 13 shows the influence of micelle grain water to dendritic cell sign express spectra.

Figure 14 shows the influence of micelle grain water to the functional status of the dabbling cerebral tissue of usefulness micelle granule medium.

You selects the embodiment explanation

Comprise that the liquid of for example alcohol, water, fuel and Zu compound thereof is by Yuan with complicated molecule structure and molecular composition. Many this arrangements can cause being formed on the big molecular array of covalent bond Yuan that has noncovalent interaction between adjacent molecule, and a little adjacent molecules of the Zhe successively noncovalent interaction by other other molecule of You reach and influence each other. Although the some big Zhen row of Zhe are more stable, its size of You Yu and can't with they ideally Yong Yu great majority use. Therefore, Zhe just need to create and provide the liquid of littler Zhen row by the quantity that reduces noncovalent interaction. The a little littler molecules of Zhe are Zai biochemical system inner penetration and being used as better. In addition, littler molecular array can provide required new features.

The chemical bond of shared electron between " covalent bond " expression Yuan that herein adopts. Between term " non-covalent bond " or " noncovalent interaction " expression Yuan without chemical bond or the interaction of common-battery. This type of noncovalent interaction comprises the interaction that ion (or electricity price) key (Zhuan from Yuan to one or more electronics of another Yuan move and form), You dipole moment, Hydrogenbond and Van der Waals power produce etc. Van der Waals power is between Zhi the Zuo Yong Yu nonpolar molecule or with the faint power between Zhi a part different piece, thereby the interim asymmetric electronic distribution of one Zu of Zhong of You Yu combines two Zus, and Zhe makes the opposite polarity of another Zu Zhong obtain induction. When the distance between Zhi the Zu during less than Van der Waals radius, its electron cloud of You Yu begins mutually to intert, and the power between Zhi them can be repelled mutually.

The method of herein mentioning is applicable to many liquid, and this liquid comprises water, alcohol, oil and fuel. The molecular composition that the one or more basic elements of liquid You such as water or Yuan (for example, hydrogen and oxygen) Zu become. The interaction of covalent bond and branch charge of the electron have formed molecule between Yuan. Hydrone has You angle or crooked figure. The angle of hydrone Zhong H-O-H key is approximately 104.5 ° to 105 °. Net dipole moment Zai Fig. 1 Zhong You explanation of hydrone. This dipole moment has formed the electrostatic force that Yun is permitted other hydrone gravitation. The people such as Pugliano, the nearlyest relation and the complicated interaction of having researched and proposed between hydrone of (Science, 257:1937,1992). Zhe studies a bit and has disclosed hydrogen bond and key effect that dioxygen interaction Zai sets up big hydrone granule Zhong. The pure water complex structure, the big Zhen row (as shown in Figure 2) that the interactional a plurality of hydrone Zu of the adjacent hydrone of You become. For example, the noncovalent interactions such as a little big Zhen row You hydrogen bond forms of Zhe form, thereby form the dipole moment molecule. Although quite stable, You studies show that a little big molecules of Zhe and are unfavorable for various chemistry and biological respinse. Therefore, embodiment Zhong of Zai the invention provides a kind of method that forms classification separation or micelle grain water, as described in Figure 3 Yue 5 hydrones of Zhi You.

The present invention is for providing the making method processed of little micelle grain liquid (for example, micelle grain hydrone), production classification Separation of Water or micelle grain water, and the various biological Zhuan conditions of Zai are processed the using method of Zhong.

Therefore, the invention provides the method for production classification separating liquid or micelle grain liquid (for example, water), comprise starting liq is forced into first pressure, reduction of blood pressure in high-speed Zhi the second pressure forms the partial vacuum pressure that causes discharging the gas of carrying secretly and form cavitation bubble then. The You cavitation bubble breaks and the hot physical reactions that provides of explosion can cause heat Zeng to add and keeps the big Zhen of liquid to be listed in together noncovalent interaction destroys Zhong and breaks. This processing method repeatedly Zhi is capable, and Zhi Zhi obtains the required physicochemical characteristic of classification separating liquid. If liquid is water, Ze processing method repeatedly Zhi is capable, and Zhi reaches Yue 140 °F (Yue 60 ℃) Zhi the temperature of water. The liquid Ying Zaike that the littler or classification that obtains separates prevents from cooling off under the Chong condition that newly the big Zhen of formation is listed as. Yong herein method Zhong, " water " or " Yuan expects water " comprises Zi Laishui, natural mineral water and through water such as the pure water of processing.

Zhi wants cavitation Yuan to be suitable for producing the enough energy of Zu big Zhen row are broken, and any known technology in this field of Ze all is used in liquid Zhong and forms cavitation phenomenon. The acoustic energy that cavitation produces provides energy for the big Zhen row of liquid are broken into littler liquid granule. For example, utilize sound transducer that required cavitation Yuan can be provided. In addition, make liquid pass through can cause cavitation with the tube of compression member, Zhe be because the Zai compression before can form high pressure, and Zai by compression member after the Zhi pressure can reduce fast. Another example is to make liquid pass through liquid pump with the form of opposite spin eddy current.

Embodiment Zhong of Zai, the liquid pressing of classification to separate becomes rotating vortex, can be formed on rotating vortex and reach the air release that will carry secretly when equaling or flow out conical nozzle near the size of atmospheric pressure and be the Xuan whirlpool of the partial vacuum pressure of cavitation bubble. This moment pressurization and decompression can make the cavitation bubble that forms the sound energy strikes ripple break and explosion. The a little shock waves of Zhe have interrupted covalent bond and non-covalent bond and the faint Zhen row key of the big Zhen row of liquid Zhong, and have formed You Yue five (5) the individual H that arrange with similar tetrahedron₂The molecular micelle grain of O or classification separating liquid (shown in Fig. 3), and distributed an electron charge for micelle grain liquid, thus liquid has had the electrolyte characteristic. Micelle grain liquid constantly circulates, Zhi Zhi forms the micelle grain fluid molecule of requirement, to reach the fixed surface tension of Zhi and electron charge, after the Zai fluid temperature was elevated to designated value with the passing of time, cavitation bubble can be emitted the power heat Zhong treated liquid. In case reach required surface tension and electron charge, micelle grain liquid just begins to cool down Zhi and adds Zhi fluid density Zeng. Required surface tension and electron charge can be measured in several ways, but the best be to detect by temperature. For example, in case liquid reaches required density, gas is general Yue to be 4 ℃ to 15 ℃, continues to pass into oxygen, and after the Zhi Zai time enough, micelle grain liquid Zhong can reach required amount of oxygen. Then, micelle grain liquor sample etc. is distributed into container or bottle Zhong, the You choosing is filled to the biggest solvent, adds a cover then, and this micelle grain liquid that adds simultaneously gas of Zai still cools off, so as along with temperature reach room temperature to the micelle grain liquid carrying of aerating for partial pressure. Partial pressure raises in the You Yu bottle, and Zhe makes a large amount of dissolved gases can be retained in solution Zhong.

The invention provides the method for Zao micelle grain processed or classification Separation of Water or liquid, for ease of explanation, described liquid represents Yong water, although the equal alternative water of the liquid of any type. For example, for Yuan material water, You Yu does not contain mineral matter comparatively speaking, so the You choosing is heated up in a steamer water Yong Zuo Yuan material with pure water or Zheng. Then, the food stage stainless steel tank that water is added Yong Yu processing. Yuan is expected that water adds can provide Yue 55 to 120psig or the liquid pump of above continuous, can form continuous current. Then, current are flowed into to form the suitable devices (referring to Fig. 4 example) of the rotating multi eddy current that reaches Yue 27 inch of mercury partial vacuum pressure, thereby reach the Zheng steam pressure that Zai water Zhong dissolves the gas of carrying secretly. The a little gases of Zhe form cavitation bubble, advance to equal or near the shared cavity of atmospheric pressure by row behind the multiple accelerating tube. The shock wave that break and the explosion of You cavitation bubble produces is by the iterative cycles of water, with the littler hydrone of big Zhen Col Break one-tenth of water. Water circulation constantly raises the temperature of water. The heat that the You cavitation bubble breaks and explosion produces is released to acoustooptical effect with energy, and the temperature of acoustooptical effect bubble estimates between Zhi 10 to 100 electron-volts of the Zai, or under 19,743,336 atmospheric pressures 2,042,033 °F. Yet the heat of generation is You submicron size only, and the very fast water that is enclosed by Zhou absorbs rapidly, emits kinetic force. Inventors have determined can utilize the Zheng string ripple of cavitation phenomenon to reach for the operation than small water molecule big Zhen Col Break, and the rising by monitoring temperature, osmotic pressure and the surface tension of can Zai processing procedure Zhong regulating water. Inventors have determined oxidation micelle grain water (Penta-hydrate^TM) ideal temperature Yue be 140 °F (Yue 60 ℃). Zhe can enter with 6 degree accelerating tubes by four strands of opposite eddy current of Yong and equal or realize that near atmospheric shared cavity back pressure should be lower than 5 pounds. As mentioned above, under the processing method that inventors also find to describe herein, the Zheng string fluctuation of heat/temperature can appear in liquid. According to required physicochemical characteristics, reprocessing method Zhi Zhi Zheng chord curve reaches required point, and when reaching this, Zai can suppress to collect and cooling liquid under the condition that big molecular array forms. For example (but being not limited to this), inventors find Zheng string heating process can occur by the water that this described method is processed. Can form high negative electrical charge Zhong the process of this water of Zai Zao processed and Zhuan moves to water. According to the water speed of operating pressure and generation, be that the voltage that 800 Zhou phases or above stack Zheng string ripple record is-350 millivolts to-1 volt by frequency. Inventors find that the 3rd Zheng chord curve peak value of temperature Zhong provides optimal values for the micelle kernel structure of water. Although having Ze to appoint, inventors do not provide the machine-processed or theoretical of operation, but it is believed that high-negative ion produces the good source that can serve as to electronics, Yong the Zuo antioxidant during consumption, also can prevent that Chong is new and form big Zhen row by the hydrone that is exposed to the negative electrostatic charge field being positioned stabilize water micelle grain and You Zhu Yu. Although be reluctant to be bound in specific theoretical Zhong, according to FTIR and NMR test data, inventors think that the high temperature that Zai cavitation phenomenon process Zhong reaches may can form the H that dissociates Zhong the Zai water₂O Yuan closes the new plasma that forms of Chong with different bonds then, thereby has generated different structures. Those of skill in the art will recognize that water of the present invention can adopt any mode to further improve. For example, after Zai forms micelle grain water, Click here it is described may be oxidized, according to Zui Zhong Yong way of water, can adopt known prior art to its further purify, seasoning, Zheng are heated up in a steamer, radiation treatment or other modification.

Another embodiment of Zai Zhong the invention provides the method for the cell performance of adjusting Zu Zhi or object. Micelle grain water (for example, oxidation micelle grain water) can be designed to induction system, to transmit aquation, oxidation, nutrition, medicine and raising general cell performance and exchange intracellular fluid body and to remove oedema. RJL Systems Bio-Electrical Impedance Analyzer model BIA101 Q Body Composition Analysis System is adopted in test^TMFinish, in the bright substantial cell of Zheng and the aquation of extracellular, only changing needs 5 minutes as a result. The male sex of the bodily form obesity of one of Zai of test 58 years old, 71.5 inches of heights, 269 pounds of body weight carries out with it. Bio-Electrical Impedance Analyzer^TMThe base-line data that reads is as follows.

Describe as following embodiment, estimate that micelle grain water of the present invention can produce wholesome effect through object consumption.Object can be any mammal (for example, horse, cattle, pig, Mus, cat, Canis animals), and is preferably human.Micelle grain water or oxidation micelle grain water (Penta-hydrate ^TM) dosage will depend on many factors of recognizing in the prior art of common modification and adjustment.This class factor comprises: age, body weight, activeness, syneresis, body fat etc.Generally speaking, 0.5 liter of oxidation micelle grain water of the present invention can provide useful result.According to expection, micelle grain water of the present invention can adopt any prior art known way to use, and comprises for example oral and intravenous in addition, mixes separately or with other activating agent, chemical compound and chemicals.Inventors estimate that also water of the present invention can be used for irrigating or the surgical incision position.Water of the present invention also can be used for treatment of infection, for example, can adopt micelle grain water (for example, oxidation micelle grain water) to carry out useful treatment by the microbial infection of anaerobism.

In another embodiment, micelle grain water of the present invention can be used for reducing the free radical level, thereby suppresses radical damage in cell.

In another embodiment, micelle grain water of the present invention can be used for removing the spot on the fabrics such as cotton.

Following examples are used for explanation but do not limit the present invention.One skilled in the art will realize that the equivalent of following examples, it comprises in the present invention.

Embodiment 1

How to make micelle grain water

Below be described as making an example of micelle grain liquid processes.One skilled in the art will realize that the alternative equivalent that the present invention includes.Therefore, following examples can not limit the present invention, but can be used as in order to understand demonstration methods of the present invention better.

With the distilled water of 325 gallons of Culligan Water brands or under 29 ℃ ambient temperature the pure water in 5 gallon bottles pour in the 316 rustless steel non-pressure water storage ponds that have a removable top cover, in order to handling.This tank links to each other with 20 inches U.S.'s filter seat of being furnished with 5 micrometer fibers filters by 21/4 inch 316 stainless steel pipe of bottom feed that can be contracted to 1 inch NPT, and this filter is used for removing the pollutant that anhydrate and may contain.The output channel of 20 inches filters is connected to Teel type 1V458316 stainless gear pump, and it is by direct drive 3HP1740 RPM3 phase Motor Drive.The output channel of 1 inch NPT of gear pump is connected to cavitation apparatus by 1 inch 316 stainless steel tube, and this stainless steel tube is equipped with one and is used for isolating and the manometric 1 inch stainless steel ball valve of pasta.It is the lasting pressure of 65psig that the output of liquid pump can produce size to cavitation apparatus.

Cavitation apparatus is not by forming with four small-sized reversing pump serpentine pipes of impeller that Teflon makes, be installed on the 316 rustless steel bases, provide common raw material water for the V458 gear pump of 65psig by tangential way by 1 pressure, flow into by the 1/4 inch aperture that is usually used in the liquid pump outlet, but be used herein to the input port that forms rotating vortex.The water that enters four serpentine pipes is discharged from the liquid pump suction inlet by the 1 inch accelerating tube that has 3/8 inch discharging opening with the form of 360 degree circles, and this discharging opening is generally the suction inlet of liquid pump serpentine pipe, but at this floss hole as this device.Four reverse feed auger pipes form water are revolved the eddy current of three-sixth turn, with the angle that is lower than centrage 5 degree water is discharged then, equal or during near atmospheric pressure accelerating tube water is entered common housing.This cavity is connected to 1 inch rustless steel discharge tube, but its feed back is to 325 gallons of tanks that fill distilled water.So far, water has passed through single treatment in device.

Above-mentioned steps can constantly repeat, and until the energy (for example, acoustic energy) that is broken by cavitation bubble and explosion produces its power heat is passed in the water, and the temperature of water reaches about 60 ℃.

Although inventors do not have obligation to explain the theory of invention, they provide following theory with interpretive mode, and are not subjected to this theory.Inventors believe that the acoustic energy that is produced by cavitation phenomenon has suppressed to keep five H in big array ₂The single tetrahedron micelle grain electrostatic bond together of O molecule, thus reduced its size also/or in water, produced local plasma, it is the different water of structure that the angle of common key is organized structure again.

Temperature is measured by the rustless steel thermometer boss by the hand-held infrared thermal detector.One skilled in the art will realize that other thermometry.In case temperature reaches 60 ℃, the liquid pump motor promptly can lock, and water begins to cool down.The heat insulation room that is equipped with the 8x8 foot of the hot air-conditioning equipment of 5,000 English is used for quickening cooling, but and nonessential.Make treating water keep static very important during cooling, try not to move.

Though chilling temperature can adopt 4 ℃, 15 ℃ enough, and according to the difference of wanting cooling water quantity, temperature is also different.In case fully be cooled to about 4 ℃ to 15 ℃, just can carry out oxidation to water.

After being water-cooled to required temperature, treating water is moved to 5 gallons of polycarbonate bottles that are used for oxidation from 325 gallons of rustless steel tanks.

Oxidation processes is that 1/4 inch ID plastic conduit by assembling plastics air diffuser charges into O under the pressure of 20psig ₂Gas, this air diffuser are used to produce tiny bubble (for example, the Lee catalog number 12522).Plastic tube passes the screw that 5 gallon bottles cover, the bottom of through bottle.Air diffuser is assemblied in the discharge of pipes end.Oxygen is depressed at the stream of 20psig and is charged into, and sees the oxygen burble to guarantee to know.At an embodiment (Penta-hydrate ^TM) in, the water Oxidation continues about five minutes, and at another embodiment (Penta-hydratePro ^TM) in, the water Oxidation continues about ten minutes.

After Oxidation finished, the 500ml PET bottle of immediately water being packed into was filled to overflowing the back and is used the press seal plastic cap seal bottle that has the insert seal packing ring.In one embodiment, bottled full with one 0.5 liter, thereby when the temperature of water rises to room temperature, it is supercharging voluntarily, makes that dissolved oxygen remains on bigger concentration under partial pressure.This step not only can keep more oxygen under dissolved state, but also can prevent water high vibration in the process of transporting.

Embodiment 2

Adopt the water of the method for the invention preparation novel unique, characterize with regard to multiple parameter.

A. conductivity

Conductivity records under the USP645 program, and this program has been specified the conductive measurement standard of the water with special nature.Except that having defined the test experimental program, USP645 has also stipulated the performance standard of conductivity measurement system, and the effectiveness of instrument and conductivity and alignment requirements.The conductivity test is by the West CoastAnalytical Service in Santa Fe Springs city, California, and Inc. carries out.

The conductivity result of the test

W/O ₂RO water micelle grain water micelle grain water

Conductivity in the time of 25 ℃ ^*

(μmhos/cm) 5.55 3.16 3.88

^*The conductivity value is the average of twice measurement.

With RO water ratio, the conductivity of observing micelle grain water reduces to be a bit larger tham half.This is quite important, and it shows that micelle grain glassware for drinking water has visibly different behavior, thereby substantially different with respect to undressed RO water.

B. fourier transform infrared spectrum (FTIR)

Water is the strong absorbent of IR spectral regions, fully characterizes through FTIR, shows to be consistent main spectral line with the vibration of O-H key when about 3000 wave numbers.This spectral line makes in the sample hydrogen bond structure distinctive.To be untreated RO sample water (sample A) and not oxidation micelle grain sample water (sample B) be placed on respectively in the chlorination silver plate, at 25 ℃ of films of analyzing every kind of liquid through FTIR.The FTIR test is by the West CoastAnalytical Service in Santa Fe Springs city, California, and Inc. adopts Nicolet Impact400D ^TMBenchtop FTIR carries out.FTIR spectrum as shown in Figure 5.

Behind the FTIR spectrum of more not oxidation micelle grain water and RO water, the result shows that clearly two kinds of sample water have multiple common trait, but notable difference is also arranged simultaneously.Micelle grain water is observed the main obvious characteristic (Fig. 5 (b)) of about 2650 wave numbers in FTIR spectrum.RO water does not have this feature (Fig. 5 (a)).This shows the behavior difference of key in the sample water, and its energy interaction changes.The result shows that inoxidized micelle grain water all is different from the RO untreated water on physical property and chemical property.

C. simulation distil

By the West Coast AnalyticalService in Santa Fe Springs city, California, Inc. carries out simulation distil to RO water with at inoxidized not oxidation micelle grain water.

The simulation distil test result

RO water is oxidation micelle grain water not

Boiling spread *

(℃) 98-100 93.2-100

* proofread and correct through air pressure.

The result shows that the boiling temperature of minimum boiling point fraction does not obviously reduce in the oxidation micelle grain sample water.Compare with the temperature of 98 ℃ of the minimum boiling point fractions of RO water, the minimum boiling point fraction of observing micelle grain water is 93.2 ℃.This shows that processing method significantly changes the composition structure of the molecular species that exists in the water that changes.Please note lessly usually than the low boiling kind, this conforms to the formation of all observed data with the micelle grain.

D. thermogravimetric analysis

In this test, a water is splashed into dsc sample dish, be drilled with the covering sealing of a pin hole in order to the precise laser mode.The sample coolant-temperature gage is every rising in 5 minutes 5 degree, until reaching final temperature.Oxidation micelle grain water and RO water do not carry out the TGA sketch plan simultaneously and describe, so that relatively.

The Analytical Products that TGA analyzes by La Canada city, California adopts TA Instruments Model TFA2950 ^TMCarry out.TGA result of the test such as Fig. 6 show.Three kinds of not three result of the tests of same water have been shown among the figure.RO sample water has been appointed as " pure water " on the TGA chart.Oxidation micelle grain water is not duplicate, comprises Super Pro 1 ^StTest and Super Pro 2 ^NdTest.The oxidation micelle grain water and the RO water that is untreated do not demonstrate significantly bigger weight saving kinetics phenomenon.Obviously, RO almost begins missing mass immediately, in about 40 ℃ of beginnings up to final temperature.Micelle grain water quality in the time of about 70 ℃ just begins to alleviate.This explanation and the RO water ratio that is untreated, the steam pressure of treated water between 40 ℃ to 70 ℃ is bigger.TGA result proves that the steam pressure of oxidation micelle grain water is not lower when reaching boiling temperature.These data show that again with RO water ratio, oxidation micelle grain water does not obviously change.These data show that once more oxidation micelle grain water does not also demonstrate more features between 75 ℃ to 100 ℃ temperature.These features can illustrate observed low boiler cut in simulation distil.

E. nuclear magnetic resonance, NMR (NMR) spectrum

The NMR test is by the Expert ChemicalAnalysis in San Diego, CA city, and Inc. adopts 600 megahertz Bruker AM500 ^TMInstrument is carried out.The NMR object of study is the micelle grain water and the RO water of aerobic or anaerobic.Result of study as shown in Figure 7. ¹⁷In the O NMR test RO water is observed expection unimodal (Fig. 7 (a)).For anaerobic micelle grain water (Fig. 7 (b)), with respect to RO water, observed unimodal displacement+54.1 hertz; For aerobic micelle grain water (Fig. 7 (c)), for RO water, unimodal displacement+49.8 hertz.Observe the displacement at NMR peak for micelle grain water and RO water.In addition, meaningfully, compare in the NMR data, micelle grain sample water is observed peak broadening with the narrower peak of sample water that is untreated.

Embodiment 4-Raman spectrum

Raman spectrum is modified quite responsive to the variation of liquid structure, characterize and distinguish micelle kernel structure and micelle grain molecular structure liquid with it.This research is based upon and obtains and handle on the basis of spontaneous Raman spectrum, can write down the type of phase transformation in 4 ℃, 19 ℃, 36 ℃ and the 75 ℃ of following aqueous waters.The meansigma methods of hydrogen bond network structure and per unit volume hydrogen bond concentration is determined, and causes characterizing the water by the distinct methods preparation, particularly distinguishes and define the water composition by the method preparation of above-mentioned manufacturing micelle grain.

Fig. 8 illustrates device used in the research.Light source is the Q-switch solid state N d:YAG laser (the Spectra Physics Corp. in mountain scene city, California (MountainView)) of two harmonic wave outputs of 1064nm, and its bifrequency can produce the wavelength of 532nm.Another harmonic oscillator is made up of ktp crystal, can buy from this Kigre company that agree (Tuscon) city of Arizona State soil.Article one, harmonic wave is 1064nm, and its pulse energy is that 200mJ, width are 10ns, and repetitive rate is 6Hz.Optical frames and translucent unit can be bought from the CVC Optics company in New Mexico Ai Baikeqi (Albuquerque) city.This spectrogrph is bought from Japanese shore pine (Hamamatsu) city, and its automatic pointing system is bought from the Newport Corporation in California Coase Lame Sa (Costa Mesa) city.The electron-optical transducer is bought from the TexasInstruments company in Houston, Texas (Houston) city.

Water is injected the unit, as subjects.The following sample water of research in the experiment: oxidation micelle grain water, not oxidation micelle grain water, Millipore (tm) distilled water, laboratory system distilled water, medical grade pair steaming water for injection, bottled commercial reverse osmosis water, and tap water (unprocessed).To feed test water by the strong ultrasonic field that pulse generator, sine-wave generator and focusing horn shape microphone produce.Laser beam is injected the unit.Signal enters spectrogrph with 90 ° angular separation, and the grid of the 2nm/mm peptizaiton equipment that provides is provided in the spectrogrph.Raman scattering spectrum is by detector measurement.

The result shows that the hydrogen bond net partial structurtes of micelle grain water change in sound field.Particularly, change and to meet that the average distance part is decreased to 2.80 dusts between oxygen atom, the cancellated ordering that strengthens with the hydrone of hydrogen bonded almost reaches hexagon ice, and this distance is 2.76 dusts among the latter.

With other sample water ratio, the granule temperature of sample that contains micelle grain water is high about 10 ℃, and this average size that shows micelle grain water is less than other sample water.And micelle grain water demonstrates more uniform granule size than other water to be formed, i.e. the micel kernel structure of homogenizing more.

II. culture medium and manufacturing thereof and using method

The present invention relates to be used for the culture media composition of biology, agricultural, pharmacy, industry and medical applications.Contain micelle grain water in these compositionss.The method of making and use culture media composition within the scope of the present invention.

Summary and definition

Unless specialize, otherwise practice of the present invention will be adopted the routine techniques in the following technical field: (1) cultivates zooblast, plant cell and tissue thereof; Microorganism, subcellular fraction, virus and phage; (2) perfusion of differentiated tissue and organ; (3) biochemistry; (4) molecular biology; (5) microbiology; (6) hereditism; (7) chemistry.These technology have detailed description in the literature.For example, see also " animal cell culture: basic technology handbook (Cultureof Animal Cells:A Manual of Basic Technique), the 4th edition, 2000, R.Ian Freshney write, and Wiley Liss Publishing publishes; " zooblast training " (Animal Cell Culture), J.W.Pollard and John M.Walker write; " plant tissue culture: theory and practice " (Plant tissue Culture:Theoryand Practice), nineteen eighty-three, Elsevier Press publishes; " culture plant cell cometabolism: commercial Application " (Plant Cell Culture Secondary MetabolismToward Industrial Application), Frank DiCosmo and MasanaruMisawa write, and CRC Press publishes; " plant tissue culture notion and laboratory operation handbook (Plant Tissue Culture Concept and Laboratory Exercises), the 2nd edition, Robert N.Trigiano and Dennis Gray write, and 1999, CRCPress published; " photobiochemistry and molecular biology " (Plant Biochemistry andMolecular Biology), the 2nd edition, Peter J.Lea and Richard C.Leegood write, and 1999, John Wiley and Sons published; " plant tissue culture experiment " (Experiments in Plant Tissue Culture), Dodds﹠amp; Roberts writes, the 3rd edition; " neuronal cell cultures: hands-on approach " (Neural Cell Culture:APractical Approach), 163 volumes, James Cohen and Graham Wilkin write; " molecular cloning: laboratory manual " (the MolecularCloning:A Laboratory Manual) that writes by people such as Maniatis; " cellular elements biology " (MolecularBiology of The Cell), people such as Bruce Alberts write, and the 4th edition, 2002, Garland Science published; " microorganism biological technology " (MicrobialBiotechnology), " using microbe basic principle " (Fundamentals of AppliedMicrobiology), Alexander N.Glazer and Hiroshi Nikaido write, nineteen ninety-five, W.H.Freeman Co. publishes; " pharmacy biotechnology " (PharmaceuticalBiotechnology), Daan J.A.Crommelin and Robert D.Sindelar write, 1997, Harwood Academic Publishers published.Relevant periodical comprises " cell tissue research " (Cell Tissue Research); " cell " (Cell); " science " (Science); " nature " (Nature); " IMMUNOLOGY KEY WORDS INDEX (Journal ofImmunology); " thymus " (Thymus); " international cell clone magazine " (InternationalJournal of Cell Cloning); " blood " (Blood); " hybridoma " (Hybridoma).

In the present invention described, the use of following term met to give a definition.

Term used herein " micelle grain culture medium " refers to contain the culture medium of micelle grain water.Modification comprises that the adjective " micelle grain " of any Aquo-composition of culture medium, medium, liquid, gel, compositions, component or composition refers to the micelle grain water in the said composition, and is dissolved in the micelle grain water or with micelle grain water and mixes.

According to " Oxford biochemistry and molecular biology dictionary " (Oxford Dictionary ofBiochemistry and Molecular Biology) (Oxford University Press, 1997) definition, term " cultivation/culture " refer to 1 (a) among the Nutrient medium (being culture medium) or on growing or be in the set of cell, tissue fragment or the organ of activated state; (b) added any culture medium of this lived material, no matter whether this class material still keeps active.2. prepare, grow or keep the practice or the process of this type of culture.3. grow, keep or prepare culture.

" cell " is the fundamental structural unit of all live organisms, is made up of the small protoplasm group that just can see with microscope usually, includes by cell membrane and/or cell wall and the extraneous Cytoplasm that separates.Eukaryotic cell is to contain the nuclear cell that wraps in the nuclear membrane.Prokaryotic cell is that the interior genomic DNA of cell is not included in the cell in the nuclear membrane.

" culture medium " refers to be exclusively used in any Nutrient medium of supporting that culture is grown or kept.Culture medium is generally by artificial preparation, and is designed for cell, tissue or the organ of particular type.They generally are made up of softish gel (being commonly referred to solid or semisolid culturemedium) or liquid, but once in a while hard solid are arranged also.

" tissue culture/culture " refers to 1. under artificial condition, grows or keeps the technology or the process of histiocyte (cell culture), whole organ (organ culture) or the part organ of animal or plant; 2. any by this type of technology growth or the living substance kept.

" tissue " refers to form any cell aggregation of carrying out one or more specific functions.

" organ " is to adapt to and/or specialization is used to carry out any part of the multicellular organisms health of one or more vital functions.

" organ culture " belongs to the category of tissue culture, and one of them organ, part organ or organ primordium after removing in animal or plant, maintain in the Nutrient medium external, keep its structure and/or function.

" organelle " is the interior any concrete structure of individual cells of unicellular organism body or multicellular organisms, and adaptation and/or specialization are used to carry out one or more vital functions.

" microorganism biological technology " refers to use cell (protokaryon or eucaryon) to produce protein, reorganization and synthetic vaccine, microbial insecticide, enzyme, polysaccharide and polyester, ethanol, aminoacid, antibiotic; Carry out organic synthesis and degraded by microorganism (and enzyme); Environmental applications comprises sewage and waste water microbiology; The microbial degradation of abnormity biomass; Reclaim and from waste water, remove in the heavy metal process and use microorganism at mineral.The microorganism biological technical scope is very extensive, the part introduction is arranged in the following books: " microorganism biological technology " (MicrobialBiotechnology), " using microbe basic principle " (Fundamentals of AppliedMicrobiology), Alexander N.Glazer and Hiroshi Nikaido write, nineteen ninety-five, W.H.Freeman Co. publishes; " pharmacy biotechnology " (PharmaceuticalBiotechnology), Daan J.A.Crommelin and Robert D.Sindelar write, 1997, Harwood Academic Publishers published.

Cell culture medium-basic principle

The basis of cell culture medium (as described below)-prepare-can buy (for example, Sigma Chemical, Invitrogen, Biomark, Cambrex, Clonetics, and more) from supplier as single component, premix composition, drying or water.The method of water preparation culture medium (" cell, organ and embryo's culture medium " (Culture Media for Cells, Organs, and Embryos), CRC Press publication, 1977 well known in the art; " zooblast: the basic document of cultivation and culture medium " (AnimalCells:Culture and Media:Essential Data), John Wiley﹠amp; Son publishes, nineteen ninety-five; " being used for culture medium, supplement and substrate preparation method that serum-free animal cell is cultivated in molecule and the cytobiology cell culture processes " (Methods for Preparationof Media, Supplements and Substrata for Serum Free Animal CellCulture in Cell Culture Methods for Molecular and Cell Biology), the 1st volume, Wiley-Liss publishes, 1984).Culture media composition of the present invention comprises micelle grain water.In order to enumerate the whole bag of tricks of cell culture, the type of cell culture processes and cell culture medium is well known in the art, below only briefly introduces.

The type of cell culture:

Primary culture is directly taken from the intact animal's tissue that cuts off.These tissues are cultivated as explant or are cultivated after the Digestion by enzyme is separated into single cell suspension.Originally these cultures are heterogeneous, are arranged by fibroblast afterwards.Generally speaking, primary culture can only continue one limited period external, and primary cell keeps many visible in vivo cell differentiation features usually during this period.

Continuous culture is made up of unicellular type.These cells can be bred the cell division or the unlimited breeding of finite population (about 50) continuously in culture.Keep certain differentiation degree.Make these cultures keep the long time, must set up cell bank.

Cultivate morphology

Cell culture or in suspension the growth (as the grumeleuse of unicellular or little free-floating) or as monolayer adherence in tissue culture flasks.Sometimes, cell culture may be grown to half attached cell that is mixed with attached cell and suspension cell.

The type of culture medium

In general, cultured cells needs gnotobasis, the nutrition supply that is suitable for growing and stable culture environment, for example, and pH value and temperature.Nowadays developed various specified basal medium types, can buy from market.They are modified, are rich in aminoacid, vitamin, fatty acid and lipid.Therefore, can obtain being suitable for supporting the culture medium of various kinds of cell type growth now.Accurate culture medium prescription derives from the concentration optimization to every kind of composition usually.

Culture medium supplier distributes the document of making and using culture medium by catalogue or its website to those skilled in the art.For example, it is " basic technology of cell culture " (Fundamental Techniques in CellCulture), " laboratory on-line operation handbook (Sigma-Aldrich company) " (ALaboratory Handbook Online (Sigma-Aldrich Company)) that the website of Sigma-Aldrich company discloses title, and following table has been enumerated example of different culture medium and using method thereof.Those skilled in the art can adopt micelle grain water to replace all or part of non-micelle grain water in the following culture medium.

Table 1: the dissimilar and use of culture medium

Balanced salt solution PBS, Hanks BSS, Earles salt

DPBS (production code member D8537/D8662)

HBSS (production code member H9269/H9394)

EBSS (production code member E2888) forms the basis of many complex mediums

Former being commissioned to train of basal medium MEM (production code member M2279) supported and the diploid cultivation.

DMEM (production code member D5671) contains the MEM improved products of higher concentration aminoacid and vitamin.Support comprises the various kinds of cell type of hybridoma.

GMEM (production code member G5154) Glasgows improves MEM, is exclusively used in the BHK-21 cell

Complex medium RPMI 1640

(production code member R0883) derives from the human leukemia cell at first.Support comprises the multiple mammalian cell of hybridoma.

Iscoves DMEM

The further enrichment improved products of (production code member 13390) DMEM is supported high-density growth

Leibovitz L-15

(production code member L5520, liquid state) is exclusively used in no CO2 environment

TC100 (production code member T3160)

Grace insecticide culture medium

(production code member G8142)

Schneider insecticide culture medium (production code member S0146) is exclusively used in the cultivation insect cell

Serum-free medium CHO (production code member C5467)

HEK293 (production code member G0791) is used for serum-free and uses

Ham F10 and derivant thereof

Ham F12 (production code member N4888)

DMEM/F12 (production code member D8062) annotates: other genes such as the necessary supplementation with insulin of these culture medium, siderophillin and epidermal growth factor.Usually, these culture medium need be used the HEPES buffering

Insect cell Sf-900II SFM, SF insecticide culture medium 2 (production code member S3902) are exclusively used in the Sf9 insect cell especially

The basis of culture medium

The solution that comprises the culture medium basis of micelle grain water is included within the range of compositions of the present invention.

Inorganic salt

Carbohydrate

Aminoacid

Vitamin

Fatty acid and lipid

Protein and peptide

Serum

Below summarized every kind of specific function that composition is carried out:

Inorganic salt not only can help to keep the Osmotic balance of cell, and also helps to regulate transmembrane potential by supply sodium, potassium and calcium ion.All these all need in cellular matrix, are used for cell adhesion or as enzyme cofactor.

Buffer system.The required pH value scope of most cells is between 7.2 to 7.4, and the monitor closely pH value is most important to reaching optimal culture condition.There are several factors can influence this optimum condition.Fibroblast needs higher pH (7.4-7.7) value, and the cell line that continues to make the transition then needs higher acid condition pH (7.0-7.4) value.Set up new cultivation and when successfully reaching, after cell inoculation, regulate the pH value particular importance immediately by one of following two kinds of buffer systems; (i) contain equilibrated " natural " buffer system of gaseous state CO2 of culture medium C O3/HCO3 and (ii) utilize the zwitterionic chemistry buffering be called HEPES (production code member H4034).

Use the culture of natural magnesium hydrogen salt/CO2 buffer system need remain in the air under the condition of CO2 content as 5-10%, provide by the CO2 incubator usually.Not only cost is lower for bicarbonate/CO2, avirulence, but also can provide other chemical benefits for cell.

HEPES (production code member H4034) has outstanding buffer capacity, and the pH value scope is between 7.2-7.4, but its cost is relatively expensive, may toxigenicity to some cell when concentration is higher.HEPES (production code member H4034) buffering culture need not to control gaseous environment.

Most of commercial culture medium are used as the pH value indicator with phenol red (production code member P3532/P0290), thereby the pH value state of culture medium can be indicated by color all the time.Usually, if its colour changed into yellow (acidity) or purpling (alkalescence) then should be changed or supplementing culture medium.

Carbohydrate.The main source of energy is the general carbohydrate that exists with the form of sugar.Used main saccharide is glucose and galactose, but some culture medium also contains maltose or fructose.In some complex medium, the sugar-containing concentration in the basal medium is that 1g/l does not wait to 4.5g/l.Contain the higher culture medium of sugar and can support the growth of wider general various cells.

Vitamin.Serum is the important source of vitamin in the cell culture.But many culture medium also are rich in vitamin, and this makes them be more suitable for wider cell line.Vitamin is the precursor of a large amount of cofactors.Many vitamin (particularly vitamin B group) all are the necessarys of cell growth and breeding, and for some cell line, vitamin B12 is indispensable.Some culture medium have also increased the content of vitamin A and E.The vitamin that is usually used in the culture medium comprises vitamin B2, vitamin B1 and biotin.

Protein and peptide.They have special importance in serum-free medium.Modal protein and peptide comprise albumin, siderophillin, fibronectin and myosin, are used for substituting by adding serum to culture medium and the albumen that exists usually.

Fatty acid and lipid.Similar with peptide to protein, because fatty acid and lipid often appear in the serum, they also have a very important role in serum-free medium.For example, cholesterol and steroid are that specialized cell is indispensable.

Trace element.Trace element comprises zinc, copper, selenium and tricarboxylic acids intermediate product etc.Selenium is antidote, helps to remove oxygen-derived free radicals.

It is very consuming time to make culture medium from basis, and the danger of pollution is arranged in processing procedure.Present most of culture medium is for the fluid medium of preparation mixed dust formulation or 10x and 1x, and is very easy to use.Culture medium commonly used is listed in culture medium supplier's the goods catalogue (for example, Sigma-Aldrich life sciences goods catalogue).

If buy the medium component of powdery or 10x culture medium, please note that reconstruct powder or the used water of dilution concentrated liquid should not contain mineral, organic or little living pollutant.And, also necessary apyrogeneity (production code member W3500, water, tissue culture's level, Sigma-Aldrich).In most of the cases, the final impedance that purifies preparation by reverse osmosis and extration resin is that the water of 16-18Mx is suitable for.After preparing, culture medium is sterilised filtration before use.Obviously, the fluid medium of directly buying 1x from supplier can need not this step.In all cases, culture medium of the present invention contains micelle grain water as component, preferred tissue culture level.Culture medium supplier (for example, Sigma-Aldrich, Invitrogen, Clonetics) and cell and cell culture supplier generally provide one or more products (medium, medium composition and cell) in the packing external member of product.The present invention includes test kit, it contains micelle grain water in himself container, or as the composition of another container in the test kit.

Serum.Serum is the complex mixture of albumin, somatomedin and growth inhibitor, and it may be one of most important composition of cell culture medium.The most frequently used serum is hyclone.The serum of other type also comprises newborn calf serum and horse serum.The quality of serum, type and concentration all may influence the cell growth, therefore, monitor serum in batches and observe the ability of its sustenticular cell growth very important.Serum can also improve the buffer capacity of culture, and this situation lower to slow growth cell or inoculum density (for example, cell clone experiment) is extremely important.

The culture medium of the present invention, its method of manufacture and use thereof that contains micelle grain water is used for following biological entities category for the application's purpose can classify as arbitrarily.Should understand this classification does not get rid of described compositions or its using method and is used for using more than a classification.

Zooblast, itself (for example, cell line etc.)

Compositions of the present invention comprises:

1. the compositions that comprises micelle grain culture medium, particularly preparation are used for the culture medium of zooblast.

2. comprise preparation and be used for the micelle grain culture medium of zooblast and the compositions of zooblast.

3. be included in the compositions of the zooblast that uses micelle grain Zooblast culture medium in the following method and prepare.

Preparing the culture medium that is used for zooblast among the present invention is used for:

1. breed, keep or preserve zooblast or its compositions.

2. separate or differentiation zooblast or its compositions.

3. preparation contains the compositions of zooblast.

In addition, the present invention is also contained preparation micelle grain Zooblast culture medium, is prepared the method for compositions that comprises micelle grain Zooblast culture medium and zooblast.Vaccine is the product example from this type of animal cell culture.

Stem cell

The compositions and methods of the invention are suitable for stem cell.Embryonic stem cell and kind system or tissue specificity stem cell are the important models of biomedical research, but in the field of this fast development, the availability of research material and accessibility regular meeting are restricted.Compounds and methods for of the present invention is used for expanding, preserving embryonic stem cell, and takes from the stem cell of originating after the birth of various bacterial strains and species.(American Studies resource center (National Center for ResearchResources); Manassas, Virginia (Manasas) US mode strain is collected (American Type Culture Collection)).Stem cell also can be taken from bone marrow, subcutaneous fat and reticular corium raised zones.The product of National Stem Cell Resource comprises: (a) the newborn stem cell of the kind of non-human embryonic stem cell, various species system or tissue specificity; Can transport by freezing bottle dry ice; That (b) selectes characterizes the reagent relevant with utilization with stem cell; This comprises antibody, nucleic probe, cDNA, genomic library and is used for site directed mutagenesis or plasmid vector that other is relevant with the stem cell purposes; (c) Kai Fa standard medium.Along with identifying and exploitation, also can obtain identifying the reagent of common denominator between stem line and the species.This comprises and is used for RT-PCR and based on the reagent of immunologic mensuration.The present invention includes micelle grain culture medium and reagent and be used for Application of stem cells, comprise the stem cell recovery.

Microorganism

Microorganism comprises actinomycetes, unicellular alga, antibacterial, fungus (yeast and mycete) and protozoacide.

Compositions of the present invention comprises:

1. the culture medium that is used for microorganism that contains micelle grain water.

2. the culture medium that contains micelle grain water and microorganism.

The culture medium of the present invention that contains microorganism is used for:

1. breed, keep or preserve microorganism or microbial composite.

2. preparation or separation contain the compositions of microorganism, comprise the compositions of using micelle grain water or containing micelle grain water.

3. separate microorganism.

In addition, the culture medium that preparation contains micelle grain water is also contained in the present invention, and preparation contains the method for compositions of culture medium and microorganism.

Carrier, itself (for example, plasmid, hybrid plasmid, cosmid, viral vector, Phage Carrier etc.)

This type of biological entities comprises the self-replicating nucleic acid molecule that can be used for nucleotide sequence or gene introducing cell; This type of nucleic acid molecules is designated as carrier, can plasmid, forms such as hybrid plasmid, cosmid, viral vector, phage vector exist.

Carrier or excipient can be used for transformation or transfection.Conversion is a process of obtaining new hereditary material by the combination of foreign DNA.Transfection is to use separated DNA or RNA hereditary information to be transferred to the process of cell.

Plasmid is the exchromosomal DNA element of independently duplicated circle.Hybrid plasmid is the plasmid that opened moving met another kind of organism DNA and reconfigured.Cosmid is the plasmid that comprises Lambda phage " cos " site.

Viral vector (for example, SV40 etc.) is to be exclusively used in plant or the animal virus of foreign DNA being introduced host cell.Phage vector (for example, Lambda phage etc.) is to be exclusively used in the bacterial virus of foreign DNA being introduced host cell.

Virus or phage

This type of biological entities comprises virus or the phage that belongs to microorganism, they are made up of the peripheral protein shell of the nucleic acid core of ribonucleic acid or DNA (deoxyribonucleic acid) (a), (b) can independently enter host microorganism, and (c) need have ribonucleic acid or DNA (deoxyribonucleic acid) host microorganism to duplicate.

Compositions of the present invention comprises:

1. preparation is used for the compositions of the micelle grain culture medium of virus or phage.

2. preparation is used for the compositions of the micelle grain culture medium of virus or phage, and said composition comprises virus or phage.

The culture medium of the present invention that contains virus or phage is used for:

1. preparation or propagative viruses or phage.

2. purified virus or phage.

3. manufacturing viral sub-units.

Breeding is subjected to the restriction with the virus multiplication relative program, and is irrelevant with the artificial hereditary material that has changed virogene type.The artificial reconstructed process of this type of hereditary material is used for gene mutation, cell fusion or genetic modification, comprise that (1) makes zooblast, plant cell or microorganisms gene mutation, (2) merge animal, plant or microbial cell, (3) by manual-induced structural change in gene or in conjunction with exogenous genetic material, make the genotype of zooblast, plant cell or microorganism produce stable, heritable change, perhaps (4) make the genotype of zooblast, plant cell or microorganism produce temporary transient the variation by in conjunction with exogenous genetic material.

Gene mutation can be that the cell DNA of the nature that caused by environmental factors or dna replication dna mistake changes, and also can be caused by physics or electrochemical conditions.Contained gene mutation process has under the situation in conjunction with foreign DNA not, the appointment of zooblast, plant cell or microbial DNA or the production process of random variation.Should be noted that by technical approach exogenous genetic material is attached to cell or microorganism that perhaps rearranging of hereditary material do not belong to gene mutation in cell or the microorganism.

For adopting the in vitro mutagenesis that outside cell or microorganism, changes the method for cloned DNA and produce not belong to gene mutation.Exogenous genetic material may contain gene synthetic by chemical method or that change.Be subjected to show one or more phenotypic characteristics of encoding by described hereditary material in conjunction with the temporary transient variation that exogenous genetic material influences.The temporary transient variation is a kind of variation of of short duration, short-term.The method that has produced the non-genomic coding change that is subjected to nucleic acid molecules (as antisensenucleic acids) influence does not belong to gene mutation.

These compositionss and method can be used for the virus of all types (being animal, plant etc.).

Plant cell or cell line, own (for example, transgenic, mutant etc.)

This type of biological entities comprises plant cell or cell line itself, and it can be that genetically modified, mutant or other obtain the product of the method for plant cell.

Compositions of the present invention comprises:

1. comprise the compositions that micelle grain water and preparation are used for the culture medium of plant cell or cell line.

2. comprise micelle grain water and prepare and be used for the culture medium of plant cell or cell line and the compositions of plant cell.

The culture medium of the present invention that contains plant cell or cell line is used for:

1. external breeding

2. keep or preserve plant cell or cell line.

3. separate or separate plant cell.

4. no matter have or not genotype to change, make plant cell be regenerated as tissue, plant part or plant itself.(New Zealand Total Lab Systems, Ltd.; For example, " the commercial Cymbidium ensifolium (L.) Sw. breeding of tissue culture: cultural method " (Commercial Propagation of Orchids inTissue Culture:Seed Flasking Methods)." Cymbidium ensifolium (L.) Sw. is cultivated handbook ABC " (Orchid Manual Basics), Kay S.Greisen writes, and 2000, U.S. Cymbidium ensifolium (L.) Sw. association (American Orchid Society) published; " plant tissue culture experimental program " (Plant Tissue Culture Protocols) can obtain from Sigma-AldrichCo. website and goods catalogue).

Subcellular fraction

Should be understood that compositions of the present invention comprises that preparation is used for the culture medium of the subcellular fraction (as organelles such as mitochondrion, microsome, chloroplasts) of microorganism, zooblast and plant.This type of culture medium is used for separating or handling subcellular fraction.The present invention also comprises the manufacture method of these culture medium.

The culture medium that is used for differentiated tissue or organ

The present invention is contained and is applicable to the differentiated tissue that comprises blood or the micelle grain culture medium of organ.This type of culture medium is used to keep differentiated tissue or organ, promptly keeps at Nutrient medium or life to keep survival condition in the culture medium.

Keep and comprise that the conservator official is making callable subsequently product (for example, hormone) or showing under the condition of active (for example, hormone is synthetic).

Therefore, the present invention includes by continous pouring liquid or the present composition in order to keep the perfusion medium with the preparation of micelle grain water of differentiated tissue or organ.United States Patent (USP) 4,879 has been introduced the solution that pours into and keep organ in 283,4,873,230 and 4,798,824 (the classifying reference herein as).D ' Alessandro AM, Kalayog 1uM, Sollinger HW, Pirsch JD, Southard JH, Belzer FO write." adopting the organ of University of Wisconsin solution to preserve current state " (Current status of organ preservation withUniversity of Wisconsin solution).Arch Pathol LabMed.1991; 115 (3): 306-310; Viaspan (r) is a kind of organ perfusion and keeps solution that by Barr Laboratories, Inc. produces, and is used for organ and tissue transplantation and active the preservation.

Compositions of the present invention comprises that preparation is used for the compositions of freezing differentiated tissue or organ, and the compositions that is used for keeping differentiated tissue or organ process by freezing method.

Compositions of the present invention comprises that preparation is used to keep blood or seminal fluid is in the compositions of physiologically active state, and is extracorporeal blood cell separation or processing method composition prepared.In addition, the present invention also comprises the compositions that is used for the artificial fertilization.

Should understand, micro-cluster compositions of the present invention comprises physiological solution or water-containing medium, they may not contain nutritional labeling, but still have features such as pH value, buffer capacity, osmotic pressure molar density, conductivity, aseptic, and be used for other physiological solution separately or jointly to keep competent cell, tissue, organ and organism.The example of physiological solution includes but not limited to Lin Ge (Ringer) solution, saline solution, buffer solution.These solution are known and be usually used in handling biomaterial, and are apparent to those skilled in the art.

Use micelle grain culture medium stimulating growth or activity

The influence of micelle grain water pair cell vigor

By the mensuration of cell membrane integrity, carry out the influence that micelle grain water pair cell vigor is determined in a research.

Macrophage is accepted by the growth medium of micelle grain water preparation and the growth medium of being prepared by distilled water (DDW).

Macrophage is taken from mice.(10mM HEPES pH7.2) is injected into the intraperitoneal of dead Mus with the Hanks solution of 2ml.Then, collect the solution that contains macrophage.Use the Hanks balanced salt solution, cell concentration is adjusted to 10 ⁶Individual cells/ml.

Generally speaking, the cell suspension of 20 microlitre sample sizes is placed on the cover glass, incubation is 45 minutes in wet cabin, uses the cell of Hanks solution Xian attachment removal at glass surface then.

(EthBr, Sigma) (FDA, Sigma) pair cell carries out double staining mensuration, to determine the integrity of cell membrane with the diacetyl fluorescein to use ethidium bromide.Used staining solution contains the EthBr of 5 mcg/ml and the FDA of 5 mcg/ml.The cell quantity that the count fine after birth damages.The method enters the ability of the cell of cell membrane damage based on EthBr.EthBr can combine with DNA.EthBr sends bright red fluorescence with after DNA combines.FDA is the permeation cell film easily, and structurally is converted into the fluorescein with bright green fluorescence.Thereby fluorescein can accumulate on the complete cell of plasma membrane, and for the damaged cell of cell membrane, fluorescein can leave cell easily.Can draw the double staining result after five minutes, can observe the unaffected cell of plasma membrane and be green fluorescence.The cell that plasma membrane the damages fluorescence that takes on a red color.

In the experiment of phase I, in the culture medium that contains EthBr and FDA to macrophage incubation 15 minutes.Thoroughly they are cleaned then, remove the free stain in the culture medium of extracellular.Then, with growth medium be replaced by 199 culture medium with the preparation of DDW or micelle grain water (199 powder culture medium, one Russia, Paneko).The counting dead cell.

In the second stage of experiment, the pair cell incubation is 230 minutes in 199 cell culture mediums of DDW or the preparation of micelle grain water.Pair cell roughly dyes then, can determine the quantity of dead cell.

Fig. 9 is the quantity assessment of the macrophage that plasma membrane damages behind the incubation in 199 cell culture mediums of DDW or the preparation of micelle grain water.Respectively after the incubation 15 minutes and 240 minutes, the percentage ratio P% of available plasma membrane damage cell represents data in different 199 cell culture mediums.The result shows, compares with the culture medium for preparing with micelle grain water, and the cell quantity that damages with cell membrane in the cell culture medium of distilled water preparation will exceed 2.6 times.This explanation is compared with the effect of the cell culture medium of preparing with DDW, prolongs or increased the life-span of cell with the cell culture medium of micelle grain water preparation.In other words, compare, with the damage of the cell culture medium plasma membrane capable of inhibiting cell of micelle grain water preparation with cell culture medium with DDW preparation.

The influence of pH value in the micelle grain water pair cell

Carry out a research and determine the influence of pH value in the micelle grain water pair cell.Obtain mouse macrophage by above-mentioned steps.In with 199 culture medium of DDW or micelle grain water preparation, after the incubation 15 minutes and 240 minutes, determine the intracellular ph value of these cells.

The intracellular ph value of macrophage adopts the fluorescence microscope (LUMAM13, LOMO, Russia) of fluorescence excitation and emission improvement system to measure based on micro-photoelectric colorimetry.

Fluorescence excitation is carried out by the photodiode of the maximum lambda=435nm of a blueness.Fluorescence is by having lambda1=520nm respectively, and the dual channel system of the interference filter of lambda2=567nm is measured when two different wave lengths simultaneously.

Fluorescence excitation and synchronized transmissions are measured and are carried out by built-in microcontroller (LA-70M4).

As among the fluorescence FDA (5 mcg/ml) of pH value indicator to macrophage incubation 15 minutes.Behind the dyestuff incubation, in the culture medium around the stain on the cell is cleaned.Then, cell is put into the culture medium of small incubation ware, and observe with water immersion objective (x40).The pH value calibration curve can demonstrate the scope of ionic condition.

At first cell incubation in the FDA dye liquor was also cleaned stain in the medium in 15 minutes around, put into 199 culture medium then with DDW or the preparation of micelle grain water.The kinetic measurement of intracellular ph value should not be less than 30 submicroscopes to be observed, and answers triplicate.Cell is incubation 230 minutes at least.Figure 10 illustration intracellular ph value changes in the macrophage after being replaced by with 199 culture medium of DDW or micelle grain water preparation dynamic (delta pHi).The x axle is the time of unit for changing behind the cell culture medium with the second.Y-axis is the variation of intracellular ph value (delta pHi).As can be seen, intracellular ph value all is about 7.15 in 199-DDW standard medium and 199 micelle grain water.In 199 micelle grain water after the incubation 15 minutes, pH value 0.16 unit that raises.After the same incubation 15 minutes, macrophage does not have obvious change in the 199-DDW culture medium.After 230 minutes, according to observation, the intracellular ph value of the cell of incubation raises 0.43 in 199 micelle grain water.The rising of the intracellular ph value of the cell of incubation can be ignored in 199-DDW.Conclusion shows, contacts with the culture medium for preparing with micelle grain water (but not " common " water) by making cell, and the intracellular ph value of cell can raise.

Another group adopts the pig embryonic kidney cell with 199 culture medium and 10% Ox blood serum incubation to experimentize, the result confirms, compare with the growth medium for preparing with light water, the growth medium for preparing with micelle grain water can make intracellular ph value raise and the enhancing cell viability.

Micelle grain water is to the influence of two kinds of human body cell growths and transfection

Carry out a series of experiments and determine that micelle grain water is to the growth of cell and the influence of transfection in the culture medium for preparing with this water.

HEP (293T) and people's dendritic cell are adopted in experiment.At AquaPhotonics from San Diego, CA (San Diego) city, the dilution spissated DMEM culture medium of 10x (the Life Technologies company in Gaithersburg, the Maryland State (Gaithersburg) city) in the micelle grain water that Inc. buys.In cell, replenish 10% hyclone (FCS).

In another experiment of carrying out at the same time, with the standard DMEM culture medium incubation cell that does not contain the preparation of micelle grain water.

At the 0th, 3,6 and 9 day, adopt the dyeing of 0.4% trypan blue (Life Technologies) pair cell, to determine the vigor of culture.

At the 1st day that cultivates, adopt calcium phosphate precipitation method (the Invitrogen company in Carlsbad, California (Carlsbad) city) the 293T cell to be carried out transfection by HIV molecular cloning (to the GFP coding).In the experiment that another group is used for contrasting, the 293T cell of cultivating in standard DMEM culture medium carries out transfection by identical HIV molecular cloning.Second day, can detect the HIV Gag p24 from HIV transfection culture with by ELISA and collect the supernatant.Find the optimum dilution degree of this method sensitive volume, can adopt titrimetry to measure by 10 times of dilutions.

Then, use the primary culture of the viral infection dendritic cell (DC) of results.Two kinds of DC cultures are maintained with in the culture medium that concentrates DMEM and 10 times of dilution preparations, and a kind of culture (being used for experiment) is with micelle grain water dilution DMEM, and another kind of culture (being used for contrast) dilutes with light water.After infected by HIV the 5th day,, can monitor the infection of unicellular level by scoring to the positive DC of GFP.

The result:

A. as shown in figure 11, survival ability test confirms, and compares with the cell culture medium of light water preparation, makes the survival ability of 293T cell improve 70% as the micelle grain water of medium preparation solvent when the 9th day of cultivating.

B. compare with control cultures, adopt when the supernatant of each culture is taken from the titration of 3log point, test in the culture duplicating of HIV in the transfection 293T cell culture and exceed three times (Figure 12 a).

C. in DMEM culture medium, cultivate DC with the preparation of micelle grain water, and DC is exposed to from the HIV of the 293T cell harvesting of cultivating among the DMEM with micelle grain water preparation, all greatly strengthened DC to " admissibility " (Figure 12 b) of HIV (compare with 3.7% data in the control medium, 35% DC is infected in the experiment culture medium).

These experiment showed, in cell transformed system, virus and primary cell, in culture medium light water are replaced with micelle grain water and can produce biological effect to these biological entities.Cell viability has strengthened 2 to 3 times; HIV duplicate and cell line and primary cell culture in the replication in vitro rate lifting is all arranged.

Micelle grain water is to the influence of the express spectra of distinctive dendritic cell sign

The purpose of this research is to monitor in the culture medium for preparing with deionized water and the difference between the characteristic DC sign express spectra in the culture medium of micelle grain water preparation.

Experimental design and result.In be diluted to the culture medium by 10 times of concentrated MEM (the Life-Technologies company in Gaithersburg, the Maryland State (Gaithersburg) city) preparation of final concentration with deionized water or micelle grain water, cultivate DC.In two kinds of culture medium, replenish cytokine IL-4 and GM-CSF (20ng/ml).Generate DC according to standard test scheme (Sallusto etc., 1994), also cultivate in the 6th day record phenotype of differentiation.Repeated phenotype test with 69 days with identical monoclonal antibody the 30th respectively.The surface marker expression adopts the cells were tested by flow cytometry of FACscan reader (California Bekton-Dickenson).

The explanation of cell surface marker:

1.DC-SIGN-M.W.～44K, cell-specific ICAM-3 receptor

Attached the paper of the function of DC-SIGN in the relevant dendritic cell.

2.CD4-main HIV gp120 receptor, MW～55K.CD4 is the anchor station of HIV envelope protein.

3.CD1a-be the analog of MHC complex in the professional antigen presenting cell, be responsible for presenting and handle lipidantigen (non-standard antigen presentation system).

4.CD80-for bringing out of T cell proliferation provides collaborative stimulation molecule from the signal 2 of antigen-presenting cell (as DC).

5.CD83-the maturation sign of dendritic cell (DC)

6.CXCR4 and CCR5-inflammation chemokine receptors

7.MHC-II-II type major histocompatibility complex.Present external source and process proteic epi-position.

As shown in figure 12, with micelle grain water as the preparation culture medium of solvent between long-time culture period, substantial variation has taken place in the expression way of observing the CD83 sign.CD83 is the sophisticated main indicator of DC.Express the DC and the immaturity of low-level CD83 and demonstration typical structure (in suspension, growing) at the 60th day, on function, prepare to accept exotic antigen.In general, at the last fortnight of differentiation, DC can show this phenotype at external (in standard medium).After further in standard medium, cultivating, the cell death that causes spontaneous maturation and mediate by apoptosis probably.In tentative phenotype experiment, detect micelle grain water (i) and preserved the immature DC phenotype and (ii) made DC existence above 2.5 months.By other sign of DC cell surface is expressed the analysis of (the most important thing is DC-SIGN and MHC II), show the phenotype preservation.According to the similarity that sign is expressed between the DC in standard and the micelle grain culture medium, this analysis explanation micelle grain culture medium has been kept the immature DC typical functions satisfactorily.The time-to-live of DC has surpassed 2.5 months, and adopts the standard culture based formulas never to observe this phenomenon before.PRELIMINARY RESULTS confirms that micelle grain water has presented the biological activity that is reflected in the modulation of DC cell surface marker.

Sum up

1. micelle grain water can be used as the solvent of delicate tissues culture experiment fully.

2. make cell contact the biological activity that has changed cell with micelle grain water, this is reflected in the modulation and the prolongation of external DC vital stage of CD83 sign.

In Figure 13, trunnion axis has reflected the different acceptor types of cell surface.Vertical axis representative reaction (the fluorescence intensity percentage ratio of the labeled monoclonal antibody that combines with special receptor).Cell is dyeed by corresponding monoclonal antibody, and signal is compared with isotype contrast (ISO percentage ratio～1.1%).

In Figure 13, the measured value of cultivating in the control medium after 6 days is represented on the Lycoperdon polymorphum Vitt hurdle.The measured value after 30 days is cultivated in the representative of black hurdle in control medium.The white hurdle is illustrated in the measured value of cultivating in the micelle grain culture medium after 60 days.Because cell culture in early apoptosis, therefore the data that do not obtain in 60 days in light water.Opposite with cell almost completely dead in standard medium, the cell of large number of viable has shown the morphology and the corresponding cell surface marker pattern of immature DC in the time of 60 days in the micro structure culture medium.Adopt micelle grain culture medium, cell viability is improved.

Micelle grain water is to being poured in the synthetic cerebrospinal fluid for preparing with distilled water and micelle grain water The influence of the functional status of cerebral tissue.

The purpose of this research is to measure the influence of various types of water to the cerebral tissue function state.Test method is the electrical signal of reaction of brain section in the infusion liquid due to the activity of record Hippocampal Neuron Cells.According to the document record, the technology of preparing of pouring into the rat brain slice of 300-450 μ m Hippocampus with synthetic cerebrospinal fluid allows cerebral tissue to keep the about 6-8 of its functional status hour.

The method that adopts is the electric nerve elementary reaction of electric current applying pulse to be tested the functional status of cerebral tissue by record. the characteristic of neuron reaction pair perfusion medium is very responsive.Aixs cylinder group's stimulation has reflected the variation of the postsynaptic cell transmembrane potential that is positioned at the measurement electrode district.The amplitude of signal is decided on stimulating the usefulness that synapse connects between aixs cylinder and the postsynaptic neuron, also decides on the irritability of postsynaptic neuron itself simultaneously.The active decline of cerebral tissue function is the neuron result that reaction reduces to the electric current applying pulse.This is directly related with the decline of summarizing amplitude.

The main advantage of this method is very easily to enter the extracellular space of cerebral tissue in the sample, can directly use the chemical substance of desired concn.And, the interference that can not prolong Repiration, heartbeat and the animal movement of Measuring Time; Under the influence that does not have anesthesia, body fluid and hormone, very easily control experiment condition.And after electrophysiology partly finished, service test was organized and is carried out fast that biochemistry is relative with structural analysis to be easier to.

The requirement of brain tissue slice survival ability.Keep the survival ability of separating brain section, should use to brain cell in medium salt form similar artificial liquid.But the composition of cerebrospinal fluid can be according to the difference of particular experiment and is different.

Use glucose as the energy substrate layer in the liquid.The pH value of liquid is controlled by bicarbonate buffer agent.The scope of osmotic pressure is 294-311mosm/l.Solution is by carbogen gas (mist that oxygen by 95% and 5% CO2 form) oxidation.Temperature range remains on 22 ℃ to 33 ℃.Because section does not have common capillary blood flow, but because the diffusion of oxygen, substrate and metabolite between culture medium and the whole tissue slice, material still can exchange.Therefore, section must be thin as far as possible, so that allow material to spread fully by sample.According to the empirical formula of calculating slice thickness, maximum should be about 600 microns, and decides on the intensity of oxidizing process.In separation process, thickness is that 100 microns surface layer section cell can damage.Pyramidal cell is approximately identical size, and therefore the degree of depth of section should be at least 300 microns.

Experiment should be carried out on the brain tissue slice of 1 months aged Wistar rats.Use etherization.The Mus brain is separated, put into cold artificial brain ridge (AC) liquid then with the distilled water preparation.AC liquid is formed: (mM): NaCl-130, KCl-3.5, NaH2 ^*PO4-1.2, MgCl2-1.3, CaCl2-2.0, NaHCO3-25.0 and glucose.The Carbogen gaseous mixture is continued to feed solution.Use the hippocampus section of 400 microns of vibratome excisions.

Then, the incubation case that contains AC liquid is put in section, temperature remains on 22-25 ℃.After the incubation case is placed 1 hour, section is transferred to the proof box that the AC flow velocity is 3-5ml/min separately.By be positioned at Shaffer relatively the bipolar tungsten electrode (200mm) of machine (nerve fiber, the end in hippocampus CAl district stimulates synapse) transmit the stimulation electric pulse (100msec, 100-400mA).Induced potential is that assembling/overall electronics that stimulates is responded, and is recorded in the CA1 district of Hippocampus by the glass microelectrode that is full of AC liquid (resistance is 0.5-1.0mW).

Carry out two groups of experiments altogether.In two groups of experiments, all with initial 100% level of standard A C liquid (A) as signal.In first group of experiment, be the solution (solution B) that contains identical salt composition and distilled water with the AC fluid exchange.In second group of experiment, with the distilled water (solution C) in the micelle grain water replacement solution B.

Make used filling system can in proof box, supply solution continuously.In 1 minute, be that the solution of 2ml is changed another kind of solution in the incubator fully with volume.The amplitude that induction is responded is a comparative characteristic.Measuring the power parameter peak swing is the negative single-phase response of induction of 30-40%, selects to stimulate duration and position.Test is one group with 10 pulses, and be 10msec blanking time.Every 2 to 10 minutes emission sets of pulses.The signal of record is prepared against by analog-digital converter digitized and preservation and is analyzed.Final Excel and the Origin software data processing of using.The analysis of statistical data adopts paired t-test to carry out.The value of P＜0.05 has been considered to significance,statistical.

The result.The Shaffer collator is upset in the CA1 district, and induction is responded behind 0.5-4msec and from the 4-6msec record.

Figure 14 has shown the local potential measured from the rat hippocampus dependency to the infusion liquid type.Trunnion axis represents to test the time after the beginning.Vertical axis is the amplitude (% is with respect to the signal of measuring in the standard A C liquid) of electronic signal.Brain section is put in mobile standard A C liquid (A), the liquid (B) or the liquid (C) with the preparation of micelle grain water with distilled water preparation.Arrow is represented the AC liquid of the standard of changing with the test medium of micelle grain water preparation.The result gets the meansigma methods of 14 sections of seven rats.

In first group of experiment, dynamic data record after preparing liquid to replace standard A C liquid with distilled water of amplitude is responded in induction.After solution is changed, can observe induction immediately and respond the amplitude rising, maximum is 128.2% (Figure 14) of the 5th minute.Amplitude can descend steadily, and after one hour, amplitude is reduced to only 31.7% of initial value.

In second group of experiment, adopt micelle grain water to replace distilled water.After preparing liquid and replace titer with micelle grain water, the amplitude that induction is responded can acutely raise immediately, reaches maximum 135.2% after 1-3 minute.Subsequently, amplitude slightly reduces, and reduces to 94.8% after reducing to 102%, 2 hour after 1 hour.

Experimental result shows, prepares liquid alternate standard AC liquid with micelle grain water, and in experimental error, the initial amplitude that does not influence the induction response continues 2 hours.Prepare liquid alternate standard AC liquid with distilled water and cause that amplitude is reduced to 31.7% (P＜0.005) after 1 hour.

When using micelle grain water, research finished after 2 hours, because there has not been solution in the testing equipment.The time that murine brain can continue to survive should remain further to be studied.When research finished, the mean amplitude of tide of the tissue in the micelle grain water still had 94.8%.

Therefore, method of the present invention comprises, contacts one section time enough by making cell with micelle grain water of the present invention or micelle grain culture media composition, stimulates or regulate the growth or the activity of cell.This method can be used for using micelle grain culture medium to strengthen and derives from zooblast, plant cell or microorganism culturing or the chemical compound of organelle cultivation or synthesizing of product.Generally speaking, comprise in the raw material by this class methods synthetic compound or product and the preparation of non-existent compositions or chemical compound.

The foregoing description shows that compositions of the present invention is useful in regulating cellular metabolism or physiological method.The active example of this class includes but not limited to, changes or regulate the differentiation state, cell of described cell to the metabolic capacity of nutrient substance, cell cycle synchronously or lack, and to the variation of the resistance of specific compound or sensitivity, intracellular ph value.Use other method of the present composition to be used in cultured cell in the culture medium, it promotes normal cell growth and division.

The biological processing technology; Industrial products by microbial treatments form

Micelle grain water of the present invention and micro-cluster compositions generally are used for the biological processing technology of small-sized, medium-sized and large-scale PROCESS FOR TREATMENT, and production method with the product recovery or separate, in inoculation and medium preparation, cultivation and the downstream.

Industry/pharmaceutical microbiology/biotechnology depends on Aquo-composition, preparation and use they method and with little, in, big/final products (" microorganism biological technology " (Microbial Biotechnology), " applied microbiology ultimate principle " (Fundamentals of Applied Microbiology) that the giant molecule form exists, Alexander N.Glazer and Hiroshi Nikaido write, nineteen ninety-five, W.H.Freeman Co.; " pharmacy biotechnology " (Pharmaceutical Biotechnology), Daan J.A.Crommelin and Robert D.Sindelar write, 1997, Harwood Academic Publishers published).Cell culture carries out in the vessel that contain suitable liquid growth medium.The cultivation of type of production scale is carried out in bioreactor usually, and it is device (ditto, Crommelin, the 3rd chapter that is used for microorganism or enzyme growth or breeding, utilizes microorganism or enzymatic synthesis compositions or chemical compound; The 250th page of Glazer).Therefore, the present invention includes the application of micro-cluster compositions in bioreactor in the biological processing technology as described here.

Compositions of the present invention comprises the Aquo-composition that adopts so far with the partially or completely alternative those skilled in the art of micro-cluster compositions.The present invention also comprises new intermediate or the final products of producing with micro-cluster compositions, and the method for using them.

Need to adopt some major product of microorganism/zooblast/plant cell biotechnology to comprise fermented juice and liquor, cheese, antibody, industrial alcohol, high fructose syrup and aminoacid, bakery yeast, steroid, vitamin, citric acid, enzyme, hormone, somatomedin, vaccine and polysaccharide glue.

Therefore, the present invention includes micro-cluster compositions and application in the following areas thereof:

1. in antibacterial, produce protein

2. in yeast, produce protein

3. produce reorganization and synthetic vaccine

4. production microbial insecticide

5. production enzyme

6. produce microbial polysaccharide and polyester

7. production ethanol

8. production aminoacid

9. production antibiotic

10. carry out organic synthesis and degraded by enzyme and microorganism

11. comprise the microbiological environmental applications of sewage and waste water; The microbial degradation of abnormity biomass; Microorganism is used for mineral and reclaims and remove heavy metal from waste water.

Micelle grain water is to the influence of mutation rate

Micelle grain water adopts in the counting human peripheral lymphocyte method of chromosomal aberration and SCE (SCE) to carry out in the research of cytogenetics level affects.In addition, by method, in cell culture, analyze during the whole cell cycle process of human lymphocyte the counting of cell quantity behind one, two, three replicative cycle.

Analysis to chromosomal aberration frequency in the human lymphocyte cultivation is one of main test of using in the research of environmental factors mutagenic activity, and by approval (" human body chromosome distortion Analysis method " (the Methods for the analysis of humanchromosome aberrations) of The World Health Organization (WHO), Buckton K.E. and Evans H.J. write, Geneva WHO, 1973, the 66th page).

Determining of SCE frequency also is to be used to one of code test of assessing mutagenicity.The method has specificity and high sensitivity (" SCE " (Sister Chromatid Exchanges), A and B part to the mutagenicity of assessment chemical compound.Compile author Tice R.R. and Hollander A.Plenum Press, New York, London, 1984).

But determine number (Bochkov R.P., Chebotarev A.N., Platonova V.I., the Debova G.A. invention authentication code 1,175,165 of the SCE that appears suddenly in the step specificity assessment cell cultivation process of SCE frequency in the human lymphocyte culture.USSR invention and discovery councils of government, 1985).

Parallel the finishing of number assessment in mid-term behind the analyzing specimen of SCE and one, two, three replicative cycle.Therefrom can determine persistent period (the Vedenkov V.G. of fissional par and cell cycle when cell fixation, Bochkov N.P., Volkov I.K., UrubkovA.R., Chebotarev A.N., Mathematical model of determinationnumber of cells passing different number of divisions inculture.Proceeding of Academy of Sciences of USSR, v.274, No1, p186-189,1984).

The assessment of mutagenicity is relatively to cultivate the frequency of SCE and chromosomal aberration in the human lymphocyte in the cell culture medium of micelle grain water and standard deionized water preparation.

Material and method

The women of 58 years old male and two 26 and 61 years old is picked up from experiment with blood.The division lymphocyte culture of peripheral blood adopts does the preparation of RPMI 1640 (Gibco) cell culture medium.Cell culture medium dry powder mixes with the sodium bicarbonate (Serva) of 25mM/ml and the HEPES (Serva) of 24mM/ml, then it is dissolved in deionized water (18Mohm/cm) (control experiment) or the micelle grain water.Then, making the cell culture based sols is that 0.22 meter membrane filter carries out degerming by the aperture.

Prepare cell culture according to the following steps: 1ml heparinization venous blood is splashed into the aseptic plastic test tube, add 0.015ml phytohemagglutinin P (Beckon﹠amp then; Dickinson), 8ml RPMI1640 culture medium (control experiment or contain micelle grain water) and 1ml hyclone (Biowest).Rock test tube, place it in temperature then and be in 37 ℃ the incubation case.Added Colchicine (Calbiochem) at fixing preceding 2 hours, ultimate density is 0.5 μ g/ml.

Cell is fixing after cultivating 48 hours, the counting chromosomal aberration.After cultivating 48 hours, add 5-bromouracil deoxyribose (high), determine the SCE in the cell to 10 μ g/ml final concentrations.Subsequently, fixed cell after 80 hours.

Before centrifugal rotation (with 1000 rev/mins speed rotations 10 minutes) back cell fixation, add 10ml 0.55% potassium chloride (37 ℃) solution, pour out the supernatant.Then, cell is suspended again and in the incubation case, shelved 10 minutes.The cell of incubation adopts the mixed liquor of methanol and glacial acetic acid (3: 1) to fix, and is cooled to-10 ℃.Cell is placed on the cooling wet glass sheet, and heating was at room temperature shelved 24 hours before dyeing at least.

Adopt azure eosin to the sample dyeing on the sheet glass, with the counting chromosomal aberration.Specimen can be determined the SCE frequency after dyeing, according to " amending method of sister chromatid differential staining " (Modified method of differential staining of sisterchromatids) that Chebotarev A.N., Selezneva T.G., Platonova V.I. write, experimental biology and medical science bulletin.V85, the 2nd phase, 242-243 page or leaf, 1978 years.

Student t check is used for determining the difference of each cell SCE average.Assess the difference of distortion frequency, the X 2 test that need in the association list case analysis, use 2 * 2 sizes.Adopt same standard when behind different number dna replication dnas, assessing the variation in the mitosis, but only be used for the form of 3 * 2 sizes.

Experimental result

SCE

Each individuality is carried out two groups of measurements.In every group, prepare two parts of specimen, analyze 25 mid-terms.The analysis showed that middle several frequencies of SCE do not have difference to two parts of specimen.And the average of SCE is significantly not different in the group.Table 1 has shown the result that SCE measures.

Table 1: the average SCE number of each cell

The donor sex, the age	Meansigma methods ± standard deviation (cell number)		Statistics
				Deionized water	Micelle grain water	Df，t，P
	The male, 58	3.25±0.189 (100)	2.87±0.183 (100)				198；1.44；0.151
The women, 26				4.46±0.272 (100)	3.47±0.190 (100)	198；2.98；0 .0032
	The women, 61	4.31±0.269 (100)	3.81±0.236 (100)				198；1.40；0.164
Comprehensively				4.01±0.145 (300)	3.38±0.120 (300)	598；3.311；0 .000 985

Data show shown in table 1 pair all individualities, compare with the standard deionized water, and when micelle grain water was used as the solvent of doing culture medium RPMI 1640, the SCE average of each cell was lower.The 2nd donor P＜0.01st, this difference is that statistics is significant.In whole group, this significant difference even higher, level is P＜0.001.Thereby SCE analyzes to have disclosed and uses micelle grain water to suppress the mutation frequency of cultured cell as solvent, compares with the standard deionized water, causes the cell culture amount of damage less.

Splitted average

The metaphase of cell division that has the level dyeing sister chromatid is with mitosis is relevant for the first time.The metaphase of cell division that has a black and bright (arlequin chromosome) chromatid is with mitosis is relevant for the second time.In these cells, half chromosomal material is a light tone, and second half is a black.Only 1/4 chromosomal material black and the cell of 3/4 light tone are with mitosis is relevant for the third time.

Average mitosis is counted computing formula:

(∑n _i·i)/(∑ni)

Consider that each postmitotic cell quantity is multiplied, cell division average computing formula:

(∑i·n _i/2 ^i-1)/(∑n _i/2 ^i-1)

In above-mentioned formula, i is the mitosis number of times, and ni is the i time mitotic cell number.Table 2 has shown the result of different mitosis ratios in the cell.

Table 2: 1st, 2 and 3 mitotic numbers

The donor sex age	The type of water	Cell number			The division average	The mitosis average	Statistics, df, x 2、P
								Mitosis
				1							2	3
The male, 58	Deionized water micelle grain water							128 75	239 220	71 92			1.58 1.75	1.87 2.04	2；14.23；0 .0008
		The women, 26	Deionized water micelle grain water	145 155	270 235	38 32	1.53 1.48				1.76 1.71	2；2.18； 0.34
The women, 61	Deionized water micelle grain water							185 198	240 224	20 15			1.42 1.38	1.63 1.58	2；1.63； 0.44
		Comprehensively	Deionized water micelle grain water	458 428	749 679	129 139	1.51 1.51				1.75 1.77	2；1.69； 0.43

Table 2 shows, and is only to first donor, faster than the split speed of cell in the culture medium for preparing with standard water with the cell in the culture medium of micelle grain water preparation.But in general, should influence not remarkable to experimental group.According to the time (32 hours) that 5-bromouracil deoxyribose exists, it can mix DNA during this period, makes chromosomal material dyeing brighter, can determine the average time of whole cell cycle process.This needs 32/1.51=21.2 hour, conforms to the data that find in the document record.

Chromosomal aberration

Similar to the SCE analysis, ANALYSIS OF CHROMOSOME ABERRATIONS divides 2 groups to experimentize to every donor.In each group, deionized water and micelle grain water are analyzed metaphase of cell division respectively 300 times.Do not obtain data from women's donor of 61 years old wherein.The analysis showed that for two groups of experiments and two individualities, to every kind of water type, the chromosomal aberration frequency does not have difference.Therefore, the aggregation of data with two groups of experiments gets up to observe.Table 3 has shown the data of chromosomal aberration frequency.

Table 3: the frequency of chromosomal aberration

The donor sex, the age	The type of water	Metaphase of cell division number	Distortion number metaphase of cell division	Distortion frequency metaphase of cell division (%)	Statistics df, x 2、P
						The male, 58	Deionized water	600	19	3.17	1；6.9； 0.0086
Micelle grain water	600	6	1.00
				The women, 26	Deionized water		600	11	1.83	1；2.28； 0.1310
Micelle grain water	600	5	0.83
					The women, 61	Deionized water	ND	ND	ND		-
Micelle grain water	ND	ND	ND
				Comprehensively		Deionized water	1200	30	2.50	1；8.96； 0.0028
Micelle grain water	1200	11	0.92

Table 3 shows, when using micelle grain water, for the women of 58 years old male and 26 years old, the frequency of distortion metaphase of cell division all significantly is inhibited or reduces.For the individuality of analyzing as a whole for, compare with the standard deionized water, less for the significance,statistical of micelle grain water distortion frequency metaphase of cell division.

Therefore, studies show that originally that (1) adopts deionized water and micelle grain water, do not observe the difference of cell cycle persistent period; (2) the SCE frequency is lower statistically in the micelle grain water; And the chromosomal aberration frequency is also lower in (3) micelle grain water.Compare with the standard deionized water, use micelle grain water to cause low mutagenic effect.

Confirm that as low SCE frequency and chromosomal aberration compare with the standard deionized water, micelle grain water has suppressed the mutation frequency in the cell culture, and hereditary material is had stablizing effect.Term used herein " hereditary material " refers to a perhaps gene, a part of gene, one group of perhaps polygenic fragment of gene on many dna moleculars fragment of dna molecular, dna fragmentation, one group of dna molecular.Hereditary material also may refer to any material from a bit of DNA to the organism whole genome.Therefore, adopt method of the present invention can suppress the frequency of genetic material mutation, but this method is included in the step that pair cell in the culture medium of the micelle grain water that contains capacity mutation inhibiting frequency is cultivated the enough time.Mutation frequency is for the biological entities such as cell, cell among the organ culture or cells in vivo in the cell in the cell culture, the tissue.According to above-mentioned detailed description, cell comprises zooblast, microorganism and plant cell.Effectively in the body or the original position cell culture culture medium that need use the micelle grain water of capacity or contain micelle grain water give the object animal or plant of multicellular organisms.Micelle grain water also can suppress the sudden change of the hereditary material in carrier, virus or the biological entities such as phage and subcellular fraction.Should be understood that micelle grain water is to realize by cultivation or any described biological entities of incubation in micelle grain water to the inhibitory action of sudden change.

The present invention also can suppress the mutation frequency under the mutagenic agent existence.Mutation frequency is for the biological entities such as cell, cell among the organ culture or cells in vivo in the cell in the cell culture, the tissue.According to above-mentioned detailed description, cell comprises zooblast, microorganism and plant cell.Micelle grain water also can suppress the sudden change as the hereditary material in the biological entities such as carrier, virus or phage.

Vitro inhibition is brought out mutationDetermine the frequency of chromosomal aberration in the human lymphocyte, in cell culture medium, add ametycin (mutagenic agent) with three kinds of different dosage preceding 24 hours of cell fixation.The control cell does not add mutagenic agent.There are 4 kinds of experiments to be provided with.Sudden change takes place before DNA is synthetic.Determine the synthetic after stain colour solid distortion of DNA, in lymphocytes culture medium, add Dioxydine with three kinds of different concentration.Always have 16 kinds of settings: control is tested, and does not contain the ametycin of mutagenic agent+3 kind of variable concentrations, the Dioxydine of+3 kinds of variable concentrations of control experiment, the ametycin of micelle grain water+3 kinds of variable concentrations, and the Dioxydine of Penta water+3 kind of variable concentrations.Be provided with and analyze 100 metaphase of cell division for every kind, or use 1600 cells.Added ametycin at fixing preceding 24 hours by three kinds of various dose, to determine the frequency of SCE (SCE).But the concentration of mutagenic agent should be lower than chromosomal aberration, and the control experiment that does not contain mutagenic agent.For standard water and micelle grain water, 4 kinds of settings are arranged respectively, therefore have 8 kinds of different settings.In every kind is provided with, analyze 25 metaphase of cell division, always have 200 cells.Result of study shows that micelle grain water has suppressed the mutation frequency under the mutagenic agent existence.

Suppress to bring out sudden change in the bodyChromosomal aberration in the counting Os Mus marrow, every kind is provided with 100 cells of analysis.Fed mice 15 days with standard water (contrast) and micelle grain water.Put to death animal preceding 24 hours and cell fixation before, be injected into ametycin (3 dosage+do not contain the contrast water of mutagenic agent).Putting to death and preceding 2 hours of cell fixation, inject Dioxydine (3 dosage+contrast water).Every group of 6 mices; Always have 96 mices or 9600 cells.Result of study shows that micelle grain water has suppressed the mutation frequency under the mutagenic agent existence in vivo.

[399056-medicine]

III. medicine, biotic influence and health treatment compositions

Summary and definition

Unless other indication is arranged, practice of the present invention will be adopted the routine techniques in following those skilled in the art's skill: (1) organic and physical chemistry; (2) biochemistry; (3) molecular biology; (4) pharmacology; (5) pharmacological treatment is learned; (6) physiology; (7) toxicology; (8) microbiology; (9) internal medicine and diagnostics.These technology have detailed description in the literature.For example, see also Maniatis etc. and write " molecular cloning: laboratory manual " (MolecularCloning:A Laboratory Manual); " pharmacy biotechnology " (PharmaceuticalBiotechnology), Daan J.A.Crommelin and Robert D.Sindelar write, 1997, Harwood Academic Publishers published; Goodman﹠amp; " pharmacological basis of treatment " of Gilman (The Pharmacological Basis ofTherapeutics), Joel G.Hardman, Lee E.Limbird, the tenth edition, calendar year 2001, McGraw Hill publishes; " basis and clinical pharmacology " (Basic﹠amp; ClinicalPharmacology), Bernard G.Katzung, the 8th edition, calendar year 2001, McGraw Hill publishes; " pharmacy dosage form and drug-supplying system " (Pharmaceutical Dosage Forms andDrug Delivery Systems), Howard C.Ansel, Loyd V.Allen, Jr., Nicholas G.Popovich, the 7th edition, 1999, Lippincott, William﹠amp; Wilkins publishes; " the gloomy internal medicine principle of Harry " (Harrison ' s Principles ofInternal Medicine), Eugene Braunwald M.D. (editor), Anthony S.Fauci M.D. (editor), Dennis L.Kasper M.D. (editor), Stephen L.Hauser M.D. (editor), Dan L.Longo M.D. (editor), J.Larry JamesonM.D. (editor).

In the present invention described, the use of following term met to give a definition.

Term " medicine " refers to be used to diagnose, alleviate, treat, cure or prevent the chemical active agent of human body or other Animal diseases.The synonym of term " medicine " has " bio-affecting agents " and " health inorganic agent ".From broadly, medicine is the material that biosystem is exerted an influence by chemical process.These materials can be to be administered to live body to make certain process in the patient is reached effective therapeutic effect or to the chemical drugs matter of the adjustment process toxigenicity effect in the parasite of infected patient.Should be understood that biological property is expressed in the cell of live body, tissue or the organ.These activating agents, material or medicine are the themes of micro-cluster compositions of the present invention.Term " pharmaceutically active ", " pharmaceutical properties " and " active component " also refer to the effect of medicine to living tissue or live body generation.

Term " biotic influence " and " health processing " comprise General Definition, particularly press U.S.'s classification manual in the United States Patent (USP) trademark office, class definition that US patent class is described and example or embodiment, the 424th class (relevant classification described herein): medicine, biotic influence and health treatment compositions are hereby incorporated by reference.According to further definition and the example of the 424th class (it is described to Click here), term and phrase " adjuvant or carrier compositions ", " fermented product ", " Plants and Animals extract or body fluid or contain plant or the material of zooblast structure " can be used as biotic influence or health treatment compositions.The present composition is according to specific structure (for example, layering tablet, capsule) definition and classification.The using method of the present composition and preparation method for compositions are at the 424th apoplexy due to endogenous wind imbody.

Plant-derived or the animal of medicine is as the side-product of growth of microorganism, by chemosynthesis, to the molecular modification of existing chemical active agent.

Medicament sources: new drug may be from various natural animals, plant or microbe-derived discovery, or synthetic at laboratory.Vegetable material is as drug-reservoir.Animal is the source of medicine, can take from its tissue or pass through its biological treatment to get.As unrestricted example, hormone substances such as thyroid extract, insulin and pituitary hormone are taken from the endocrine gland of cattle, sheep and pig.Trimester of pregnancy mare urine in contain a large amount of estrogen.Fermented product is the compositions of the microorganism that exists in antibacterial or the one-celled plants such as yeast, mycete or fungus or derives from this, (Glazer and Nikaido now are widely known by the people, " microorganism biological technology " (Microbial Biotechnology), " using microbe basic principle " (Fundamentals of Applied Microbiology), calendar year 2001, W.H.Freeman and Company publishes).

Exercise question " medical science pharmacology " refers to be used to prevent, diagnose and treat the science of the material of disease.

Medicinal and diagnosis composition of the present invention comprises that also biological technology products is (with reference to " pharmacy dosage form and drug-supplying system " (Pharmaceutical Dosage Forms and Drug DeliverySystems), Howard C.Ansel, Loyd V.Allen, Jr., Nicholas G.Popovich, the 7th edition, 1999, Lippincott, William﹠amp; Wilkins publishes, and sees the 18th chapter).

Term " prodrug " need after using to refer to metabolism biological to transform to produce the chemical compound of required active constituents of medicine.

Term used herein " micro-cluster compositions " refers to contain the compositions of micelle grain water.Modified biological influences the compositions of agent, health inorganic agent, adjuvant or carrier or the adjective of its composition " micelle grain " and refers to micelle grain water in the compositions, and promptly it is dissolved in, mixes or otherwise in conjunction with micelle grain water.

" cell " is the fundamental structural unit of all live organisms, is made up of the small protoplasm group that just can see with microscope usually, includes by cell membrane and/or cell wall and the extraneous Cytoplasm that separates.Eukaryotic cell is to contain the nuclear cell that is included in the nuclear membrane.Prokaryotic cell is that genomic DNA is not included in the cell in the intracellular nucleic film.

" tissue " refers to form the cell aggregation of carrying out one or more specific functions.

" organ " is to adapt to and/or specialization is used to carry out any part of the many cells organic-biological body health of one or more vital functions.

Compositions of the present invention

Compositions of the present invention is a micelle grain water composition.These compositionss comprise micelle grain water and one or more and are selected from bio-affecting agents, health inorganic agent and adjuvant or the carrier compositions one or more activating agent.

Summary of the invention

Micelle grain water composition described herein comprises one or more and is selected from bio-affecting agents, health inorganic agent and adjuvant or the carrier compositions one or more activating agent.The biological nature of health inorganic agent comprises that (a) prevents, alleviates, treats or cure the unusual and pathological condition of live body; (b) keep, promote, reduce, limit or destroy the physiological organism function; (c) by in vivo test diagnosis physiological conditions or state; (d) control or protect environment or live body by attracting, disable, suppress, kill, modify, repel or hinder animal or microorganism.The health inorganic agent is optional from the activating agent that is used for deodorant, protection, decoration or ornamenting health.

The dosage form of composition of the present invention can be liquid, ointment, cream, gel, dispersion, powder, granule, capsule, tablet, and transdermal delivery device.In either case, these compositionss all can be Pharmaceutical compositions.

The method of using the present composition disclosed herein, these methods comprise the step that described compositions is applied to live body, or these compositions are applied to cell, tissue and organ outward.

On the other hand, the present invention also provides the preparation method for compositions, and these methods comprise the step that micelle grain water and one or more is selected from one or more activating agent merging combination in bio-affecting agents, health inorganic agent and adjuvant or the carrier compositions.

The prescription ultimate principle

The every kind of specific drug products that contains medicine or health inorganic agent all has the prescription of its uniqueness.Except the active treatment composition, pharmaceutical formulation also comprises many non-treatments or pharmaceutical composition.By using, prescription reaches its unique composition and physical property.Pharmaceutical composition comprises materials such as filler, thickening agent, solvent, suspending agent, tablet coating and disintegrating agent, stabilizing agent, antibiotic antiseptic, flavoring agent, coloring agent and sweetener.

Compositions all in the prescription all should be compatible on physics and chemical property, comprises active therapeutic agent, pharmaceutical composition and packaging material.

Pharmaceutical composition: definition and type

Medicine to be made dosage form or Pharmaceutical composition, need pharmaceutical composition, be also referred to as adjuvant or carrier in this area.For example, in the process of preparation medical solution, the aesthetics, the interpolation antiseptic that use one or more dissolution with solvents medicines, use flavoring agent and sweetener to regulate the taste of medicine, interpolation coloring agent raising medicine can prevent growth of microorganism, add stabilizing agents such as antioxidant and chelating agen and can prevent that medicine from decomposing.In the process of preparation tablet; usually can add that diluent or filler can increase the medicine volume, binding agent can make powdered drug and medical substance is bonding, antiplastering aid or lubricant help to make the tablet course of processing more smoothly, disintegrating agent can promote tablet disintegrate after dispensing, coating can improve stability of drug, or the control disintegrating agent can strengthen outward appearance.Because utilized the basis of medicine, ointment, cream and suppository are its property feature.Therefore for every kind of dosage form, ingredient has formed the principal character of product, and physical aspect, structure, stability, taste and general shape.

As everyone knows, pharmaceutical composition has many primary categories.For repeating no more, details about classification and example, please refer to monograph " pharmacy dosage form and drug-supplying system " (PharmaceuticalDosage Forms and Drug Delivery Systems), Howard C.Ansel, LoydV.Allen, Jr., Nicholas G.Popovich, the 7th edition, 1999, Lippincott, William﹠amp; Wilkins publishes.Please note the design of the 3rd chapter one dosage form especially: pharmacy and prescription points for attention.Table 3.3 in this reference content provides the non-restrictive example and the example thereof of pharmaceutical composition.Micelle grain composition of the present invention comprises the Aquo-composition of pharmaceutical composition and/or excipient.

The reader it shall yet further be noted that and " comprises in the pharmacy excipient handbook (Handbook of PharmaceuticalExcipients) and to have told about the monograph that kind more than 200 is used for the excipient of pharmacy dosage form preparation.The data that comprises in every piece of monograph has: non-patent, chemistry and trade name; Experiment and chemical formula and molecular weight; Pharmacy specification and chemistry and physical property; Incompatibility and interaction with other experiment material and medicine; The control state; And the application in pharmaceutical formulation or technology.

Dosage form

Except that the liquid dosage form of micro-cluster compositions, micro-cluster compositions of the present invention relates to the on-liquid dosage form (as described below) that contains micelle grain water.Referring to " pharmacy dosage form and drug-supplying system " (Pharmaceutical Dosage Forms and Drug Delivery Systems), HowardC.Ansel, Loyd V.Allen, Jr., Nicholas G.Popovich, the 7th edition, 1999, Lippincott, William ﹠amp; Wilkins publishes.

Solid dosage forms and improvement release administration system

Powder and granule

Capsule and tablet

Improve release dosage form and drug-supplying system

Semisolid and transdermal drug delivery system

Ointment, cream and gel

Transdermal drug delivery system

Medicinal implant

Suppository and implant

Liquid dosage form generally includes solution and disperse system.Sterile formulation and drug-supplying system comprise parenteral dosage form, biological preparation, ophthalmic solution and suspension.

Novel and senior dosage form, drug-supplying system and device comprise and are used to radiopharmaceuticals and the liposome diagnosing and treat.

As mentioned above, the micro-cluster compositions drug-supplying system of being made up of machinery, electronics and computer assembly that generally is used for injecting drug use is also contained in the present invention.The method that micro-cluster compositions is administered in the live body comprises the step of using machinery, electronics or computer module, is also contained in the row of the scope of the invention.The example of these medical treatment device auxiliary elements contain comprise iontophoresis, phonophoresis, dialysis, implantable pump, fluorine carbon boost pump, intravenous injection control measuring device and infusion pump drug-supplying system (referring to the 19th chapter, Ansel).Offeing medicine to the common drug-supplying system of vascular system comprises injection syringe, syringe needle or device, carries conduit, liquid combination container or the tube for transfusion of liquid between device and/or health or tissue.

The solvent for injection and the excipient that contain micelle grain water

The most frequently used solvent of large-scale injection manufacturer is the water for injection that meets the USP standard.Injection preferably adopts aqueous excipient, makes water in producing injectable product.Micelle grain water example comprises:

Meet the USP standard pure water, meet the USP standard sterile water for injection, meet the bacteriostatic water for injection of USP standard.

Meet the US standard sodium chloride injection, meet the USP standard bacteriostateic sodium chloride inj, meet the ringer's inj of USP standard, and the lactic acid ringer's inj that meets the USP standard.

Bio-affecting agents and health inorganic agent

Micro-cluster compositions of the present invention comprises the compositions of bio-affecting agents and health inorganic agent." bio-affecting agents " and " health inorganic agent " is the material that possible have following biology or medicinal property.These activating agents, material or medicine are the components of micro-cluster compositions of the present invention.Should be understood that on cell, tissue or the organ of biological nature live body and express.These biologies used herein or the term of medicinal property and standard medical dictionary are (for example, " Dao Lanshi medical science dictionary " (Dorland ' s Medical Dictionary)) consistent (for example with usage in the monograph, " pharmacological basis of treatment " (The Pharmacological Basis ofTherapeutics), Joel G.Hardman, Lee E.Limbird, the tenth edition, calendar year 2001, McGraw Hill publishes; " basis and clinical pharmacology " (Basic﹠amp; ClinicalPharmacology), Bernard G.Katzung, the 8th edition, calendar year 2001, McGraw Hill publishes; " pharmacy dosage form and drug-supplying system " (Pharmaceutical Dosage Forms andDrug Delivery Systems), Howard C.Ansel, Loyd V.Allen, Jr., Nicholas G.Popovich, the 7th edition, 1999, Lippincott, William﹠amp; Wilkins publishes.)

Though the health inorganic agent has drug influence, the primary implication that is used for " the health inorganic agent " of the object of the invention refers to be locally applied to live body and is used for the activating agent of deodorant, protection, decoration or ornamenting health.

Generally, the biological nature of bio-affecting agents and health inorganic agent comprises:

A. prevent, alleviate, treat or cure the unusual and pathological condition of live body;

B. keep, promote, reduce, limit or destroy the physiological organism function;

C. by in vivo test diagnosis physiological conditions or state;

D. control or protect environment or live body by attracting, disable, suppress, kill, modify, repel or hinder animal or microorganism.

The health inorganic agent includes but not limited to, toothpaste; Black preparation is covered or shone in the part with the sun or radiation; Repair first or pedicure compositions, live and send out or the skin-active bleach; Skin-active coloring agent (for example, lip pomade); Anti-diaphoretic or perspiration deodorizer; Live and send out or scalp care compositions; The body part preparation (for example, cosmetics for skin) that comprises the solid synthetic organic polymer.

The treatment classification of bio-affecting agents

Following classification of drug (indefiniteness) comes from Goodman﹠amp; " pharmacological basis of treatment " of Gilman (The Pharmacological.Basis of Therapeutics), Joel G.Hardman, Lee E.Limbird, the tenth edition, calendar year 2001, McGraw Hill publishes, for this paper theme is hereby incorporated by reference.Micro-cluster compositions of the present invention comprises the medicine with one or more following medicinal activities.

Act on the medicine of synapse and neural effector joint area

These medicaments influence unify neurotransmission in the somatic movement nervous system of autonomic nervous system.Include muscarinic receptor agonist and antagonist: anticholinesterase; Act on the medicament of neuromuscular junction and autonomic ganglion; Catecholamine, sympathomimetic drug, and adrenergic aceptor antagonist; 5-hydroxy tryptamine (serotonin) receptor stimulating agent and antagonist.

Act on central nervous system's medicine

This comprises anesthetic,general and local anesthetic; Treatment gas (oxygen, carbon dioxide, nitric oxide and helium); Hypnotic and tranquilizer; Ethanol; The medicine of treatment melancholia, anxiety disorder, psychosis, mental sickness such as manic; The medicine of treatment epilepsy; The medicine of treatment central nervous system degenerative disease; The class opioid analgesics; The medicine of medicine addiction and drug dependence.

Autacoid; The inflammation pharmacotherapy

This comprises histamine, bradykinin and antagonist thereof; The lipid autacoid of deriving: eicosanoid and platelet activating factor; The medicine that adopts in antipyretic-antalgic agent and antiinflammatory and the gout treatment; The medicine that adopts in the treating asthma.

Influence the medicine of kidney and cardiovascular function

This comprises diuretic; Vassopressin and other act on the medicine that kidney water is retained; Feritin and angiotensin; The medicine of treatment myocardial ischemia; Antihypertensive and the hypertensive medicine of treatment; The medicine of treatment heart failure; Anti-arrhythmic agents; The medicine of treatment hypercholesterolemia and dyslipidemia.

Influence the medicine of gastrointestinal function

This comprises the medicine of gastric acidity controlling agent and treatment gastric ulcer and gastroesophageal reflux disease; Gastrointestinal motility promoter and the medicament that is used for irritable bowel syndrome; Be used to suffer from diarrhoea, the medicament of constipation and inflammatory bowel; The medicament that is used for biliary tract and pancreatic diseases.

The chemotherapy of parasitic infection

This comprises the medicament of protozoan infection chemotherapy such as being used for malaria, amebiasis, giardiasis, trichomoniasis, african trypanosomiasis, leishmaniasis and treatment vermination;

The chemotherapy of microbial diseases

This comprises the medicament of antimicrobial such as sulfonamides, trimethoprim-sulfamethoxazole, quinolinones and treatment urinary tract infection; Penicillin, cephalosporin and other beta-lactam antibiotic; The aminoglycoside medicine; Protein synthesis inhibitor; Pulmonary tuberculosis, bird type tuberculosis compound bacteria catch and the leprosy chemotherapy in the medicine that uses.In addition, also comprise antifungal, antiviral agent and antiretroviral agent.

The chemotherapy of neoplastic disease

This comprises alkylating agent, chlormethine, aziridine and methyl melamine class medicine; Alkylsulfonate; Nitroso ureas; Folacin; Pyrimidine analogue; Natural drugs such as vinblastine, paclitaxel, epipodophyllotoxin; Camptothecin analogues; Antibiotic such as dactinomycin, daunomycin, amycin, jaundice element; Bleomycin, mitomycin; Platinum coordination complex; Hydroxyurea; Procarbazine; Adrenocorticosteroids; Aminoglutethimide and other aromatase inhibitor; Antiestrogen (for example, tamoxifen); Gonadotropin releasing hormone analogues; Antiandrogen; Biological response modifiers such as interleukin, granulocyte colony-stimulating factor, granulocyte/M-CSF; Monoclonal antibody.

Be used for immunoregulatory medicine

This comprises immunosuppressant, toleragen and immunostimulant.This type of medicine comprises based on the vaccine from the antibody compositions of immunoglobulin to the antibody purification compositions to monoclonal antibody combination.

Act on blood and hemopoietic organ's medicine

This comprises hemopoietic agent such as somatomedin, minerals and vitamins; Anticoagulant, thrombolytics and antiplatelet drug.

Hormone and hormone antagonist

This comprises pituitary hormone and hypothalamic releasing factor thereof; Thyroid and antithyroid drug; Estrogen and progestogen; Androgen; Thyroliberin; Adrenocorticosteroids and synthetic analogues thereof; Synthetic and the function inhibitor of adrenocortical hormone; Insulin, oral hypoglycemic; Act on calcification and osteoporotic medicament: calcium, phosphate, parathyroid hormone, vitamin D, calcitonin.

Vitamin

This comprises water soluble vitamins: compound vitamin B and ascorbic acid; Fatsoluble vitamin: vitamin A, K and E.

The medicament of treatment dermatosis; The treatment of eye disorders agent

Use the approach of the present composition

For the micro-cluster compositions of the present invention that is applied to live body, be to be used for part or systemic effect according to compositions, those skilled in the art can utilize and select different dosing ways.Method of the present invention comprises according to above-mentioned medical science or the treatment activity is used for the treatment of or the present composition of diagnostic purpose.Method comprise by in oral, Sublingual, parenteral, skin surface (part), transdermal, conjunctiva, ophthalmic, intranasal, ear, the respiratory tract, approach such as rectum, vagina, the urethra step of using or carrying pharmaceutical composition.According to method and the standard described in common and the training medical science standard textbook, the technical staff of treatment and diagnostic field can find the guide of using the present composition.

Treated in vitroAlternatively, the biological nature of micro-cluster compositions of the present invention can be applied to and be expressed on isolated cells, tissue and the organ.Assessment, the mammalian cell that screening and processing are cultivated, tissue and organ are gene therapies, stem-cell therapy (for example, navel blood stem cell is transplanted), move connect or transplanted cells/tissue (for example, hemopoietic tissue), tumor medical science (for example, host/graft/tumor interacts) and reproductive medicine is (for example, embryo culture) common scheme in (" from blood and bone marrow transplantation x: the tenth international symposium's summary " (AutologousBlood and Marrow Transplantation X:Proceedings of the TenthInternational Symposium), editor Karel A.Dicke and ArmandKeating, May calendar year 2001; " blood commentary " (Bloodline Reviews); " blood and bone marrow transplantation commentary " (Blood and Marrow Transplantation Reviews); " cell in vitro treatment " (Ex Vivo Cell Therapy), Klaus Schindhelm and Robert Nordon write).The purpose of external treatment is the biological function that replaces, repairs or strengthen damaged tissues or organ.External process comprises from patient or donor collecting cell, and manipulation in vitro is with the treatment potential that improves harvesting and intravenous fluids subsequently.

The method for preparing the present composition

The method for preparing micro-cluster compositions of the present invention comprises the step with one or more bio-affecting agents, health inorganic agent, adjuvant or carrier compositions and combination of micelle grain water or preparation.The guide of the relevant preparation present composition, those skilled in the art can be referring to the standard monograph of chemistry, clinical chemistry, pharmaceutical chemistry, pharmacology, dispensatory.

Diagnosis composition

Compositions of the present invention comprises diagnosis composition (" Mosby diagnosis and laboratory test handbook (Mosby ' s Manual of Diagnostic and Laboratory Tests), Kathleen Deska Pagana, Timothy James Pagana write); The inventive method is included in (being in-vivo diagnostic or in vivo test) of carrying out in the live body or uses micelle grain diagnosis composition in external or stripped diagnostic techniques of carrying out.The micro-cluster compositions that contains contrast medium that is used for the diagnostic roentgenology method is also included within the scope of the invention.Diagnostic reagent and its preparation method (Sigma Aldrich Co.; Worthington Biochemical Corporation; Wako Chemicals USA) and using method be widely known by the people in this area.The present invention includes the test kit that contains micro-cluster compositions.

IV. food or edible material and beverage: processing, compositions and goods

Summary and definition

Unless otherwise stated, the present invention uses general food technology, Food Chemistry, food processing, the organic and Measurement for Biochemistry in the skill of this area.In documents and materials, describe the technology that is used for Food ﹠ Drink in detail.For example, see also Potter, " Food Science " that N.N. and Hotchkiss, J.H. write (Food Science), the 5th edition, 1998, AspenPublishers published; " Food Chemistry " that Belitz, H.D. and Grosch, W. write (Food Chemistry), second edition, 1999, Springer published; " food: composition chemistry " that T.P.Coultate writes (The Chemistry of Its Components), the 4th edition, 2002, Royal Society of Chemistry published; " Food Chemistry " that Owen R.Fennema writes (Food Chemistry), the third edition, 1996, MarcelDekker, Inc. publishes; " characteristic of water among the food ISOPOW6 " that David S.Reid edits (The Properties of Water in Foods ISOPOW6), 1998; " food properties handbook (Food PropertiesHandbook), nineteen ninety-five, Culinary and Hospitality IndustryPublications Services publication that Shafiur Rahman writes; Brennan, J.G., Butters, J.R. etc. write, nineteen ninety, " food engineering operation " (Food Engineering Operations), Chapman and Hall publishes; Heldman, D.R. and Hartel, R.W. writes, and 1997, " principle of food " (Principles of Food Processing), Chapman and Hall publishes; " agricultural, food and biological engineering encyclopedia " (Encyclopedia of Agricultural, Food, and BiologicalEngineering), 2003, the editor: Dennis R.Heldman, Marcel Dekker, Inc. publishes; " food configuration-foundation and assessment " (Food Structure-Creationand Evaluation), 1987, J.R.Mitchell and J.M.V.Blanshard write, and Woodhead Publishing Ltd. publishes; " amorphous food and pharmacy system " (Amorphous Food and Pharmaceutical Systems), 2002, H.Levine edited, and RSC Publlshers publishes; Ruan, Roger and Chen, " water in food and the biomaterial " that Paul L. writes (Water in Foods and BiologicalMaterials), " magnetic nuclear resonance method " (A Nuclear Magnetic ResonanceApproach), 1998, Culinary and Hospitality IndustryPubl.Services published; Jose M.Aguilera and Stanley, " the micro structure principle of food processing and engineering " that David W. writes (Microstructural Principlesof Food Processing and Engineering), second edition, 2000, Culinaryand Hospitality Industry Publ.Services published; " functional characteristic of food super large molecule " (Functional Properties of Food Macromolecules), editor J.R.Mitchell and D.A.Ledward, 1986, Elsevier AppliedScience Publ. published; " stage in the food transforms " (the PhaseTransitions in Foods) that Roos, Y.H. write, nineteen ninety-five, Academic Press publishes.Can be from English American Technical Publishers, Ltd., Hitchin, Herts., SG40SX find at the place numerous Food Science of kind and technical manual.Online structure and the behavior of at large having introduced water of http://www.s bu.a c.uk/water/ comprises the effect of water in the food molecule hydration.The introducing of list of references cited herein or patent can be used as and replenishes the relevant instruction of the disclosure of invention.

The present invention relates to take place with structuring water (structured water) food or the eatable material and the beverage of hydration.On the one hand, the present invention includes food or eatable material and beverage that contains structuring water and structure component or the additive that participates in the food of preparation structure or non-structure.

On the other hand, present invention resides in utilization structure water in the Foods or drinks processing, comprise by composition or product coming in contact structuring water and at least a food processing system, and the step of hydration food processing system.

The present invention relates to utilization structure water and food material is handled or finished to the structural group compound.Particularly present invention resides in the method for the various water of utilization structure on that water rises, include but not limited to these effects of listing in the following table:

The effect of table-water in Food ﹠ Drink

Effect	Humidity range	The mechanism of action	Affected mass property
				Solvent	All-the eliminating bound water	Dissolving	All
Reaction medium	All-the eliminating bound water	Promote chemical change	All
				Reactant	All	Hydrolytic reagent	Taste, quality
Antioxidant	Low	The peroxide of hydration and precipitation of gold metal catalyst, bonding protein and carbohydrate and functional group, the reorganization of promotion free radical.	Taste, color, quality, nutritive value
				Prooxidant	In	Reduce viscosity, the activeness of augmenting response thing and catalyst.The expansion solid matrix exposes catalytic surface and oxidable group	Taste, color |, quality, nutritive value
Structure-intramolecularly	All	Keep the complete of protein molecule	Quality and the characteristic that influenced by enzyme
				Structure-intermolecular	Low	With surface group hydrogen bonded and the macromolecular crosslink sites hydrogen bonded on the macromole	The viscosity quality-in dehydrated food
Structure-intermolecular	Neutralization is high	Influence Emulsion structure (for example, mating surface lipid).Influence gel formation polysaccharide and protein interactions and conformation.	The rheological behavior of Emulsion and the texture characteristic of gel.

As following " Food Science " third edition of writing from Fennema reprint out form proposed, the classification of water-solute interaction type is further characterized the solute hydration of structuring water in food processing.

The classification of table 3 water-solute interaction type

Type	Example	With the interaction strength that water-the water hydrogen bond is compared a
			Dipole ion dipole-dipole hydrophobic hydration hydrophobic interaction	The charged group water of water-You on ionized water-You machine molecule-protein N H water-protein C O water-side chain OH water+Rc→ R (hydration) R (hydration)+R (hydration) → R 2(hydration)+H 20	Bigger bMuch lower about equally (Δ G＞0) can't compare d(＞hydrophobic interaction; (Δ G＜0)

^aAbout 12-25kJ/mol.

^bBut it is more a lot of than covalent single bond weak strength.

^cR is an alkyl.

^dHydrophobic interaction is that entropy drives, and the interaction of dipole-ion and dipole-dipole is the enthalpy driving.

Structurize product that any physical form is provided and compositions that the present invention is total, it is used for being consumed by the oral cavity of the mankind or animal is all or part of.In addition, the present invention also comprises the structuring water that exists with any physical form.

The present invention can be used for food engineering, Food Chemistry and biological food and learn the field.

Food engineering comprises food manufacturing, processing, packing and storage.Be similar to the effect of water, the method for structuring water and compositions thereof and the processed food that comprises the structuring water hydration process of introducing herein is applicable to the fluid machinery in the extruding and mix, the dough rheology, mix the prediffusion of flavoring agent, understand the mechanism of intumescence in the extruding, the both macro and micro structure of food, cure and microwave processing, the heat during mixing is cured and the synchronous transmission of quality, ice crystal size control in thin film technique and the refrigerating process, the steam jet impact cures, promote healthy by processed food, food wastes and non-staple food utilization, improve packing and intelligent packaging for guaranteeing microbial safety.

Food Chemistry applied chemistry technology, notion and law are determined the kind and the quantity of molecule in the food, its physical attribute and chemical change in manufacturing and storage process.Structuring water and compositions thereof are extensive with the method subject range of the processed food that comprises the hydration process of introducing herein, from analyzing food composition to the molecular mobility of measuring amorphous solid; Lipid, carbohydrate and proteinic chemical change, process technology for example sprays, controls antimicrobial or ice crystal nucleus albumen; Spectrum, machinery and thermal technology be qualitative noncrystalline matter, noncrystal, solid physical attribute how, thereby changes its chemistry and physical attribute, even shelf-life and stability.

Definition

Appear in the patentability subject categories described herein various, but be not included in the implication of " technology " term in the following vocabulary, and generally acknowledge usually or commonly used identical.

Term " water ", " structuring water " and " micelle grain water " are used interchangeably." structuring " subject matter of an invention and scope are informed and similar (U.S. Patent number 6,521,248) in the deutero-implication of this its application context by term " water ".

Term " food/food " and " edible " synonym of using can exchange use herein.

No matter every kind of composition that uses in the food processing system or additive are naturally occurring product or the synthetic product of producing, and as the part of edible compositions, or handle edible compositions, or disclosed or claim and will be considered to edible into edible.

Food or eatable material are included in the beverage that extensively defines in the classification 426.For instance, but be not limited thereto, subclass 590 comprises the liquid of drinking or adds the concentrate that hydrous material forms the liquid of drinking.Comprise subclass 569 (forming foamy beverage) and 580 (beverage containing milk).Subclass 592 comprises and contains alcoholic acid product in the theme.From the US patent class handbook that United States Patent and Trademark Office obtains, can find each beverage that divides apoplexy due to endogenous wind tabulation and definition thereof and relate to the classification of subject area.

The compositions that contains structuring water is called " micro-cluster compositions " herein.The expression compositions modified in adjective " micelle grain ", and () noun for example, material, additive, composition shows in the compositions of modification and contains micelle grain water, as the result who is carried out hydration to small part by structuring water.Abbreviation MCW represents structuring water.

On the one hand, the food processing system comprises food material or the inherent structure in preparing burden resolves into the degree that has nothing in common with each other, thereby relate to the chemistry of all aspect-food of food and physical characteristic with and the packing and the sale of the processing of composition, food and production, food, it represents the constituent in the food processing system.But the food inner structure forms for food quality-quality, the fragrance that distributes, nutritional labeling availability, moisture moves and growth of microorganism-be subjected to, stability and the influence and the decision of decomposing.Every kind of composition that uses in the food processing system or additive, no matter be the naturally occurring product or the product of synthetic,, or handle edible compositions as the part of edible compositions, or it is disclosed or claim and be to be considered to edible by edible.Food processing comprises and converts raw material and batching to consumer food or edible product.Food processing comprises that all change raw material or animal material or change into safe, edible and all operations of tasty food more.Another target of food processing is to improve storage or shelf life.

Hydration chemistry in the food processing

The present invention relates to utilization structure water in food processing.With the combined water of carbohydrate, lipid and protein, be one of food main component.Therefore, the present invention relates to the method for implementation structure water and carbohydrate, lipid and protein bound in the food processing system.

The hydration characteristic of water partly rely on it bunch the collection (water-bound and behavior comprise the effect of water in food molecule hydration, and detailed online explanation is arranged on http://www.sbu.ac.uk/water/; Simultaneously in " characteristic of water among the food ISOPOW6 " (The Properties of Water in Foods ISOPOW6) that David S.Reid edits, explanation is arranged also).Structuring water determines its three dimensional structure to the hydration characteristic of biomacromolecule (particularly protein and nucleic acid), thereby determines its function in solution.

Utilization structure water can improve quality, mixing and flow behavior and function in food processing.MCW also relates to many and interaction other food composition.These interactions have and help confuse molecule, make it be in amorphous state, for example, and low moisture food.In amorphous food system, gamma transition is distinctive temperature range, surpasses the firm material softening of this scope and begins to show tough characteristic.This variation is the material specific change of physical state from " glassy " mechanicalness solid to " rubber-like " viscous fluid of a kind of temperature, time (or frequency) and compositing dependence.

MCW makes the amorphous materials plasticizing, strengthens crystallization.The plastication of MCW causes that molecular mobility increases, and helps to arrange molecule, and may strengthen enzymatic reaction.The effectiveness of MCW is a factor of enzymatic reaction speed in the amorphous food system of influence.MCW can significantly plastify food material.Increase the amount of MCW, the water activity of material also can be higher.Plasticizer is used to improve the flexibility and the machinability of polymer, together to also reducing viscosity.

Enzymatic reaction causes the harmful variation in the low moisture food usually.The speed of this variation is relevant with the variation of physical state, for example gamma transition.Water with common form and structured form is most important food material plasticizer.Water and other plasticizer also can influence the speed of enzymatic reaction.Even reduced moisture, comprise the also very easy influence that is subjected to crystallization of food system of carbohydrate (for example sugar).In the crystallisation procedure does, the water of absorption is discharged in the food material, and the moisture in the food system is changed, and may influence the speed of enzymatic reaction.Water is as the effect of plasticizer, and is to keep the quality of low moisture food system and the key factor of shelf life as the speed of food matter structure function effect enzymatic reaction.

The physical state of food system depends on the amount of water and other plasticizer and the interaction of molecules type that relates to all the components.

" water combination " and " hydration " are meant that water is with various toughness and the hydrophilic or bonded trend of hydrophobic substance.Hydrophilic solute (hydrophilic solute or structure are for example arranged) is stronger or suitable with the strength ratio water-water mutual effect of water mutual effect, and hydrophobic solute (hydrophobic solute or structure are for example arranged) only with the faint interaction of water, its intensity is far below water-water mutual effect.

Known in this field about determining molecular species (comprising food) hydration and hydration method (" the food properties handbook (FoodProperties Handbook) that Shafiur Rahman writes to the food quality influence, nineteen ninety-five, Culinary and HospitalityIndustry Publications Services publishes).

The hydrogen bonded site of water contention intramolecularly and intermolecular hydrogen bonding is the main determining factor of carbohydrate, protein and lipid conformation.

Water is to the effect of protein structure

Hydration is very important for proteinic three dimensional structure and activity.In fact, azymia activity under the situation of hydropenia.In solution, they have the conformation motility, comprise hydration status miscellaneous, and this is in crystallization or can't not see in the aqueous environment.Balance between these states is looked water in its microenvironment ActiveAnd decide; The degree of freedom of water and protein hydration for example.Therefore, strong reactivity water (the high density water that for example contains many weak bendings and/or fracture hydrogen bond) helps the protein conformation of the strong hydration of needs, help ' desiccant ' conformation relatively than low activity water (low density water that for example, contains many strong moisture hydrogen bonds of intramolecularly).

Protein Folding depends on controls some PolysaccharideIn the same factor that forms of bonding land; It is the incompatibility between low density water (LDW) and the hydrophobic surface that drives this group formation hydrophobic core.In addition, water changes as discharging essential hydrogen bond as lubricant.Hydrone can the bridging carbonylic oxygen atom and the ammonia proton of different peptide bonds, thus the formation that promotes the peptide hydrogen bond with separate.Interior molecules motion in the protein is that biological activity makes so, and it extremely relies on the plasticizing rate that is subjected to the decision of hydration intensity.Therefore, it is folding that internal water can make protein take place, and the interaction of the protein chain by cooperation is discharged from hydrophobic central core when finally extruding.It is very important for balance flexibility and inflexible enzymatic activity to demonstrate enzyme equilbrium position on every side.

Form the very nonpolarity base of high surface area owing to be subjected to the adverse effect and the water of entropy minimizing, make protein folding be subjected to the driving of hydrophobic interaction.In the protein denaturation process, water is not only for proteinic correct folding very important, and also very important to keeping this structure.Folding or separate folding Gibbs free be because protein folding/separate the joint effect of folding and hydration change.

Peptide and protein work in foam, gel, emulsifying, fragrance precursor material, perfume compound and use as enzyme.These characteristics come from aminoacid and proteinic physicochemical characteristics.As mentioned above, proteinic hydration plays an important role to proteinic function, comprises protein bound composition of food, gelling, expansion, dough making, emulsifying and foaming.The catalytic activity of enzyme and the control of enzyme reaction need possess the knowledge of protein hydration and moisture microenvironment aspect.The crucial enzyme of food processing is comprised oxidoreductase, transferring enzyme, hydrolytic enzyme, lyases, isomerase and ligase.The water activity plays pivotal role in the regulatory enzyme reaction.

Even in low moisture food, although the water activity is very low, changing still can appear in enzyme.Owing to these reactions take place, thereby the product stability of stored are reduced.Water plays a different role in food system: (1) water can be used as second kind of substrate.As everyone knows, the stereochemical structure of multiple interaction partners decision protein function characteristic plays Stabilization, these effects comprise between polar group and polar group and water between hydrogen bond, the hydrogen bond relevant around the protein molecule with water-bound; (2) thus change the material of protein structure as the division hydrogen bond; (3) as the dissolving media that promotes the reactant diffusion.

In short, the activity of enzyme is decided on the interaction between water and enzyme, water and substrate and water and the substrate.Also may comprise simultaneously the interaction between substrate and substrate and substrate and the enzyme.

At last, in the low moisture system, carry out the water of catalyzing enzyme reaction needed some, to promote the animal migration and the diffusibility of reaction.According to the characteristics of enzyme and the dissolubility and the molecular size of substrate, can change the quantity of water.

Enzymatic reaction comprises reacting to each other of enzyme and substrate, and at this moment, water is usually both as solvent, again as second kind of substrate.The hydrolysis of sucrose needs the water-disintegrable key contacts of invertase and sucrose.If dewatering system must add water to recover the activity of enzyme.Therefore, composition must possess animal migration.Water should spread by system, and enzyme can apply certain animal migration with approaching water-disintegrable key, and perhaps substrate need move to the active site of enzyme.The speed of enzymatic reaction should depend on movement velocity, and this viewing system matrix structure and deciding.For example proved in memory fluid, to have polysaccharide, can cause polysaccharide chain to interweave, and the diffusion of restriction hydrone.

In the water limited systems, can think that animal migration is restricted.The activity of enzyme will be decided on its compactness to substrate.Therefore, the enzyme that should distribute by this way is so that it is effective around substrate.The intersolubility difference also can reduce response speed, because it can reduce intermolecular interaction.Composition, structure and comprise the physical state and the dynamic characteristic of the environmental condition decision systems of water content, temperature and pH value.

Whitaker (" the zymetology principle of Food Science " (Principles of Enzymologyfor the Food Sciences), 1994, second edition, Marcel Dekker, Inc. publishes; " Food Chemistry " that Fennema, O.R. write (Food Chemistry) the 7th chapter, the third edition, 1996, Marcel Dekker, Inc. published) effect of water to enzymatic activity be described.Water in all enzymic catalytic reactions, play a part at least four important: (1) unfolded protein, (2) be as the transhipment medium of substrate and enzyme, the ionization of prototropic change group in (3) proteinic hydration and (4) enzyme active sites.

The nucleic acid hydration

Hydration is very important to the conformation and the effectiveness of nucleic acid.Because the suitable electron distributions of diffusion, make to keep bigger and stronger hydration around the phosphate group, and base since its revise and decide to the hydrogen bonded ability, its hydration on every side is more orderly and more lasting.Because the tactical rule of DNA keeps hydrate water with cooperation mode along the Double helix in major groove and the ditch.The cooperative nature of this hydration helps double-helical annealing and untwists.

Nucleic acid have many groups can with the water hydrogen bonded (be ribose 02 ') and unpaired base site because the excess oxygen atom, RNA has hydration greatly than DNA.In DNA, base participates in the hydrogen bond pairing.Even yet these groups, except the theheterocyclic nitrogen atom (pyrimidine N3 and purine N1) of hydrogen bonded, also further hydrogen bonded is connected to water in major groove or the ditch.This type of solvent interaction partners hydration environment is very important, so its identification around nucleic acid directly helps the DNA conformation.

The water activity

The water activity has become exceedingly useful instrument in Food Science and technology.It is very useful when the zone mapping of relevant moisture dynamic transfer and growth of microorganism, physical change and chemical reaction.Control water activity is for reaching required food stability and predicting that the shelf life of product is very crucial in food processing system.

The water activity, a _w, be the characteristic of water in material.In 1970 mid-nineties 90s, the water activity becomes the principal element of control food spoilage, mainly is to be used for moisture minimizing, dried foods, medicine and biosystem.According to the study, rotten general fashion, i.e. the chemical reaction of physics and physicochemical change, growth of microorganism, aqueous and lipid all is subjected to the influence of the total moisture content of the thermodynamic activity of water and system.The chemical potential difference of water between two systems, this can cause exchange of moisture, exceeds certain chemical potential, as the molecular weight of system, when abundant water occurring, can cause physics and chemical reaction.

Overstate from function and sense organ the angle food wanted or the physical arrangement of biological product are often because moisture increase or lose and cause the water activity change to change.For example, the powder caking is the result owing to the metamict crystals shape transfer of saccharide and oligosaccharide, and this occurs when the active increase of water surpasses glass transition point.This caking influences powder dissolving or free-flow, and state-transition can cause the oxidation of volatile los or sealing lipid.If the moisture content increase causes the water activity to be elevated to a limit, surpass gamma transition once more, then cookies, do the heart, for example chrisps and frumentum breakfast will be not crisp.On the contrary, owing to lack the relation of water, the water activity reduces, and dried Fructus Vitis viniferae and other dry fruit can hardening.Therefore, be surrounded by sugar-coat on the dried Fructus Vitis viniferae in frumentum breakfast or other dry fruit,, thereby prevent moisture loss with the loss rate or the use glycerol minimizing water activity of the moisture that slows down.The remaining water transport of these measures inhibition dried Fructus Vitis viniferae is in frumentum, and therefore, the flexibility of the fragility of maintenance frumentum and fruit material is being put the chemical potential driving force in front people.Finally, as a _wIncrease equally, because the fast development of rubbery state makes packaging film improve greatly the permeability of oxygen and water evaporation.

As physical chemical phenomenon, the production of microorganism and death also are subjected to the active influence of water.Through proving that repeatedly every kind of microorganism all has a kind of critical water activity, is lower than it and just can't grows.For example, aspergillus parasiticus is not grown when active being lower than certain water, and can be suppressed below being created in high slightly water activity from the flavacin (a kind of doxycycline) of this microorganism.In order to stop growth or Production of Toxin, just must stop the main enzymatic reaction in the microbial cell.Therefore, reduce the water activity and suppress these biochemical reaction, like this can be on the whole the function of restriction micro-organisms successively.For spore, the water activity is low more, and killing property of heat resistanceheat resistant is just strong more.

Microbiologically stabilised dry food is generally defined as the food that the water activity is lower than a certain prescribed level, can grow in this no known microorganism below horizontal.

The water activity has been proved to be the kinetics that can influence many chemical reactions.Except lipid oxidation reaction, its speed when the water activity is very low reduces and outside increasing, other chemical reaction velocity all increases with the active increase of water usually with water is active.

When water and solute and surface interaction, can not carry out other hydration.The available balance water gaging of material hydrates effect described in term ' water activity '; Unit amount is represented pure water, and zero representation does not have hydrone fully.It and Food Chemistry and preserve closely bound up.

The active variation of water may cause that water moves in food composition.Contain the active food that contains the pond of the different water of active macroscopic view of different water or microcosmic, the migration to the active low zone of water from the active high zone of water of time of origin and temperature dependent water will be tended to, this is the useful properties that is usually used in processed with salt fish and cheese, but under other situation, may have catastrophic sensory results.The active this variation of water may cause the migration of water in food composition.The food that the water activity is lower trends towards obtaining moisture, and the active food with high of those water trends towards drying out.

Because active the hanging down of water can prevent growth of microorganism (prolongation shelf life), cause for example crispness generation significant change and change chemical reaction and (increase hydrophobic lipophilic reaction of texture characteristic, but reduce the reaction of hydrophilic restriction water diffusion) speed, so control water activity (rather than water content) is very important in the grocery trade.

In food technology, can determine free moisture with term water activity.The water activity is defined in the ratio of saturated vapor pressure under water vapor pressure in the sealing chamber that is placed with food and the uniform temp.Therefore the active indication discovery of water unbound water can be used as solvent or participates in the degree of destructive chemistry and microbial reaction.

The food a that very easily addles _w＞0.95.Be lower than a _w=0.91 o'clock, the growth of most antibacterials was restricted; Equally, be lower than a _w=0.87 o'clock, most yeast stopped growing, and are lower than a _w＞0.80 o'clock, most mycetes stopped growing.The absolute extremes of growth of microorganism approximately is a _w=0.6.When needs produce a _wDuring＜0.96 solute concentration very high (common＞1 mole), solute (surface interaction when water content is low) can effectively used a _wThe structuring of ken internal control controlling the water circulation.

The processing of numerous food product storage is attempted eliminating the purpose that addles by reducing the availability of microorganism to water thereby reach.Reduce the quantity of free moisture or unbound water, also can reduce other unnecessary chemical change, can be used for handling the food of storage.The method that is used for reducing the amount of food unbound water comprises various technology, for example concentrates, dehydration and freezing.These processing methods need intensive energy expenditure usually, do not have economic benefit.

By controlling the stability that the water activity can successfully realize food, the transfer of prediction moisture between the multicomponent food system predicts that steam comprises the final water activity of dissolving type by the transfer and the prediction blending constituent of packaging for foodstuff.

Molecular mobility.Molecular mobility (Mm) is a kind of method that develops in Food Science recently, is used to the storage stability that how to change food by freezing and oven dry is described, and be water activity (a _w) a kind of alternative and compensation process of notion.

Food material can not form crystal structure mostly.Become crystal, the molecule in the solution must be placed an existing lattice, and a bit as a seven-piece puzzle, it only can same direction be placed.Molecule rotates in solution and is crooked, but all moisture leave stop with activity before, must guarantee that they form crystal and want enough soon.When relatively slow micromolecule crystal carries out drying operation, might form: the sucrose of megacryst and salt.But, move slowly macromole or fast drying operation can't provide time enough for crystalline formation, in fact, in most cases can't form crystal.On the contrary, solution becomes gets very sticking, and picture rubber at last becomes.If remove more juicy, it is more and more sticking that rubber becomes, and until arriving a certain critical point, stops fully moving, and material can be considered a kind of glass.Glassy and elastomeric material is described to amorphous solid.Freezingly be regarded as and dry very similar process.Water crystallization Cheng Chunbing, it does not carry out the dissolving of food material.When food product refrigeration becomes the ice form, with food storage day by day the dehydration environment in.

In all cases, main parameter is the ability that molecule mobility-molecule moves.The molecule mobility increases along with temperature (molecule heat energy is many more, move fast more) and micromolecule concentration (common water is as molecular level lubricant or plasticizer).Baking step has reduced water content, has also reduced the molecule mobility of solute simultaneously.And the freezing water content (form of ice crystals) that also reduced, cooling reduces the heat energy of food molecule in addition, thereby has reduced their mobility.

Molecule mobility and its viscosity of material is inversely proportional to (if liquid is thick more, the molecule mobility is poor more), the speed of viscous effects diffusion limited reaction.If two intermolecular reacting, molecule at first must collisions, overcome the activation energy barrier after possessing enough heat energy, react.

Two kinds of technical methods that food is processed into vitreousness are freezing and oven dry.The molecule mobility is an a kind of novel method of understanding water role in food spoilage, is to a _wThe useful of method replenished.In general, use the analysis meeting of molecule mobility relatively good in diffusion limited reaction, frozen food and physical change, it is also more suitable to understand crispness and viscosity, and a _wThe food and the non-diffusion limited that then are more suitable for drying are processed.List some in the following table and depended on ambulant food properties of molecule and behavioral trait:

Table 7 is subjected to some food quality and the behavioral trait (diffusion-restricted that comprises in the product of amorphous region changes) of molecule mobility decision

Do or half-dried food	Frozen food
		The volatile matter escape that food cracking during the sugar flower oven dry of the chocolate Zhong of flow behavior and Nian crystallization and Zai crystallization is done and the structure of Zhi ground freeze-drying second (desorb) the stage Zhong of the food containing water such as Zhong is caved in and is sealed in solid amorphous matrix Zhong, the gelatification of enzymatic activity Maillard reaction starch	The contraction (part of foam-like frozen dessert is caved in) of caving in of the structure of water translocation (ice crystal, form the ice in the packing) lactose crystn (frozen dessert Zhong " sandy ") enzymatic activity lyophilization (first) stage Zhong amorphous phase

The hot deactivation of the cracking microbial spore of baked foods when bakery product wears out cooling owing to the starch degeneration

Gamma transition and water activity: the physical property of rubber and vitreousness and food stability

Phase transformation and attitude become.Phase transformation is that the state of material changes, and occurs in well-defined transformation temperature-fusing (solid-state to liquid)-crystallization (liquid state is to solid-state)-vaporization (liquid to gaseous state)-concentrated (gaseous state is to liquid).Many materials comprise that food all is noncrystal, but can show the characteristic of solid or liquid.Amorphous material is an amorphous material, and for example its molecule is a random alignment.The normally supercooled liquid or solid of amorphous material.Supercooled liquid is commonly referred to " rubber ", fixedly is commonly referred to " glass ".Surpass certain temperature range, the conversion between supercooled liquid and the solid state will take place, this conversion is called as " gamma transition ".

Gamma transition is comparatively typical in inorganic and organic amorphous material, comprises this as sugar and proteinic food composition.When surpassing the temperature range of gamma transition, many properties of materials can change.

The water plasticizingWater is most important solvent, disperse medium and plasticizer in biological and food system.Plasticizing and modulating action thereof in the temperature range of gamma transition are the key technologies of synthetic polymer technology, and in the middle of this, plasticizer is defined as incorporating into polymeric material, with machinability, flexibility or the easy autgmentability that increases material.The glass transition temperature of quality, volume or the molfraction of relevant water is depended in plastication usually.Reduce glass transition temperature when increasing water content, can observe the water plasticizing process, so also can improve the detectability that transforms.Carbohydrate and protein all can significantly be plastified by water, and for example water suppresses the temperature of gamma transition as softening agent.The glass transition temperature of water is about-135 ℃, for example solid amorphous attitude water.When water content was high, gamma transition was near the temperature of water.When surpassing the conversion temperature scope, the detectability of gamma transition is usually along with increasing water content-reduce to transform width-increases thermal capacity variation and increasing.

Gamma transition in the food

Understand gamma transition and very important for the state and the behavior of food in prediction processing, dispensing and the storage with the relation of physicochemical change.

The gamma transition curve is a key factor of understanding the food physical change.For instance, in the cereals system of processing, recognize that the quality variation in the frumentum system can be relevant with gamma transition, know the state diagram of cereals, control is processed and environmental condition then, so that obtain the required state of food, in dispensing and storage, kept very important simultaneously.

The solid amorphous state of fat-free food is very typical in low moisture and frozen food.Typically noncrystalline, glassy or rubber-like food is-freeze the solid that contracts in the crystal sugar-free-flowing powder-frozen food of the fruit done and vegetable-compression dessert and breakfast frumentum-hard.The gamma transition of food material can change from thermal capacity, observe from mechanical qualitative change with from dielectric property changes.The temperature range of gamma transition depends on the food composition that food material-molecular weight is little, for example sugared, when surpassing about 20 ℃ of temperature range, represented the big food composition of gamma transition-molecular weight clearly, for example protein and starch, the gamma transition of its performance is very broad.

The glass transition temperature scope is the particular community of every kind of material.

CarbohydrateSugar has gamma transition clearly.The glass transition temperature of sugar raises along with the increase of molecular weight.

ProteinAmorphism protein is important structure biopolymer.Amorphism protein is the important constituent of cereals, for example Testa Tritici in the bread.Because the variation of thermal capacity and conversion range are less, thereby proteinic gamma transition is difficult to determine with the throughput full-boiled process usually.

Refrigeration materialFreeze in the refrigerating process and can cause the freezing contraction of solute.The program of freezing contraction depends on solute and temperature.Under the low temperature condition, the solute of freezing contraction and the water vitrify that does not freeze, for example material comprises crystalline ice state and amorphous state, glassy solute state.Some solute can crystallization, and for example the sugar of NaCl solution-freezing contraction and food change vitrification usually.Very likely, temperature factor is depended in the gamma transition that freezing contraction solution shows when initial concentration, is higher than this temperature, and the ice fusing has a beginning temperature.

During concerning between definition water content and chemical reaction velocity, polymer science provides gamma transition and the active theory of water, explain the texture characteristic of food system and the variation that takes place at food processing and storage, for example viscosity, conglomeration, softening and sclerosis.Food is the complex mixts that lipoid, polysaccharide, sugar, protein etc. constitute, and is present in each different state.Influencing possibility part difference on the water content of gamma transition.

For instance, if amorphous material exists with vitreousness, then it can be very hard and frangible, and for example for frumentum, it represents friable product.In rubbery state, this material is soft and very flexible, and for fried snack or corn, it represents underbaked defective mode.

Therefore, the gamma transition theory provides a method more clearly, and to understand when water content increases, the crisp frumentum or the physics of dessert and quality change.Quality is the important organoleptic feature of many cerealss, lacks necessary quality and can diminish product quality and reduce shelf life.If the water activity breaks bounds, then the crispness of refined salt soda cracker, puffed rice, popcorn volume, expanded rice cake and potato block is just not enough.Crispness is the feature of key between starch molecule, when low amounts of water occurring, just forms little crystalloid zone.These zones need firmly to split, thereby provide crisp quality for food.Surpass certain water activity, water may interrupt these keys, allows starch molecule to slip over mutually when chewing.The sound that the frumentum dessert of doing crisp felt to take place when chewing causes that it reduces with the active enhancing of water.Conversion from the vitreousness to the rubbery state can be explained the disappearance of crispness well.

Caking is another feature relevant with gamma transition.When sugar in solution and when being the drying shrinkage state, it is in the non-crystalline glass state, powder free-flow.When reaching sufficiently high moisture or temperature, material enters rubbery state.In rubbery state, the non-crystallizable sugars of doing is easy to rapid crystallization, and this is that diffusion velocity increases owing to surpass certain temperature, and certain condition causes occurring tedious caking, thereby suppresses to flow automatically.Caking is followed the characteristic by steam moistened particles step.

The selection of batching and the rank of plasticizer, for example water influences the glass transition temperature of food with the little branch that becomes of other molecular weight.In general, when the polymer molecule amount in the homologous series increased, glass transition temperature also raise.Increase the temperature that plasticizer can reduce gamma transition.

The effect of water to spreading in the food system

The operational approach of numerous food product processing and the stability of foodstuff storing are subjected to the influence of the diffusion property of food system, any barrier (packing or coating) that they comprise the direct environment in food itself, the packing and are used for food.Water content and these diffusion properties of " water activity " appreciable impact by to the food plasticizing and/or to polymer packaging, influence the glass transition temperature of composition, and water also can be as the internal forwarding medium sometimes.

Term used herein " additive " is meant material or mixture of substances, be mainly used in the purpose beyond its nutritive value, and be added in the food with relatively small amount and produce with (1) or improve required characteristic (2) or suppress unwanted characteristic and (3) become the part of food or cambic.(compare with following composition, it can be additive in some cases).

Term used herein " basic ingredient " is meant the main component (except that the water that adds) of compositions, is considered to essential part, and people determine compositions by it usually.Usually, basic ingredient has constituted the major part of compositions, and for example chocolate milk-milk is basic ingredient.Providing under the situation of most percentage compositions, the composition that promptly constitutes whole compositions (except the water of interpolation) 50% is considered to basic ingredient.Can be by analogous components being sued for peace determine 50%, for example lactose, milk surum and butterfat all are newborn derivants.

" carbohydrate " is meant a kind of chemical compound, and the monomer whose unit comprises at least five carbon atoms, and product, and wherein the carbon skeleton of carbohydrate unit is not damaged.Corresponding to the pure and mild acid of carbohydrate, for example sorbitol ascorbic acid or mannonic acid are not considered to be carbohydrate.

Term " is done " and is meant anhydrous or anhydrous relatively product fully, under normal environmental condition, comprise such characteristic, but unnecessary have every specific character, as free-flow, sense of touch does, sticking, no adhesion strength, granule, powder, fritter, thin slice, flour, coarse powder, microgranule, spherolite, in small, broken bits etc.

Term " ferment " is meant any enzyme or any active organism that can cause or change fermentation.

Term " composition " is meant the ingredient (normally mainly forming) of the mixture that constitutes food.

Composition or additive do not comprise packaging material, container, paper products etc., or any other is regarded as the material that is not suitable for eating.But in some cases, additive can be a composition.

" isolating fat of triglyceride or oil " is meant without any the fat of its plant that is derived from or animal tissue or oil (as giving a definition).

" packing " be meant totally-enclosed by eatable material by solid material, surround or fully around commercial assembly.

" tissue " is meant the material (with respect to extract, it is considered to lack primary cellularity) that comprises a certain amount of initial animal or plant.Comprise the material of handling through shred, cut, smash to pieces, grind, roll, section etc. in this term.

" fat of triglyceride or oil " is meant the esters (for example, comprising the monocarboxylic acid and the carbonyl bonding of the chain that do not rupture of at least 7 carbon) of glycerol and higher fatty acids, and wherein three available glycerol hydroxy functional groups are by identical or different fatty monocarboxylic acid esterification.Triglyceride is the main component of natural fat and oils.

The present invention includes can following classification according to water food or edible product:

Table I is classified to food according to water content and suitable physico-chemical process type

Physical state	Product example	Physical chemistry is handled
			Rare molten solution/dispersion half weak solution/dispersion, (high-moisture) solid particle, (high-moisture), (medium moisture), (low moisture)	Beverage, soup beans, the fruit jelly fish, vegetable, meat, ice cream preserve, the sausage dried food and nuts, frumentum	Epuilibrium thermodynamics is referring to Henry's law polymer chemistry, chain entanglement, collosol and gel conversion biophysical chemistry, colloid science material science materialogy, glass/rubber transforms

Below discuss and introduced the Food Science those skilled in the art and be used for synthetic food and the employed physical and chemical principle of composition.

Colloid and rheology

Colloid is the spread condition of the small-particle of a phase (decentralized photo) in the second, continuous phase.Colloid extensively exists in food.Colloidal research comes down between the decentralized photo surface and physical interaction research between continuous phase and the decentralized photo.Rheology is the research to when distortion material.

Numerous food product is colloid and characteristic complexity, has continuous phase, exists with the form of true solution, and a plurality of decentralized photos are arranged.Milk has and comprises polysaccharide in the aqueous solution, electrolytes and proteinic continuous phase and comprise liquid fat and the decentralized photo of solid-state protein matter.The table gel-type

Type	Decentralized photo	Continuous phase	Example
				Aerosol, flue dust	Liquid	Gas	Flue dust
Mist, vaporific aerosol	Solid	Gas	The gas of breathing out
				Foam	Gas	Liquid	Whipped cream, play foamy egg.
Emulsion	Liquid	Liquid	Milk, mayonnaise
				Colloidal sol, colloidal solution, gel, beans	Solid	Liquid	Muddy medicated beer, milk, gel, tomato sauce
Solid foam	Gas	Solid	Ice cream, meringue

Emulsion and surface activity.

Emulsion is that decentralized photo and continuous phase all are the colloids of liquid, is the most frequently used food gel-type.In this food, generally include oil phase and water and these two types:

● oil-in-water (o/w) emulsion, decentralized photo are oil

● Water-In-Oil (w/o) emulsion, decentralized photo are oil.

Phase in the emulsion can exchange by the method that is called phase inversion.The common example of phase inversion is the butter manufacturing in the food, and it changes into butter by the processing method in concentrating and being stirred in butterfat.In case obtain enough oil concentrations, stirring can change into the oil-in-water emulsion of butterfat the water-in-oil emulsion of butter.In the course of processing,, further strengthen the concentration of oil by eliminating more waters as buttermilk.In short, the more stable form of concentrated decision.

Emulsifying agent and stabilizing agent

The process that forms emulsion generally includes vigorous agitation, so that oil is broken into droplet.Add the formation that emulsifying agent helps emulsion, quicken to smash process by reducing interfacial tension, so emulsifying agent surfactant normally.Emulsifying agent commonly used comprises detergent, glycerol monostearate and lecithin.

In case formation emulsion just must be kept, function of stabilizer that Here it is.It is owing between the molecule hydrophilic segment electrostatic interaction takes place that emulsifying agent can play a part stable.But this is also not enough, still needs to add stabilizing agent.By in system, adding or providing macromolecule can realize stability.They have two effects.

They form a layer on the oil droplet surface, prevent that oil droplet from converging, thereby form steric hindrance barrier.Undissolved protein, for example the casein in the milk is often fulfiled this function.

They can be dissolved in the continuous phase, strengthen viscosity.For example, the polysaccharide in the food is through being usually used in this purposes.As long as add a spot of polysaccharide glue, for example xanthan gum and antler glue just can strengthen viscosity significantly.

Colloidal decomposition comprises that particle is flocked together by gravitational the influence, forms bigger particle.According to the precise characteristics of this processing method, this processing method has different terms.

● cohesion is the loose combination of particle, and it relatively easily breaks, the phase redispersion

● condensing is stronger bonded particle assembly.Because interparticle gravitation is strong more a lot of than cohesion, therefore agglomerative decentralized photo is not easy redispersion.

● coalescent is to merge at particle to form single bigger particle.

Some is loose a little in preceding two definition, and these two terms are used interchangeably sometimes.In general, if take place that total surface free energy descends as a result the time use cohesion.

Coalescent

Coalescent is to merge two particles to form single bigger particle.Main difference is that floccule body and condensed particles keep unique feature, but coalescent situation is not like this.

Coalescently can have liquid and solids, but fluid-borne particles normally.This processing method comprises the continuous phase thin film between diluent particles, till discharge all continuous phases, two particles merge.

Ostwald ripening

If decentralized photo has tangible solubility in any continuous phase, may be known as the phenomenon of Ostwald ripening.Owing to be subjected to capillary the influence, the solubility of small-particle is strong than macroparticle usually.Thereby the macroparticle trend is grown up with the infringement small-particle.If the course of processing is enough fast, colloid can be unstable.On the other hand, this course of processing is controlled the generation that very helps emulsion.In frozen food, in the long-term storage process,, cause disorganization because big ice crystal body trend is grown up with the infringement small-particle, cause food spoilage.

Gel

Gel is that the particle interphase interaction in decentralized photo is enough strong, forms in the time of can forming stiff mesh.In this case, colloid is equivalent to solid, flexibly alleviates the reaction of shear stress with this.In fact, gel is made up of the continuous flocculate of filling whole system.

With regard to based on high molecular gel, existing has the molecular domains of attraction-often with the form of hydrogen bond or the form by certain ion stability to other molecule.Based on the gel of flocculate, the result is characteristic such as solid three-dimensional netted thing equally.

The expansion of gel

The cancellated formation of 3-D that constitutes gel causes continuous phase to be trapped in the gel.Under many situations, continuous phase in the solution and floccule net are taken on semipermeable membrane.As a result, osmosis takes place, gel expands.Can offset inflationary spiral by external pressure, required pressure is called as the bulbs of pressure.It can reach very high value.For example, timber wedge is driven in rock and soak wood and can cause enough bulbs of pressure so that rock splits.

Hydrocolloid

Hydrocolloid is the hydrophilic polymer in vegetable, animal, microorganism or synthetic source, comprises many hydroxyls usually, can be polyelectrolyte.Their natural existence or adding are with the functional characteristic of control food containing water.In these characteristics, the most important thing is viscosity (comprising multiviscosisty and gelation) and water combination, but other numerous characteristics is also very remarkable, comprise emulsion stability, anti-stagnant ice recrystallization and organoleptic feature.

Food is very complicated material, it and the multifactor functional several different hydrocolloids of needs that cause together of hydrocolloid, wherein the most important thing is: alginate, araboxylan, carrageenan, carmellose, cellulose, gel, beta glucosan, guar gum, arabic gum, locust bean gum, pectin, starch, xanthan gum.

Every kind of hydrocolloid all is made up of similar mixture, but is not identical, and molecule and separate sources, preparation method, heat treatment and food product environment (for example salt content, pH value and temperature) all can influence the physical characteristic that they show.Introduce Utopian structure through regular meeting when describing hydrocolloid, but should remember that they are natural products (or derivant) of band structure, are subjected to the influence of enzyme reaction at random, and not exclusively rely on genetic code.They are made of the molecular mixing body of different molecular weight, and the neither one molecule is identical on conformation with any other molecule, or even (except the cellulose) structurally also inequality.

The hydrocolloid mixture shows a kind of like this complexity of the non-additive feature that just has recently, and these are interpreted as a science, but not technology.Have a high potential aspect the polysaccharide structures functional knowledge that closes at hydrocolloid mixture with water-bound.The special parameter of necessary every kind of application of scrutiny is noted required influence (for example quality, flowability, burn into water content, stability, viscosity, coherency, adaptability, elasticity, extensibility, process time, machining accuracy) and type, source, grade and the structural heterogeneity of noting hydrocolloid.

All hydrocolloids With the water mutual effect, reduce its diffusibility and stability.In general, neutral water colloid solubility is lower, and the solubility of polyelectrolyte is higher.By direct hydrogen bond or water-bound or a large amount of but comprise intermolecular or the intramolecularly space in can control such water.Reacting to each other of hydrocolloid and water depended on Hydrogen bondIn conjunction with, therefore, The water clusterFormation depend on temperature and pressure equally.Same situation have contrary balance between entropy loss and enthalpy obtain, but this process is subjected to dynamics Controlling, possibly can't obtain optimum network.Hydrocolloid can be showed far-ranging conformation in solution because with the link of polymeric chain can Potential energy ViewRotation relatively freely in the paddy.Surface big, the more condensed hydrocolloid of structure is static basically, can impel the extensive structuring of ambient water.Water prevents in conjunction with influencing quality and process characteristic Take off Water shrinks, have great economic worth.Particularly, hydrocolloid can provide the water that increases other food composition motility (plasticizing).They also can influence the formation and the growth of ice crystal, therefore can bring into play special influence on the quality of frozen food.Some hydrocolloid, for example Locust bean gum GlueWith Xanthan gum, the time may form stronger gel at the freezing ice of separating, because water is removed (Cheng Bing) when freezing, causes and force combination.

Hydrocolloid usually can influence the deadweight flow behavior of water significantly, most hydrocolloids be used to strengthen viscosity (referring to Rheology), by the stability that prevents to precipitate, is separated, foam collapse and crystallization improve food.Viscosity usually with concentration, temperature and Shear strain rateVariation and change, on certain complicated meaning, depend on the material of hydrocolloid and other existence.The mixture of hydrocolloid can be worked in coordination with and be weakened viscosity with strengthening viscosity or antagonism.

Many hydrocolloids also are gels, the multiple texture characteristic of may command.Gel be show like solid property contain the aqueous water net, its characteristic intensity depends on concentration, hardness and fragility depend on the structure that hydrocolloid has.Hydrocolloid shows elasticity and viscous characteristics, can not in time untie when the polymer that twines to occur elasticity when allowing to flow.The hydrocolloid mixture is synergism mutually, in conjunction with precipitate, gel or form incompatible Binary systemSuch phase limitations affect viscosity and elasticity.Hydrocolloid has considerable purposes, can be used for many others, when comprising (a) high temperature PseudoplasticityMake (flowability under the shear stress), by cooling concentration mixing and processing are more prone to, (b) liquefaction of the heating after the cooling gelization (c) adds Thermogelling and makes structure more fastening (Thermogelling), (d) multiphase system comprises the processing and the Stabilization of rete.

The characteristic of these hydrocolloids is to determine by its architectural feature with the mode of water mutual effect.For example:

● when intramolecularly or intermolecular hydrogen bond have enough attraction to the hydrogen bond (salt formation sometimes) of water, hydrocolloid becomes gel in the time of can overcoming entropy.Usually, hydrocolloid demonstrates delicate balance between hydrophobicity and hydrophilic.The hydrocolloid that enlarges when higher concentration is chaotic gradually, similar molecule can twine (form helical form cross district) lacking under the situation of hydrogen bond mutually, but heterogeneous and reduce the hydrophobic surface of getting in touch by reducing conformation with water as far as possible, thus for more positive in other place, more advantageously use and discharge it.In this case, need to form the link (for example if the helical form bonding land needs complicated spiral usually) of minimum number, overcome entropic effect and form stable link.At this moment, the bonding land reacts to each other to remove moisture along with the variation of time is slowly grown, and syneresis (as in some fruit jam and fruit jelly) may take place.

● polysaccharide hydrocolloids mainly takes stable emulsion by strengthening viscosity, also can be used as emulsifying agent, and according to reports, its emulsifying capacity is mainly by following due to (pollute or intrinsic) protein portion.Particularly, as long as suitable pH value and ionic strength state continue, the electrostatic interaction between ionized water colloid and the protein will cause significant emulsification, thereby improves stability greatly.Proteinic degeneration may improve emulsifying capacity and stability.

● owing to hindering crystallization but help dissolved structural heterogeneity reason, the hydrocolloid mixture can prevent to assemble voluntarily when high concentration.Hydrocolloid can help fatty emulsive food composition with other and be harmonious, and the viscosity of glutelin is rolled into a ball or influenced to the stabilised milk protein molecule.

● the granular size of hydrocolloid and distribution thereof are the important parameters relevant with hydration rate and emulsifying capacity.

● electronegative hydrocolloid changes the counter ion type and the concentration (comprising pH value and ionic strength effect) of its architectural characteristic; For example, when acidity was high, electric charge disappeared, and the molecule expansion tails off.

● physical characteristic can be depending on the control that concentration is subjected to thermodynamics or kinetics (operation record and environment).Particularly these characteristics can change with no change mode or swing mode in time.

● different hydrocolloid preferences Low-density or high density water, other hydrocolloid then demonstrates the compatibility to two kinds of situations.Because more hydrogen bond forms in the polymolecular, so hydrocolloid becomes more hydrophobic, and this can change the partial structurtes of water.The mixing water colloid is liked different environment product excluded volume Liao, and is influential to mutual valid density, thus influence Rheology

● in vitreousness, conformation change is strict the inhibition, but the water that hydrocolloid keeps can be used as plasticiser (permission molecular motion), by the hydrogen bond between the saboteur, significantly reduces the temperature of gamma transition.

Natural gum and starch: controlled humidity behavior

Understand the interaction mechanism of water in the food, and using polysaccharides class how, for example natural gum and starch are controlled these interactions, make the designer take measures to improve the quality of products and prolong the pot-life.

This typical example is the dough/pasta of baked product.Here, water is not only the solvent of activation reaction and/or yeast fermentation, also is simultaneously the processing aid that allows the glutelin growth, can form mixing, coherent mass (dough/pasta), can and bake its setting afterwards.Starch and natural gum itself are component of polymer, need water to activate as plasticiser.

The polysaccharide that natural gum and starch are made up of the molecule straight chain.Natural gum has a functional group at this chain end, and starch has various branches on this chain.Accurate structure is looked the source of material and is changed to some extent.

In immovable shape, both sides' absorption water expands in solution, and as gentle viscosifier.Be subjected to the activation of heat and/or mechanism, glue and amylum grain all reorganize.This is that two kinds of materials begin to show different places.The hydrated gums molecule has affinity each other, can form gel.On the other hand, starch continues as the individual molecule with enhancing thickening properties.Each brood lac way of act with starch is different, and feasible even more variation of the modification of stock become possible (for example pre-gelatinized starch and cold expansion gel).

The flavoring agent composition.

The water activity is represented important variable, influences the speed of many flavoring agent composition chemical reactions.In the aqueous system of complexity, the constituted mode of food substrate presents with the flavoring agent organoleptic indicator quite important for local flavor.

In the food containing water system, polysaccharide and protein normally determine the main component of food configuration.When other less molecule occurring, for example during aromatic compound, for the ease of observed result, the hydration of these macromolecular components is quite important.Therefore the mode that comprises volatile compound in food system will determine that local flavor presents, and the flavoring agent organoleptic indicator that provides for consumer of decision and the profile of product.

Comprise the physical-chemical reaction of flavoring agent composition-no matter be between the flavoring agent, or between the non-flavoring ingredients of flavoring agent and food and the environment-broadly be defined as " flavoring agent reacts to each other ".These react to each other influences the quality of flavoring agent in the food, quantity, stability and final organoleptic indicator.Flavoring agent mainly is the synthetic of taste and abnormal smells from the patient, and with outward appearance and quality, the sense organ that comprises food is accepted standard.

Term " artificial flavoring " is meant the flavoring agent that those add food to or are made up of the non-existent chemical compound of occurring in nature.Naturally occurring flavoring agent, perhaps those are called " natural flavouring " by the flavoring agent that heating, ageing or fermentation form.For the synthetic natural flavouring that adds in the food, adopt label " with natural identical " flavoring agent.

Flavoring agent of fruit is to be specific purposes manufacturing and synthetic.The purpose of product designer is to select the best flavoring agent of effect in the food product environment of chemical reaction.This target to be successfully realized, the interaction of flavoring agent must be understood.

Physics and chemical flavoring agent interact and continue to take place in growth, harvesting, processing, storage and the process of consumption of food.Interaction is the reason owing to various types of chemical bonds: covalent bond, hydrogen bond, hydrophobic bond and comprise the formation of coordination compound.The interact physics aspect of the most normal measurement of flavoring agent is bonding, distribution and release.Bonding is meant the volatility of flavoring agent and involatile constituent is absorbed on the composition of food substrate.Distribute and to have described flavoring agent in food relevant moisture, liquid state or the gaseous state distribution situation in mutually with packing.Human sense organ accepter is called " release " to effective this point of flavoring agent.Product is depended in optimization to flavoring agent release time, because food is chewed fully, need the long time, and beverage only needs in mouth to stop several seconds.

According to the chemical constitution of flavoring agent and the chain length of fatty acid, flavoring agent can be distinguished their difference between oil and water.In lipopenic food, flavoring agent discharges the influence that is subjected to this distribution, because aqueous system has higher equilibrated steam pressure than the flavorant in the lipid system.Volatile matter discharges sooner from aqueous system and dissipates, and causes human sensory flavoring agent organoleptic effects less.

The owned few fragrance of protein, but particularly under the situation of thermal denaturation, they are in conjunction with some volatile flavoring agent compositions.Because the combination of hydrophobic interaction and hydrogen bond is reversible, resemble at ketone, under Hydrocarbon and the situation based on the flavoring agent of alcohol.Covalent bond is normally irreversible, for example formation of schiff base (acetaldehyde and amino).Influencing protein bound to some factor of volatile matter is: the existence of temperature, pH value, concentration and water.According to heat treated length and degree, protein can be in conjunction with more or less flavoring agent composition.In protein of dairy product, several flavoring agent compositions, for example vanillin, benzaldehyde and d-limonene in the solution that comprises whey protein or sodium caseinate, reduce reaching 50%.The combination of protein flavoring agent can reduce the influence with desirable flavoring agent, brings sensory receptors with not desirable flavoring agent.It is stink and the combining of soybean protein that the protein flavoring agent of the confirmation of broad research interacts.

Carbohydrate has several important enhancing taste functions.Its envelop of function is extensive, can be used as sweetener; Browning reaction participates in agent; Fat substituted dose; Viscosifier; And agent is surrounded in seasoning.Sugar is by the carrier of the chemical combination in interaction of the physics in the Aquo System and the dry ingredient as flavouring agent.The structure of the carbohydrate that molecule is bigger (as starch and cyclodextrin) can form the hydrophobic region of the mechanism that includes as similar flavouring agent chemical compound, hydrophobic chemical.The flavoring agent molecule that is fit to these hydrophobic regions is called " guest molecule ".Because at the chemical reaction that does not take place between starch and the guest molecule except that hydrophobic attraction, so these interactions are highly reversible.This interaction has formed the basis that the flavoring agent molecule surrounds.

Polysaccharide, particularly hydrocolloid and gellant are to some extent in conjunction with the flavoring agent composition.When the concentration of flavoring agent kept constant, the level of polysaccharide can increase, fragrance sense and mouthfeel reduction, result's more toughness that becomes.For example, when the viscosity of guar gum or cmc soln strengthened, the sugariness of sucrose can reduce.

Carbohydrate also can change the volatility of aromatic compound.When comparing, add monosaccharide and disaccharide and can increase volatility, add polysaccharide and can reduce volatility with the flavoring agent chemical compound in the aqueous solution.In the food system particular importance of using fat substituted dose, this is that a little less than interacting between the carbohydrate that contains the hydrophobic flavor immunomodulator compounds, the speed that volatile matter discharges is faster because when fat content is low to carbohydrate to volatile influence.

Food matrix is made up of protein, carbohydrate and fat usually, so and the interaction of flavoring agent be everlasting and take place between two kinds or the above chemical compound.Reducing sugar and aminoacid react in the Maillard reaction (being also referred to as nonenzymatic browning), can generate the volatile matter and the brown stain product of fragrance, thereby form flavoring agent in the food heat treatment process, as chocolate, coffee, barbecue, bread food and caramel.Decide on the amino acid whose value volume and range of product that can be used for participating in reaction by the quantity of these flavoring agents of forming of reaction and type.In conjunction with the Oxidation of Fat and Oils reaction, Maillard reaction can generate the flavoring agent chemical compound after carbonyl compound (from sugar or greasy decomposition) and amine or the heating of thio-alcohol material react.Flavoring agent reaction in the fabricated food substrate takes place seldom separately, and is subjected to the influence of reactant, intermediate products and other reactor product.

The interaction of flavoring agent and packing is caused by three kinds of factors: the moving of packing or food composition; Gas, water and organic vapo(u)r infiltrate packing; And sunlight direct projection.

Prevent that flavoring agent from reducing or the interaction of degraded comprises processing influence (heat, pH value); Environmental factors (steam, oxygen); And reduce to minimum with the chemical reaction of food substrate.

The flavoring agent mouthfeel is relevant with the mode that discharges (reservation) fragrance from food system.Flavoring agent discharges the characteristic and the concentration of looking flavoring agent chemical compound in the food, and is decided by the mouthfeel that the reaction between main component in the food and flavoring agent chemical compound causes.Factor of food composition and structure (as the super large molecule occurring) and way of dining have determined the degree that mouthfeel and flavoring agent discharge.The relative maintenance or the processing of flavoring agent in determining the course of processing, deposit and edible process in the selectivity of specific compound when discharging, the chemical combination characteristic of the flavoring agent chemical compound relevant with main food composition, they are at the allotment ratio of different phase, and the knowledge of the structure organization of food substrate is quite important to the practice of foodstuff flavouring.

Main mechanism may appear in the flavoring agent dispose procedure, (i) combination of the specificity of aromatic molecules and the (ii) embedding of these molecules in substrate.Some aromatic molecules may combine with protein or polysaccharide generation specificity.In addition, because protein and polysaccharide can enter air by food to fragrance and impact, thereby can influence the release of fragrance.Therefore in compound Aquo System, the mode of food substrate structure is quite important to flavoring agent release and mouthfeel thereof.

The mechanism that may exist different control flavoring agents to discharge in the food system.Be subjected to the system viscous effects diffusing phenomenon, to combine with the non-specific binding of one of macromolecular components or specificity may be the interaction of flavoring agent molecule in the food substrate.

The food processing general introduction

Food processing is a generality term, and it refers to be used for human consumption's Food ﹠ Drink and all activities that animal is used the preparation food production.The sector is defined as food and like product by standard industry classification (SIC) 20.

Food processing is tended to the inherent structure in raw-food material or the composition is subdivided into different parts, therefore it relates to the packing and the marketing of the processing of the chemistry of the various aspects-food of food and composition thereof and physical characteristic, food and production, food, and these all are the ingredients of food processing system.Food quality-quality, taste, nutrition effectiveness, water translocation and growth of microorganism-be subjected to the influence of formation, stability and the textural classification of food, and determine by them.

Food processing relates to raw material and composition is converted to consumer's food or edible product.Food processing comprises the plant of not process processing or animal material change or changes into safety, edible and more all behaviors of tasty food.Raising is deposited or storage life is another target of food processing.

The purpose of food processing is to produce balanced diet composition, food pollution-free, attraction consumer aspect color, taste and quality can be provided.

Food processing also drives a series of flavoring agent chemical reaction, and mouthfeel also depends on the releasing degree of flavor compounds in edible process.Pass between flavor compounds forms in the structure of food, machinery and physicochemical characteristics and the flavoring agent mouthfeel and the course of processing ties up on the part degree and is determined by hydration.

The food processing operation relates to one or more ambient temperature processing, machining, high-temperature process, K cryogenic treatment, sweat, and various post-treatment step.

The ambient temperature processing comprises cleaning and classification, removes the peel; Pulverize, cut and mill; Mix, merge and be shaped.These are the preparation of subsequent operation normally.

Physical separation comprises filtering, centrifugal filtration; Press and squeeze and extract; Divided thin film from.These are usually directed to obtain specific part from raw material.

High-temperature process has two main purposes: make food reach safety criterion by pasteurization and sterilization; Food is cooked ripe, can be changed flavoring agent, quality, trophism.Each processing procedure can realize two kinds of functions simultaneously.High-temperature process comprises sterilization and pasteurize; Blanch; Cure and toast; Fried; Microwave and infrared heating.

The purpose of blanch is as dehydration, sterilization, refrigerated pretreatment.Heat is enough to make enzyme to lose activity, but can't cook food, and undertreatment is with to cross the result who handles the same poor.

Curing and toast basic identical, is dry heat in hot-air.Cure and be generally used for dough products.Baking is often referred to meat, nut and vegetable.Make the surface that processes material carry out the chemical change of color and taste.Heat the effect of nutritious aspect, make the food ratio be easier to edible and digestion, but in this process, may cause losing of vitamin.

Fried is to cook with deep fat.Its objective is the cating quality (taste, quality) that improves food.Fried effect to cure similar.Because it is deep fat directly contacts with food, fried usually than toasting or curing faster.

Microwave and infrared heating utilize electromagnetic radiation.Microwave heating uses short wavelength radiation.Waveform frequency conforms to the hydrone frequency of natural vibration.Infrared ray is the radiation that only exceeds the spectrum visible region.Energy apparent temperature, surface characteristic, shape and decide.

K cryogenic treatment can make growth of microorganism slow down, but can't kill microorganisms.To a certain extent, temperature is low more, and storage life is long more.When being lower than-10 ℃, all microorganisms all stop growing, but some residual enzyme still keeps active.Therefore, cooling and refrigerated major function are to deposit and prolong storage life.

Fermentation has a lot of effects, comprise preservations, improve nutritional quality, the raising digestibility, to healthy and helpful.Fermented food has many kinds, comprises the meat of milk product, fermentation and vegetable, beverage, bread etc.

Post processing processing comprises packing and deposits.The purpose of these operations comprises protection, displaying, prolongs storage period.In order to prolong storage life, now just updating air ambient, generally be to reduce oxygen content and improve nitrogen content.

Packaging material

Main packaging material comprise metal, paper and cardboard, glass, and polymer.The metal that is widely used in packaging for foodstuff has iron and steel (being common in the canned food iron sheet), and the aluminum that is used for three kinds of main food applications such as beverage can, paper tinsel container, aerosol can.

Canned food is made

The canned food of producing mainly contains two kinds of forms.The three-chip type canned food comprises latasuture and two independent end sockets of winding up sheet metal, welding.The two-piece type canned food, a side and an end are made of flat board, do not have seam.End socket is sealed by two seals, fully by machine-building.Canned food inside scribbles " enamel " usually, prevents contaminated food products.

Paper and cardboard

Can use various other paper of level.Kraft paper is harder, is commonly used for paper bag.Vegetable parchment is the paper through the peracid special handling, and quality more closely, smoothly.Unbleached sulphite paper is more light and soft than kraft paper, is commonly used for paper bag and sweet wrapper.Greaseproof is by the unbleached sulphite paper manufacturing, and its paper fiber thoroughly changes, and quality is tightr.Can be grease proofing.Cotton paper is soft resilient paper, shields.

Aseptic packaging

Aseptic packaging is to prevent to pack under the aseptic condition of recontaminate the process of sterile chamber after to food sterilization.It is different with sterilization in container and food separate disinfectant packing.

Aseptic process

Number of processes is less to mean that the food processing number of times is less, and this can reduce the loss of vitamin and taste.Because packing need not heating, therefore can the wide selection packing.But, in packaging process, must guarantee aseptic.Aseptic process can make food prolong storage life at normal temperatures, improves food quality.

The polymer that is used for packaging for foodstuff

Polymer is based on the macromole that is derived from micromolecular repetitive.They can be natural (for example, polysaccharide), also can synthetic.They have various characteristics, can be used for packaging for foodstuff.The example of polymer has polyethylene, low density polyethylene (LDPE) (LDPE), high density polyethylene (HDPE) (HDPE), polypropylene, polystyrene, polrvinyl chloride (PVC), polyethylene, terephthalonitrile (PET), Merlon, polyamide (nylon), cellulose (cellulose acetate, cellophane).

Polymer can be divided into the thermoplastic of heating and melting; Also can be divided into the thermosetting substance that adds thermal decomposition.

Unit operations in the food

Evaporation

Evaporation is to evaporate the water and the process of concentrated liquid by heating.In food processing, adopt evaporation that multiple use is arranged:

■ usually before drying waits some other program to the food pre-concentration, or reduce cost of transportation

■ improves the preservation quality by reducing operations such as water activity, for example makes fruit jam.

■ production has genuine food, for example, and condensed milk, fruit juice.

The heat of evaporation is provided by condensed steam usually.Therefore, this processing procedure comprises latent heat from the vapor transmission to the evaporation water.The food evaporation is carried out under vacuum usually.This can reduce the boiling temperature of liquid, thereby reduces the hot injury to food.Therefore, need be in evaporation short stay a period of time.The most general evaporation type is a film type, provides steam to the test tube outside, and liquid spreads in thin film by the inner surface of one group of test tube.Have two types thin-film evaporator, rise film and falling liquid film.This needs highly concentrated, adopts multiple-effect evaporation then.This is included in the several stages by carrying out evaporation operation as the steam that produces in the step of next stage heating steam.This can save a large amount of steam.

Dry

Dry or the dehydration of food is for removing the moisture in the food, moisture is reduced to low-down level (being usually less than 5%wt).It is to prolong storage life that food is carried out exsiccant purpose, thereby method is almost reduced to water activity the activity of zero inhibition growth of microorganism and enzyme.General dry run is to the food heating, and through after this step, food can't recover usually, does not for example have enzymatic browning and decomposition of vitamin and protein denaturation.Unless careful control treatment condition, otherwise drying may cause serious negative effect to nutritive value of food.

Dry run

Drying normally heats solid in air, makes water evaporates in air.Dry run may need to write down the chart of moisture and time.Moisture is finally reduced to certain steady state value.This is called equilibrium moisture content.

Dry mechanism

Constant rate of drying appears when solid material is covered by one deck water fully.

Drying is from the water layer surface evaporation, and speed is controlled by the temperature and the moisture of dry air purely.Behind the moisture that has evaporated capacity, water layer no longer covers solid surface, and this moment, moisture can shift out from the solid interior diffusion, could evaporate from the surface of solids then.Under these circumstances, along with the reduction of interior moisture content, moisture diffusion to the speed on surface also can reduce, thereby evaporation rate also reduces thereupon.

Dry rate and time

At constant rate period, dry rate is controlled by surface evaporation, and the function of speed of the latter's to be heat be transferred to wet surface of solids.

Extract

Solid-liquid extracts or leaching is to separate two kinds of solid processes, and method is solid mixture to be separated with a kind of solid dissolvable and the undissolvable solvent of another kind of solid contacts.This method is widely used in the recovery vegetable oil, the caffeine that also is used for instant tea and instant coffee and removes coffee.Extraction can be carried out in batches, also can carry out continuously.The most general method be by to solvent extraction with absorb similar mode and use Continuous Countercurrent Extraction.

Food additive and food configuration

These " accessory " adding ingredients are important making food in good to eat and attractive in the food, but do not have nutritive value usually.Though these additives of the natural existence of possibility still will add them in the food to, usually to guarantee controlled and stable properties in the food.Additive can influence rheological characteristic and quality, colloid property, color (comprising the brown stain of food) and the taste of food.

Food additive itself is not consumed as food usually, not as the typical composition of food and consume yet.Additive is incorporated in the food, can changes the characteristic (comprising processing characteristics) of food in some mode.Food additive and food contaminant should be made a distinction.

Pollutant are the nonconforming materials that appear in the food, can't remove (owing to chemistry or cost reason) fully.On the other hand, additive is specially to add the material that specific use is arranged.

Food additive is used for following purpose:

1. the nutritional quality that keeps food.

2. improve food preservation quality or stability, reduce the loss.

3. the method with sincerity makes food attract consumer more.

4. important miscellaneous function is provided in food processing.

As everyone knows, have many manufacturers illegally to use additive deception consumer at present, the fact of defective composition or illegal processing and treatment technology is used in concealment.

The primary categories of food additive comprises

The E numbering	Additive types
		E1xx	Coloring agent
E2xx	Antiseptic
		E3xx	Antioxidant, emulsifying agent, stabilizing agent and thickening agent
E4xx	Sweetener
		E5xx	Mineral salts
E6xx	Flavour enhancer
		E9xx	Food wax and glazing agent

Natural and synthetic additive

Reality is from plant or animal origin separates (using its broad sense) or the additive that appears at plant or the animal extracts can be described as natural additive.If certain chemicals of certain additive and occurring in nature on chemical property identical but actual be synthetic by chemical mode, can be described as with natural identical.Synthetic additive is not present in occurring in nature, must adopt fermentation or other biological technique method synthetic.

The present invention comprises following theme, described in US patent class handbook 426 classes.According to described in class definition (and related compound, method and product classification system) and the patentable theme such as US patent class handbook 426 classes of wherein classifying, it draws at this and is reference to the understanding of wherein classification, definition and the example of regulation.

A. structuring (micelle grain) edible product or compositions

1. be regarded as the product or the compositions of food traditionally, and the product or the compositions that contain the natural material (being plant or animal tissue) that is regarded as food traditionally; As milk, cheese, Fructus Mali pumilae, bread, living face, bacon, whiskey etc.).

2. known have or disclosed product or compositions with Nutrition.

3. open or be called as edible, or perfect to carrying out such as the edible product of above-mentioned (1) or (2) or another kind of edible product, modify, handle or unite use with it, thereby become the part of edible composition or product or product or the compositions that is edible form with not edible formal transformation.

4. edible own, or be used to prepare the mixture of the enzyme of suitable product of food or edible material or compositions.

5. be used for making the food or compositions related products or the compositions that contain the active microorganism that can strengthen or improve the intestinal Digestion, for example, acidophilic lactobacillus milk etc.

6. the edible product or the compositions that have architectural feature.

7. be used to strengthen the composite inorganic element or the mineral of nutrition.

8. edible bait.

B. The edibility food product that combines with non-foodstuff it typically is:

1. the product of above-mentioned A or compositions combine with packaging structure, not edible outer package, liner or substrate, preserved material bag etc.

2. with the chemical compound with (A.1-3) identical function of the combined thing of eatable material not.

3. drinkable water in packing.

4. chewing gum and chewing gum base itself.

C. Seasoning and sweetening compositions

1. wherein at least a composition is not the flavoring compositions of carbohydrate type material.

2. the sweetening compositions that wherein at least a composition is a non-carbohydrate type material.

D. Product or the compositions of above-mentioned A-C offered animal by the oral cavity method.

F. The chemical compound through port that will have identical function with compositions or the product of above-mentioned A-CThe chamber Offer the method for animal.

G. In conjunction with butchering operation with improving from product, chemical compound or the ferment of the food of live animal preparation The plain method of handling this living animal, or take out food product from living animal and handle taking-up then The method of food, or butchering the laggard line operate of operation.

H. Product or the method for compositions of A-C handled or improved in preparation.

I. The special-purpose drying single use that is equipped with food and does not have the structure of cooperating with the food device specially Steeper or storage.

J. Handle or improve the compositions and the using method of food material.

The field reader understands under the present invention, and the description of aforementioned preferred embodiment details is interpreted as restriction the present invention never in any form.The reader understands other embodiment that can implement to drop within the scope of the invention, and this is determined by following claim and legal equivalent thereof.

Claims (40)

1. the culture medium that contains micelle grain water.

2. the medium that contains micelle grain water.

3. the culture that contains micelle grain water.

4. the method for preparing culture medium comprises the dissolving or the step of mixotrophism material and micelle grain water.

5. the method for preparing medium comprises dissolving or mixes the step that one or more are selected from the medium composition of inorganic salt, mineral, carbohydrate, aminoacid, vitamin, fatty acid and lipid, protein and peptide class and serum.

6. the method for preparing culture comprises the step that cell, tissue, organ, subcellular fraction, virus, phage or carrier are contacted with the culture medium that contains micelle grain water.

7. contain the application of culture medium in cell culture of micelle grain water.

8. contain the application of medium in cell culture of micelle grain water.

9. contain the application of culture in cell culture of micelle grain water.

10. contain micelle grain water culture medium, contain the culture of micelle grain water and contain one or more application in the microorganism biological technology in the medium of micelle grain water.

11. strengthen the method for cell survival, be included in the culture medium that contains micelle grain water, contain the culture of micelle grain water and contain the step of cultivating described cell in the medium of micelle grain water one or more.

But 12. strengthen the method for cell, tissue or organ viability, comprise with described cell, tissue or organ with the culture medium that contains micelle grain water, contain the culture of micelle grain water and contain one or more steps of cultivating in the medium of micelle grain water.

13. contain micelle grain water culture medium, contain the culture of micelle grain water and contain one or more application in organ, tissue or cell transplantation in the medium of micelle grain water.

14. contain micelle grain water culture medium, contain the culture of micelle grain water and contain one or more application in transfection in the medium of micelle grain water.

15. contain micelle grain water culture medium, contain the culture of micelle grain water and contain one or more application in the results stem cell in the medium of micelle grain water.

16. contain micelle grain water culture medium, contain the culture of micelle grain water and contain in the medium of micelle grain water one or more stem cell biological learn or the clone in application.

17. test kit comprises the culture medium that contains micelle grain water, contains the culture of micelle grain water and contains in the medium of micelle grain water one or more.

18. suppress the method for genetic material mutation frequency, described method comprises the step that described hereditary material is cultivated with the medium that contains micelle grain water, wherein said hereditary material is arranged in biological entities.

19. micelle grain water, it comprises one or more and is selected from bio-affecting agents, health inorganic agent and adjuvant or the carrier compositions one or more activating agent.

20. being selected from, the compositions of claim 19, wherein said bio-affecting agents have the one group of activating agent that is selected from following biological nature:

A. prevent, alleviate, treat or cure the unusual and pathological condition of live body;

B. keep, promote, reduce, limit or destroy the physiological organism function;

C. by in vivo test diagnosis physiological conditions or state;

D. control or protect environment or live body by attracting, disable, suppress, kill, modify, repel or hinder animal or microorganism.

21. the compositions of claim 19, wherein said health inorganic agent is selected from the activating agent that is intended to be used for deodorization, protection, decoration or ornamenting health.

22. the compositions of claim 19, wherein said bio-affecting agents or described health inorganic agent are selected from fermented product, plant and animal extract, body fluid or contain plant or the material of zooblast structure.

23. the compositions of claim 19, its dosage form is selected from liquid, ointment, cream, gel, dispersion, powder, granule, capsule, tablet and transdermal delivery device.

24. the compositions of claim 19, it is a Pharmaceutical composition.

25. the compositions of claim 19, wherein said bio-affecting agents or health inorganic agent are selected from: the medicine that acts on synapse and neural effector joint area; Act on central nervous system's medicine; The autacoid or the medicine of treatment inflammation; Influence the medicine of kidney and cardiovascular function; Influence the medicine of gastrointestinal function; The chemotherapeutic agent that is used for parasitic infection; The chemotherapeutic agent that is used for microbial diseases; The chemotherapeutic agent that is used for neoplastic disease; Be used for immunoregulatory medicine; Act on blood and hemopoietic organ's medicine; Hormone and hormone antagonist; Vitamin; Be used for the treatment of dermopathic medicament; And the medicament that is used for treatment of eye disorders.

26. the compositions of claim 19 also comprises drug-supplying system.

27. use the method for compositions of claim 19, comprise described compositions is applied to live body or the stripped step that is applied to cell, tissue or organ.

28. the method for claim 27, wherein step of applying comprises the use drug-supplying system.

29. the method for claim 27, wherein said method are diagnostic method.

30. the method for compositions of preparation claim 19, it comprises the combined step of activating agent that micelle grain water and one or more is selected from bio-affecting agents, health inorganic agent and adjuvant or the carrier compositions one or more.

31. the method for at least a composition and product in the hydrated food prod system of processing, described method comprise that the micelle grain water with capacity contacts the step of enough time with at least a described composition with product, thereby form at least a micelle grain composition and micelle granule product.

32. the method for claim 31, wherein said product is selected from:

(a) edible product or compositions,

(b) comprise the edible food product that micelle grain water and non-food combination of materials form,

(c) flavoring compositions and

(d) sweetening compositions.

33. the method for claim 31, wherein said composition comprise that one or more are selected from following group one or more composition: aminoacid, peptide class, protein, lipid, carbohydrate, aromatic substance, vitamin, mineral, and food additive.

The food that the living animal handled through micelle grain composition and/or micelle granule product is made 34. the method for claim 32, wherein said edible product are served as reasons, described treatment step further be selected from following step and combine:

A. butcher operation

B. take out food product from living animal, handle then the food that takes out and

C. butcher operation, then slaughter product is carried out post processing.

35. contain the edible product or the compositions of micelle grain water.

36. the edible product of claim 35 or compositions, it also comprises non-food material.

37. contain the flavoring compositions of micelle grain water.

38. contain the sweetening compositions of micelle grain water.

39. use micelle grain food product or method for compositions by the oral cavity to animal or human's class, described method comprises to the mankind or animal feeds with the food product that contains micelle grain water or the step of compositions.

40. the method for claim 39, wherein said micelle granule product or compositions are selected from:

(a) contain the edible product or the compositions of micelle grain water,

(b) contain the edible food product that micelle grain water and non-food combination of materials form,

(c) contain micelle grain water flavoring compositions and

(d) contain the sweetening compositions of micelle grain water.

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