CN105779114A - Method for extraction of rice bran with reverse micelle solvent - Google Patents
Method for extraction of rice bran with reverse micelle solvent Download PDFInfo
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- CN105779114A CN105779114A CN201410822843.XA CN201410822843A CN105779114A CN 105779114 A CN105779114 A CN 105779114A CN 201410822843 A CN201410822843 A CN 201410822843A CN 105779114 A CN105779114 A CN 105779114A
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Abstract
The invention provides a treatment method of rice bran. The method includes: crushing rice bran, then mixing the crushed rice bran with a reverse micelle solution, performing ultrasonic extraction, separating the layered liquid to obtain an aqueous phase and an oil phase, and conducting treatment respectively so as to obtain rice bran oil and phytic acid.
Description
Technical field
A kind of method that the present invention relates to reverse micelle solvent extraction Testa oryzae, concrete, relate to a kind of method extracting Testa oryzae oil and effective ingredient from Testa oryzae.
Background technology
China's Oryza glutinosa annual production, at about 1.8～200,000,000 tons, accounts for the 38% of Asia, accounts for the 34% of world's Oryza glutinosa annual production, occupy first place in the world.Testa oryzae is the side-product in paddy processing, China's Testa oryzae yield more than more than 1,400 ten thousand tons, at present except China's Testa oryzae is used for except small part extracting Testa oryzae oil, mostly as low value-added feedstuff, have even as waste material, the ratio effectively utilized also less than 20%, serious waste of resources.The deep processing of Testa oryzae and comprehensive utilization are to utilizing grain resource fully, promoting that the development of Grain Industry has profound significance.
Rice bran protein has fabulous economy and practicality.At present, the main method extracting rice bran protein has: alkaline process, enzyme process and multiple method combination etc..Alkali process cost is low, but there is the shortcomings such as pH value height, the easy degeneration of rice bran protein of preparation and extraction ratio be low.It is gentleer that enzyme process prepares rice bran protein reaction condition, and gained nutrient protein is worth high, but relative alkaline process, its process costs is higher.With defatted rice bran for raw material in CN201110102062.X, first adopting alkali to soak, adopt compound enzyme (cellulase, phytase, compound protease) to extract albumen afterwards, remaining residue is rice bran dietary fiber.With defatted rice bran for raw material in CN201210081904.2, first adopt alpha-amylase to remove starch as fermentable raw material, adopt acid soak to extract phytic acid afterwards, again through basic protein enzyme extraction albumen, finally obtaining dietary fiber, protein extracting ratio is 10.3%, and purity is 57.5%.With full fat rice bran for raw material in CN201110004840.1, first adopt normal hexane extraction fat.Then alkali extraction and acid precipitation obtains rice-bran crude fiber and rice bran protein, crude oil yield 11.3%, crude fibre yield 53.2%, albumen yield 26.5%.US0104607 discloses a kind of method of Enzymatic Extraction Testa oryzae oils and fats, albumen and fiber, enzymolysis is carried out with water after being mixed by Testa oryzae, enzymatic hydrolysate is divided into oil phase, aqueous phase and precipitation, and aqueous phase dries and obtains rice bran protein concentrate, precipitates and arrives rice bran protein and rice bran-fiber by what alkali extraction and acid precipitation extracted.Extracting method many employings acid-hatching of young eggs of current phytic acid in rice bran, such as CN201310138197.0, needs substantial amounts of acid solution in extraction process, there is no the correlational study adopting reverse micelle simultaneously to extract Testa oryzae oils and fats and phytic acid at present.
Reverse micelle refers to and is scattered in continuous organic solvent medium, and including the aggregation of the nano-scale of the surfactant of hydrone kernel, reverse micelle is also referred to as reversed micelle or reverse micelle.Usually, surfactant is soluble in water, can form aggregation when concentration exceedes critical micelle concentration, and this aggregation belongs to normal micelle.If be dissolved in nonpolar organic solvent by surfactant and make its concentration exceed critical micelle concentration, aggregation contrary to the above can be formed, i.e. reverse micelle.Inverse micelle abstraction is a kind of novel isolation technics, there is technique simple, process need not add substantial amounts of bronsted lowry acids and bases bronsted lowry, saving water resource, main agents used can recycle, and extracts cost low, environmentally safe, extraction conditions is gentle, and therefore product purity advantages of higher has good prospects for commercial application.
Traditional plant protein extracting method complex process, energy resource consumption is big, and in extraction process, the addition of a large amount of soda acids is easily caused protein denaturation, very easily causes the wasting of resources.Therefore, the method lacking active ingredient in a kind of high efficiency extraction Testa oryzae fat in prior art.
Reverse micelle method is except being used for extracting phytoprotein, it is also possible to for the separation of oils and fats.The research that current reverse micelle extracts at vegetable protein and oils and fats, mainly at Semen sojae atricolor, Semen arachidis hypogaeae, Semen Allii Tuberosi, Fructus Tritici aestivi etc., there is no the application in Testa oryzae component is extracted.
Summary of the invention
The first object of the present invention is in that to obtain a kind of reverse micelle solvent, this reverse micelle solvent has the abstraction function of excellence for Testa oryzae, current inverse micellar solution is primarily adapted for use in Semen sojae atricolor, Semen arachidis hypogaeae, Semen Allii Tuberosi, Fructus Tritici aestivi etc. what vegetable protein and oils and fats extracted, but inverse micellar solution disclosed in it is not all suitable for Testa oryzae, therefore, for this problem, the present invention proposes a kind of inverse micellar solution suitable in Testa oryzae component.
The first inverse micellar solution disclosed in this invention, wherein reverse micelle solvent includes organic solvent, also includes containing buffer solution and alkyl sulfate, and in described inverse micellar solution, the content of alkyl sulfate is 10g/L～150g/L, it is preferred to 50g/L～100g/L；The pH of described inverse micellar solution is 8.0～11.0.Preferably, being 0.001%～0.5% alkaline protease possibly together with weight content in described inverse micellar solution, this content is preferably 0.1～0.3%.Inventor have found that, this inverse micellar solution is a kind of inverse micellar solution being particularly suited for extracting Testa oryzae.Preferably, in described inverse micellar solution, the mol ratio Wo value of water and surfactant is 10～40, it is preferred to 20～40.
The second inverse micellar solution disclosed by the invention, wherein reverse micelle solvent includes organic solvent, also includes containing buffer solution and alkyl sulfate, and in described inverse micellar solution, the content of alkyl sulfate is 10g/L～150g/L, it is preferred to 50g/L～100g/L；The pH of described inverse micellar solution is 1.0～5.0.Inventor have found that, this inverse micellar solution is a kind of inverse micellar solution being particularly suited for extracting Testa oryzae.Preferably, in described inverse micellar solution, the mol ratio Wo value of water and surfactant is 10～40, it is preferred to 20～40.
In inverse micellar solution as above, described organic solvent is selected from alkane alcohol, alkane, carbon tetrachloride, aromatic hydrocarbon equal solvent, it is preferred that described organic solvent is selected from alkane and alkane alcohol.Wherein said alkyl sulfate is carbon number is the alkyl sulfate of 6 to 20, described alkane solvent is selected from 5 carbon alkanes, 6 carbon alkanes, 7 carbon alkanes, 8 carbon alkanes, 5 carbocyclic ring alkane, 6 carbocyclic ring alkane, 7 carbocyclic ring alkane, 8 carbocyclic ring alkane or its homologue, alkane alcoholic solvent is selected from the low carbon chain alcohol that carbon number is less than 10, preferably one or more in methanol, ethanol, propanol, propylene glycol, butanol, amylalcohol, hexanol, capryl alcohol, enanthol, nonyl alcohol, the volume ratio of described alkane solvent and alkane alcoholic solvent is 1:1～10:1.
A second aspect of the present invention discloses a kind of method simultaneously extracting Testa oryzae oil and rice bran protein, its method includes: is undertaken Testa oryzae pulverizing and sieves with 50～200 orders, it is preferably 60～100 orders to sieve, Testa oryzae after being pulverized, Testa oryzae by 50～200 orders, preferably the Testa oryzae of 60～100 orders carries out ultrasonic extraction after mixing with upper the first described inverse micellar solution, mixed proportion is 0.01～0.2g Testa oryzae/mL inverse micellar solution, it is preferably 0.01～0.1g Testa oryzae/1mL inverse micellar solution, ultrasonic extraction more than 0.1 hour, it is preferably 0.1～2 hour, liquid is obtained after being then peeled off solid, it is 8.0～11.0 by being adjusted to pH after addition buffer in described liquid, carry out ultrasonic extraction, layering obtains aqueous phase and oil phase.Described aqueous phase and oil phase are respectively containing rice bran protein and Testa oryzae oil.Described aqueous phase is permeated, obtains protein.
The invention also discloses a kind of method simultaneously extracting Testa oryzae oil and phytic acid, its method comprises the following steps: carry out ultrasonic extraction after being mixed with the second inverse micellar solution as above by Testa oryzae, described Testa oryzae is 50～200 orders, it is preferably 60～100 orders, Testa oryzae is mixed with inverse micellar solution, its ratio is 0.01～0.2g Testa oryzae/mL inverse micellar solution, it is preferably 0.01～1.0g Testa oryzae/1mL inverse micellar solution, ultrasonic extraction more than 0.1 hour, it is preferably 0.1～2 hour, liquid is obtained after being then peeled off solid, described liquid is added buffer, it is adjusted to pH1.0～5.0, layering obtains oil phase and aqueous phase.Obtained oil phase and aqueous phase contain Testa oryzae oil and phytic acid respectively.Being mixed with alkali salt by described aqueous phase, then pH value is adjusted to 6～9, obtains phytate solid.
Method as above, after wherein said oil phase breakdown of emulsion, removes solvent and obtains Testa oryzae oil.
Method as above, the wherein said solid obtained adds water, preferably, its solid-liquid mass ratio is 1:5～1:10, temperature 30～60 DEG C, regulating to pH2.0～5.0, extract 1～5h, the clear liquid being filtrated to get adds alkali salt and mixes, it is adjusted to pH6.0～9.0 again, obtains phytate solid.
A third aspect of the present invention, also discloses a kind of reverse micelle solvent as above and is processing or extracting the purposes in Testa oryzae.
Detailed description of the invention
The present inventor is through extensive and deep research, by improving component and the pH value of micellar solution, for the shortcoming that Testa oryzae can't efficiently be processed by current inverse micellar solution, the present invention proposes the inverse micellar solution of a kind of effective extraction Testa oryzae and the corresponding method extracted or process Testa oryzae thereof.Complete the present invention on this basis.
The technology design of the present invention is as follows:
Instant invention overcomes the defect of prior art, propose a kind of processing method utilizing inverse micelle abstraction Testa oryzae, the inverse micellar solution that this technology utilization is unique, the particular surface in inverse micellar solution is lived agent and specific pH value, Testa oryzae is processed, by the effective ingredient in Testa oryzae, having efficiently separated such as Testa oryzae oil, protein and phytic acid etc. and reclaimed, processing method is simple, effect is obvious, expand the purposes of Testa oryzae, therefore, further add the added value of Testa oryzae.
In the present invention, term " contains " or " including " represents that various composition can be applied in mixture or the compositions of the present invention together.Therefore, term " mainly by ... composition " and " by ... form " be included in term " containing " or " including ".
Hereinafter it is described in detail to various aspects of the present invention:
Raw material
In the present invention, the Testa oryzae handled by reverse micelle derives from rice, for obtaining according in the prior art being currently known.Testa oryzae is the mixture in brown rice pearling process by the cortex (including peel, seed coat, megarchidium layer and aleurone) under stone roller and a small amount of rice germ and small brokens bits.
Reverse micelle solvent
In inverse micellar solution described herein, containing organic solvent, possibly together with buffer solution and alkyl sulfate.
Described organic solvent is selected from alkane, alkane alcohol, carbon tetrachloride, arene and their homologue, organic solvent in the application is preferably alkane solvent and alkane alcoholic solvent, wherein organic solvent refers to and can make what solvent used under room temperature, it it is at room temperature liquid organic compound, wherein the non-limitative example of alkane includes: selected from 5 carbon alkanes, 6 carbon alkanes, 7 carbon alkanes, 8 carbon alkanes, 5 carbocyclic ring alkane, 6 carbocyclic ring alkane, 7 carbocyclic ring alkane, 8 carbocyclic ring alkane or its homologue, concrete, pentane can be selected from, isopentane, normal hexane, neohexane, heptane, isoheptane, octane, isobutyltrimethylmethane., normal octane, hexamethylene or its homologue；Described alkane alcoholic solvent is the alcohols material that can use as solvent under room temperature, its non-limitative example includes being selected from the low carbon chain alcohol that carbon number is less than 10, and its non-limitative example includes one or more in methanol, ethanol, propanol, propylene glycol, butanol, amylalcohol, hexanol, capryl alcohol, enanthol, nonyl alcohol or its alcohols homologue.Homologue in the application, is that structure is similar, difference 1 or several " CH on composition2" organic compound of atomic group.The volume ratio of described alkane solvent and alkane alcoholic solvent is 1:1～10:1, and the effect in reaction system of the described alkane solvent is identical, and therefore these solvents all can be replaced.
Described buffer solution is from the buffer solution of various routines, such as the buffer system of weak acid class, such as phosphate system, Fructus Citri Limoniae acids buffer system, glycine-HCI buffer, phthalic acid-hydrochloride buffer system etc..Buffer system in the application uses as regulating pH value, it is possible to adding buffer and make the pH of system be maintained at certain scope, it act as pH and is adjusted to certain scope, and regulates the Wo value of inverse micellar solution accordingly.
Also including the alkyl sulfate that carbon number is 6 to 20 in described inverse micellar solution, it is the acid obtained after alkanes substance carries out sulfonation with sulphuric acid.Its non-limitative example includes: sodium lauryl sulphate (SDS), sodium hexadecyl sulfate, ten alkyl sodium sulfates etc..Inventor finds in an experiment, adopt the surfactant of alkyls, compared to other type of surfactant, have, for the extraction efficiency of Testa oryzae, a phytic acid being obviously improved and can extracting in Testa oryzae simultaneously, and this result has been inventor has been surprisingly found that in an experiment.Therefore, the inverse micellar solution in the application is particularly suitable for the application of Testa oryzae, relative to the process object in the present invention, has prominent applicable performance.
In a specific embodiment of the present invention, the reverse micelle solvent in a kind of inverse micellar solution a includes alkane and alkane alcohol, also includes containing buffer solution and alkyl sulfate, and in described reverse micelle, the content of alkyl sulfate is 50g/L～100g/L；The pH of described inverse micellar solution is 8.0～11.0, and the mol ratio Wo value of water and surfactant is 10～40, and the weight content of described alkaline protease is 0.001%～0.5%, it is preferred to 0.1～0.3%.When adopting described inverse micellar solution that Testa oryzae is processed, it is possible to be used as to extract Testa oryzae oil and protein simultaneously.The both products obtained have the productivity of excellence.
In a specific embodiment of the present invention, the reverse micelle solvent in a kind of inverse micellar solution a ' includes alkane and alkane alcohol, also includes containing buffer solution and alkyl sulfate, and in described reverse micelle, the content of alkyl sulfate is 50g/L～100g/L；In described inverse micellar solution, the addition of slow buffer solution is 0.01L/L-0.20L/L, and the pH of described described buffer solution is 8.0～11.0, and the weight content of described alkaline protease is 0.001%～0.5%, it is preferred to 0.1～0.3%.The mol ratio Wo value of the described water in inverse micellar solution and surfactant is 10～40.When adopting described inverse micellar solution that Testa oryzae is processed, it is possible to be used as to extract Testa oryzae oil and protein simultaneously.The both products obtained have the productivity of excellence.
In a specific embodiment of the present invention, the reverse micelle solvent in a kind of inverse micellar solution b includes alkane and alkane alcohol, also includes containing buffer solution and alkyl sulfate, and in described reverse micelle, the content of alkyl sulfate is 50g/L～100g/L；The pH of described inverse micellar solution is 1.0～5.0, and the mol ratio Wo of water and surfactant is 10～40.When adopting described inverse micellar solution that Testa oryzae is processed, it is possible to be used as to extract Testa oryzae oil and phytic acid simultaneously.The both products obtained have the productivity of excellence.
In a specific embodiment of the present invention, the reverse micelle solvent in a kind of inverse micellar solution b ' includes alkane and alkane alcohol, also includes containing buffer solution and alkyl sulfate, and in described reverse micelle, the content of alkyl sulfate is 50g/L～100g/L；In inverse micellar solution, the addition of buffer solution is 0.01L/L-0.20L/L, and the pH of described buffer solution is 1.0～5.0.In this inverse micellar solution, the mol ratio Wo value of water and surfactant is 10～40.When adopting described inverse micellar solution that Testa oryzae is processed, it is possible to be used as to extract Testa oryzae oil and phytic acid simultaneously.The both products obtained have the productivity of excellence.
Extraction step
The above-mentioned inverse micellar solution that utilizes in the present invention can process Testa oryzae efficiently.
In the extracting process of the present invention, above-mentioned inverse micellar solution is mixed with Testa oryzae, after ultrasonic extraction, separate solid, the liquid obtained adds buffer solution, then carries out ultrasonic extraction, obtain oil phase and aqueous phase.Separate oil phase and aqueous phase, after oil phase is carried out breakdown of emulsion, carry out solution is distilled off, obtain Testa oryzae oil.Above-mentioned breakdown of emulsion, can adopt in this area conventional method to process, and as added demulsifier or carrying out freezing and carry out breakdown of emulsion, described organic solvent includes the breakdown of emulsion solvents such as acetone, methanol, ethanol.In one embodiment of the invention, the ethanol water adopting the aqueous solution such as 20～70% of ethanol carries out breakdown of emulsion.
The mixed solution that described buffer solution is made up of weak acid and salt, weak base and salt thereof, can offset to a certain extent, alleviate the impact on solution acidity of additional strong acid or highly basic, is the known solution of a kind of routine in this area.Being used in the present invention can be the buffer solution of various routine, such as the buffer system of weak acid class, such as phosphate system, Fructus Citri Limoniae acids buffer system, glycine-HCI buffer, Acetic acid-sodium acetate buffer, phthalic acid-hydrochloride buffer system etc..Buffer system in the application uses as regulating pH value, it is possible to adds buffer and makes the pH of system be maintained at certain scope.As among the process step of the present invention, it is necessary to the pH using buffer solution is 1～5, or pH is the buffer of 8～11.In a preferred embodiment of the present invention, the described electrolyte content in buffer solution is 0.05～3mol/L, it is preferred to 0.05～2mol/L, and described electrolyte is selected from the chlorate of potassium or sodium.
In a specific embodiment of the present invention, Testa oryzae adds inverse micellar solution a or a ' as above mix, the inverse micellar solution a of every milliliter adds Testa oryzae 0.01～0.2g, it is preferably 0.01～0.1, namely inverse micellar solution is 0.01～0.2g/mL with the ratio of Testa oryzae, it is preferred to 0.01～0.1g/mL, after being extracted by described mixture ultrasound wave, separate solid, obtain liquid.Add buffer as above in a liquid, make pH be adjusted to about 8～11.Then, carry out ultrasonic extraction, obtain oil phase and aqueous phase, wherein adopt method as above that oil phase carries out process and obtain Testa oryzae oil, and aqueous phase is easily separated albumen and processes, obtain rice bran protein.
The method separating albumen in aqueous phase includes dialysis, ultrafiltration, salting-out separation, adsorption chromatography etc..
In the application another embodiment, Testa oryzae adds inverse micellar solution b or b ' as above mix, the inverse micellar solution b of every milliliter adds Testa oryzae 0.01～0.2g, it is preferably 0.01～0.1g, namely Testa oryzae is 0.01～0.2g/mL with the mixed proportion of inverse micellar solution, it is preferred to 0.01～0.1g/mL, after being extracted by described mixture ultrasound wave, separate solid, obtain liquid.Add buffer in a liquid, make pH be adjusted to about 1～5, then carry out ultrasonic extraction, obtain oil phase and aqueous phase.Adopt method as above that oil phase is carried out process and obtain Testa oryzae oil, and to aqueous phase, being transferred to pH by alkaline-earth metal salt treatment is 3～5, it is preferably pH4～5, again solution aqueous slkali is adjusted to pH6.0～9.0, obtain phytic acid alkali salt, it is preferred that aqueous phase magnesium slaine or calcium slaine are processed.The non-limitative example of the alkali salt of magnesium or calcium includes: magnesium chloride, calcium chloride, calcium hydroxide, magnesium hydroxide, magnesium carbonate, calcium carbonate, magnesium bicarbonate, calcium bicarbonate or its mixture.It was found by the inventors of the present invention that after the inorganic salt of magnesium or calcium is added described aqueous phase, described inorganic salt can produce precipitation with phytic acid, precipitates out with the form of phytic acid magnesium or phytic acid calcium.
The mixed proportion of above-mentioned Testa oryzae and inverse micellar solution is preferably 0.02～0.08g/mL.
Heretofore described ultrasonic extraction is the abstraction technique of a kind of routine, its principle is that the strong cavitation utilizing Ultrasonic Radiation pressure to produce is answered effect, mechanical vibration, disturbance effect, high acceleration, emulsifying, spreads, smashed and the multistage effects such as stirring action, increase molecular motion of material frequency and speed, increase solvent penetration power, thus accelerating target component to enter solvent, promote the carrying out extracted.Those skilled in the art can select suitable power for Testa oryzae is carried out ultrasonic extraction.The ultrasound wave that power is 100～500W adopted in the present invention extracts.
Summary
The method of the present invention may include that and carries out ultrasonic extraction by adding Testa oryzae in the inverse micellar solution containing organic solvent and alkyl sulfate, and the liquid obtained after the extract separation solid obtained is divided into oil phase and aqueous phase.Reclaiming respectively of the oil phase that obtains and aqueous phase, can obtain Testa oryzae oil, protein and phytic acid.
Advantage
The method of the present invention processes Testa oryzae, has reached good effect, has specifically had the advantage that
1. improve the technique that Testa oryzae elder generation solvent extraction oils and fats carries out rice bran protein extraction again, overcome rice bran protein serious degenerative in solvent extracting process so that the problem that cannot extract, improve the extraction ratio of rice bran protein, rice bran protein extraction rate reached is to 40%, even up to more than 60%.Utilizing the Testa oryzae slag that reverse micelle obtains after extracting Testa oryzae is raw material, extracts further and obtains phytic acid calcium and rice bran-fiber, comprehensively utilize Testa oryzae, improves the value of Testa oryzae.
2. the present invention utilizes reverse micelle to extract Testa oryzae oils and fats and phytic acid simultaneously, obtains Testa oryzae oil and low phytic acid rice bran meal simultaneously, solves the nutritional labeling problem in the application of rice bran meal.
3. present invention process is simple, agents useful for same reusable edible, extracts cost low, environmentally safe.
As no specific instructions, the various raw materials of the present invention all can pass through to be commercially available；Or the conventional method according to this area prepares.Unless otherwise defined or described herein, the same meaning that all specialties used herein are familiar with scientific words and those skilled in the art.In addition any method similar or impartial to described content and material all can be applicable in the inventive method.
Other aspects of the present invention, due to this disclosure, are apparent to those skilled in the art.
Below in conjunction with specific embodiment, the present invention is expanded on further.Should be understood that these embodiments are merely to illustrate the present invention rather than restriction the scope of the present invention.The experimental technique of unreceipted actual conditions in the following example, generally measures according to national standard.If there is no corresponding national standard, then according to general international standard, normal condition or according to manufacturer it is proposed that condition carry out.Unless otherwise indicated, otherwise all of number is weight portion, and all of percentage ratio or ratio are weight percentage or weight ratio.
Unless otherwise defined or described herein, the same meaning that all specialties used herein are familiar with scientific words and those skilled in the art.In addition any method similar or impartial to described content and material all can be applicable in the inventive method.
Heretofore described Testa oryzae is with high-quality fresh rice bran for raw material, through cleaning, screening, extruding, dries, pulverizes and obtains.Extrusion process adopts RicebranTechnologies company of U.S. stabilization equipment, and High Temperature High Pressure (130 DEG C, 276kPa), the extruding thickness of sample obtained is thin.
Basic protein enzyme source and kind are Novi's letter alkaline protease Alcalase2.4LFG (subtilisin), nominal vigor 2.4AU-A/g.
The detection method of the mol ratio Wo of water and surfactant is shown in: the mensuration karr. Fei Xiufa of moisture in GB/T6283-2008 chemical products；
The source of SDS is Tianjin Kermel Chemical Reagent Co., Ltd., AR level.
Hyperacoustic power is 240W.
The preparation of the buffer used in following embodiment: KH2PO4-Na2HPO4The configuration of buffer solution: take the Na of the 1/15mol/L of 500mL2HPO4Solution, adds the KH of 1/15mol/L2PO4Regulate pH to 8.0,8.68,7.0 respectively, then press 0.1mol/L or 1mol/L concentration addition KCL mix homogeneously；C6H8O7-Na2HPO4The configuration of buffer solution: take the 0.2mol/LC of 500mL6H8O7Solution, adds 0.2mol/LNa2HPO4Solution regulate respectively pH to 2.5,4.5.
Embodiment 1
The configuration of SDS (dodecyl sodium sulfate) inverse micellar solution: weigh 30gSDS, joins in 500mL isobutyltrimethylmethane ./n-octyl alcohol (5:1) solvent, adds the KH of the 0.1mol/LKCL of 70mL2PO4-Na2HPO4Buffer solution (pH=8.68), and add 1.425mL alkaline protease, then supersonic oscillations are completely dissolved to SDS, solution is transparent is then reverse micelle, and to make the pH of inverse micellar solution be 8.68, Wo values is 30, the content of SDS is 52.6g/L, and alkaline protease content is 0.25%.
Testa oryzae is crossed 80 mesh sieves and is then mixed with the ratio of 0.05g/mL with the inverse micellar solution configured, ultrasound assisted extraction 1h, after extraction centrifugal (3000r/min, 10min), is centrifuged and obtains forward extraction liquid and Testa oryzae slag.Forward extraction liquid is added isopyknic 1.0mol/LKCl buffer (KH2PO4-Na2HPO4Buffer, pH=8), ultrasound on extracting certain time, then above-mentioned solution is centrifuged (3000r/min, 10min), is divided into oil phase and aqueous phase biphase.Aqueous phase is dry after dialysis (bag filter molecular weight 7000Da) 48h obtains rice bran protein, rice bran protein extraction ratio 69.90%.Oil phase adds 70% alcoholic solution of 2.5 times of volumes, stirs breakdown of emulsion 1h, and centrifugal (3000r/min, 10min) layering, upper oil phase rotary evaporation obtains Rice bran crude oil, Rice bran crude oil extraction ratio 80.0% after removing organic solvent.
Being added to the water (solid-liquid weight/volume 1:9) by Testa oryzae slag, acid leaching extraction phytic acid (is adjusted to pH=3.0), keeps 4h at 50 DEG C, is centrifuged (3000r/min, 10min), obtains supernatant and rice bran-fiber after extraction.Supernatant carries out two-step precipitation, with 10%Ca (OH)2It is adjusted to pH4.5, then is adjusted to pH7.5 with 10%NaOH, be then centrifuged (3000r/min, 10min) and obtain phytic acid calcium precipitation, obtain phytic acid calcium, phytic acid calcium yield 9.65% after drying.
Embodiment 2
The configuration of SDS inverse micellar solution: weigh 30gSDS, joins in 500mL isobutyltrimethylmethane ./n-octyl alcohol (5:1) solvent, adds the KH of the 0.1mol/LKCL of 70mL2PO4-Na2HPO4Buffer solution (pH=8.68), then supersonic oscillations are completely dissolved to SDS, and solution is transparent is then reverse micelle, the pH of inverse micellar solution is 8.68, Wo values be 30, SDS content be 52.6g/L.
Testa oryzae is crossed 80 mesh sieves and is then mixed with the ratio of 0.05g/mL with the inverse micellar solution configured, ultrasound assisted extraction 1h, after extraction centrifugal (3000r/min, 10min), is centrifuged and obtains forward extraction liquid and Testa oryzae slag.Forward extraction liquid is added isopyknic 1.0mol/LKCl buffer (KH2PO4-Na2HPO4Buffer, pH=8), ultrasound on extracting certain time, then above-mentioned solution is centrifuged (3000r/min, 10min), is divided into oil phase and aqueous phase biphase.Aqueous phase is dry after dialysis (bag filter molecular weight 7000Da) 48h obtains rice bran protein, rice bran protein extraction ratio 59.86%.Oil phase adds 70% alcoholic solution of 2.5 times of volumes, stirs breakdown of emulsion 1h, and centrifugal (3000r/min, 10min) layering, upper oil phase rotary evaporation obtains Rice bran crude oil, Rice bran crude oil extraction ratio 75.8% after removing organic solvent.
Comparative example 1
The configuration of SDS inverse micellar solution: weigh 30gSDS, joins in 500mL isobutyltrimethylmethane ./n-octyl alcohol (5:1) solvent, adds the KH of the 0.1mol/LKCL of 70mL2PO4-Na2HPO4Buffer solution (pH=7.0), and add 1.425mL alkaline protease, then supersonic oscillations are completely dissolved to SDS, solution is transparent is then reverse micelle, and to make the pH of inverse micellar solution be 7.0, Wo values is 30, the content of SDS is 52.6g/L, and alkaline protease content is 0.25%.
Testa oryzae is crossed 80 mesh sieves and is then mixed with the ratio of 0.05g/mL with the inverse micellar solution configured, ultrasound assisted extraction 1h, after extraction centrifugal (3000r/min, 10min), is centrifuged and obtains forward extraction liquid and Testa oryzae slag.Forward extraction liquid is added isopyknic 1.0mol/LKCl buffer (KH2PO4-Na2HPO4Buffer, pH=8), ultrasound on extracting certain time, then above-mentioned solution is centrifuged (3000r/min, 10min), is divided into oil phase and aqueous phase biphase.Aqueous phase is dry after dialysis (bag filter molecular weight 7000Da) 48h obtains rice bran protein, rice bran protein extraction ratio 3.02%.Oil phase adds 70% alcoholic solution of 2.5 times of volumes, stirs breakdown of emulsion 1h, and centrifugal (3000r/min, 10min) layering, upper oil phase rotary evaporation obtains Rice bran crude oil, Rice bran crude oil extraction ratio 76.5% after removing organic solvent.
Comparative example 2
Weigh 30gSDS, join in 500mL isobutyltrimethylmethane ./n-octyl alcohol (5:1) solvent, add the KH of the 0.1mol/LKCL of 70mL2PO4-Na2HPO4Buffer solution (pH=7.0), then supersonic oscillations are completely dissolved to SDS, and solution is transparent is then reverse micelle, and the content making the pH of inverse micellar solution to be 7.0, Wo values be 30, SDS is 52.6g/L.
Testa oryzae is crossed 80 mesh sieves and is then mixed with the ratio of 0.05g/mL with the inverse micellar solution configured, ultrasound assisted extraction 1h, after extraction centrifugal (3000r/min, 10min), is centrifuged and obtains forward extraction liquid and Testa oryzae slag.Forward extraction liquid is added isopyknic 1.0mol/LKCl buffer (KH2PO4-Na2HPO4Buffer, pH8), ultrasound on extracting certain time, then above-mentioned solution is centrifuged (3000r/min, 10min), is divided into oil phase and aqueous phase biphase.Aqueous phase is dry after dialysis (bag filter molecular weight 7000Da) 48h obtains rice bran protein, rice bran protein extraction ratio 1.25%.Oil phase adds 70% alcoholic solution of 2.5 times of volumes, stirs breakdown of emulsion 1h, and centrifugal (3000r/min, 10min) layering, upper oil phase rotary evaporation obtains Rice bran crude oil, Rice bran crude oil extraction ratio 75.5% after removing organic solvent.
Embodiment 3
Weigh 30gSDS, join in 500mL isobutyltrimethylmethane ./n-octyl alcohol (5:1) solvent, add the NaOH solution (pH=11) of the 0.1mol/LKCL of 70mL, then supersonic oscillations are completely dissolved to SDS, solution is transparent is then reverse micelle, the content making the pH of inverse micellar solution to be 11.0, Wo values be 30, SDS is 52.6g/L.
Testa oryzae is crossed 80 mesh sieves and is then mixed with the ratio of 0.05g/mL with the inverse micellar solution configured, ultrasound assisted extraction 1h, after extraction centrifugal (3000r/min, 10min), is centrifuged and obtains forward extraction liquid and Testa oryzae slag.Forward extraction liquid is added isopyknic 1.0mol/LKCl buffer (KH2PO4-Na2HPO4Buffer, pH=8), ultrasound on extracting certain time, then above-mentioned solution is centrifuged (3000r/min, 10min), is divided into oil phase and aqueous phase biphase.Aqueous phase is dry after dialysis (bag filter molecular weight 7000Da) 48h obtains rice bran protein, rice bran protein extraction ratio 58.50%.Oil phase adds 70% alcoholic solution of 2.5 times of volumes, stirs breakdown of emulsion 1h, and centrifugal (3000r/min, 10min) layering, upper oil phase rotary evaporation obtains Rice bran crude oil, Rice bran crude oil extraction ratio 74.3% after removing organic solvent.
Comparative example 3
The configuration of CTAB (cetyl trimethylammonium bromide) inverse micellar solution: weigh 30gCTAB, joins in 500mL isobutyltrimethylmethane ./n-octyl alcohol (5:1) solvent, adds the KH of the 0.1mol/LKCL of 70mL2PO4-Na2HPO4Buffer solution (pH=8.68), and add 1.425mL alkaline protease, then supersonic oscillations are completely dissolved to CTAB, solution is transparent is then reverse micelle, and to make the pH of inverse micellar solution be 8.68, Wo values is 30, the content of CTAB is 52.6g/L, and alkaline protease content is 0.25%.
Testa oryzae is crossed 80 mesh sieves and is then mixed with the ratio of 0.05g/mL with the inverse micellar solution configured, ultrasound assisted extraction 1h, after extraction centrifugal (3000r/min, 10min), is centrifuged and obtains forward extraction liquid and Testa oryzae slag.Forward extraction liquid is added isopyknic 1.0mol/LKCl buffer (KH2PO4-Na2HPO4Buffer, pH=8), ultrasound on extracting certain time, then above-mentioned solution is centrifuged (3000r/min, 10min), is divided into oil phase and aqueous phase biphase.Aqueous phase is dry after dialysis (bag filter molecular weight 7000Da) 48h obtains rice bran protein, rice bran protein extraction ratio 35.55%.Oil phase adds 70% alcoholic solution of 2.5 times of volumes, stirs breakdown of emulsion 1h, and centrifugal (3000r/min, 10min) layering, upper oil phase rotary evaporation obtains Rice bran crude oil, Rice bran crude oil extraction ratio 56.5% after removing organic solvent.
Embodiment 4
The configuration of SDS inverse micellar solution: weigh 30gSDS, joins in 500mL isobutyltrimethylmethane ./n-octyl alcohol (5:1) solvent, adds the KH of the 0.1mol/LKCL of 70mL2PO4-Na2HPO4Buffer solution (pH=8.68), and add 1.425mL alkaline protease, then supersonic oscillations are completely dissolved to SDS, solution is transparent is then reverse micelle, and to make the pH of inverse micellar solution be 8.68, Wo values is 30, the content of SDS is 52.6g/L, and alkaline protease content is 0.25%.
Testa oryzae is crossed 80 mesh sieves and is then mixed with the ratio of 0.11g/mL with the inverse micellar solution configured, ultrasound assisted extraction 1h, after extraction centrifugal (3000r/min, 10min), is centrifuged and obtains forward extraction liquid and Testa oryzae slag.Forward extraction liquid is added isopyknic 1.0mol/LKCl buffer (KH2PO4-Na2HPO4Buffer, pH=8), ultrasound on extracting certain time, then above-mentioned solution is centrifuged (3000r/min, 10min), is divided into oil phase and aqueous phase biphase.Aqueous phase is dry after dialysis (bag filter molecular weight 7000Da) 48h obtains rice bran protein, rice bran protein extraction ratio 45.10%.Oil phase adds 70% alcoholic solution of 2.5 times of volumes, stirs breakdown of emulsion 1h, and centrifugal (3000r/min, 10min) layering, upper oil phase rotary evaporation obtains Rice bran crude oil, Rice bran crude oil extraction ratio 75.4% after removing organic solvent.
As seen from the above, when using the technical scheme in the present invention, it is possible to be obviously improved the extraction ratio of the oils and fats in Testa oryzae and protein.When using alkaline protease, effect is more preferably.
Embodiment 5
The configuration of SDS inverse micellar solution: weigh 30gSDS, joins in 500mL isobutyltrimethylmethane ./n-octyl alcohol (5:1) solvent, adds the C of 70mL6H8O7-Na2HPO4Buffer solution (pH=2.5), then supersonic oscillations are completely dissolved to SDS, and solution is transparent is then reverse micelle, and the content making the pH of inverse micellar solution to be 2.5, Wo values be 30, SDS is 52.6g/L.
Testa oryzae is crossed 80 mesh sieves and is then mixed with the ratio of 0.05g/mL with the inverse micellar solution configured, ultrasound assisted extraction 1h, after extraction centrifugal (3000r/min, 10min), is centrifuged and obtains forward extraction liquid and Testa oryzae slag.Forward extraction liquid is added isopyknic 1.0mol/LKCl buffer (C6H8O7-Na2HPO4Buffer, pH=2.5), ultrasound on extracting certain time, then above-mentioned solution is centrifuged (3000r/min, 10min), is divided into oil phase and aqueous phase biphase.Oil phase adds 70% alcoholic solution of 2.5 times of volumes, stirs breakdown of emulsion 1h, centrifugal layering, and upper oil phase rotary evaporation obtains Rice bran crude oil after removing organic solvent, and Rice bran crude oil extracts 81.2%.Aqueous phase is with 10%Ca (OH)2Adjust pH4.5, then be adjusted to pH7.5 with 10%NaOH, be then centrifuged (3000r/min, 10min) and obtain phytic acid calcium precipitation, obtain phytic acid calcium, phytic acid extraction ratio 60.92% after drying.Testa oryzae slag drying obtains low phytic acid rice bran meal, phytic acid content 4.00% in rice bran meal.
Embodiment 6
The configuration of SDS inverse micellar solution: weigh 30gSDS, joins in 500mL isobutyltrimethylmethane ./n-octyl alcohol (5:1) solvent, adds the C of 70mL6H8O7-Na2HPO4Buffer solution (pH=4.5), then supersonic oscillations are completely dissolved to SDS, and solution is transparent is then reverse micelle, and the content making the pH of inverse micellar solution to be 4.5, Wo values be 30, SDS is 52.6g/L.
Testa oryzae is crossed 80 mesh sieves and is then mixed with the ratio of 0.05g/mL with the inverse micellar solution configured, ultrasound assisted extraction 1h, after extraction centrifugal (3000r/min, 10min), is centrifuged and obtains forward extraction liquid and Testa oryzae slag.Forward extraction liquid is added isopyknic 1.0mol/LKCl buffer (C6H8O7-Na2HPO4Buffer, pH4.5), ultrasound on extracting certain time, then above-mentioned solution is centrifuged (3000r/min, 10min), is divided into oil phase and aqueous phase biphase.Oil phase adds 70% alcoholic solution of 2.5 times of volumes, stirs breakdown of emulsion 1h, centrifugal layering, and upper oil phase rotary evaporation obtains Rice bran crude oil after removing organic solvent, and Rice bran crude oil extracts 81.2%.Aqueous phase is with 10%Ca (OH)2Adjust pH4.5, then be adjusted to pH7.5 with 10%NaOH, be then centrifuged (3000r/min, 10min) and obtain phytic acid calcium precipitation, obtain phytic acid calcium, phytic acid extraction ratio 55.71% after drying.Testa oryzae slag drying obtains low phytic acid rice bran meal, phytic acid content 4.53% in rice bran meal.
Embodiment 7
The configuration of SDS inverse micellar solution: weigh 30gSDS, joins in 500mL isobutyltrimethylmethane ./n-octyl alcohol (5:1) solvent, adds the C of 70mL6H8O7-Na2HPO4Buffer solution (pH=2.5), then supersonic oscillations are completely dissolved to SDS, and solution is transparent is then reverse micelle, and the content making the pH of inverse micellar solution to be 2.5, Wo values be 30, SDS is 52.6g/L.
Testa oryzae is crossed 80 mesh sieves and is then mixed with the ratio of 0.11g/mL with the inverse micellar solution configured, ultrasound assisted extraction 1h, after extraction centrifugal (3000r/min, 10min), is centrifuged and obtains forward extraction liquid and Testa oryzae slag.Forward extraction liquid is added isopyknic 1.0mol/LKCl buffer (C6H8O7-Na2HPO4Buffer, pH2.5), ultrasound on extracting certain time, then above-mentioned solution is centrifuged (3000r/min, 10min), is divided into oil phase and aqueous phase biphase.Oil phase adds 70% alcoholic solution of 2.5 times of volumes, stirs breakdown of emulsion 1h, centrifugal layering, and upper oil phase rotary evaporation obtains Rice bran crude oil after removing organic solvent, and Rice bran crude oil extracts 74.5%.Aqueous phase is with 10%Ca (OH)2Adjust pH4.5, then be adjusted to pH7.5 with 10%NaOH, be then centrifuged (3000r/min, 10min) and obtain phytic acid calcium precipitation, obtain phytic acid calcium, phytic acid extraction ratio 45.84% after drying.Testa oryzae slag drying obtains low phytic acid rice bran meal, phytic acid content 5.54% in rice bran meal.
From the foregoing, it will be observed that when using the technical scheme in the present invention, it is possible to extract Testa oryzae oils and fats and phytic acid simultaneously, and both extraction ratios are all higher, reached to extract the effect of active ingredient in Testa oryzae simultaneously.
The foregoing is only presently preferred embodiments of the present invention, it is not limited to the substantial technological context of the present invention, the substantial technological content of the present invention is broadly to be defined in the right of application, any technology entities that other people complete or method, if defined with the right of application is identical, also or the change of a kind of equivalence, all it is covered by being considered among this right.
The all documents mentioned in the present invention are incorporated as reference all in this application, are individually recited as reference such just as each section of document.In addition, it is to be understood that after the foregoing having read the present invention, the present invention can be made various changes or modifications by those skilled in the art, these equivalent form of values fall within the application appended claims limited range equally.
Claims (10)
1. an inverse micellar solution, it is characterised in that described reverse micelle solvent includes organic solvent, also includes buffer solution and alkyl sulfate, and in described inverse micellar solution, the content of alkyl sulfate is 10g/L～150g/L, it is preferred to 50g/L～100g/L；The pH of described inverse micellar solution is 8.0～11.0, it is preferred that also comprise the alkaline protease that weight content is 0.001～0.5% in described inverse micellar solution, it is preferred to 0.1～0.3%.
2. an inverse micellar solution, it is characterised in that described reverse micelle solvent includes organic solvent, also includes buffer solution and alkyl sulfate, and in described inverse micellar solution, the content of alkyl sulfate is 10g/L～150g/L, it is preferred to 50g/L～100g/L；The pH of described inverse micellar solution is 1.0～5.0.
3. inverse micellar solution as claimed in claim 1 or 2, it is characterized in that, described alkyl sulfate is carbon number is the alkyl sulfate of 6 to 20, and/or, described organic solvent is selected from alkane and alkane alcohol, described alkane solvent preferably is selected from 5 carbon alkanes, 6 carbon alkanes, 7 carbon alkanes, 8 carbon alkanes, 5 carbocyclic ring alkane, 6 carbocyclic ring alkane, 7 carbocyclic ring alkane, 8 carbocyclic ring alkane and/or its homologue, and/or, alkane alcohol is selected from the low carbon chain alcohol that carbon number is less than 10, it is preferably methanol, ethanol, propanol, propylene glycol, butanol, amylalcohol, hexanol, capryl alcohol, enanthol, one or more in nonyl alcohol, and/or the volume ratio of described alkane solvent and alkane alcoholic solvent is 1:1～10:1；And/or water is 10～40 with the mol ratio Wo value of surfactant in described inverse micellar solution, it is preferable to 20～40.
4. the method simultaneously extracting Testa oryzae oil and rice bran protein, it is characterised in that:
Ultrasonic extraction is carried out with the inverse micellar solution described in claim 1 after being mixed by Testa oryzae, preferably, described Testa oryzae is 50～200 order Testa oryzaes, it is more preferably the Testa oryzae of 60～100 orders, mixed proportion is 0.01～0.2g Testa oryzae/mL inverse micellar solution, it is preferably 0.01～0.1g Testa oryzae/mL inverse micellar solution, ultrasonic extraction more than 0.1 hour, it is preferably 0.1～2 hour, liquid is obtained after being then peeled off solid, being 8.0～11.0 by being adjusted to pH after addition buffer in described liquid, carry out ultrasonic extraction, layering obtains aqueous phase and oil phase.
5. the method simultaneously extracting Testa oryzae oil and phytic acid, it is characterised in that comprise the following steps:
Ultrasonic extraction is carried out with the inverse micellar solution described in claim 2 after being mixed by Testa oryzae, preferably, described Testa oryzae is 50～200 order Testa oryzaes, more preferably the Testa oryzae of 60～100 orders, mixed proportion is 0.01～0.2g/1mL inverse micellar solution, it is preferably 0.01～0.1g Testa oryzae/1mL inverse micellar solution, ultrasonic extraction more than 0.1 hour, it is preferred to 0.1～2 hour, liquid is obtained after being then peeled off solid, described liquid being added buffer, is adjusted to pH1.0～5.0, layering obtains oil phase and aqueous phase.
6. the method as described in claim 4 or 5, it is characterised in that after described oil phase breakdown of emulsion, removes solvent and obtains Testa oryzae oil.
7. method as claimed in claim 4, it is characterised in that described aqueous phase is easily separated albumen and processes, obtain protein.
8. method as claimed in claim 5, it is characterised in that being mixed with alkali salt by described aqueous phase, then pH value is adjusted to 6～9, obtains phytate solid, and described alkali salt preferably is selected from calcium slaine or magnesium slaine.
9. method as claimed in claim 4, it is characterized in that, the described solid obtained adds water regulate to pH2.0～5.0, preferably, solid-to-liquid ratio is 1:5～1:10, and more preferably in extracting 1～5h at temperature 30～60 DEG C, the clear liquid being filtrated to get adds alkali salt and mixes, it is adjusted to pH6.0～9.0 again, obtains phytate solid.
10. reverse micelle solvent as claimed in claim 1 or 2 is processing or is extracting the purposes in Testa oryzae.
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| CN109238815A (en) * | 2018-09-19 | 2019-01-18 | 上海市计量测试技术研究院 | A kind of pre-treating method for oil content in detection carbon slag |
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