CN111772188A - Liver polypeptide for protecting liver and improving liver function and composition thereof - Google Patents
Liver polypeptide for protecting liver and improving liver function and composition thereof Download PDFInfo
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- CN111772188A CN111772188A CN202010645508.2A CN202010645508A CN111772188A CN 111772188 A CN111772188 A CN 111772188A CN 202010645508 A CN202010645508 A CN 202010645508A CN 111772188 A CN111772188 A CN 111772188A
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- liver
- temperature
- extract
- turmeric
- polypeptide
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Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J3/00—Working-up of proteins for foodstuffs
- A23J3/04—Animal proteins
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J3/00—Working-up of proteins for foodstuffs
- A23J3/30—Working-up of proteins for foodstuffs by hydrolysis
- A23J3/32—Working-up of proteins for foodstuffs by hydrolysis using chemical agents
- A23J3/34—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes
- A23J3/341—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes of animal proteins
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/38—Other non-alcoholic beverages
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/385—Concentrates of non-alcoholic beverages
- A23L2/39—Dry compositions
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/17—Amino acids, peptides or proteins
- A23L33/18—Peptides; Protein hydrolysates
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C45/00—Preparation of compounds having >C = O groups bound only to carbon or hydrogen atoms; Preparation of chelates of such compounds
- C07C45/78—Separation; Purification; Stabilisation; Use of additives
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- Nutrition Science (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Mycology (AREA)
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- Proteomics, Peptides & Aminoacids (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
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- Botany (AREA)
- Medicines Containing Plant Substances (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
The present invention discloses a liver polypeptide for protecting liver and improving liver function and a composition thereof, which comprises a liver polypeptide hydrolysate and a turmeric extract, wherein the liver polypeptide hydrolysate and the turmeric extract are mixed in a specific ratio, so that the composition does not feel fishy smell peculiar to the liver hydrolysate and has better flavor and taste when being taken. The composition is easy to drink and has high safety.
Description
Technical Field
The invention relates to the technical field of liver polypeptide products, in particular to liver polypeptide and a composition thereof for protecting and improving liver functions.
Background
The liver polypeptide is hydrolysate of liver of pig, cattle, etc., and has effects of improving liver function, and promoting ethanol metabolism in vivo when taking alcohol. Therefore, liver polypeptide is used for health care products and nutritional supplements (nutraceuticals) for regulating blood lipid, regulating immunity, and assisting in the protection of chemical liver injury, in addition to preventing and alleviating alcoholism, and is particularly provided in the form of beverages such as beverages and oral liquids which are easy to take in daily life.
However, since the liver polypeptide is produced from animal liver as a basic material, there is a problem that the fishy smell of animal liver tissue remains, and if the liver polypeptide is used as a drink as it is, the drink is difficult to drink because of the characteristic fishy smell of the liver polypeptide.
Although a method for preparing liver polypeptide has been proposed so far, for example, chinese patent 201711310781.4 discloses a liver extract freeze-dried food capable of improving liver function of the elderly, no means for removing fishy smell of food containing liver polypeptide has been disclosed.
The Curcuma longa extract is obtained by extracting Curcuma plants of Zingiberaceae with solvent, and optionally heating and/or reducing pressure to volatilize the solvent and drying. The plant material comprises rhizome of Curcuma rhizome, Curcuma zedoaria, Curcumae rhizoma, Curcuma zedoaria, Curcuma mauritiana, Curcuma xanthorrhiza and/or Curcuma wenyujin of Curcuma of Zingiberaceae, and can be in original shape or appropriate size or shape, or sliced or pulverized form. These plant materials may be suitably dried or may be fresh or alive. The method for extracting the turmeric extract from the plant material is not particularly limited. The extraction solvent may be water, hot water, or a hydrophilic organic solvent, or a mixed solvent of water and a hydrophilic organic solvent, and is preferably alcohol, water, or a mixed solvent of alcohol and water. The mixing ratio of the hydrophilic organic solvent and water is not particularly limited, and is, for example, preferably in the range of 5:95 to 90:10, more preferably in the range of 20:80 to 50:50, by weight. The content of the turmeric extract used in the present invention is measured by mixing the turmeric extract with ethyl acetate, removing ethyl acetate from the supernatant obtained by centrifugal separation, and then measuring the sample with High Performance Liquid Chromatography (HPLC) using a liquid dissolved in methanol.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides the liver polypeptide which is easy to drink, safe and used for protecting the liver and improving the liver function and the composition thereof.
The present inventors have unexpectedly found that by mixing a liver polypeptide and a turmeric extract in a specific ratio, the fishy smell peculiar to the liver polypeptide can be reduced. The liver polypeptide composition has good flavor and taste when taken orally.
The invention provides a preparation method of the liver polypeptide, which comprises the following steps:
(1) taking fresh or thawed animal liver, detecting shape, smell and volatile basic nitrogen, and determining to be qualified,
(2) cell disruption: cutting animal liver, removing foreign matters such as fascia, blood vessel, fat, etc., and washing with water; primarily crushing the cleaned liver; then cell disruption is carried out to prepare cell disruption solution,
(3) and (3) protease hydrolysis: adding proteolytic enzyme, hydrolyzing,
(4) virus inactivation: after the hydrolysis is finished, the hydrolysate is subjected to high-temperature virus inactivation treatment,
(5) filtering or centrifuging to obtain liver polypeptide.
Wherein the animal liver in the step (1) is collected from the livers of healthy livestock qualified by quarantine of legal quarantine departments, and comprises a freshly collected spleen and an animal liver which is frozen and stored at the temperature of below-20 ℃ immediately after collection. When the animal liver is in a unfrozen state, the animal liver is observed by naked eyes, the animal liver is bright in color, the spleen is basically complete in shape, no odor or peculiar smell exists, and the volatile basic nitrogen is less than or equal to 0.30g/kg, preferably less than or equal to 0.15g/kg, and most preferably less than or equal to 0.10 g/kg.
Wherein, the cell crushing operation in the step (2) can adopt common physical crushing modes such as a grinder, a homogenizer, a ball mill, ultrasonic waves, repeated freeze thawing and the like; preferably, the combination of repeated freezing and thawing and mechanical disruption is performed at a temperature of less than-40 deg.C, preferably less than-80 deg.C.
Wherein the protease hydrolysis according to claim 4 in step (3) may or may not be carried out, and preferably the hydrolysis is carried out. The protease used for hydrolysis includes all protease preparations allowed in food industry processing, such as pepsin, pancreatin, chymotrypsin, papain, etc.
Wherein, the virus in the step (4) is inactivated at the temperature of not less than 60 ℃, preferably not less than 100 ℃, and more preferably not less than 115 ℃.
The liver polypeptide prepared by the method has the excellent characteristics shown in the table, and the quality of the liver polypeptide is strictly controlled by detection technologies such as Kjeldahl nitrogen, high liquid chromatography, atomic absorption and the like. Wherein the proportion of peptides with relative molecular weight lower than 10kDa is higher than 95%; total fat and cholesterol are zero; the content of harmful heavy metals is strictly controlled and is lower than the national standard.
The invention provides a preparation process for extracting curcumin from turmeric, which comprises the following steps:
(1) selection of raw materials: selecting fresh root of Curcuma rhizome of Zingiberaceae, selecting rhizome with dark color, plumpness, no cold injury, no rot, and no pest, cleaning, removing mud, and removing root hair.
(2) The turmeric is frozen and dried at low temperature, and the operation steps are as follows: firstly, boxing: directly spreading rhizome of Curcuma rhizome on the plate layer of the low temperature freeze dryer; pre-freezing: reducing the temperature of the plate layer to below-30 ℃, and keeping the temperature for at least 120 min; thirdly, refrigerating by a condenser of the low-temperature freeze dryer until the temperature is lower than minus 50 ℃ and the vacuum degree of the box body is lower than 0.50 mbar; fourthly, sublimation for one time: sublimating for at least 10 hours at the temperature of the plate layer not higher than-5 ℃ under the condition that the vacuum degree of the box body is 0-0.5 mbar, and directly removing most of water in the turmeric through sublimation; secondary drying, namely raising the temperature of the plate layer to 20-50 ℃ and the vacuum degree of the box body to 0-0.5 mbar, raising the temperature for at least 3 hours, and performing secondary drying on the turmeric; discharging the box: and (4) after drying, quickly filling the dried turmeric rhizomes into a sealed container or a dryer.
(3) Crushing turmeric: and (3) putting the dried turmeric rhizome into a grinder, a wall breaking machine, a ball mill or a grinder, and grinding the turmeric rhizome into powder with the particle size of less than 100 mu m.
(4) Extraction of curcumin: taking 1 part of turmeric, adding 2-10 parts of organic solvent, heating, refluxing and extracting for at least 0.5 hour, and filtering; repeating the above operation for at least 2 times, mixing extractive solutions, and stirring.
(5) Vacuum concentrating the extracting solution to obtain an extract: vacuum concentrating the extractive solution, and removing most water to obtain extract.
(6) Drying the extract: the extract is frozen and dried at low temperature, and the method comprises the following specific steps: firstly, boxing: after the extract is placed in a tray, the extract is spread on a plate layer of a low-temperature freeze dryer or the extract is directly spread on the plate layer; pre-freezing: reducing the temperature of the plate layer to below-40 ℃, and keeping the temperature for at least 120 min; thirdly, the temperature of the freezer of the low-temperature freeze dryer is reduced to be below 50 ℃ below zero, and the vacuum degree of the box body is below 0.40 mbar; fourthly, sublimation for one time: controlling the temperature of the plate layer to be not higher than-10 ℃, controlling the vacuum degree of the box body to be 0-0.4 mbar, sublimating for at least 10 hours, and directly removing most of water in the extract through sublimation; secondary drying, namely increasing the temperature of the plate layer to 20-40 ℃, and the vacuum degree of the box body to 0-0.4 mbar, drying for at least 3 hours again, and performing secondary drying on the extract; discharging the box: and (5) after drying, quickly filling the dried extract into a sealed container or a dryer to obtain the curcumin.
Wherein the rhizome of Curcuma rhizome can be used directly or after slicing.
Wherein, the raw material turmeric rhizome can be directly put into a low-temperature freeze dryer for low-temperature freeze drying; or boiling or steaming until the core is penetrated, drying in the sun, storing, washing or soaking for 10-60 min with a large amount of water (based on soaking turmeric) before use, and then freezing and drying at low temperature by a low-temperature freeze dryer.
Wherein, after the turmeric rhizome is frozen and dried at low temperature, the drying weight loss is not higher than 5 percent when the detection is carried out at 105 ℃.
Wherein, the turmeric is pulverized by a pulverizer, a wall breaking machine, a ball mill or a grinder.
Wherein the organic solvent comprises: one or more of ethanol, acetone, ethyl acetate, n-hexane, isopropanol, No. 6 light gasoline, methanol, etc. The concentration of the organic solvent is 50-99% during extraction.
The curcumin extraction method is not limited to reflux extraction of an organic solvent, and extraction modes such as soaking extraction, percolation extraction and the like are also within the protection range of the application, and the curcumin extraction method is characterized in that the extraction temperature is not higher than 80 ℃.
Wherein, the organic solvent can be recycled after being used, the dosage of the organic solvent is reduced, and the concentration of the organic solvent is 50 to 99 percent in the extraction process, so the solvent recovery difficulty and the recovery cost are reduced.
Wherein the concentration temperature of vacuum concentration should not exceed 70 deg.C.
Wherein, the water content of the curcumin obtained after drying is not higher than 4 percent when detected at 105 ℃; the main components are curcumin, demethoxycurcumin and bisdemethoxycurcumin, and the bisdemethoxycurcumin: demethoxycurcumin: the ratio of curcumin is close to 1: 1: 3.
the method fully utilizes the characteristics of a low-temperature freeze drying process, the dried turmeric has low moisture, is loose and crisp, has lower requirements on a crushing mode and machinery, is easy to crush to smaller particle size, is loose and porous in the crushed particles, is favorable for solvent to enter the particle content, and improves the yield of the turmeric, and the yield of the effective component curcumin can reach more than 1.90 percent by utilizing the extraction of the method. The extracted curcumin is a natural extract, has the characteristics of low water content, low organic solvent residue, looseness and easy dissolution, can be used in the fields of food production and medicine, and can be further prepared into health-care food, food additives and pharmaceutical preparations.
The invention provides a liver polypeptide for protecting and improving liver functions and a composition thereof.
Wherein the weight ratio of the liver polypeptide to the turmeric extract is 100: 0.025-100: 5.55.
Wherein the weight ratio of the liver polypeptide to the turmeric extract is 100: 0.2625-100: 5.25.
Wherein the composition is in liquid form and has a pH of 4.0 or less.
Wherein the composition is in a solid form, which is formulated with water into a liquid prior to drinking, the liquid having a pH of 4.0 or less.
The invention provides a liver polypeptide composition for protecting and improving liver functions, which contains liver polypeptide and turmeric extract, wherein the weight ratio of the liver polypeptide to the turmeric extract is 100: 0.25, the composition is in a liquid form, and the pH value is 3.
Wherein the liver polypeptide is a liver protein hydrolysate obtained by hydrolyzing the liver of animals such as cattle and pigs with protease, or a liver extract directly extracted.
Wherein the composition further comprises carbonic acid and a gelling agent.
Wherein the gelling agent is starch, agar and thickener.
Wherein the composition further comprises all food additives allowed to be added in food industry processing, such as nutrition enhancer including one or more of vitamins (such as vitamin A, vitamin D, nicotinic acid, etc.), amino acids (8 essential amino acids: lysine, tryptophan, phenylalanine, methionine, threonine, isoleucine, leucine, valine, non-essential amino acids glutamic acid, alanine, glycine, aspartic acid, cystine, proline, serine, tyrosine, etc.), minerals (calcium, iron, zinc, etc.), DHA, taurine, casein, etc. permitted by national food safety standards; the antioxidant comprises one or more of tocopherol, tea polyphenol, calcium lactate, disodium ethylene diamine tetraacetate and the like which are allowed to be used in national food safety standards; the thickening agent comprises one or more of carrageenan, sodium carboxymethylcellulose, beta-cyclodextrin, hydroxypropyl distarch phosphate, frank gum, xanthan gum and the like which are allowed to be used in national food safety standards; the sweetener comprises one or more of sucrose, xylitol, honey, neotame, calcium cyclamate, maltitol, thaumatin and other permitted in national food safety standards; the acidity regulator comprises one or more of fumaric acid, phosphoric acid, malic acid, lactic acid, carbonic acid and salts thereof which are allowed to be used in national food safety standards; the emulsifier comprises one or more of caprylic acid, capric acid glyceride, sodium stearoyl lactylate, sucrose fatty acid ester and the like which are allowed to be used in national food safety standards; the preservative comprises one or more of benzoic acid and sodium salt thereof, -polylysine hydrochloride, sorbic acid and potassium salt thereof which are allowed to be used in national food safety standards; the food essence or spice comprises natural spice, synthetic spice, and extract of natural spice.
The invention also provides a method for preparing the liquid composition, which comprises the following steps: preparing liver polypeptide and turmeric extract into an aqueous solution, wherein the weight ratio of the liver polypeptide to the turmeric extract is 100: 0.025-100: 5.55, optionally adding food additives, adjusting the pH to below 4.0, heating to 65-100 ℃ for sterilization, filling into a beverage container, and sealing; or adjusting the pH to be below 4.0, filtering, filling into a sterilizable container, and sterilizing at 115-135 deg.C to obtain a liquid product meeting the commercial aseptic requirement.
Examples of the liquid beverage container include a polyethylene terephthalate (PET) container, a metal can, a composite container of a metal foil and a plastic film, and a glass bottle, and the form of the container is not particularly limited. The capacity of the container is not particularly limited, but is, for example, 10ml to 500ml, preferably 10ml to 50 ml. The means for containing the liquid composition in the container is arbitrary.
The invention also provides a method for preparing the solid composition, which comprises the following steps: preparing the liver polypeptide and the turmeric extract into an aqueous solution, wherein the weight ratio of the liver polypeptide to the turmeric extract is 100: 0.025-100: 5.55, optionally adding food additives, adjusting the pH to be below 4.0, heating to 65-100 ℃ for sterilization, drying to prepare solid powder, filling the solid powder into a beverage container, sealing, and preparing the solid beverage for drinking by water brewing; or making liver polypeptide and Curcuma rhizome extract into powder, optionally adding food additive according to the mixing ratio of liver polypeptide and Curcuma rhizome extract of 100: 0.025-100: 5.55, adjusting pH to below 4.0, sealing in a beverage container, and making into solid beverage which can be directly drunk after being mixed with water.
The beverage container includes a polyethylene terephthalate (PET) container, a metal can, a composite container of a metal foil and a plastic film, and a glass bottle, and the form of the container is not particularly limited. The capacity of the container is not particularly limited, but is, for example, 2g to 500g, preferably 2g to 20 g. The means of containing the solid composition in the container is arbitrary.
The invention also provides the application of the composition in preventing and relieving alcoholism, regulating blood fat and regulating immunity, and the application in health care products and nutritional auxiliary agents with auxiliary protection effect on chemical liver injury.
Wherein the composition is provided in the form of a beverage such as a drink and an oral liquid in daily life.
Detailed Description
Example 1 preparation of liver polypeptide
(1) Fresh or thawed animal liver is taken, and observed by naked eyes, the color and luster is bright, the spleen form is basically complete, no odor or peculiar smell exists, and the volatile basic nitrogen is 0.08 g/kg.
(2) Cell disruption: cutting animal liver, removing foreign matters such as fascia, blood vessel, fat, etc., and washing with water; primarily crushing the cleaned liver; then crushing the cells by a grinder to prepare cell crushing liquid,
(3) and (3) protease hydrolysis: adding pepsin, hydrolyzing,
(4) virus inactivation: after the hydrolysis is finished, the hydrolysate is subjected to virus inactivation treatment at the high temperature of 100 ℃,
(5) filtering or centrifuging to obtain liver polypeptide.
Example 2 preparation of liver polypeptide
(1) Fresh or thawed animal liver is taken, and observed by naked eyes, the color and luster is bright, the spleen form is basically complete, no odor or peculiar smell exists, and the volatile basic nitrogen is 0.05 g/kg.
(2) Cell disruption: cutting animal liver, removing foreign matters such as fascia, blood vessel, fat, etc., and washing with water; primarily crushing the cleaned liver; then cell disruption is carried out by a homogenizer to prepare cell disruption solution,
(3) and (3) protease hydrolysis: adding pancreatin, hydrolyzing,
(4) virus inactivation: after the hydrolysis is finished, the hydrolysate is subjected to virus inactivation treatment at the high temperature of 110 ℃,
(5) filtering or centrifuging to obtain liver polypeptide.
EXAMPLE 3 preparation of liver Polypeptides
(1) Fresh or thawed animal liver is taken, and observed by naked eyes, the color and luster is bright, the spleen form is basically complete, no odor or peculiar smell exists, and the volatile basic nitrogen is 0.07 g/kg.
(2) Cell disruption: cutting animal liver, removing foreign matters such as fascia, blood vessel, fat, etc., and washing with water; primarily crushing the cleaned liver; then the cell is crushed by combining the repeated freezing and thawing with the mechanical crushing to prepare cell crushing liquid, the temperature of the repeated freezing and thawing is-40 ℃,
(3) and (3) protease hydrolysis: adding papain, carrying out hydrolysis,
(4) virus inactivation: after the hydrolysis is finished, the hydrolysate is subjected to virus inactivation treatment at the high temperature of 115 ℃,
(5) filtering or centrifuging to obtain liver polypeptide.
Example 4 preparation of liver Polypeptides
(1) Fresh or thawed animal liver is taken, and observed by naked eyes, the color and luster is bright, the spleen form is basically complete, no odor or peculiar smell exists, and the volatile basic nitrogen is 0.03 g/kg.
(2) Cell disruption: cutting animal liver, removing foreign matters such as fascia, blood vessel, fat, etc., and washing with water; primarily crushing the cleaned liver; then the cell is crushed by combining the repeated freezing and thawing with the mechanical crushing to prepare cell crushing liquid, the temperature of the repeated freezing and thawing is-80 ℃,
(3) and (3) protease hydrolysis: adding chymotrypsin for hydrolysis,
(4) virus inactivation: after the hydrolysis is finished, the hydrolysate is subjected to virus inactivation treatment at the high temperature of 115 ℃,
(5) filtering or centrifuging to obtain liver polypeptide.
Example 5 preparation of liver Polypeptides
(1) Fresh or thawed animal liver is taken, and observed by naked eyes, the color and luster is bright, the spleen form is basically complete, no odor or peculiar smell exists, and the volatile basic nitrogen is 0.03 g/kg.
(2) Cell disruption: cutting animal liver, removing foreign matters such as fascia, blood vessel, fat, etc., and washing with water; primarily crushing the cleaned liver; then cell disruption is carried out by a mode of combining optimized repeated freezing and thawing with mechanical disruption to prepare cell disruption solution, the temperature of repeated freezing and thawing is-60 ℃,
(3) and (3) protease hydrolysis: adding chymotrypsin for hydrolysis,
(4) virus inactivation: after the hydrolysis is finished, the hydrolysate is subjected to virus inactivation treatment at the high temperature of 110 ℃,
(5) filtering or centrifuging to obtain liver polypeptide.
Example 6 preparation of liver Polypeptides
(1) Fresh or thawed animal liver is taken, and observed by naked eyes, the color and luster is bright, the spleen form is basically complete, no odor or peculiar smell exists, and the volatile basic nitrogen is 0.02 g/kg.
(2) Cell disruption: cutting animal liver, removing foreign matters such as fascia, blood vessel, fat, etc., and washing with water; primarily crushing the cleaned liver; then a ball mill is used for cell disruption to prepare cell disruption liquid,
(3) and (3) protease hydrolysis: adding pepsin, hydrolyzing,
(4) virus inactivation: after the hydrolysis is finished, the hydrolysate is subjected to virus inactivation treatment at the high temperature of 110 ℃,
(5) filtering or centrifuging to obtain liver polypeptide.
Example 7 characterization of liver Polypeptides
The quality of the product is strictly controlled by detection technologies such as Kjeldahl nitrogen determination, high liquid chromatography, atomic absorption and the like. The detection methods of the indexes in the following table are respectively described in the national food safety standard of the latest version, such as GB5009.5-2016 food safety standard food protein determination. The results are shown in Table 1.
TABLE 1 examination of hepatic polypeptide Properties
Example 8 preparation of turmeric extract
(1) Selection of raw materials: the raw material is fresh rhizome of Curcuma rhizome of Zingiberaceae, selecting rhizome with dark color, plumpness, no cold injury, no rot, and no pest, cleaning, removing mud, and removing root hair.
(2) The turmeric is frozen and dried at low temperature, and the operation steps are as follows: firstly, boxing: directly spreading rhizome of Curcuma rhizome on the plate layer of the low temperature freeze dryer; pre-freezing: reducing the temperature of the plate layer to-40 ℃, and preserving the heat for 120 min; thirdly, refrigerating the low-temperature freeze dryer condenser to the temperature of-60 ℃ and the vacuum degree of the box body of 0.40 mbar; fourthly, sublimation for one time: sublimating for 10 hours at the temperature of the plate layer of-5 ℃ and the vacuum degree of the box body of 0.5mbar to ensure that most of water in the turmeric is directly removed by sublimation; secondary drying, namely raising the temperature of the plate layer to 20 ℃, vacuumizing the box body for 0.5mbar, and raising the temperature for 3 hours again to perform secondary drying on the turmeric; discharging the box: and (4) after drying, quickly filling the dried turmeric rhizomes into a sealed container or a dryer.
(3) Crushing turmeric: pulverizing dried Curcuma rhizome into powder with particle size of 90 μm in pulverizer.
(4) Extraction of curcumin: taking 1 part of turmeric, adding 2 parts of ethanol, heating, refluxing and extracting for 0.5 hour, and filtering; repeating the above operation for 2 times, mixing extractive solutions, and stirring.
(5) Vacuum concentrating the extracting solution to obtain an extract: vacuum concentrating the extractive solution, and removing most water to obtain extract.
(6) Drying the extract: the extract is frozen and dried at low temperature, and the method comprises the following specific steps: firstly, boxing: after the extract is placed in a tray, the extract is spread on a plate layer of a low-temperature freeze dryer or the extract is directly spread on the plate layer; pre-freezing: reducing the temperature of the plate layer to-50 ℃, and preserving the heat for 120 min; thirdly, the temperature of a freezer of the low-temperature freeze dryer is reduced to-60 ℃, and the vacuum degree of the box body is less than 0.30 mbar; fourthly, sublimation for one time: controlling the temperature of the plate layer at-10 deg.C, controlling the vacuum degree of the box body at 0.4mbar, sublimating for 10 hr, and removing most water in the extract by sublimation; secondary drying, namely increasing the temperature of the plate layer to 20 ℃, ensuring the vacuum degree of the box body to be 0.4mbar, drying for 3 hours again, and performing secondary drying on the extract; discharging the box: and (4) after drying, quickly filling the dried extract into a sealed container or a dryer to obtain the turmeric extract.
Example 9 preparation of turmeric extract
(1) Selection of raw materials: the raw material is fresh rhizome of Curcuma rhizome of Zingiberaceae, selecting rhizome with dark color, plumpness, no cold injury, no rot, and no pest, cleaning, removing mud, and removing root hair.
(2) The turmeric is frozen and dried at low temperature, and the operation steps are as follows: firstly, boxing: directly spreading rhizome of Curcuma rhizome on the plate layer of the low temperature freeze dryer; pre-freezing: reducing the temperature of the plate layer to-50 ℃, and preserving the heat for 150 min; thirdly, refrigerating the low-temperature freeze dryer condenser to the temperature of-70 ℃ and the vacuum degree of the box body of 0.30 mbar; fourthly, sublimation for one time: sublimating for 12 hours at the temperature of the plate layer of-15 ℃ and the vacuum degree of the box body of 0.3mbar to ensure that most of water in the turmeric is directly removed by sublimation; secondary drying, namely raising the temperature of the plate layer to 25 ℃, vacuumizing the box body for 0.4mbar, raising the temperature for drying for 3.5 hours again, and carrying out secondary drying on the turmeric; discharging the box: and (4) after drying, quickly filling the dried turmeric rhizomes into a sealed container or a dryer.
(3) Crushing turmeric: pulverizing dried Curcuma rhizome into powder with particle size of 80 μm.
(4) Extraction of curcumin: taking 1 part of turmeric, adding 3 parts of acetone, heating, refluxing and extracting for 0.8 hour, and filtering; repeating the above operation for 3 times, mixing extractive solutions, and stirring.
(5) Vacuum concentrating the extracting solution to obtain an extract: vacuum concentrating the extractive solution, and removing most water to obtain extract.
(6) Drying the extract: the extract is frozen and dried at low temperature, and the method comprises the following specific steps: firstly, boxing: after the extract is placed in a tray, the extract is spread on a plate layer of a low-temperature freeze dryer or the extract is directly spread on the plate layer; pre-freezing: reducing the temperature of the plate layer to-50 ℃, and preserving the heat for 120 min; thirdly, the temperature of a freezer of the low-temperature freeze dryer is reduced to-60 ℃, and the vacuum degree of the box body is less than 0.30 mbar; fourthly, sublimation for one time: controlling the temperature of the plate layer at-10 deg.C, controlling the vacuum degree of the box body at 0.4mbar, sublimating for 10 hr, and removing most water in the extract by sublimation; secondary drying, namely increasing the temperature of the plate layer to 20 ℃, ensuring the vacuum degree of the box body to be 0.4mbar, drying for 3 hours again, and performing secondary drying on the extract; discharging the box: and (4) after drying, quickly filling the dried extract into a sealed container or a dryer to obtain the turmeric extract.
Example 10 preparation of test beverages
The liver polypeptide prepared in example 1 and the turmeric extract prepared in example 8 were weighed according to the weight shown in table 2, mixed, and dissolved in 100g of water to obtain a total amount of 100g of aqueous solution. The pH was then adjusted with citric acid, the adjusted pH being shown in table 1. The pH values in the table represent the values determined at a sample temperature of 20 ℃.
Table 2 compounding scale of test beverages
Example 11 sensory index evaluation
The sensory index was evaluated by selecting 5 healthy men and women aged 20 to 50 as a test group. Each test beverage was ingested, and the "fishy smell peculiar to the liver" item was evaluated on 5 grades (0: very weak 4: very strong) of 0 to 4 (table 3 below). As a result, it was found that the composition without turmeric extract had strong liver odor and was not acceptable as a drink; when the weight ratio of the liver polypeptide to the turmeric extract is 100: 0.025-100: 5.55, especially when the weight ratio of the liver polypeptide to the turmeric extract is 100: 0.25 and the pH is 3, the taste is the best.
TABLE 3 sensory index evaluation results
Example 12 Effect of liver Polypeptides (product of example 4) on blood lipid levels and liver function in diabetic hyperlipidemic mice
1 test animal:
40 female mice with the feed Kunming and the weight of 25-30 g are provided by Guangzhou experimental animal center. The cage is fed by moving rack cages, 4-6 cages are fed, and the cage is cleaned once a day to keep the interior of the cage clean. The laboratory is kept quiet and ventilated, the room temperature is 20-25 ℃, and the humidity is 70 +/-15%. The rat basal feed is provided by Xinjiang experimental animal center, and the total heat of the feed is 15.5 kJ/g. The high-fat feed is prepared by self and comprises the following components: 37.5% of commercially available fine flour, 20% of lard, 10% of cane sugar, 7% of egg yolk, 25% of cholesterol and 0.5% of ox-gall salt are fully mixed and baked for later use.
2, test method:
2.1 test grouping: after the Wistar rats are adaptively fed for 1 week, 30 rats are taken and fed with high-fat feed for 4 weeks, tail vein injection Alloxan (ALX) is carried out for 60mg/kg, after 1 week, animals are randomly and averagely divided into 3 groups after fasting blood glucose is detected to be more than or equal to 11.1 mmol/L: the model group (no drug treatment), the example group (gavage administration of 300mg/kg of the product of example 4) and the positive control group (gavage administration of Xuezhikang 400mg/kg), and 10 other rats were used as a blank group, and were fed with normal diet, and the tail vein was injected with an equal volume of physiological saline without injection of ALX.
2.2 method: the animals of the above groups were continuously gazed for 4 weeks, and after 1h fasting for 14h from the last gavage, the animals were sacrificed, serum was separated, and Triglyceride (TG), Total Cholesterol (TC), high low density lipoprotein cholesterol (HDL-C), and low density lipoprotein cholesterol (LDL-C) were measured.
3, statistical analysis:
the experimental data were analyzed using SPSS16.0 software, and the mean between groups was compared using one-way analysis of variance, Mann-Whitney test.
4. Results
TABLE 4 Effect (x. + -. s, mmol/L) on the blood lipid levels in DM hyperlipidemia model mice
| Group of | TC | TG | HDL-C | LDL-C |
| Blank space | 2.35±0.14 | 1.55±0.12 | 0.97±0.07 | 1.15±0.14 |
| Model set | 3.95±0.54** | 2.34±0.37** | 1.31±0.12** | 1.95±0.21** |
| Group of embodiments | 2.82±0.07# | 1.77±0.16# | 0.96±0.07# | 1.27±0.26## |
| Positive control group | 2.67±0.94 | 1.82±0.25# | 0.93±0.32# | 1.31±0.43# |
Note: p < 0.05, P < 0.01 compared to blank; compared with the model group, # P < 0.05, # P < 0.01.
The results show that compared with a blank group, the serum contents of TG, TC, HDL-C and LDL-C in the mice of the model group are all increased remarkably. Compared with the model group, the traditional Chinese medicine composition prepared in the gastric perfusion example 4 and the positive control group medicament are remarkably reduced in TG, TC, HDL-C and LDL-C after 4 weeks, which shows that the traditional Chinese medicine composition and the positive control group medicament have remarkable effects of reducing blood lipid level in serum, and the example 4 group has more remarkable effects.
Claims (10)
1. A method for preparing liver polypeptide for protecting liver and improving liver function comprises the following steps:
(1) taking fresh or thawed animal liver, detecting shape, smell and volatile basic nitrogen, and making it be qualified,
(2) cell disruption: cutting animal liver, removing foreign matters such as fascia, blood vessel, fat, etc., and washing with water; primarily crushing the cleaned liver; then cell disruption is carried out to prepare cell disruption solution,
(3) and (3) protease hydrolysis: adding proteolytic enzyme, hydrolyzing,
(4) virus inactivation: after the hydrolysis is finished, the hydrolysate is subjected to high-temperature virus inactivation treatment,
(5) filtering or centrifuging to obtain liver polypeptide.
2. The method according to claim 1, wherein the animal liver in step (1) is collected from healthy livestock qualified for quarantine by the legal quarantine department, and comprises freshly collected spleen and animal liver stored frozen at a temperature of-20 ℃ or lower immediately after collection, and has a volatile basic nitrogen of 0.30g/kg or less, preferably 0.15g/kg or less, and most preferably 0.10g/kg or less.
3. The method according to claim 1, wherein the cell disruption in step (2) is performed by a conventional physical disruption method such as a grinder, homogenizer, ball mill, ultrasonic wave, repeated freeze-thaw, etc.; preferably, the combination of repeated freezing and thawing and mechanical disruption is performed at a temperature of less than-40 deg.C, preferably less than-80 deg.C.
4. The method according to claim 1, wherein the proteolytic enzyme in the step (3) is pepsin, pancreatin, chymotrypsin, papain, or the like.
5. The method according to claim 1, wherein the virus is inactivated in step (4) at a temperature of not less than 60 ℃, preferably not less than 100 ℃, and more preferably not less than 115 ℃.
6. The method according to claim 1, wherein the liver polypeptide is prepared to have excellent properties as shown in the following, wherein the relative molecular weight of the peptide of less than 10kDa is more than 95%; total fat and cholesterol are zero; the content of harmful heavy metals is strictly controlled and is lower than the national standard,
7. A preparation process for extracting curcumin from Curcuma rhizome comprises the following steps:
(1) selection of raw materials: selecting fresh root of Curcuma rhizome of Zingiberaceae, selecting rhizome with dark color, plumpness, no cold injury, no rot, and no plant diseases and insect pests, cleaning, removing mud, removing root hair,
(2) the turmeric is frozen and dried at low temperature, and the operation steps are as follows: firstly, boxing: directly spreading rhizome of Curcuma rhizome on the plate layer of the low temperature freeze dryer; pre-freezing: reducing the temperature of the plate layer to below-30 ℃, and keeping the temperature for at least 120 min; thirdly, refrigerating by a condenser of the low-temperature freeze dryer until the temperature is lower than minus 50 ℃ and the vacuum degree of the box body is lower than 0.50 mbar; fourthly, sublimation for one time: sublimating for at least 10 hours at the temperature of the plate layer not higher than-5 ℃ under the condition that the vacuum degree of the box body is 0-0.5 mbar, and directly removing most of water in the turmeric through sublimation; secondary drying, namely raising the temperature of the plate layer to 20-50 ℃ and the vacuum degree of the box body to 0-0.5 mbar, raising the temperature for at least 3 hours, and performing secondary drying on the turmeric; discharging the box: after drying, quickly filling dried rhizome of Curcuma into a sealed container or a dryer,
(3) crushing turmeric: pulverizing dried Curcuma rhizome into powder with particle diameter less than 100 μm in pulverizer, wall breaking machine, ball mill or grinding machine,
(4) extraction of curcumin: taking 1 part of turmeric, adding 2-10 parts of organic solvent, heating, refluxing and extracting for at least 0.5 hour, and filtering; repeating the above operation for at least 2 times, mixing extractive solutions, stirring,
(5) vacuum concentrating the extracting solution to obtain an extract: vacuum concentrating the extractive solution, removing most water to obtain extract,
(6) drying the extract: the extract is frozen and dried at low temperature, and the method comprises the following specific steps: firstly, boxing: after the extract is placed in a tray, the extract is spread on a plate layer of a low-temperature freeze dryer or the extract is directly spread on the plate layer; pre-freezing: reducing the temperature of the plate layer to below-40 ℃, and keeping the temperature for at least 120 min; thirdly, the temperature of the freezer of the low-temperature freeze dryer is reduced to be below 50 ℃ below zero, and the vacuum degree of the box body is below 0.40 mbar; fourthly, sublimation for one time: controlling the temperature of the plate layer to be not higher than-10 ℃, controlling the vacuum degree of the box body to be 0-0.4 mbar, sublimating for at least 10 hours, and directly removing most of water in the extract through sublimation; secondary drying, namely increasing the temperature of the plate layer to 20-40 ℃, and the vacuum degree of the box body to 0-0.4 mbar, drying for at least 3 hours again, and performing secondary drying on the extract; discharging the box: drying, quickly filling the dried extract into a sealed container or a dryer to obtain curcumin,
wherein, the selected rhizome of the turmeric can be directly used or can be combined after being sliced, wherein, the raw material rhizome of the turmeric can be directly frozen and dried at low temperature by a low-temperature freeze dryer; or boiling or steaming until the turmeric is soaked, drying in the sun, storing, washing or soaking for 10-60 min with a large amount of water (based on soaking the turmeric) before use, and then performing low-temperature freeze drying by using a low-temperature freeze dryer, wherein after the turmeric rhizome is subjected to low-temperature freeze drying, the drying weight loss is not higher than 5 percent when detected at 105 ℃, the turmeric rhizome is crushed, and the crushing machinery adopts a crusher, a wall breaking machine, a ball mill or a grinder, wherein the organic solvent comprises: one or more of ethanol, acetone, ethyl acetate, n-hexane, isopropanol, No. 6 light gasoline, methanol, etc. The concentration of the organic solvent is 50-99% during extraction, wherein the extraction method of the curcumin is not limited to reflux extraction of the organic solvent, and the extraction modes such as soaking extraction, percolation extraction and the like are also within the protection range of the application, and the curcumin is characterized in that the extraction temperature is not higher than 80 ℃, wherein the organic solvent can be recycled after being used, wherein the concentration temperature of vacuum concentration is not higher than 70 ℃, wherein the moisture of the curcumin obtained after drying is not higher than 4% when detected at 105 ℃; the main components are curcumin, demethoxycurcumin and bisdemethoxycurcumin, and the bisdemethoxycurcumin: demethoxycurcumin: the ratio of curcumin is close to 1: 1: 3.
8. a liver polypeptide and a composition thereof for protecting and improving liver function, the composition comprises the liver polypeptide and a turmeric extract, characterized in that the liver polypeptide is prepared according to the preparation method of claim 1, the turmeric extract is prepared according to the preparation method of claim 7, and the weight ratio of the liver polypeptide to the turmeric extract is 100: 0.025-100: 5.55, preferably 100: 0.2625-100: 5.25.
9. A liver polypeptide and its composition for protecting and improving liver function, the composition contains liver polypeptide and Curcuma rhizome extract, wherein the weight ratio of liver polypeptide and Curcuma rhizome extract is 100: 0.25, the composition is in liquid form, and pH is 3.
10. The composition further comprises all food additives allowed to be added in the food industry processing, including fortifiers, antioxidants, thickeners, sweeteners, acidity regulators, emulsifiers, preservatives and flavors.
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Cited By (1)
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