CN114941020A - Preparation method and application of donkey-hide gelatin polypeptide - Google Patents
Preparation method and application of donkey-hide gelatin polypeptide Download PDFInfo
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- CN114941020A CN114941020A CN202210768126.8A CN202210768126A CN114941020A CN 114941020 A CN114941020 A CN 114941020A CN 202210768126 A CN202210768126 A CN 202210768126A CN 114941020 A CN114941020 A CN 114941020A
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- hide gelatin
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- 108010010803 Gelatin Proteins 0.000 title claims abstract description 96
- 229920000159 gelatin Polymers 0.000 title claims abstract description 96
- 239000008273 gelatin Substances 0.000 title claims abstract description 96
- 235000019322 gelatine Nutrition 0.000 title claims abstract description 96
- 235000011852 gelatine desserts Nutrition 0.000 title claims abstract description 96
- 229920001184 polypeptide Polymers 0.000 title claims abstract description 57
- 102000004196 processed proteins & peptides Human genes 0.000 title claims abstract description 57
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 57
- 238000002360 preparation method Methods 0.000 title claims abstract description 22
- 239000000243 solution Substances 0.000 claims abstract description 35
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 29
- 239000000843 powder Substances 0.000 claims abstract description 27
- 238000004321 preservation Methods 0.000 claims abstract description 25
- 239000008367 deionised water Substances 0.000 claims abstract description 20
- 229910021641 deionized water Inorganic materials 0.000 claims abstract description 20
- 238000003756 stirring Methods 0.000 claims abstract description 18
- 238000002347 injection Methods 0.000 claims abstract description 8
- 239000007924 injection Substances 0.000 claims abstract description 8
- 239000002775 capsule Substances 0.000 claims abstract description 6
- 238000004140 cleaning Methods 0.000 claims abstract description 6
- 239000003814 drug Substances 0.000 claims abstract description 6
- 239000008187 granular material Substances 0.000 claims abstract description 6
- 238000002156 mixing Methods 0.000 claims abstract description 6
- 229940100688 oral solution Drugs 0.000 claims abstract description 6
- 239000006188 syrup Substances 0.000 claims abstract description 6
- 235000020357 syrup Nutrition 0.000 claims abstract description 6
- 239000003826 tablet Substances 0.000 claims abstract description 6
- 229940098465 tincture Drugs 0.000 claims abstract description 6
- 238000009777 vacuum freeze-drying Methods 0.000 claims abstract description 6
- 241001122767 Theaceae Species 0.000 claims abstract 2
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 30
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 24
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 16
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 claims description 16
- 238000001816 cooling Methods 0.000 claims description 15
- 239000006228 supernatant Substances 0.000 claims description 10
- 238000000034 method Methods 0.000 claims description 9
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 claims description 8
- 102000057297 Pepsin A Human genes 0.000 claims description 8
- 108090000284 Pepsin A Proteins 0.000 claims description 8
- 108010052008 colla corii asini Proteins 0.000 claims description 8
- 235000019253 formic acid Nutrition 0.000 claims description 8
- 238000010438 heat treatment Methods 0.000 claims description 8
- 239000012528 membrane Substances 0.000 claims description 8
- 229910052757 nitrogen Inorganic materials 0.000 claims description 8
- 238000002414 normal-phase solid-phase extraction Methods 0.000 claims description 8
- 229940111202 pepsin Drugs 0.000 claims description 8
- 238000001914 filtration Methods 0.000 claims description 7
- 230000003213 activating effect Effects 0.000 claims description 5
- 238000000861 blow drying Methods 0.000 claims description 5
- 238000004090 dissolution Methods 0.000 claims description 5
- 239000003480 eluent Substances 0.000 claims description 5
- 230000000694 effects Effects 0.000 claims description 4
- 102000004190 Enzymes Human genes 0.000 claims description 3
- 108090000790 Enzymes Proteins 0.000 claims description 3
- 238000005119 centrifugation Methods 0.000 claims description 3
- 229940088598 enzyme Drugs 0.000 claims description 3
- 239000002994 raw material Substances 0.000 abstract description 2
- 238000000605 extraction Methods 0.000 abstract 1
- 239000012535 impurity Substances 0.000 abstract 1
- 241000283074 Equus asinus Species 0.000 description 2
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 2
- 244000269722 Thea sinensis Species 0.000 description 2
- 208000031971 Yin Deficiency Diseases 0.000 description 2
- 230000000740 bleeding effect Effects 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 210000004072 lung Anatomy 0.000 description 2
- 235000015091 medicinal tea Nutrition 0.000 description 2
- 208000011580 syndromic disease Diseases 0.000 description 2
- 235000013616 tea Nutrition 0.000 description 2
- MTCFGRXMJLQNBG-REOHCLBHSA-N (2S)-2-Amino-3-hydroxypropansäure Chemical compound OC[C@H](N)C(O)=O MTCFGRXMJLQNBG-REOHCLBHSA-N 0.000 description 1
- 239000004475 Arginine Substances 0.000 description 1
- 206010010904 Convulsion Diseases 0.000 description 1
- 206010011224 Cough Diseases 0.000 description 1
- LEVWYRKDKASIDU-QWWZWVQMSA-N D-cystine Chemical compound OC(=O)[C@H](N)CSSC[C@@H](N)C(O)=O LEVWYRKDKASIDU-QWWZWVQMSA-N 0.000 description 1
- 241000283086 Equidae Species 0.000 description 1
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 1
- 239000004471 Glycine Substances 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- PMMYEEVYMWASQN-DMTCNVIQSA-N Hydroxyproline Chemical compound O[C@H]1CN[C@H](C(O)=O)C1 PMMYEEVYMWASQN-DMTCNVIQSA-N 0.000 description 1
- ONIBWKKTOPOVIA-BYPYZUCNSA-N L-Proline Chemical compound OC(=O)[C@@H]1CCCN1 ONIBWKKTOPOVIA-BYPYZUCNSA-N 0.000 description 1
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 description 1
- ODKSFYDXXFIFQN-BYPYZUCNSA-P L-argininium(2+) Chemical compound NC(=[NH2+])NCCC[C@H]([NH3+])C(O)=O ODKSFYDXXFIFQN-BYPYZUCNSA-P 0.000 description 1
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 description 1
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 1
- HNDVDQJCIGZPNO-YFKPBYRVSA-N L-histidine Chemical compound OC(=O)[C@@H](N)CC1=CN=CN1 HNDVDQJCIGZPNO-YFKPBYRVSA-N 0.000 description 1
- KDXKERNSBIXSRK-YFKPBYRVSA-N L-lysine Chemical compound NCCCC[C@H](N)C(O)=O KDXKERNSBIXSRK-YFKPBYRVSA-N 0.000 description 1
- AYFVYJQAPQTCCC-GBXIJSLDSA-N L-threonine Chemical compound C[C@@H](O)[C@H](N)C(O)=O AYFVYJQAPQTCCC-GBXIJSLDSA-N 0.000 description 1
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 description 1
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 1
- 239000004472 Lysine Substances 0.000 description 1
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 description 1
- 206010037660 Pyrexia Diseases 0.000 description 1
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 description 1
- 208000013738 Sleep Initiation and Maintenance disease Diseases 0.000 description 1
- AYFVYJQAPQTCCC-UHFFFAOYSA-N Threonine Natural products CC(O)C(N)C(O)=O AYFVYJQAPQTCCC-UHFFFAOYSA-N 0.000 description 1
- 239000004473 Threonine Substances 0.000 description 1
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 description 1
- 235000004279 alanine Nutrition 0.000 description 1
- 235000001014 amino acid Nutrition 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 1
- 235000003704 aspartic acid Nutrition 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- OQFSQFPPLPISGP-UHFFFAOYSA-N beta-carboxyaspartic acid Natural products OC(=O)C(N)C(C(O)=O)C(O)=O OQFSQFPPLPISGP-UHFFFAOYSA-N 0.000 description 1
- 238000004061 bleaching Methods 0.000 description 1
- 239000005018 casein Substances 0.000 description 1
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 description 1
- 235000021240 caseins Nutrition 0.000 description 1
- 230000036461 convulsion Effects 0.000 description 1
- 229960003067 cystine Drugs 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 230000006735 deficit Effects 0.000 description 1
- PMMYEEVYMWASQN-UHFFFAOYSA-N dl-hydroxyproline Natural products OC1C[NH2+]C(C([O-])=O)C1 PMMYEEVYMWASQN-UHFFFAOYSA-N 0.000 description 1
- 208000017574 dry cough Diseases 0.000 description 1
- 235000013922 glutamic acid Nutrition 0.000 description 1
- 239000004220 glutamic acid Substances 0.000 description 1
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 description 1
- 229960002591 hydroxyproline Drugs 0.000 description 1
- 206010022437 insomnia Diseases 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 235000013930 proline Nutrition 0.000 description 1
- 235000004400 serine Nutrition 0.000 description 1
- 235000008521 threonine Nutrition 0.000 description 1
- FGMPLJWBKKVCDB-UHFFFAOYSA-N trans-L-hydroxy-proline Natural products ON1CCCC1C(O)=O FGMPLJWBKKVCDB-UHFFFAOYSA-N 0.000 description 1
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- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/01—Hydrolysed proteins; Derivatives thereof
- A61K38/012—Hydrolysed proteins; Derivatives thereof from animals
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/08—Solutions
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
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Abstract
The invention discloses a preparation method and application of donkey-hide gelatin polypeptide, comprising the following steps: s1, cleaning and airing the donkey-hide gelatin, crushing the aired donkey-hide gelatin by a traditional Chinese medicine ultrafine grinder, putting the donkey-hide gelatin powder into a heat preservation tank, adding deionized water into the heat preservation tank, and mixing and stirring the donkey-hide gelatin powder and the deionized water in the heat preservation tank by a stirring device. The invention is convenient for dissolving the donkey-hide gelatin in water and preparing the dissolved solution, improves the extraction percentage of the donkey-hide gelatin, has higher utilization rate of raw materials, can effectively extract and separate impurities from samples, can ensure the stability of the solution, is convenient for separating and obtaining the donkey-hide gelatin polypeptide better, can prepare donkey-hide gelatin polypeptide oral preparations such as donkey-hide gelatin polypeptide oral solution, decoction, syrup, medicinal liquor, decocted extract, tincture, extract, tea, tablets, capsules, granules and the like by utilizing the donkey-hide gelatin polypeptide, can also prepare donkey-hide gelatin polypeptide injection, or prepare donkey-hide gelatin polypeptide powder injection by vacuum freeze drying, and has very wide application.
Description
Technical Field
The invention relates to the technical field of donkey-hide gelatin polypeptides, and particularly relates to a preparation method and application of a donkey-hide gelatin polypeptide.
Background
Colla Corii Asini is a block prepared by bleaching and unhairing donkey skin of animal donkey of Equidae, and is composed of ossein, and hydrolyzed to obtain various amino acids such as lysine, arginine, histidine, cystine, tryptophan, hydroxyproline, aspartic acid, threonine, serine, glutamic acid, proline, glycine, and alanine. Has effects in replenishing blood, nourishing yin, moistening lung, and stopping bleeding. Mainly treats blood deficiency syndromes, bleeding syndromes, lung yin deficiency and dry cough, fever and yin impairment, vexation and insomnia, yin deficiency and wind-movement, and limb convulsion. At present, the research on polypeptide products mainly focuses on casein and hydrolyzed soybeans, the research on the polypeptide of donkey-hide gelatin is not much, the problems of low utilization rate of raw materials and the like exist in the preparation of a donkey-hide gelatin polypeptide solution, and the pH value cannot be effectively kept balanced in the preparation process, so that a preparation method and application of the donkey-hide gelatin polypeptide are provided.
Disclosure of Invention
The invention aims to solve the defects in the prior art and provides a preparation method and application of donkey-hide gelatin polypeptide.
In order to achieve the purpose, the invention adopts the following technical scheme:
a preparation method of donkey-hide gelatin polypeptide comprises the following steps:
s1, cleaning and airing the donkey-hide gelatin, crushing the aired donkey-hide gelatin by a traditional Chinese medicine superfine crusher, putting the donkey-hide gelatin powder into a heat preservation tank, adding deionized water into the heat preservation tank, mixing and stirring the donkey-hide gelatin powder and the deionized water in the heat preservation tank by a stirring device, and heating the heat preservation tank at the same time, so as to dissolve the donkey-hide gelatin powder;
s2, cooling to room temperature after the dissolution is finished in the step S1, centrifuging the dissolved solution through a centrifugal machine, collecting supernatant of the dissolved solution to obtain donkey-hide gelatin extract, adjusting the pH of the donkey-hide gelatin extract with NaOH solution, performing enzymolysis with pepsin in water bath, cooling after the enzymolysis is finished, centrifuging through a centrifugal machine after the cooling, taking the supernatant, separating and purifying through a solid phase extraction column, activating with methanol and deionized water in advance, eluting with methanol, collecting eluent, blow-drying with nitrogen, dissolving with formic acid solution, and filtering through a filter membrane to obtain the donkey-hide gelatin polypeptide.
Preferably, the weight of the deionized water in the step S1 is 10-15 times of the weight of the donkey-hide gelatin powder, the heating temperature of the heat preservation tank in the step S1 is 80-90 ℃, and the stirring time of the stirring device in the step S1 is 25-30 min.
Preferably, the concentration of the NaOH solution in the step S2 is 8%, the pH of the donkey-hide gelatin extract in the step S2 is maintained at 8, the pH is adjusted every 1 hour in the enzymolysis process in the step S2, and the enzyme activity is inactivated after 4 hours of enzymolysis.
Preferably, the water bath temperature in the step S2 is 45 ℃, and the concentration of the pepsin is 6 x 10 4 U/g-8×10 4 U/g, the concentration of the formic acid solution is 2%, the centrifugation time of the centrifuge in the step S2 is 8min, and the nitrogen temperature in the step S2 is 50 ℃.
Preferably, the parameter of the solid phase extraction column is C18, and the parameter of the filter membrane is 0.45 μm.
Preferably, the donkey-hide gelatin polypeptide obtained in the step S2 is prepared into a donkey-hide gelatin polypeptide solution, or is subjected to vacuum freeze drying to obtain donkey-hide gelatin polypeptide powder.
The application of the preparation method of the donkey-hide gelatin polypeptide comprises the step of preparing donkey-hide gelatin polypeptide oral preparations, such as donkey-hide gelatin polypeptide oral solution, decoction, syrup, medicinal liquor, decoction, tincture, extract, medicinal tea, tablets, capsules, granules and the like, or preparing donkey-hide gelatin polypeptide injection, or preparing donkey-hide gelatin polypeptide powder injection through vacuum freeze drying.
Compared with the prior art, the invention has the beneficial effects that:
the donkey-hide gelatin polypeptide oral preparation can be prepared by using the donkey-hide gelatin polypeptide, such as donkey-hide gelatin polypeptide oral solution, decoction, syrup, medicinal liquor, decoction, tincture, extract, tea, tablets, capsules, granules and the like, can also be prepared, and can be widely applied.
Detailed Description
The technical solutions in the embodiments of the present invention are clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments.
Example 1:
a preparation method of donkey-hide gelatin polypeptide comprises the following steps:
s1, cleaning and airing the donkey-hide gelatin, crushing the aired donkey-hide gelatin by a traditional Chinese medicine superfine crusher, putting the donkey-hide gelatin powder into a heat preservation tank, adding deionized water into the heat preservation tank, mixing and stirring the donkey-hide gelatin powder and the deionized water in the heat preservation tank by a stirring device, and heating the heat preservation tank at the same time, so as to dissolve the donkey-hide gelatin powder;
s2, cooling to room temperature after the dissolution is finished in the step S1, centrifuging the dissolved solution through a centrifugal machine, collecting supernatant of the dissolved solution to obtain donkey-hide gelatin extract, adjusting the pH of the donkey-hide gelatin extract with NaOH solution, performing enzymolysis with pepsin in water bath, cooling after the enzymolysis is finished, centrifuging through the centrifugal machine after cooling, taking the supernatant, separating and purifying through a solid phase extraction column, activating with methanol and deionized water in advance, eluting with methanol, collecting eluent, blow-drying with nitrogen, dissolving with formic acid solution, and filtering through a filter membrane to obtain the donkey-hide gelatin polypeptide.
Example 2:
a preparation method of donkey-hide gelatin polypeptide comprises the following steps:
s1, cleaning and airing the donkey-hide gelatin, crushing the aired donkey-hide gelatin by a traditional Chinese medicine superfine crusher, putting the donkey-hide gelatin powder into a heat preservation tank, adding deionized water into the heat preservation tank, mixing and stirring the donkey-hide gelatin powder and the deionized water in the heat preservation tank by a stirring device, and heating the heat preservation tank at the same time, so as to dissolve the donkey-hide gelatin powder;
s2, cooling to room temperature after the dissolution is finished in the step S1, centrifuging the dissolved solution through a centrifugal machine, collecting supernatant of the dissolved solution to obtain donkey-hide gelatin extract, adjusting the pH of the donkey-hide gelatin extract with NaOH solution, performing enzymolysis with pepsin in water bath, cooling after the enzymolysis is finished, centrifuging through the centrifugal machine after cooling, taking the supernatant, separating and purifying through a solid phase extraction column, activating with methanol and deionized water in advance, eluting with methanol, collecting eluent, blow-drying with nitrogen, dissolving with formic acid solution, and filtering through a filter membrane to obtain the donkey-hide gelatin polypeptide.
The weight of the deionized water in the step S1 is 10-15 times of that of the donkey-hide gelatin powder, the heating temperature of the heat preservation tank in the step S1 is 80-90 ℃, the stirring time of the stirring device in the step S1 is 25-30min, the concentration of the NaOH solution in the step S2 is 8%, the pH of the donkey-hide gelatin extracting solution in the step S2 is kept to be 8, the pH is adjusted every 1h in the enzymolysis process in the step S2, the enzyme activity is inactivated after 4h of enzymolysis, the water bath temperature in the step S2 is 45 ℃, and the concentration of the pepsin is 6 multiplied by 10 4 U/g-8×10 4 U/g, concentration of the formic acid solution is 2%, and centrifugation time of the centrifuge in the step S2The time is 8min, the nitrogen temperature in the step S2 is 50 ℃, the parameters of the solid phase extraction column are C18, and the parameters of the filtering membrane are 0.45 mu m.
Example 3:
a preparation method of donkey-hide gelatin polypeptide comprises the following steps:
s1, cleaning and airing the donkey-hide gelatin, crushing the aired donkey-hide gelatin by a traditional Chinese medicine superfine crusher, putting the donkey-hide gelatin powder into a heat preservation tank, adding deionized water into the heat preservation tank, mixing and stirring the donkey-hide gelatin powder and the deionized water in the heat preservation tank by a stirring device, and heating the heat preservation tank at the same time, so as to dissolve the donkey-hide gelatin powder;
s2, cooling to room temperature after the dissolution is finished in the step S1, centrifuging the dissolved solution through a centrifugal machine, collecting supernatant of the dissolved solution to obtain donkey-hide gelatin extract, adjusting the pH of the donkey-hide gelatin extract with NaOH solution, performing enzymolysis with pepsin in water bath, cooling after the enzymolysis is finished, centrifuging through a centrifugal machine after the cooling, taking the supernatant, separating and purifying through a solid phase extraction column, activating with methanol and deionized water in advance, eluting with methanol, collecting eluent, blow-drying with nitrogen, dissolving with formic acid solution, and filtering through a filter membrane to obtain the donkey-hide gelatin polypeptide.
The donkey-hide gelatin polypeptide obtained in the step S2 can be used for preparing donkey-hide gelatin polypeptide solution or can be used for obtaining donkey-hide gelatin polypeptide powder by a vacuum freeze drying method.
The application of the preparation method of the donkey-hide gelatin polypeptide comprises the step of preparing donkey-hide gelatin polypeptide oral preparations, such as donkey-hide gelatin polypeptide oral solution, decoction, syrup, medicinal liquor, decoction, tincture, extract, medicinal tea, tablets, capsules, granules and the like, or preparing donkey-hide gelatin polypeptide injection, or preparing donkey-hide gelatin polypeptide powder injection through vacuum freeze drying.
The donkey-hide gelatin polypeptide oral preparation can be prepared by using the donkey-hide gelatin polypeptide, such as donkey-hide gelatin polypeptide oral solution, decoction, syrup, medicinal liquor, decoction, tincture, extract, tea, tablets, capsules, granules and the like, can also be prepared, and can be widely applied.
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be considered to be within the technical scope of the present invention, and the technical solutions and the inventive concepts thereof according to the present invention should be equivalent or changed within the scope of the present invention.
Claims (7)
1. The preparation method of the donkey-hide gelatin polypeptide is characterized by comprising the following steps:
s1, cleaning and airing the donkey-hide gelatin, crushing the aired donkey-hide gelatin by a traditional Chinese medicine superfine crusher, putting the donkey-hide gelatin powder into a heat preservation tank, adding deionized water into the heat preservation tank, mixing and stirring the donkey-hide gelatin powder and the deionized water in the heat preservation tank by a stirring device, and heating the heat preservation tank at the same time, so as to dissolve the donkey-hide gelatin powder;
s2, cooling to room temperature after the dissolution is finished in the step S1, centrifuging the dissolved solution through a centrifugal machine, collecting supernatant of the dissolved solution to obtain donkey-hide gelatin extract, adjusting the pH of the donkey-hide gelatin extract with NaOH solution, performing enzymolysis with pepsin in water bath, cooling after the enzymolysis is finished, centrifuging through the centrifugal machine after cooling, taking the supernatant, separating and purifying through a solid phase extraction column, activating with methanol and deionized water in advance, eluting with methanol, collecting eluent, blow-drying with nitrogen, dissolving with formic acid solution, and filtering through a filter membrane to obtain the donkey-hide gelatin polypeptide.
2. The method of claim 1, wherein the weight of the deionized water in step S1 is 10-15 times of the weight of the colla corii asini powder, the heating temperature of the heat-preserving tank in step S1 is 80-90 ℃, and the stirring time of the stirring device in step S1 is 25-30 min.
3. The method of claim 1, wherein the concentration of the NaOH solution in step S2 is 8%, the pH of the E-jiao extract in step S2 is maintained at 8, the pH is adjusted every 1h in the enzymolysis process in step S2, and the enzyme activity is inactivated after 4h of enzymolysis.
4. The method of claim 1, wherein the temperature of the water bath in step S2 is 45 ℃, and the pepsin concentration is 6 x 10 4 U/g-8×10 4 U/g, the concentration of the formic acid solution is 2%, the centrifugation time of the centrifuge in the step S2 is 8min, and the nitrogen temperature in the step S2 is 50 ℃.
5. The preparation method of the donkey-hide gelatin polypeptide according to claim 1, characterized in that the parameter of the solid phase extraction column is C18, and the parameter of the filtration membrane is 0.45 μm.
6. The method of claim 1, wherein the donkey-hide gelatin polypeptide obtained in step S2 is processed into a donkey-hide gelatin polypeptide solution, or vacuum freeze-dried to obtain donkey-hide gelatin polypeptide powder.
7. A method for preparing colla Corii Asini polypeptide according to any one of claims-6, comprising preparing colla Corii Asini polypeptide oral preparation such as colla Corii Asini polypeptide oral solution, decoction, syrup, medicated liquor, soft extract, tincture, extract, medicated tea, tablet, capsule, granule, or colla Corii Asini polypeptide injection, or vacuum freeze drying to obtain colla Corii Asini polypeptide powder for injection.
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| CN116649580A (en) * | 2023-04-11 | 2023-08-29 | 济宁学院 | Preparation method of small molecule donkey-hide gelatin polypeptide product |
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| CN102101884A (en) * | 2009-12-18 | 2011-06-22 | 济南瑞安药业发展有限公司 | Preparation method and use of donkey-hide gelatin polypeptide |
| CN102321716A (en) * | 2011-08-18 | 2012-01-18 | 江中药业股份有限公司 | Preparation method of donkey-hide gelatin oligopeptide |
| US20140315819A1 (en) * | 2011-12-29 | 2014-10-23 | Shandong Dong-E E-Jiao Co., Ltd. | Active small-molecule donkey-hide gelatin mixture and preparation method and application thereof |
| CN205518037U (en) * | 2016-01-28 | 2016-08-31 | 北京本草方源药业有限公司 | Novel traditional chinese medicine superfine pulverizer |
| CN110790819A (en) * | 2019-10-25 | 2020-02-14 | 福州海关技术中心 | Donkey-hide gelatin polypeptide and preparation method thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102101884A (en) * | 2009-12-18 | 2011-06-22 | 济南瑞安药业发展有限公司 | Preparation method and use of donkey-hide gelatin polypeptide |
| CN102321716A (en) * | 2011-08-18 | 2012-01-18 | 江中药业股份有限公司 | Preparation method of donkey-hide gelatin oligopeptide |
| US20140315819A1 (en) * | 2011-12-29 | 2014-10-23 | Shandong Dong-E E-Jiao Co., Ltd. | Active small-molecule donkey-hide gelatin mixture and preparation method and application thereof |
| CN205518037U (en) * | 2016-01-28 | 2016-08-31 | 北京本草方源药业有限公司 | Novel traditional chinese medicine superfine pulverizer |
| CN110790819A (en) * | 2019-10-25 | 2020-02-14 | 福州海关技术中心 | Donkey-hide gelatin polypeptide and preparation method thereof |
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| CN116649580A (en) * | 2023-04-11 | 2023-08-29 | 济宁学院 | Preparation method of small molecule donkey-hide gelatin polypeptide product |
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