CN109337952A - A kind of extracting method of snake slough polypeptide - Google Patents
A kind of extracting method of snake slough polypeptide Download PDFInfo
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- CN109337952A CN109337952A CN201811278380.XA CN201811278380A CN109337952A CN 109337952 A CN109337952 A CN 109337952A CN 201811278380 A CN201811278380 A CN 201811278380A CN 109337952 A CN109337952 A CN 109337952A
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- C07K1/16—Extraction; Separation; Purification by chromatography
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- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/36—Extraction; Separation; Purification by a combination of two or more processes of different types
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Abstract
The invention discloses a kind of extracting method of snake slough polypeptide, step includes: snake slough pretreatment；Alcohol extracting is added ethanol solution and extracts, filters while hot, obtain ethanol extract and filter residue；Water and alkali protease enzymatic hydrolysis are added into filter residue, obtains enzymolysis liquid for enzymatic hydrolysis；Water mentions, and water is added into enzymolysis liquid, and filtering takes supernatant；Concentration merges ethanol extract and supernatant, is concentrated into certain volume, obtains polypeptide concentrate；Purifying, ammonium sulfate, which is added, into polypeptide concentrate makes protein saltout, and precipitating is taken after centrifugation, then through gel column desalination, obtain polypeptide solution after purification, then be freeze-dried to get snake slough polypeptide.The easy to operate of this method, extraction cost are low, can quickly and effectively extract snake slough polypeptide, the recovery rate of polypeptide is higher, and percent hydrolysis is lower.
Description
Technical field
The present invention relates to animal polypeptide extractive technique field, in particular to a kind of extracting method of snake slough polypeptide.
Background technique
Polypeptide is the compound as made of three or three or more amino acid molecular dehydrating condensations, in the market common polypeptide
There are animal polypeptide and plant polypeptide, the chemical combination that animal polypeptide and plant polypeptide are composed of a variety of amino acid by peptide bond
Object, the type of amino acid contains needed by human 8 in animal polypeptide more than the type of amino acid in plant polypeptide in animal polypeptide
Kind amino acid, coincide, easily by human consumption compared with the trophic structure of the mankind.
Snake slough also known as imperial clothing, are the cuticula under snake sloughs off naturally during the growth process.Contain egg very rich in snake slough
The nutritional ingredients such as white matter, fatty acid, sterol, polypeptide and microelement, medicinal peace is strong, not only wind-dispelling but also anti-inflammatory, no impairment of yin
Fire, the cold and cool disadvantage upset one's stomach are helped, it is significant in efficacy, securely and reliably, there is good effects of antiinflammation and bacteriostasis, can be widely used for treating a variety of
Active chronic inflammation.China's snake drug is abundant, and many kinds of, studies in China personnel are more to the research of snakeskin, but to snake slough
Chemical constitution study is less, and then less for the research of snake slough polypeptide.Currently, there is researcher by adding water high-temp extracting to obtain
To the hydrolyzate with snake slough polypeptide, the hydrolyzate which mentions contains polypeptide and other water-soluble substances, snake slough polypeptide recovery rate compared with
It is low.Wu Yuqiong etc. is extracted the albumen in snake slough with biologic enzymolysis method, and this method can be suitable for small-scale extraction, work as needs
When a large amount of albumen extracted in snake slough, the enzyme amount used is big, and extraction cost is high；And in the snake slough residue after this method extraction also
It containing a small amount of albumen, extract the albumen in snake slough residue can not completely, cause the albumen in snake slough that cannot access rationally
It utilizes, reduces its utilization rate and recovery rate.
In conclusion the extraction snake slough polypeptide how to be simple and efficient, improves the utilization rate of snake slough polypeptide, extraction cost is reduced
It is current urgent problem to be solved.
Summary of the invention
The technical problem to be solved by the present invention is overcoming the deficiencies of the prior art and provide a kind of easy to operate, production cost
Extracting method low, high-efficient, that extensive extraction snake slough polypeptide can be suitable for.
The technical solution adopted by the present invention to solve the technical problems is:
A kind of extracting method of snake slough polypeptide, which comprises the following steps:
1) snake slough pre-processes；
2) alcohol extracting: ethanol solution is added and extracts, filters while hot, obtains ethanol extract and filter residue；
3) it digests: water and alkali protease enzymatic hydrolysis being added into filter residue, obtains enzymolysis liquid；
4) water mentions: water being added into enzymolysis liquid, filters, takes supernatant；
5) it is concentrated: merging ethanol extract and supernatant, be concentrated into certain volume, obtain polypeptide concentrate；
6) purify: ammonium sulfate is added into polypeptide concentrate makes protein saltout, and takes precipitating after centrifugation, then through gel column desalination,
Obtain polypeptide solution after purification；
7) polypeptide solution obtained in step 6) is freeze-dried to get snake slough polypeptide.
Ethyl alcohol can dissolve alcohol-soluble or water-soluble part in snake slough polypeptide, and then improve the extraction of snake slough polypeptide
Rate.Polypeptide solubility in hot solution is bigger than in cold soln, therefore, needs to filter while hot after having been extracted with ethyl alcohol, improves filtering
Efficiency and polypeptide recovery rate.
Further, the pretreated step of the snake slough are as follows: collect snake slough, clean and remove impurity, dry, crush, obtain
Snake slough powder.
Further, the drying temperature is 60-80 DEG C, and degree of grinding is 100 mesh.
To prevent high temperature from be denaturalized snake slough polypeptide, this method selects drying temperature for 60-80 DEG C.
In order to expand the contact of snake slough with solvent and enzyme, then the method that this method is ground using drying crosses 100 purposes point
Analysis sieve.Snake slough reduction ratio is thinner, during extraction, increases its contact area with solvent, makes mentioning for snake slough polypeptide
It takes more sufficiently, enzymolysis speed is faster, and hydrolysis result is better.
Further, the solid-liquid ratio of snake slough and ethyl alcohol is 1:20-1:30 in the step 2, and the concentration of ethyl alcohol is 30-60%,
Ethyl alcohol Extracting temperature is 50-80 DEG C, and ethyl alcohol extraction time is 2-4h.
Further, the water in the step 3) is that 1:3-1:5 is added into filter residue by solid-liquid ratio.
Further, the amount that alkali protease is added in every gram of filter residue is 1000~3000U, and the temperature of the enzymatic hydrolysis is
40-60 DEG C, the pH of enzymatic hydrolysis is 7-9, and the time of enzymatic hydrolysis is 3-6h.
Alkali protease is added by the quality of filter residue, acts on protease sufficiently with snake slough, it in a relatively short period of time will be residual
Remaining snake slough sufficiently digests in slag, improves the utilization rate of raw material and the recovery rate of snake slough polypeptide.
When alkali protease is digested within the scope of suitable temperature and pH, alkali protease is made to keep higher work
Property, can efficiently, quickly by the proteolysis in snake slough be polypeptide；Suitable enzymolysis time is controlled, to guarantee that snake slough fills
Divide enzymatic hydrolysis, guarantees the utilization rate and yield of raw material.
Further, the solid-liquid ratio for the water being added in the filter residue and step 4) is 1:10-1:15, mentioning in the step 4)
Taking temperature is 60-80 DEG C, extraction time 2-4h.
The ratio for increasing solvent extracts snake slough polypeptide more abundant, extracts under the conditions of 60-80 DEG C, can increase solvent
To the solubility of snake slough polypeptide, but temperature be not easy it is excessively high, it is excessively high to make polypeptide loss of activity.
Further, the temperature being concentrated in the step 5) is 60-80 DEG C, and the volume after concentration is the 1/2-1/3 of original volume.
Reduced pressure can remove a large amount of moisture in snake slough polypeptide solution, increase the precipitation of snake slough polypeptide, keep freezing dry
The volume decline of polypeptide solution, reduces the workload in freezing dry process during dry, improves the efficiency that polypeptide extracts.Subtract
Press the temperature of concentration unsuitable excessively high, to guarantee the activity of polypeptide.
Concentration process may be that the methods of ultrafiltration, nanofiltration, reverse osmosis carry out film filtering and concentrating, and film filtering and concentrating is according to need
The molecular size range for filtering or retaining is different, and suitable aperture is selected to be retained.Ultrafiltration nanofiltration is in retention peptide molecule
Meanwhile other small molecular weight impurities in supernatant can be removed, the purity of polypeptide product is improved, while can fully ensure that polypeptide point
The activity of son.
Further, the concentration of the ammonium sulfate is 60-85%.
The salt ion of high concentration can compete hydrone with protein in protein solution, to destroy protein surface
Hydration shell reduces its solubility, is allowed to be precipitated out from solution.Ammonium sulfate is with solubility is big, temperature coefficient is small and is not easy
The advantages of making protein denaturation, ammonium sulfate is added in the snake slough polypeptide after reduced pressure can reduce the dissolution of snake slough polypeptide
Degree promotes snake slough polypeptide to be precipitated, and obtained polypeptide impurities content is minimum, improves the recovery rate and purity of snake slough polypeptide.
Further, the step of gel column desalination are as follows: it is heavy to be dissolved in water by precipitating for 1:20-30 with the solid-liquid ratio of water
It forms sediment, with sephadex G25 gel column desalination, collects separating liquid to get polypeptide solution after purification.Using gel column by snake slough
Ammonium sulfate in polypeptide solution is separated, and polypeptide solution is further purified.
Further, the temperature of the freeze-drying is 60-80 DEG C.
Freeze-drying process can largely keep the active function of polypeptide, and the structure for reducing peptide molecule changes.
In freezing dry process, the volume of solution is excessive, and the moisture after freezing in the middle part of material is not easy to diffuse to the surface, to be not easy to do
It is dry complete, therefore, before freeze-drying, the volume of solution should be reduced as far as possible, while increasing the surface area of solution, make to be lyophilized
Moisture is easier diffusive evaporation afterwards.
Beneficial effects of the present invention: a kind of extracting method of snake slough polypeptide of the present invention, this method are first adopted using snake slough as raw material
With traditional alcohol extracting method, most of polypeptide in snake slough is extracted, since part snake slough polypeptide is not easy to be taken out by alcohol extracting,
Remain to this part snake slough polypeptide in filter residue, recycles biologic enzymolysis method to digest snake slough filter residue, reduce and directly adopt biology
Enzymatic isolation method digests usage amount and the action time of snake slough, to reduce extraction cost, increase the efficiency of snake slough polypeptide extraction；
It is extracted in snake slough by biologic enzymolysis method and is not easy the polypeptide portion taken out by alcohol extracting, increase the utilization of snake slough polypeptide residue,
Improve the recovery rate of snake slough polypeptide；The polypeptide that may do not digested in enzymolysis liquid further finally is extracted with water extraction, further
Improve the recovery rate of snake slough polypeptide；The reaction condition of the extracting method is mild, at low cost, easy to operate, the snake slough polypeptide of extraction
Degree of hydrolysis it is low, can be applied to industrialized production, have very high economic value.
Detailed description of the invention
Fig. 1-is the process flow diagram that the present invention extracts snake slough polypeptide.
Specific embodiment
The invention will be further described with reference to embodiments, described in embodiment, be only present pre-ferred embodiments and
, it is not intended to limit the scope of the present invention, therefore according to the technical essence of the invention to above embodiments institute
Any subtle modifications, equivalent variations and modifications made, all of which are still within the scope of the technical scheme of the invention.
Embodiment 1
(1) snake slough is removed into impurity, washed with water completely, 60 DEG C of drying crushed 100 mesh；
(2) 60% ethanol solution is added by solid-liquid ratio 1:20,2h is extracted at 60 DEG C, is filtered while hot, takes filtrate, and rushed with water
Three times, merging filtrate and eluent obtain ethanol extract to filter wash slag, spare；
(3) alkali protease of water and 2500U/g immobilization is added by solid-liquid ratio 1:3 into filter residue, and adjusts pH value to 8.4,
4h is digested at 42 DEG C, after having digested, obtains enzymolysis liquid；
(4) it is that water is added into enzymolysis liquid by 1:15 according to the solid-liquid ratio of filter residue and water, 2h is extracted at 65 DEG C, after the completion of extraction,
It filters while hot, takes supernatant, it is spare；
(5) merge ethanol extract and supernatant, be concentrated under reduced pressure under conditions of 65 DEG C, when being concentrated into the 1/2 of original volume, is added
68% ammonium sulfate makes protein coagulation, is centrifuged 10min under the gravity of 1200G, takes precipitating, and with the sulfuric acid of isoconcentration
Ammonium salt solution cleaning precipitating, precipitating is transferred in beaker, is that 1:30 is dissolved in water precipitating by precipitating and the solid-liquid ratio of water, use
Sephadex G25 gel column removes the ammonium sulfate in solution, collects separating liquid, is freeze-dried at 60 DEG C more to get snake slough
Peptide.
Embodiment 2
The present embodiment difference from example 1 is that: be added alkali protease amount be 2000U/g, other steps and reality
It is consistent to apply example 1.
Embodiment 3
The present embodiment difference from example 1 is that: be added alkali protease amount be 1500U/g, other steps and reality
It is consistent to apply example 1.
Embodiment 4
The present embodiment difference from example 1 is that: be added alkali protease amount be 2000U/g, ammonium sulfate
Concentration is 30%, other steps and embodiment 1 are consistent.
Comparative example 1
(1) snake slough is removed into impurity, washed with water completely, 60 DEG C of drying crushed 100 mesh；
(2) 60% ethanol solution is added by solid-liquid ratio 1:20,2h is extracted at 60 DEG C, is filtered while hot, takes filtrate, and rushed with water
Three times, merging filtrate and eluent obtain ethanol extract to filter wash slag；
(3) ethanol extract is concentrated under reduced pressure under conditions of 65 DEG C, when being concentrated into the 1/2 of original volume, 68% ammonium sulfate is added
Solution makes protein coagulation, is centrifuged 10min under the gravity of 1200G, takes precipitating, and cleaned and sunk with the ammonium sulfate of isoconcentration
It forms sediment, precipitating is transferred in beaker, be that 1:30 is dissolved in water precipitating by precipitating and the solid-liquid ratio of water, with sephadex G25 gel
Column removes the ammonium sulfate in solution, collects separating liquid, and freeze-drying is at 60 DEG C to get snake slough polypeptide.
Comparative example 2
(1) snake slough is removed into impurity, washed with water completely, 60 DEG C of drying crushed 100 mesh；
(2) into snake slough powder by solid-liquid ratio 1:3 be added water and 2500U/g immobilization alkali protease, and adjust pH value to
8.4,4h is digested at 42 DEG C, after having digested, obtains enzymolysis liquid；
(3) it is that water is added into enzymolysis liquid by 1:15 according to the solid-liquid ratio of filter residue and water, 2h is extracted at 65 DEG C, after the completion of extraction,
It filters while hot, takes supernatant, it is spare；
(4) merge enzymolysis liquid and supernatant, be concentrated under reduced pressure under conditions of 65 DEG C, when being concentrated into the 1/2 of original volume, be added 68%
Ammonium sulfate make protein coagulation, be centrifuged 10min under the gravity of 1200G, take precipitating, and molten with the ammonium sulfate of isoconcentration
Liquid cleaning precipitating, precipitating is transferred in beaker, is that 1:30 is dissolved in water precipitating by precipitating and the solid-liquid ratio of water, is used sephadex
G25 gel column removes the ammonium sulfate in solution, collects separating liquid, and freeze-drying is at 60 DEG C to get snake slough polypeptide.
Comparative example 3
(1) snake slough is removed into impurity, washed with water completely, 60 DEG C of drying crushed 100 mesh；
(2) 60% ethanol solution is added by solid-liquid ratio 1:20,2h is extracted at 60 DEG C, is filtered while hot, takes filtrate, and rushed with water
Three times, merging filtrate and eluent obtain ethanol extract to filter wash slag, spare；
(3) alkali protease of water and 2500U/g immobilization is added by solid-liquid ratio 1:3 into filter residue, and adjusts pH value to 8.4,
4h is digested at 42 DEG C, after having digested, obtains enzymolysis liquid；
(4) merge ethanol extract and enzymolysis liquid, be concentrated under reduced pressure under conditions of 65 DEG C, when being concentrated into the 1/2 of original volume, is added
68% ammonium sulfate makes protein coagulation, is centrifuged 10min under the gravity of 1200G, takes precipitating, and with the sulfuric acid of isoconcentration
Ammonium salt solution cleaning precipitating, precipitating is transferred in beaker, is that 1:30 is dissolved in water precipitating by precipitating and the solid-liquid ratio of water, use
Sephadex G25 gel column removes the ammonium sulfate in solution, collects separating liquid, precipitates obtained by freeze-drying at 60 DEG C to obtain the final product
Snake slough polypeptide.
Comparative example 4
(1) snake slough is removed into impurity, washed with water completely, 60 DEG C of drying crushed 100 mesh；
(2) 60% ethanol solution is added by solid-liquid ratio 1:20,2h is extracted at 60 DEG C, is filtered while hot, takes filtrate, and rushed with water
Three times, merging filtrate and eluent obtain ethanol extract to filter wash slag, spare；
(3) it is that water is added into ethanol extract by 1:15 according to the solid-liquid ratio of filter residue and water, 2h is extracted at 65 DEG C, extracts and completes
Afterwards, it filters while hot, takes supernatant, it is spare；
(4) merge ethanol extract and supernatant, be concentrated under reduced pressure under conditions of 65 DEG C, when being concentrated into the 1/2 of original volume, is added
68% ammonium sulfate makes protein coagulation, is centrifuged 10min under the gravity of 1200G, takes precipitating, and with the sulfuric acid of isoconcentration
Ammonium salt solution cleaning precipitating, precipitating is transferred in beaker, is that 1:30 is dissolved in water precipitating by precipitating and the solid-liquid ratio of water, use
Sephadex G25 gel column remove solution in ammonium sulfate, collect separating liquid, at 60 DEG C be freeze-dried to get for snake slough it is more
Peptide.
The present invention carries out the yield, purity and degree of hydrolysis of the snake slough polypeptide extracted in embodiment 1-4 and comparative example 1-4
Measurement, measuring method are as follows:
The measurement of snake slough polypeptide yield: it combines and is measured with formol titration using trichloroacetic acid (TCA) precipitation method.It takes
10mL enzymolysis liquid adds 10mL20% (g/g) TCA, stirs evenly and is centrifuged 10min at 4800r/min after standing half an hour, it is resulting on
Clear liquid is denoted as supernatant TCA, respectively with the total nitrogen content and ammoniacal nitrogen in Kjeldahl's method and formol titration measurement supernatant
Content, then peptide nitrogen quantity and peptide yield are calculated as the following formula:
The total peptide nitrogen quantity of the sample=sample TCA nitrogen pool-total ammoniacal nitrogen amount of supernatant TCA,
The total peptide nitrogen content of peptide yield=sample/raw material total protein nitrogen content × 100%.
The measurement of snake slough Purity: with bovine serum albumin(BSA) (BSA) for standard items, the BSA solution of various concentration is made.
5mL coomassie brilliant blue staining solution dyeing 5min is separately added into 4 test tubes.Using the solution in the 1st test tube as blank pair
According to measuring absorbance of the solution at 595nm in remaining 3 test tube.With protein quality concentration (gL-1) it is abscissa, it inhales
Luminosity is ordinate, carries out linear regression analysis.Protein concentration is 0~100 μ gL-1In range, equation of linear regression Y=
(6.6154X+0.01 r=0.999).The result shows that linear relationship is good, it can be used to carry out the measurement of protein content.Point
Other extraction embodiment and the resulting snake slough molecular peptide of comparative example measure absorbance as test sample at 595nm, and according to standard
Curve calculates protein content, calculates protein quality score/%.
The measurement of snake slough polypeptide degree of hydrolysis: using ninhydrin colour developing hair measurement-NH2Number.
Degree of hydrolysis is calculated using formula formula:
In formula: AhTotal free-NH in different time enzymolysis liquid2Number (mmol)；A0Intrinsic free-NH in material protein2Number
(mmol)；AAlwaysTotal free-NH after material protein strong acid hydrolysis2Number (mmol).
Polypeptide yield, purity, the measurement result of degree of hydrolysis are as shown in table 1 in embodiment 1-4 and comparative example 1-4:
1 snake slough polypeptide yield of table, purity, degree of hydrolysis
The degree of hydrolysis for the snake slough polypeptide that embodiment and comparative example is extracted is shown in Table 1.
As seen from the above table, high, purity is high, polypeptide using extracting method extraction snake slough polypeptide recovery rate obtained of the invention
Degree of hydrolysis it is lower.
Claims (10)
1. a kind of extracting method of snake slough polypeptide, which comprises the following steps:
1) snake slough pre-processes；
2) alcohol extracting: ethanol solution is added and extracts, filters while hot, obtains ethanol extract and filter residue；
3) it digests: water and alkali protease enzymatic hydrolysis being added into filter residue, obtains enzymolysis liquid；
4) water mentions: water being added into enzymolysis liquid, filters, takes supernatant；
5) it is concentrated: merging ethanol extract and supernatant, be concentrated into certain volume, obtain polypeptide concentrate；
6) purify: ammonium sulfate is added into polypeptide concentrate makes protein saltout, and takes precipitating after centrifugation, then through gel column desalination,
Obtain polypeptide solution after purification；
7) polypeptide solution obtained in step 6) is freeze-dried to get snake slough polypeptide.
2. a kind of extracting method of snake slough polypeptide as described in claim 1, which is characterized in that the pretreated step of snake slough
Are as follows: snake slough is collected, impurity is cleaned and remove, is 60-80 DEG C of drying in temperature, then be crushed to 100 mesh, obtains snake slough powder.
3. a kind of extracting method of snake slough polypeptide as claimed in claim 1 or 2, which is characterized in that in the step 2 snake slough with
The solid-liquid ratio of ethyl alcohol is 1:20-1:30, and the concentration of ethyl alcohol is 30-60%, and ethyl alcohol Extracting temperature is 50-80 DEG C, ethyl alcohol extraction time
For 2-4h.
4. a kind of extracting method of snake slough polypeptide as described in claim 1, which is characterized in that the water in the step 3) presses feed liquid
Than being added for 1:3-1:5 into filter residue.
5. a kind of extracting method of snake slough polypeptide as claimed in claim 4, which is characterized in that alkalinity is added in every gram of filter residue
The amount of protease is 1000~3000U, and the temperature of the enzymatic hydrolysis is 40-60 DEG C, and the pH of enzymatic hydrolysis is 7-9, and the time of enzymatic hydrolysis is 3-
6h。
6. a kind of extracting method of snake slough polypeptide as described in claim 1, which is characterized in that be added in the filter residue and step 4)
The solid-liquid ratio of water be 1:10-1:15, the Extracting temperature in the step 4) is 60-80 DEG C, extraction time 2-4h.
7. a kind of extracting method of snake slough polypeptide as described in claim 1, which is characterized in that the temperature being concentrated in the step 5)
It is 60-80 DEG C, the volume after concentration is the 1/2-1/3 of original volume.
8. a kind of extracting method of snake slough polypeptide as described in claim 1, which is characterized in that the concentration of the ammonium sulfate is
60-85%。
9. a kind of extracting method of snake slough polypeptide as claimed in claim 8, which is characterized in that the step of the gel column desalination
Are as follows: it by precipitating and the solid-liquid ratio of water is that the 1:20-30 precipitating that is dissolved in water with sephadex G25 gel column desalination is collected and separated
Liquid is to get polypeptide solution after purification.
10. a kind of extracting method of snake slough polypeptide as described in claim 1, which is characterized in that the temperature of the freeze-drying is
60-80℃。
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111850077A (en) * | 2020-08-04 | 2020-10-30 | 西安惠普生物科技有限公司 | Preparation method of periplaneta americana polypeptide solution and periplaneta americana polypeptide solution |
| CN114250261A (en) * | 2021-12-28 | 2022-03-29 | 河南省国德科果蔬研究院有限公司 | Extraction method of red date kernel polypeptide |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102277405A (en) * | 2011-08-02 | 2011-12-14 | 唐作安 | Preparation method of snakeskin active peptide |
-
2018
- 2018-10-30 CN CN201811278380.XA patent/CN109337952A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102277405A (en) * | 2011-08-02 | 2011-12-14 | 唐作安 | Preparation method of snakeskin active peptide |
Non-Patent Citations (1)
| Title |
|---|
| 陈红红等: "银环蛇蛇蜕的化学成分研究Ⅱ. 脂肪酸和氨基酸组分", 《分析测试学报》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111850077A (en) * | 2020-08-04 | 2020-10-30 | 西安惠普生物科技有限公司 | Preparation method of periplaneta americana polypeptide solution and periplaneta americana polypeptide solution |
| CN114250261A (en) * | 2021-12-28 | 2022-03-29 | 河南省国德科果蔬研究院有限公司 | Extraction method of red date kernel polypeptide |
| CN114250261B (en) * | 2021-12-28 | 2023-08-22 | 河南省国德科果蔬研究院有限公司 | Method for extracting red date pit polypeptide |
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