CN103969367A - Identification method of arca subcrenata protein - Google Patents
Identification method of arca subcrenata protein Download PDFInfo
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Abstract
The invention relates to an identification method of arca subcrenata protein, which is characterized in that gel chromatography column high performance liquid chromatography is used. The invention provides a specificity identification method of arca subcrenata protein polypeptide, which has the benefits that the product quality is effectively ensured during the production and the use of an arca subcrenata preparation, and further, the curative effect of drugs is ensured.
Description
Technical field
The present invention relates to medicine detection method, be specifically related to a kind of discrimination method of blood clam protein, belong to field of pharmaceutical technology.
Background technology
Blood clam meat is containing rich in protein, vitamin and the necessary amino acid of human body, just there is edible, medical value from ancient times, " medical center bun will " note " blood clam meat bushing blood, loose hemostasis, relieving restlessness are sobered up, the dissolving phlegm of broken knot ", " Japan hanako materia medica " note " blood clam meat benefit color ".In recent years, research report shows that blood clam has antitumor action, has effect of removing interior free yl, improving immunity of organisms.Blood clam meat preparation has good therapeutic action to kinds cancers such as lung cancer, kidney, cancer of the stomach, liver cancer, and security is good.
Be one of main bioactive ingredients of blood clam according to bibliographical information blood clam protein, and be rich in the essential amino acid of needed by human body, 8 kinds of essential amino acids account for 36.5% of total amino acid.Antineoplaston has very high active anticancer.The content assaying method of existing document plant animal protein is more, but to the specificity report of plant animal protein seldom, more has no the research report to blood clam protein specificity discrimination method.Cannot in producing and using, effectively ensure product quality, and then pharmaceutical effectiveness can not be guaranteed.
Summary of the invention
The object of the invention is to overcome the deficiencies in the prior art, a kind of specificity discrimination method of blood clam protein and peptide is provided, make blood clam preparation in producing and using, effectively ensure product quality, and then ensure pharmaceutical effectiveness.
The present invention is achieved in that
A discrimination method for blood clam protein and peptide, is characterized in that using gel chromatographic columns high performance liquid chromatography, and concrete steps are as follows:
Chromatographic condition and system flexibility: the gel chromatographic columns that is filling agent with hydrophilic spherical superpolymer; Taking the phosphate buffer of 0.01-0.10mol/L as mobile phase; Detecting wavelength is 280 nm; Column temperature: 20-40 DEG C; Flow velocity: 0.1ml-1ml/min.Number of theoretical plate calculates and should be not less than 3000 by protein molecular weight 100000.
Prepare blood clam polypeptide: get blood clam preparation porphyrize, take the sample containing the about 5-20g of blood clam extract, add the 10-15 times of ultrasonic extraction of water 20-40 min, centrifugal 5-15 min, get upper strata liquid, filter, filtrate is mark 50-70%(w/v by volume) add ammonium sulfate, 0～4 DEG C of placement is spent the night, centrifugal, get precipitation, the 5-10ml that adds water dissolves, taking the ammonium bicarbonate soln of 0.5-2% as dislysate, the dialysis membrane that is 3500 with molecular cut off, fully dialysis under 0～4 DEG C of condition (checking with the BaCl2 solution of 0.5-2% whether sulfate ion dialyses completely), solution in dialysis membrane is transferred in 20-100ml measuring bottle, water constant volume, 0～4 DEG C of preservation, obtain blood clam polypeptide test sample.
Measure: get blood clam polypeptide need testing solution 10-100 μ l, injection liquid chromatography, measures, and to obtain final product.
In blood clam polypeptide collection of illustrative plates, have a sharp-pointed characteristic peak " peak S " in retention time 15.6min left and right, peak S retention time difference should be within ± 5%.
The discrimination method of above-mentioned a kind of blood clam polypeptide, chromatographic condition and system flexibility are optimized for: the gel chromatographic columns that is filling agent with hydrophilic spherical superpolymer; Taking the phosphate buffer of 0.05 mol/L as mobile phase; Detecting wavelength is 280 nm; Column temperature: 25 DEG C; Flow velocity: 0.4ml/min.Number of theoretical plate calculates and should be not less than 5000 by protein molecular weight 100000.
The discrimination method of above-mentioned a kind of blood clam polypeptide, prepare blood clam polypeptide and be optimized for:
Get blood clam preparation porphyrize, take containing the about 10-12g of blood clam extract, add the 10-12 times of ultrasonic extraction of water 30-35 min, centrifugal 8-12 min, get upper strata liquid, filter, filtrate is mark 60-65%(w/v by volume) add ammonium sulfate, 0～4 DEG C of placement is spent the night, centrifugal, get precipitation, the 10ml that adds water dissolves, ammonium bicarbonate soln taking 1% is dislysate, the dialysis membrane that is 3500 with molecular cut off, fully dialysis under 0～4 DEG C of condition (checking with the BaCl2 solution of 0.8-1% whether sulfate ion dialyses completely), solution in dialysis membrane is transferred in 25-50ml measuring bottle, water constant volume, 0～4 DEG C of preservation, obtain blood clam polypeptide test sample.
The discrimination method of above-mentioned a kind of blood clam polypeptide, prepare blood clam polypeptide and be further optimized for:
Blood clam preparation porphyrize is got in the preparation of blood clam extract solution, take containing blood clam extract 10g, add 12 times of ultrasonic extraction 30 min of water, centrifugal 10 min, get upper strata liquid, filter, filtrate is mark 60%(w/v by volume) add ammonium sulfate, 0～4 DEG C of placement is spent the night, centrifugal, get precipitation, the 10ml that adds water dissolves, ammonium bicarbonate soln taking 1% is dislysate, the dialysis membrane that is 3500 with molecular cut off, fully dialysis under 0～4 DEG C of condition (the BaCl2 solution with 1% checks whether sulfate ion dialyses completely), solution in dialysis membrane is transferred in 50ml measuring bottle, water constant volume, 0～4 DEG C of preservation, obtain blood clam polypeptide test sample.
The discrimination method of above-mentioned a kind of blood clam polypeptide, measures to be optimized for and gets blood clam polypeptide need testing solution 10-50 μ l, and injection liquid chromatography is measured.
The discrimination method of above-mentioned a kind of blood clam polypeptide, measures to be further optimized for and gets blood clam polypeptide test sample solution 20 μ l, and injection liquid chromatography is measured.
The discrimination method of above-mentioned a kind of blood clam polypeptide, can be used for the discriminating of the various preparations of blood clam extract.
The discrimination method of above-mentioned a kind of blood clam polypeptide, can be used for the discriminating of stalwart blood clam, mud blood clam, flower clam, Scallop Extract and various preparations thereof.
Beneficial effect of the present invention is: a kind of discrimination method of blood clam polypeptide is provided, can accurately differentiates the polypeptide in the sources such as the nearly source of blood clam species chief blood clam, mud blood clam, scallop, flower clam.
Below by test example, further illustrate beneficial effect of the present invention.
key instrument and reagent
Shimadzu-20A high performance liquid chromatograph, UV-detector; Chromatographic column: TSK GEL G4000 PW
xL(7.8mm × 300mm); Mobile phase: the phosphate buffer of 0.05 mol/l; 25 DEG C of column temperatures; Flow velocity 0.4ml/min; Detect wavelength 280nm; Number of theoretical plate is calculated as 5000 by protein molecular weight 100000.
test method and result
2.1 methodprepare respectively blood clam meat, blood clam meat extract, blood clam extract capsule, blood clam extract tablet, blood clam extract pill, blood clam extract particles, stalwart blood clam meat, mud blood clam meat, flower clam meat, scallop polypeptide.
Get respectively dry blood clam, stalwart blood clam, mud blood clam, flower clam, scallop, pulverize, cross 60 mesh sieves, add the water of 8 times of amounts, heating is extracted 2 times, each 45 minutes, merge extract, filter, get filtrate, being concentrated into relative density is 1.10～1.15(60 DEG C) time, below 80 DEG C, drying under reduced pressure, pulverizes, and makes respectively blood clam, stalwart blood clam, mud blood clam, flower clam, Scallop Extract.
Get respectively blood clam extract capsule content
,blood clam extract tablet
,blood clam extract pill
,blood clam extract particles is pulverized, and obtains respectively blood clam extract capsule
,blood clam extract tablet
,blood clam extract pill
,blood clam extract particles powder.
Prepare respectively polypeptide, get respectively blood clam meat extract, blood clam extract capsule powder, blood clam extract tablet powder, blood clam extract pill powder, blood clam extract particles powder, chief blood clam meat extract, mud blood clam meat extract, flower clam meat extract, scallop extract 10 g, add 12 times of ultrasonic extraction 30 min of water, centrifugal 10 min, get upper strata liquid, filter, filtrate is mark 60%(w/v by volume) add ammonium sulfate, 0～4 DEG C of placement is spent the night, centrifugal, get precipitation, the 10ml that adds water dissolves, and the ammonium bicarbonate soln taking 1% is dislysate, the dialysis membrane that is 3500 with molecular cut off, fully dialysis under 0～4 DEG C of condition (the BaCl2 solution with 1% checks whether sulfate ion dialyses completely), is transferred to solution in dialysis membrane in 50ml measuring bottle water constant volume, 0～4 DEG C of preservation, obtains respectively blood clam meat, blood clam meat extract, blood clam extract capsule, blood clam extract tablet, blood clam extract pill, blood clam extract particles, chief blood clam meat, mud blood clam meat, flower clam meat, scallop polypeptide test sample.
Get respectively blood clam meat
,blood clam meat extract
,blood clam extract capsule
,blood clam extract tablet
,blood clam extract pill
,blood clam extract particles, stalwart blood clam meat, mud blood clam meat, flower clam meat, scallop polypeptide need testing solution, injection liquid chromatography, measures.
result
The collection of illustrative plates of homopolypeptide not the results are shown in Table 1.
Table 1 is the collection of illustrative plates result of homopolypeptide not
Note: t
r: retention time.
Medicinal material, blood clam extract found that, in blood clam meat, blood clam extract and formulation samples HPLC figure thereof, all there is sharp-pointed characteristic peak " peak S " (average retention time 15.593min in retention time 15.6min left and right, RSD:1.3%), peculiar by blood clam polypeptide sample, the specificity that can be used for blood clam meat, blood clam extract and preparation polysaccharide is differentiated; Under the sampling amount prerequisite consistent with processing procedure, in stalwart blood clam chromatogram (Fig. 5), there is not peak S.Under the sampling amount prerequisite consistent with processing procedure, the peak height of other chromatograms in mud blood clam, flower clam and scallop spectrogram (Fig. 6～Fig. 8) within the scope of retention time corresponding to peak S is well below peak S, point out the protein component relevant to peak S content in these three kinds of medicinal materials few, therefore also ignore.In sum, peak S is the specificity chromatographic peak that blood clam control medicinal material and Haishengsu-active substance extracted from marine living things ball are different from other medicinal material.
Brief description of the drawings
Fig. 1: blood clam meat HPLC collection of illustrative plates: in figure, peak S is the characteristic peak of blood clam protein value polypeptide, the retention time at S peak is 15.587min.
Fig. 2: blood clam extractive HPLC collection of illustrative plates: in figure, peak S is the characteristic peak of blood clam protein and peptide, the retention time at S peak is 15.590min.
Fig. 3: blood clam extract capsule HPLC collection of illustrative plates: in figure, peak S is the characteristic peak of blood clam protein and peptide, the retention time at S peak is 15.602min.
Fig. 4: stalwart blood clam HPLC collection of illustrative plates: figure Zhongkui blood clam is without blood clam protein and peptide characteristic peak.
Fig. 5: mud blood clam HPLC collection of illustrative plates: in figure, mud blood clam is without blood clam protein and peptide characteristic peak.
Fig. 6: flower clam HPLC collection of illustrative plates: in figure, flower clam is without blood clam protein and peptide characteristic peak.
Fig. 7: scallop HPLC collection of illustrative plates: in figure, scallop is without blood clam protein and peptide characteristic peak.
Below in conjunction with embodiment, the invention will be further described.
embodiment 1
Chromatographic condition and system flexibility: by Shimadzu-20A high performance liquid chromatograph, UV-detector; Chromatographic column: TSK GEL G4000 PW
xL(7.8mm × 300mm); Mobile phase: the phosphate buffer of 0.01 mol/l; 22 DEG C of column temperatures; Flow velocity 0.1ml/min; Detect wavelength 280nm, number of theoretical plate is calculated as 5000 by protein molecular weight 100000.
Prepare blood clam extract: get blood clam meat, fragmentation, crosses 40 mesh sieves, adds 1.5 times of water gagings, extract 4 hours 10 DEG C of following stirrings, extracting liquid filtering, centrifugal.Get above-mentioned centrifugate freeze-drying, obtain blood clam extract.
Prepare blood clam polypeptide: get blood clam extract appropriate, porphyrize, take 10g, add 12 times of ultrasonic extraction 35 min of water, centrifugal 8 min, get upper strata liquid, filter, filtrate is mark 60%(w/v by volume) add ammonium sulfate, 0～4 DEG C of placement is spent the night, centrifugal, get precipitation, the 10ml that adds water dissolves, ammonium bicarbonate soln taking 0.5% is dislysate, the dialysis membrane that is 3500 with molecular cut off, fully dialysis under 0～4 DEG C of condition (the BaCl2 solution with 0.8% checks whether sulfate ion dialyses completely), solution in dialysis membrane is transferred in 25ml measuring bottle, water constant volume, 0～4 DEG C of preservation, obtain blood clam polypeptide test sample.
Get blood clam polypeptide need testing solution 5 μ l, injection liquid chromatography, measures, and records collection of illustrative plates.
In collection of illustrative plates, there is a sharp-pointed characteristic peak " peak S " at retention time 15.323min.
embodiment 2
Chromatographic condition and system suitability test: by Shimadzu-20A high performance liquid chromatograph, UV-detector; Chromatographic column: TSK GEL G4000 PWXL(7.8mm × 300mm); Mobile phase: the phosphate buffer of 0.05 mol/l; 35 DEG C of column temperatures; Flow velocity 0.5ml/min; Detect wavelength 280nm, number of theoretical plate is calculated as 8000 by protein molecular weight 100000.
Prepare blood clam extract: get dry blood clam meat, pulverize, cross 80 mesh sieves, add the water of 10 times of amounts, heating is extracted 2 times, each 1 hour, merge extract, filter, get filtrate, being concentrated into relative density is 1.10～1.15(60 DEG C) time, below 80 DEG C, drying under reduced pressure, pulverizes, and obtains blood clam extract.
Prepare polypeptide: get tablet porphyrize prepared by blood clam extract, take 6g, containing the about 5g of blood clam extract, add the ultrasonic extraction of 50ml water 20min, centrifugal 10min, get upper strata liquid, filter, filtrate is mark 50%(w/v by volume) add ammonium sulfate, 0～4 DEG C of placement is spent the night, centrifugal, get precipitation, the 5ml that adds water dissolves, ammonium bicarbonate soln taking 1.2% is dislysate, the dialysis membrane that is 3500 with molecular cut off, fully dialysis under 0～4 DEG C of condition (the BaCl2 solution with 1% checks whether sulfate ion dialyses completely), solution in dialysis membrane is transferred in 50ml measuring bottle, water constant volume, 0～4 DEG C of preservation, obtain blood clam polypeptide test sample.
Get blood clam polypeptide need testing solution 20 μ l, injection liquid chromatography, measures, and records collection of illustrative plates.
In collection of illustrative plates, there is a sharp-pointed characteristic peak " peak S " at retention time 15.847min.
embodiment 3
Chromatographic condition and system suitability test: by Shimadzu-20A high performance liquid chromatograph, UV-detector; Chromatographic column: TSK GEL G4000 PWXL(7.8mm × 300mm); The phosphate buffer of mobile phase: 0.1mol/l; 40 DEG C of column temperatures; Flow velocity 1ml/min; Detect wavelength 280nm, number of theoretical plate is calculated as 7000 by protein molecular weight 100000.
Get fresh blood clam meat, fragmentation, crosses 40 mesh sieves, adds 1 times of water gaging, extract 4 hours 10 DEG C of following stirrings, and extracting liquid filtering, centrifugal.Get above-mentioned centrifugate freeze-drying, obtain blood clam extract.
Prepare polypeptide: get capsule 's content porphyrize prepared by blood clam extract, take 5.5g, containing the about 5g of blood clam extract, add the 12 times of ultrasonic extraction of water 40min, centrifugal 12min, get upper strata liquid, filter, filtrate is mark 70%(w/v by volume) add ammonium sulfate, 0～4 DEG C of placement is spent the night, centrifugal, get precipitation, the 8ml that adds water dissolves, ammonium bicarbonate soln taking 1.5% is dislysate, the dialysis membrane that is 3500 with molecular cut off, fully dialysis under 0～4 DEG C of condition (the BaCl2 solution with 1.8% checks whether sulfate ion dialyses completely), solution in dialysis membrane is transferred in 100ml measuring bottle, water constant volume, 0～4 DEG C of preservation, obtain blood clam polypeptide test sample.
Get blood clam polypeptide need testing solution 40 μ l, injection liquid chromatography, measures, and records collection of illustrative plates.
In collection of illustrative plates, there is a sharp-pointed characteristic peak " peak S " at retention time 15.652min.
embodiment 4:
According to the blood clam method for preparing extractive index blood clam extract in embodiment 3.
Prepare polypeptide: get granule porphyrize prepared by blood clam extract, take 22g, containing the about 20g of blood clam extract, add the 15 times of ultrasonic extraction of water 30min, centrifugal 15min, get upper strata liquid, filter, filtrate is mark 65%(w/v by volume) add ammonium sulfate, 0～4 DEG C of placement is spent the night, centrifugal, get precipitation, the 6ml that adds water dissolves, ammonium bicarbonate soln taking 2% is dislysate, the dialysis membrane that is 3500 with molecular cut off, fully dialysis under 0～4 DEG C of condition (the BaCl2 solution with 2% checks whether sulfate ion dialyses completely), solution in dialysis membrane is transferred in 100ml measuring bottle, water constant volume, 0～4 DEG C of preservation, obtain blood clam polypeptide test sample.
Get blood clam polypeptide need testing solution 10 μ l, injection liquid chromatography, measures, and records collection of illustrative plates.
In collection of illustrative plates, there is a sharp-pointed characteristic peak " peak S " at retention time 15.247min.
embodiment 5:
Prepare blood clam extract according to the blood clam method for preparing extractive in embodiment 3.
Prepare polypeptide: get pill porphyrize prepared by blood clam extract, take 11g, containing the about 10g of blood clam extract, add the 13 times of ultrasonic extraction of water 1min, centrifugal 25min, get upper strata liquid, filter, filtrate is mark 55%(w/v by volume) add ammonium sulfate, 0～4 DEG C of placement is spent the night, centrifugal, get precipitation, the 7ml that adds water dissolves, ammonium bicarbonate soln taking 0.8% is dislysate, the dialysis membrane that is 3500 with molecular cut off, fully dialysis under 0～4 DEG C of condition (the BaCl2 solution with 0.5% checks whether sulfate ion dialyses completely), solution in dialysis membrane is transferred in 50ml measuring bottle, water constant volume, 0～4 DEG C of preservation, obtain blood clam polypeptide test sample.
Get blood clam polypeptide need testing solution 10 μ l, injection liquid chromatography, measures, and records collection of illustrative plates.
In collection of illustrative plates, there is a sharp-pointed characteristic peak " peak S " at retention time 15.692min.
Claims (8)
1. a discrimination method for blood clam protein and peptide, is characterized in that using gel chromatographic columns high performance liquid chromatography, and concrete steps are as follows:
Chromatographic condition and system flexibility: the gel chromatographic columns that is filling agent with hydrophilic spherical superpolymer; Taking the phosphate buffer of 0.01-0.10mol/L as mobile phase; Detecting wavelength is 280 nm; Column temperature: 20-40 DEG C; Flow velocity: 0.1ml-1ml/min; Number of theoretical plate calculates and should be not less than 3000 by protein molecular weight 100000;
Prepare blood clam polypeptide: get blood clam preparation porphyrize, take the sample containing the about 5-20g of blood clam extract, add the 10-15 times of ultrasonic extraction of water 20-40 min, centrifugal 5-15 min, get upper strata liquid, filter, filtrate is mark 50-70%(w/v by volume) add ammonium sulfate, 0～4 DEG C of placement is spent the night, centrifugal, get precipitation, the 5-10ml that adds water dissolves, taking the ammonium bicarbonate soln of 0.5-2% as dislysate, the dialysis membrane that is 3500 with molecular cut off, fully dialysis under 0～4 DEG C of condition (checking with the BaCl2 solution of 0.5-2% whether sulfate ion dialyses completely), solution in dialysis membrane is transferred in 20-100ml measuring bottle, water constant volume, 0～4 DEG C of preservation, obtain blood clam polypeptide test sample,
Measure: get blood clam polypeptide need testing solution 10-100 μ l, injection liquid chromatography, measures;
In blood clam polypeptide collection of illustrative plates, have a sharp-pointed characteristic peak " peak S " in retention time 15.6min left and right, peak S retention time difference should be within ± 5%.
2. according to the discrimination method of a kind of blood clam polypeptide of claim 1, it is characterized in that chromatographic condition and system flexibility: the gel chromatographic columns that is filling agent with hydrophilic spherical superpolymer; Taking the phosphate buffer of 0.05 mol/L as mobile phase; Detecting wavelength is 280 nm; Column temperature: 25 DEG C; Flow velocity: 0.4ml/min; Number of theoretical plate calculates and should be not less than 5000 by protein molecular weight 100000.
3. according to the discrimination method of a kind of blood clam polypeptide of claim 1, it is characterized in that preparing blood clam polypeptide is: get blood clam preparation porphyrize, take containing the about 10-12g of blood clam extract, add the 10-12 times of ultrasonic extraction of water 30-35 min, centrifugal 8-12 min, get upper strata liquid, filter, filtrate is mark 60-65%(w/v by volume) add ammonium sulfate, 0～4 DEG C of placement is spent the night, centrifugal, get precipitation, the 10ml that adds water dissolves, ammonium bicarbonate soln taking 1% is dislysate, the dialysis membrane that is 3500 with molecular cut off, fully dialysis under 0～4 DEG C of condition (checking with the BaCl2 solution of 0.8-1% whether sulfate ion dialyses completely), solution in dialysis membrane is transferred in 25-50ml measuring bottle, water constant volume, 0～4 DEG C of preservation, obtain blood clam polypeptide test sample.
4. it is characterized in that preparing blood clam polypeptide according to the discrimination method of a kind of blood clam polypeptide of claim 1 or claim 3 is: get blood clam preparation porphyrize, take containing blood clam extract 10 g, add 12 times of ultrasonic extraction 30 min of water, centrifugal 10 min, get upper strata liquid, filter, filtrate is mark 60%(w/v by volume) add ammonium sulfate, 0～4 DEG C of placement is spent the night, centrifugal, get precipitation, the 10ml that adds water dissolves, ammonium bicarbonate soln taking 1% is dislysate, the dialysis membrane that is 3500 with molecular cut off, fully dialysis under 0～4 DEG C of condition (the BaCl2 solution with 1% checks whether sulfate ion dialyses completely), solution in dialysis membrane is transferred in 50ml measuring bottle, water constant volume, 0～4 DEG C of preservation, obtain blood clam polypeptide test sample.
5. according to the discrimination method of a kind of blood clam polypeptide of claim 1, it is characterized in that getting blood clam polypeptide need testing solution 10-50 μ l, injection liquid chromatography, measures.
6. according to the discrimination method of a kind of blood clam polypeptide of claim 1 or claim 6, it is characterized in that getting blood clam polypeptide need testing solution 20 μ l, injection liquid chromatography, measures.
7. according to the discrimination method of a kind of blood clam polysaccharide of claim 1 or claim 2 or claim 3 or claim 4 or claim 5 or claim 6 or claim 7, it is characterized in that the discriminating for blood clam extract and various preparations thereof.
8. according to the discrimination method of a kind of blood clam polypeptide of claim 1 or claim 2 or claim 3 or claim 4 or claim 5 or claim 6 or claim 7, it is characterized in that the discriminating for stalwart blood clam, mud blood clam, flower clam, Scallop Extract and various preparations thereof.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104825496A (en) * | 2015-04-19 | 2015-08-12 | 青岛大学 | Scapharca granosa extract and application thereof in medicine for promoting postpartum uterine contraction |
| CN110618229A (en) * | 2018-06-20 | 2019-12-27 | 成都康弘生物科技有限公司 | Non-reducing peptide map analysis method of protein |
-
2014
- 2014-05-06 CN CN201410187610.7A patent/CN103969367B/en active Active
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104825496A (en) * | 2015-04-19 | 2015-08-12 | 青岛大学 | Scapharca granosa extract and application thereof in medicine for promoting postpartum uterine contraction |
| CN110618229A (en) * | 2018-06-20 | 2019-12-27 | 成都康弘生物科技有限公司 | Non-reducing peptide map analysis method of protein |
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