CN106198783A - A kind of LC-MS detection method of Colla Corii Asini - Google Patents

A kind of LC-MS detection method of Colla Corii Asini Download PDF

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CN106198783A
CN106198783A CN201610490202.8A CN201610490202A CN106198783A CN 106198783 A CN106198783 A CN 106198783A CN 201610490202 A CN201610490202 A CN 201610490202A CN 106198783 A CN106198783 A CN 106198783A
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corii asini
colla corii
solution
acetonitrile
detection method
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国锦琳
叶茂
童芯锌
曹秀君
彭成
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Chengdu University of Traditional Chinese Medicine
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Chengdu University of Traditional Chinese Medicine
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/04—Preparation or injection of sample to be analysed
    • G01N30/06—Preparation
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/04—Preparation or injection of sample to be analysed
    • G01N2030/042—Standards
    • G01N2030/045—Standards internal

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  • Life Sciences & Earth Sciences (AREA)
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Abstract

The invention discloses the LC-MS detection method of a kind of Colla Corii Asini.Each peptides separation that Colla Corii Asini hydrolyzes effectively is detected by the method for the present invention, and by extracting ion m/z 536.7653, m/z 765.9070, m/z 834.9236, can effectively differentiate certified products Colla Corii Asini and adulterant, thus the quality of effective monitoring Colla Corii Asini.

Description

A kind of LC-MS detection method of Colla Corii Asini
Technical field
The present invention relates to the detection method of medical material, be specifically related to the LC-MS detection method of Colla Corii Asini.
Background technology
Colla Corii Asini is the skin of the Chordata Mammalia equine species donkey dry blob of viscose through decocting, concentrating.Medicinal go through Existing more than 2000 year of history, from ancient times to the present, all as nourshing Yin and drynsessmoistening prescription, the good medicine of tonifying blood and arresting bleeding.But, existing market occurs in that many The adulterant made with other animal skins or mixing skin, brings confusion to medical market, therefore, how to differentiate that the true and false of Colla Corii Asini is non- The most important.
Providing the discrimination method of a kind of Colla Corii Asini on Chinese Pharmacopoeia, the method is using the content of several amino acids as Colla Corii Asini Criteria of quality evaluation.But, when Colla Corii Asini comprises the miscellaneous hide glue such as Corii Bovis seu Bubali, Corii Sus domestica, due to its composition, preparation technology and genuine piece pole It is similar, to differentiating to bring the biggest difficulty.
Summary of the invention
For solving the problems referred to above, the invention provides the LC-MS detection method of a kind of Colla Corii Asini, it comprises the following steps:
(1) prepare reference substance solution: take Colla Corii Asini control medicinal material, enzyme hydrolysis, obtain the enzyme hydrolysis solution of Colla Corii Asini, as comparison Product solution；
(2) prepare need testing solution: take Colla Corii Asini test sample, according to the method that step (1) is identical, obtain test sample Enzyme hydrolysis solution, as need testing solution；
(3) respectively reference substance solution and need testing solution are injected HPLC-TOF/MS LC-MS instrument to detect:
Chromatographic condition is as follows:
Chromatographic column: C18 chromatographic column；
Flowing phase: mobile phase A is 0.1% aqueous formic acid, and Mobile phase B is acetonitrile solution；
Condition of gradient elution is:
0～5min, 5% acetonitrile；5～8min, 5%～10% acetonitrile；8～32min, 10%～20% acetonitrile；32～ 38min, 20%～95% acetonitrile；38～38.1min, 95%～5% acetonitrile；
Mass Spectrometry Conditions is as follows:
Ion source: electric spray ion source；
Detection mode: cation reaction monitoring；
(3) the extraction ion flow chromatography figure of test sample with reference substance is compared, i.e. can detect that Colla Corii Asini is for sample The quality condition of product；Described extraction ion flow chromatography figure be charge-mass ratio be the extraction ion flow chromatography of 536.7,765.9 and 834.9 In figure any one or more.
Further, in described chromatographic condition, chromatographic column is Agilent peptide post-C18 chromatographic column, specification be 2.1mm × 100mm, 2.7 μm.
Further, in described chromatographic condition, column temperature is 40 DEG C.
Further, in described Mass Spectrometry Conditions, capillary voltage is 4kV.
Further, in described Mass Spectrometry Conditions, nitrogen drying stream amount is 10.0L/min,
Further, in described Mass Spectrometry Conditions, atomization gas temperature is 350 DEG C.
Further, in described Mass Spectrometry Conditions, atomisation pressure is 40Psi.
Further, in described Mass Spectrometry Conditions, fragmentation voltage is 150V.
Further, the method for described enzyme hydrolysis is as follows:
Take sample powder, add NH4HCO3Solution dissolves, centrifugal, takes supernatant, with the membrane ultrafiltration of 10kDa, takes trapped fluid, Hydrolyze with trypsin solution, after hydrolysis, with the membrane ultrafiltration of 3kDa, take trapped fluid, obtain enzyme hydrolysis solution.
Further, described NH4HCO3The concentration of solution is 1%；The temperature of hydrolysis is 37 DEG C, and the time of hydrolysis is 12h.
Present invention also offers the discrimination method of a kind of Colla Corii Asini, the method be by detection Colla Corii Asini protease hydrolysate in special Property peptide fragment or a combination thereof carry out differentiating, described specificity peptide fragment selected from molecular weight be 1071.5222,1529.7994 or The peptide fragment of 1667.8294, described molecular weight is by the calculated molecular weight of the deconvolution of first mass spectrometric.
Further, the specificity peptide fragment in described detection Colla Corii Asini protease hydrolysate or the method for a combination thereof, before using The LC-MS detection method stated.
Result of the test shows, in the method for the present invention, can effectively be detected by Colla Corii Asini hydrolysis peptides separation, extracts Ion m/z 536.7653, m/z 765.9070, m/z 834.9236 all can effectively differentiate the Colla Corii Asini sample collected in market.And And, the enzymatic hydrolysis process in the present invention, also in terms of obtaining peptide fragment, show good repeatability.
Obviously, according to the foregoing of the present invention, according to ordinary technical knowledge and the customary means of this area, without departing from Under the present invention above-mentioned basic fundamental thought premise, it is also possible to make the amendment of other various ways, replace or change.
The detailed description of the invention of form by the following examples, remakes the most specifically the foregoing of the present invention Bright.But this should not being interpreted as, the scope of the above-mentioned theme of the present invention is only limitted to Examples below.All based on foregoing of the present invention The technology realized belongs to the scope of the present invention.
Accompanying drawing explanation
Fig. 1～4 is followed successively by the chromatogram of gradient elution program 1～4.
Fig. 5～7 is followed successively by the total ion current figure of pig skin gelatinum enzymatic hydrolysate, animal glue enzymolysis, Colla Corii Asini enzymolysis.
Fig. 8 is enzymolysis pig skin gelatinum, enzymolysis animal glue, enzymolysis Colla Corii Asini 13.4min first mass spectrometric figure, wherein, A: enzymolysis Corii Sus domestica Glue；B: enzymolysis animal glue；C: enzymolysis Colla Corii Asini.
Fig. 9 is enzymolysis Colla Corii Asini 13.4min m/z 536.7653 deconvolution figure.
Figure 10 is enzymolysis pig skin gelatinum, enzymolysis animal glue, enzymolysis Colla Corii Asini 27.8min first mass spectrometric figure, wherein, A: enzymolysis Corii Sus domestica Glue；B: enzymolysis animal glue；C: enzymolysis Colla Corii Asini.
Figure 11 is enzymolysis Colla Corii Asini 27.8min m/z 765.9070 deconvolution figure.
Figure 12 is enzymolysis pig skin gelatinum, enzymolysis animal glue, enzymolysis Colla Corii Asini 35.8min first mass spectrometric figure, wherein, A: enzymolysis Corii Sus domestica Glue；B: enzymolysis animal glue；C: enzymolysis Colla Corii Asini.
Figure 13 is enzymolysis Colla Corii Asini 35.8min m/z 834.9236 deconvolution figure.
Figure 14-24 is the total ion current figure of 11 groups of Colla Corii Asini samples.
Figure 25-35 is the chromatogram under 11 groups of Colla Corii Asini sample extraction ion m/z 536.7.
Figure 36-46 is the chromatogram under 11 groups of Colla Corii Asini sample extraction ion m/z 765.9.
Figure 47-57 is the chromatogram under 11 groups of Colla Corii Asini sample extraction ion m/z 834.9.
Detailed description of the invention
Sample: early stage enzymatic hydrolysate, including enzymolysis Corii Sus domestica, enzymolysis Corii Bovis seu Bubali, enzymolysis donkey skin and enzymolysis Donga Colla Corii Asini.
Reagent: acetonitrile, water, methanol fly your (Chinese) company of generation, chromatographically pure purchased from U.S.'s match is silent；Formic acid is purchased from Chengdu section dragon Chemical reagent factory, analytical pure.
Instrument: Agilent 1200 type high performance liquid chromatograph: Agilent company of the U.S. online degasser G1322A type, G1311A type quaternary pump, G315C1260 type DAD detector；LC-MS instrument (HPLC-TOF/MS): 1200SL series HPLC system System associating Agilent 6210A liquid chromatography mass time of-flight mass spectrometer；Agilent SB-C18 chromatographic column (3.0mm × 100mm, 1.8 μm) 1Z2N3L type；Agilent SB-C18 chromatographic column (3.0mm × 100mm, 1.8 μm) 4Z5N6L type； Angilent 300A type C18 chromatographic column (4.6mm × 250mm, 5 μm)；Agilent peptide post-C18 chromatographic column (2.1mm × 100mm,2.7μm)；DP-D501 plain edition freezer dryer (Depew, Wuxi instrument manufacturing company limited)；Tian Jinjin rises GM- 0.20 type oil-free diaphragm vacuum pump.
The detection method of embodiment 1 present invention
Sample preparation condition is: weigh 1g sample powder, adds 40mL 1%NH4HCO3Solution is placed in 50mL conical flask, Putting into after sealing in water-bath with 60 DEG C of heating 30min, take out sample solution after dissolving, 8000r/min is centrifuged 20min, takes Clear liquid is placed in 8000r/min in the super filter tube of 10kDa and is centrifuged 40min by small molecular weight impurity removing.Then above-mentioned centrifuge tube is filtered Protein sample on layer takes out, and adds 1mg/mL trypsin solution (1%NH4HCO3Preparation).Again by above-mentioned solution at 37 DEG C of water-baths Middle sustained response 12h.Draw above-mentioned sample after having reacted respectively in 3kDa super filter tube, be centrifuged 50min with 3000r/min.This Sample can remove unnecessary trypsin, terminates endonuclease reaction.Finally by the filtrate collection to of above-mentioned centrifuge tube bottom In Ep pipe, it is placed in refrigerator 4 DEG C and saves backup.
Mass Spectrometry Conditions: all samples flies in 1200SL Series HPLC System associating Agilent 6210A liquid chromatography mass Carry out under row time mass spectrum (HPLC-TOF/MS), use cation (ESI+) pattern to detect.Capillary voltage (capillary voltage) 4.0kV, it is 10.0L/min that nitrogen is dried gas (Nitrogen drying gas) flow, atomization Temperature (drying gas temperature) is 350 DEG C；Atomisation pressure (Nebulizer) is set to 40Psi；Fragmentation voltage (Fragmentor) it is set to 150V.
LC-MS condition: chromatographic column is Agilent peptide post-C18 chromatographic column (2.1mm × 100mm, 2.7 μm).Select 0.1% aqueous formic acid, as mobile phase A, selects acetonitrile as Mobile phase B.Gradient 0～5min, 5% acetonitrile；5～8min, 5%～10% acetonitrile；8～32min, 10%～20% acetonitrile；32～38min, 20%～95% acetonitrile；38～38.1min, 95%～5% acetonitrile.Flow velocity 0.30mL/min.Column temperature 30 DEG C.Sample size 1.0 μ L.With this understanding collagen protein is carried out liquid Matter combination analysis.
In the case of extraction ion charge-mass ratio (m/z) is 536.7 (double charges), 765.9 (double charges), 834.9 (double charges), If sample presents the obvious chromatographic peak consistent with control medicinal material and miscellaneous peak content is less, then prove certified products ass-hide glue.Otherwise, If in the case of charge-mass ratio (m/z) is 536.7 (double charges), 765.9 (double charges), 834.9 (double charges), sample does not present with right The obvious chromatographic peak consistent according to medical material and miscellaneous peak content are more, then prove adulterant or mix adulterant Colla Corii Asini.
Embodiment 2 condition of gradient elution of the present invention and the screening of diversity peptide fragment
One, the screening of condition of gradient elution
Sample is Colla Corii Asini enzymatic hydrolysate, and chromatographic column selects Angilent 300A type C18 chromatographic column (4.6mm × 250mm, 5 μ M), detection wavelength is 230nm, and using mobile phase A is 0.1% aqueous formic acid, and Mobile phase B is acetonitrile elution samples, gropes liquid Phase condition, the part gradient elution program of screening is as follows.
(1) gradient elution program 1
0～38min, 5%～95% acetonitrile, flow velocity 0.80mL/min；Column temperature 30 DEG C, sample size 10 μ L.Result such as Fig. 1 institute Show.
(2) gradient elution program 2
Mobile phase A is 0.1% aqueous formic acid, and Mobile phase B is acetonitrile.0～20min, 5% acetonitrile；20～30min, 5% ～10% acetonitrile；30～100min, 10%～95% acetonitrile；Flow velocity 0.80mL/min；Column temperature 30 DEG C, sample size 10 μ L.Result As shown in Figure 2.
(2) gradient elution program 3
Mobile phase A is 0.1% aqueous formic acid, and Mobile phase B is acetonitrile.0～20min, 5% acetonitrile；20～30min, 5% ～10% acetonitrile；30～50min, 10%～20% acetonitrile；50～100min, 20%～95% acetonitrile；Flow velocity 0.80mL/min； Column temperature 30 DEG C, sample size 10 μ L.Result is as shown in Figure 3.
(2) gradient elution program 4
Mobile phase A is 0.1% aqueous formic acid, and Mobile phase B is acetonitrile.0～20min, 5% acetonitrile；20～30min, 5% ～10% acetonitrile；30～50min, 10%～20% acetonitrile；50～80min, 20%～50% acetonitrile；80～100min, 50% ～95% acetonitrile；Flow velocity 0.80mL/min；Column temperature 30 DEG C, sample size 10 μ L.Result is as shown in Figure 4.
By comparing, Fig. 4 can present optimal separation effect.Though high performance liquid chromatography can be by polypeptides with donkey hide gelatin base after enzymolysis This separation, but effect is poor, it is impossible to obtain the preferable finger printing of separating degree.But its result is LC-MS technology (HPLC- TOF-MS) separation condition provides basis.Therefore inventor uses LC-MS technology (HPLC-MS) to enter collagen protein further Row separates, to seek diversity peptide fragment.
Two, the screening of diversity peptide fragment
(1) total ion current figure comparison
Select 0.1% aqueous formic acid as mobile phase A, select acetonitrile as Mobile phase B.LC-MS gradient condition For: 0～5min, 5% acetonitrile；5～8min, 5%～10% acetonitrile；8～32min, 10%～20% acetonitrile；32～38min, 20%～95% acetonitrile；38～38.1min, 95%～5% acetonitrile.Flow velocity 0.30mL/min；Column temperature 30 DEG C, sample size 1.0 μ L.
Owing to LC-MS detector have employed fine vacuum, being different from efficient liquid phase, Gradient program the most here is at sieve It is optimized on the basis of the Gradient program 4 that choosing obtains and obtains.
With this understanding pig skin gelatinum enzymatic hydrolysate, animal glue enzymatic hydrolysate, Colla Corii Asini enzymatic hydrolysate are carried out LC-MS and divide Analysis, obtains total ion current figure the most as illustrated in figs. 5-7.
As seen from the figure, different skin class collagen protein enzymatic hydrolysate LC-MS total ion current figures are the most similar.This is mainly Being caused by the homology of collagen protein, quantity proportion in collagen sequences with characteristic peptide fragment is little, need to enter One step amplifies difference.
(2) first mass spectrometric comparison
Choose in three groups of samples containing the time point of differential fragment: 13.4min, 27.8min, 35.8min, in the case of taking this Sample carry out first mass spectrometric contrast, below figure.Choose high response value specificity mass-to-charge ratio (m/z) fragments of peptides in Colla Corii Asini and do uncoiling Long-pending calculating, then comparison early stage bioinformatics software preanalysis result.
Enzymolysis pig skin gelatinum, enzymolysis animal glue, enzymolysis Colla Corii Asini 13.4min first mass spectrometric figure are as shown in Figure 8.Choose now enzymolysis In Colla Corii Asini, high response value specificity m/z 536.7653 does deconvolution calculating, and result is as shown in Figure 9.
Enzymolysis pig skin gelatinum, enzymolysis animal glue, enzymolysis Colla Corii Asini 27.8min first mass spectrometric figure are as shown in Figure 10.Choose now enzyme Solving high response value specificity m/z 765.9070 in Colla Corii Asini and do deconvolution calculating, result is as shown in figure 11.
Enzymolysis pig skin gelatinum, enzymolysis animal glue, enzymolysis Colla Corii Asini 35.8min first mass spectrometric figure are as shown in figure 12.Choose now enzyme Solving high response value specificity m/z 834.9236 in Colla Corii Asini and do deconvolution calculating, result is as shown in figure 13.
The comparison result of above-mentioned first mass spectrometric figure shows, pig skin gelatinum after enzymolysis, animal glue, Colla Corii Asini total ion current figure and Similar, only exist small variations.In the Colla Corii Asini that there are differences specificity peptide fragment mass-to-charge ratio be 536.7653,765.9070, 834.9236, do deconvolution calculating and wait until that specific peptide segment molecule amount is 1071.5222,1529.7994,1667.8294.
Therefore, the big peptide fragment of aforementioned molecular weight as identifying that Colla Corii Asini is true and false or can mix the index composition of puppet.Deconvolution Result is as shown in table 1 below.
Table 1
Simulating enzymolysis with bioinformatics software Peptide Cutter, ExPASy PeptideMass software system is carried out Pancreatin enzyme action, in the present invention, only molecular weight is 1071.5222 close to bioinformatics in donkey skin I-type collagen alpha-1 chain Software enzymolysis specificity peptide fragment, can do different glue class source Collagenase from other 2 molecular weight simultaneously as appraisal basis Hydrolysis products is verified, preferentially chooses specific molecular weight.
Three, confirmatory experiment
Take off the medical material stating 11 groups of different batches, detect according to the method for the present invention
A:1 Donga Colla Corii Asini；B:2 good fortune glue；C:3 stone peak Colla Corii Asini；D:4 stone peak Colla Corii Asini；E:5 Tongrentang Colla Corii Asini； F:6 field rain Colla Corii Asini；G:7 field rain Colla Corii Asini；H:8 A Gu Colla Corii Asini；I:9 A Gu Colla Corii Asini；J:10 tribute true hall Colla Corii Asini；K:11 Number tribute true hall Colla Corii Asini.
Figure 14-24 is the total ion current figure of aforementioned sample.
Figure 25-35 is to extract the chromatogram under ion m/z 536.7.
Figure 36-46 is to extract the chromatogram under ion m/z 765.9.
Figure 47-57 is to extract the chromatogram under ion m/z 834.9.
Empirical tests, extract that ion m/z 536.7, m/z 765.9, m/z 834.9 can effectively differentiate to collect 11 groups Ah Glue sample, extraction ion m/z 536.7, m/z 765.9, m/z 834.9 can specifically differentiate Colla Corii Asini.
In sum, each peptides separation of Colla Corii Asini is effectively detected by the method for the present invention, and by extracting ion M/z 536.7653, m/z 765.9070, m/z 834.9236, can effectively differentiate certified products Colla Corii Asini and adulterant, thus effectively supervise The quality of control Colla Corii Asini.Meanwhile, the enzymatic hydrolysis process in the present invention, also in terms of obtaining peptide fragment, show good repeatability.

Claims (10)

1. the LC-MS detection method of a Colla Corii Asini, it is characterised in that: it comprises the following steps:
(1) prepare reference substance solution: take Colla Corii Asini control medicinal material, enzyme hydrolysis, obtain the enzyme hydrolysis solution of Colla Corii Asini, molten as reference substance Liquid；
(2) prepare need testing solution: take Colla Corii Asini test sample, according to the method that step (1) is identical, obtain the enzyme water of test sample Solve solution, as need testing solution；
(3) respectively reference substance solution and need testing solution are injected HPLC-TOF/MS LC-MS instrument to detect:
Chromatographic condition is as follows:
Chromatographic column: C18 chromatographic column；
Flowing phase: mobile phase A is 0.1% aqueous formic acid, and Mobile phase B is acetonitrile solution；
Condition of gradient elution is:
0～5min, 5% acetonitrile；5～8min, 5%～10% acetonitrile；8～32min, 10%～20% acetonitrile；32～38min, 20%～95% acetonitrile；38～38.1min, 95%～5% acetonitrile；
Mass Spectrometry Conditions is as follows:
Ion source: electric spray ion source；
Detection mode: cation reaction monitoring；
(4) the extraction ion flow chromatography figure of test sample with reference substance is compared, i.e. can detect that Colla Corii Asini test sample Quality condition；Described extraction ion flow chromatography figure is in the extraction ion flow chromatography figure that charge-mass ratio is 536.7,765.9 and 834.9 Any one or more.
Detection method the most according to claim 1, it is characterised in that: in described chromatographic condition, chromatographic column is Agilent peptide Post-C18 chromatographic column, specification is 2.1mm × 100mm, 2.7 μm.
Detection method the most according to claim 2, it is characterised in that: in described chromatographic condition, column temperature is 40 DEG C.
4. according to the detection method described in any one of claim 1-3, it is characterised in that: in described Mass Spectrometry Conditions, capillary tube electricity Pressure is 4kV.
Detection method the most according to claim 4, it is characterised in that: in described Mass Spectrometry Conditions, nitrogen drying stream amount is 10.0L/min, atomization gas temperature is 350 DEG C, and atomisation pressure is 40Psi.
Detection method the most according to claim 5, it is characterised in that: in described Mass Spectrometry Conditions, fragmentation voltage is 150V.
7. according to the detection method described in any one of claim 1-6, it is characterised in that: the method for described enzyme hydrolysis is as follows, takes Sample powder, adds NH4HCO3Solution dissolves, centrifugal, takes supernatant, with the membrane ultrafiltration of 10kDa, takes trapped fluid, with trypsin solution Hydrolysis, after hydrolysis, with the membrane ultrafiltration of 3kDa, takes filtered solution, obtains enzyme hydrolysis solution.
Detection method the most according to claim 7, it is characterised in that: described NH4HCO3The concentration of solution is 1%；Hydrolysis Temperature is 37 DEG C, and the time of hydrolysis is 12h.
9. the discrimination method of a Colla Corii Asini, it is characterised in that: the method is by the specific peptide in detection Colla Corii Asini protease hydrolysate Section or a combination thereof carry out differentiating, described specificity peptide fragment selected from molecular weight be 1071.5222,1529.7994 or The peptide fragment of 1667.8294, described molecular weight is by the calculated molecular weight of the deconvolution of first mass spectrometric.
Discrimination method the most according to claim 9, it is characterised in that: the specificity in described detection Colla Corii Asini protease hydrolysate Peptide fragment or the method for a combination thereof, use the LC-MS detection method described in any one of claim 1-8.
CN201610490202.8A 2016-06-27 2016-06-27 A kind of LC-MS detection method of Colla Corii Asini Pending CN106198783A (en)

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CN106589063A (en) * 2016-12-13 2017-04-26 山东省食品药品检验研究院 Donkey-source characteristic peptide group and application process thereof in qualitative detection of donkey skin and donkey-hide gelatin
WO2022028371A1 (en) * 2020-08-04 2022-02-10 Hong Kong Baptist University Peptide markers for authenticating ejiao and related gelatins
CN115015409A (en) * 2022-05-23 2022-09-06 山东省食品药品检验研究院 Method for establishing LCMS (liquid Crystal display System) characteristic spectrum of donkey-hide gelatin polypeptide and quality evaluation method thereof

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CN106589114A (en) * 2016-12-13 2017-04-26 山东省食品药品检验研究院 Porcine-derived characteristic peptide and application processes thereof in pigskin and colla corii asini qualitative detection
CN106589063A (en) * 2016-12-13 2017-04-26 山东省食品药品检验研究院 Donkey-source characteristic peptide group and application process thereof in qualitative detection of donkey skin and donkey-hide gelatin
CN106589063B (en) * 2016-12-13 2020-06-19 山东省食品药品检验研究院 Donkey-derived characteristic peptides and application process thereof in qualitative detection of donkey skin and donkey-hide gelatin
WO2022028371A1 (en) * 2020-08-04 2022-02-10 Hong Kong Baptist University Peptide markers for authenticating ejiao and related gelatins
CN115015409A (en) * 2022-05-23 2022-09-06 山东省食品药品检验研究院 Method for establishing LCMS (liquid Crystal display System) characteristic spectrum of donkey-hide gelatin polypeptide and quality evaluation method thereof

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