CN103630620A - Method for detecting deer-derived ingredients in glue type traditional Chinese medicines and products thereof - Google Patents

Method for detecting deer-derived ingredients in glue type traditional Chinese medicines and products thereof Download PDF

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CN103630620A
CN103630620A CN201310516759.0A CN201310516759A CN103630620A CN 103630620 A CN103630620 A CN 103630620A CN 201310516759 A CN201310516759 A CN 201310516759A CN 103630620 A CN103630620 A CN 103630620A
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deer
glue
amino acid
chinese medicine
class chinese
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CN103630620B (en
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周祥山
秦玉峰
尤金花
田守生
嵇传良
郭尚伟
段小波
史兆松
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Shandong Dong E E Jiao Co Ltd
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Abstract

The invention provides a method for detecting deer-derived ingredients in glue type traditional Chinese medicines and products thereof. The method provided by the invention comprises the step of detecting by using the difference between the specific nucleotide sequence or amino acid sequence of a genome of a red deer and the specific nucleotide sequence or amino acid sequence of a genome of a spotted deer, wherein the specific amino acid sequence is shown in SEQ ID No. 1. By adopting the method provided by the invention, whether the glue type traditional Chinese medicines contain the deer-derived ingredients or not can be detected rapidly, so as to distinguish the authenticity; although polypeptides of collagen in the glue type traditional Chinese medicines are subjected to certain hydrolysis and destruction, the deer-derived ingredients in the glue type traditional Chinese medicines and products thereof can be identified on the basis that differential polypeptides in a main ingredient, namely collagen, of the glue type traditional Chinese medicines are high in content and little in damaged extent.

Description

The detection method of deer derived component in a kind of glue class Chinese medicine and goods thereof
Technical field
The present invention relates to a kind of detection method of deer derived component, be specifically related to the detection method of deer derived component in a kind of glue class Chinese medicine and goods thereof, belong to Chinese medicine detection field.
Background technology
Deer horn, ossified angle or the fine and soft angle base coming off spring in next year afterwards of saw of animal in deer family red deer (Cervus elaphus Linnaeus) or sika deer (Cervus nipponTemminck), belongs to traditional rare Chinese medicine.According to the Chinese Pharmacopoeia of latest edition, record, taking deer horn can warm kidney yang, strengthening the bones and muscles, promoting circulation of blood detumescence.The deer horn glue that the deer horn of take is made as raw material on whole Market of Chinese Materia Medica in occupation of important position.
According to version Chinese Pharmacopoeia in 2010, record, deer horn glue is to take deer horn as raw material, through decocting, boils, concentrates the solid gum of making, and property is sweet, salty, warm, returns kidney, Liver Channel.Can warm filling liver kidney, beneficial intensive culture blood, the waist knee acid being mainly used in due to kidney deficiency and liver is cold, impotence and seminal emission, consumptive disease is thin thin, metrostaxis, the hematuria of having blood in stool, cloudy subcutaneous ulcer swells and ache.Along with improving constantly of living standard, similar with donkey-hide gelatin, deer horn glue also becomes one of first-selection that people nourish.
Because deer horn self is famousr and precious, medical value is remarkable, and venison is also usually edible for people for cook various kinds tonic, and the quantity of animal in deer family is reducing, and sika deer is classified country-level focused protection animal as, and red deer is classified national secondary as and lays special stress on protecting animal.Therefore, the supply of deer horn raw material is very limited, and price is high, has also directly had influence on the production of deer horn glue.On deer horn glue market, have some counterfeit and shoddy goods, it makes raw material is not deer horn, but is replaced by the assorted skin of other animals, broken bone, comprises pigskin, ox-hide, the skin of animals died of illness even, dirty skin, rotten skin etc.Once such counterfeit and shoddy goods are shaped, outward appearance, color and luster, smell all to certified products deer horn glue and similar, true and false difficulty is distinguished.Take these adulterants not only without any curative effect, also probably harmful to health, consequence is very serious.
Glue class Chinese medicine forms gelatin substance by skin, bone, first-class the boiling of animal, and comprises donkey-hide gelatin, colla carapacis et plastri testudinis, deer horn glue etc., has the traditional Chinese medicine of typical national characters.Glue class Chinese medicine is general to be concentrated and forms through long-time thermophilic digestion, principal ingredient production technology very close and different manufacturers there are differences, and adopts the methods such as infrared, near infrared, X-diffraction to carry out the true and false to glue class Chinese medicine sterling and differentiates the normal situations such as judgement is inaccurate that exist.The discriminating of deer derived component in glue class Chinese medicine, existing scholar utilizes mass spectrometry to detect deer derived component glue class Chinese medicine from the angle of polypeptide, and result is accurately and reliably.At present, also not to the method for tracing to the source in the goods of glue class Chinese medicine and identifying.
In glue class Chinese medicine, principal ingredient is collagen polypeptide, the collagen, amino acid sequence of different animals there are differences, the otherness polypeptide of same species stable existence can be used as the characteristic polypeptide of these species, by digestion with restriction enzyme, utilize mass spectrum completely likely accurately glue class Chinese medicine and goods thereof to be traced to the source and identified and content detection.
Summary of the invention
The present invention is directed to the problems referred to above, the detection method of deer derived component in a kind of glue class Chinese medicine and goods thereof is provided, the method is simple to operate, and characteristic is strong, highly sensitive, can be used for the evaluation of deer derived component in glue class Chinese medicine and goods thereof.
Technical scheme of the present invention is as follows:
First, the invention provides the detection method of deer derived component in a kind of glue class Chinese medicine and goods thereof, it utilizes the difference in red deer and sika deer genome with specificity nucleotide sequence or amino acid sequence to detect, and described specific amino acid is as shown in SEQ.ID No1.
Preferably, described glue class Chinese medicine is selected from donkey-hide gelatin, oxhide gelatin, colla carapacis et plastri testudinis or deer horn glue.
Preferably, the described choosing of glue class Chinese herbal product food, health products or medicine that freely corresponding glue class Chinese medicine is made.
Particularly, the method comprises the steps:
(1) choose specific amino acid or amino acid sequence in red deer and sika deer genome, described specific amino acid is as shown in SEQ.ID No1;
(2) glue class Chinese medicine or its goods are dissolved, or extract polypeptide constituents dissolving wherein, add restriction enzyme to carry out enzyme and cut;
(3) put into subsequently LC-MS instrument, take negative sample to be detected as matrix, add step (1) gained amino acid sequence or deer horn glue sterling in contrast, select parent ion m/z435.0 and the daughter ion thereof of this polypeptide to monitor.
If it is consistent with reference substance to detect the retention time of this ion, and its daughter ion is consistent with the daughter ion of reference substance, and described glue class Chinese medicine or its goods contain deer derived component; If this ion consistent with reference substance retention time, does not contain deer derived component.
Preferably, described in step (2), dissolving reagent used is that mass percent is 1%, the NH of pH value 8.0 4hCO 3solution; Described restriction enzyme is trypsase.
Adopt method of the present invention, can in fast detecting glue class Chinese medicine, whether contain deer derived component, thereby minute evident, although collagen polypeptide has certain hydrolysis and destruction in glue class Chinese medicine, but the present invention is based on the otherness polypeptide in its principal ingredient collagen, its content is high, and the degree that is damaged is very little, can identify the deer derived component in glue class Chinese medicine and goods thereof.
The invention still further relates to polypeptide, its amino acid sequence is Gly-Glu-Ala-Gly-Pro-Gln-Gly-Pro-Arg, and the composition being comprised of described polypeptide and respective carrier.In addition, also comprise the application in deer derived component in detecting glue class Chinese medicine and goods thereof of described polypeptide and composition thereof.
Accompanying drawing explanation
Fig. 1 selects the mass spectrogram of ion pair m/z435.0 → 319.2,554.3,611.3 monitorings under synthetic polypeptide and sterling glue multiple-reaction monitoring scan pattern;
Fig. 2 selects the mass spectrogram of ion pair m/z435.0 → 319.2,554.3,611.3 monitorings under synthetic polypeptide and the epoxy glue multiple-reaction monitoring scan pattern that adds 5% deer horn glue;
Fig. 3 is the mass spectrogram that synthetic polypeptide and deer horn glue goods (the celestial oral liquid of tortoise deer two of take is example) are selected ion pair m/z435.0 → 319.2,554.3,611.3 monitorings under multiple-reaction monitoring scan pattern.
Embodiment
Below in conjunction with specific embodiment, further describe the present invention, advantage and disadvantage of the present invention will be more clear along with description.But embodiment is only exemplary, scope of the present invention is not formed to any restriction.It will be understood by those skilled in the art that lower without departing from the spirit and scope of the present invention and can the details of technical solution of the present invention and form be modified or be replaced, but these modifications and replacement all fall within the scope of protection of the present invention.
The acquisition of specificity peptide chain in the present invention:
1 material and reagent
Material: sterling glue comprises donkey-hide gelatin, horse skin glue, oxhide gelatin, pig skin gelatin, colla carapacis et plastri testudinis, deer horn glue, decocts and obtains with donkey hide, horse skin, ox-hide, pigskin, tortoise plastron, deer horn respectively.
Reagent: ammonium bicarbonate (analyzing pure), trypsase (sequence is pure, purchased from National Institute for Food and Drugs Control), synthetic polypeptide Gly-Glu-Ala-Gly-Pro-Gln-Gly-Pro-Arg.
The detection pre-treatment of 2 samples
(1) preparation of glue sample enzymolysis solution
Get 1.0g glue sample to be measured in 500mL measuring bottle, add a small amount of 1%NH 4hCO 3solution, the ultrasonic sample that makes dissolves completely, uses 1%NH 4hCO 3solution (pH8.0) is diluted to scale, shakes up, and filtering with microporous membrane, precision measures the trypsin solution 100 μ L that subsequent filtrate 1mL adds 2mg/mL, 37 ℃ of constant temperature enzymolysis 6h.
(2) preparation of synthetic polypeptide control sample enzymolysis solution
Take a certain amount of synthetic polypeptide, add 1%NH 4hCO 3solution (pH8.0), shakes up, and be mixed with concentration and be about 0.3 μ g/mL, filtering with microporous membrane, standby.
The discovery of 3 polypeptide
(1) discovery of ion
Get 5 μ L enzymolysis solution and put into the detection of LC-MS instrument.Liquid-phase condition: C 18reverse-phase chromatographic column (2.1mm * 100mm, 1.8 μ m), mobile phase A is 0.1% formic acid solution [volume fraction], Mobile phase B is acetonitrile, flow velocity 0.3mL/min; Gradient elution: 0~40min, 2%~50% Mobile phase B (the percentage here represents from 0 minute to the 40th minute, flow B by 2% linear gradient be 50%, mobile phase A fades to 50% by 98%).Mass spectrum condition: electron spray positive ion mode (ESI +) carry out one-level and entirely sweep, sweep limit m/z400-m/z1000.
From the mass spectrum of glue sample separately, entirely sweep figure and extract ion m/z435.0, by contrast, find to contain this quasi-molecular ions in 4.42min left and right only has deer horn glue, and all do not have in donkey-hide gelatin, horse skin glue, oxhide gelatin, pig skin gelatin and colla carapacis et plastri testudinis.Detection and checking by multiple batches of sample show that above-mentioned conclusion is correct.Illustrate: can, by detecting the ion m/z435.0 in deer horn glue, determine whether to contain deer derived component.
(2) amino acid sequence of polypeptide is tentatively inferred
Utilize triple quadrupole bar mass spectrum, the above-mentioned ion of finding out is carried out to daughter ion scanning, confirm this ion band double charge, by sequence assembly, infer the partial sequence that polypeptide.Partial sequence by inference, at NCBI(http: //www.ncbi.nlm.nih.gov/) database carries out sequence retrieval, feature in conjunction with it (has in deer, and all do not have in donkey, horse, ox, pig or tortoise, and be about 869.0 by molecular weight after tryptic digestion), find out the possible sequence Gly-Glu-Ala-Gly-Pro-Gln-Gly-Pro-Arg of this polypeptide.According to the CID fracture theory of peptide chain, calculate the possible daughter ion of this polypeptide as follows:
a b c" Res: x y″ z
30.0 58.0 77.1 Gly - - -
159.1 187.1 206.1 Glu 835.4 811.4 792.4
230.1 258.1 277.2 Ala 706.3 682.4 663.3
287.1 315.1 334.2 Gly 635.3 611.3 592.3
384.2 412.2 431.2 Pro 578.3 554.3 535.3
512.2 540.2 559.3 Gln 481.2 457.3 438.2
569.3 597.3 616.3 Gly 353.2 329.2 310.2
666.3 694.3 713.4 Pro 296.1 272.2 253.1
- - - Arg 199.1 175.1 156.1
Can entirely sweep coincideing in figure with the daughter ion of peculiar ion m/z435.0 in deer horn glue.
(3) sequence is determined
Amino acid sequence by inference, entrusts polypeptide Synesis Company (the biochemical company limited of gill) to carry out the synthetic of this polypeptide.Then the daughter ion that utilizes LC-MS instrument to synthesize polypeptide is swept entirely, and in its testing conditions and glue sample, the daughter ion of corresponding polypeptide is swept term harmonization entirely.By contrasting ion appearance time, the daughter ion of synthetic polypeptide and deer horn glue, entirely sweep figure, find that both fit like a glove, thereby determine that this ion m/z435.0 is polypeptide Gly-Glu-Ala-Gly-Pro-Gln-Gly-Pro-Arg.
Embodiment 1
1 material and reagent
Material: sterling glue comprises donkey-hide gelatin, horse skin glue, oxhide gelatin, pig skin gelatin, colla carapacis et plastri testudinis, deer horn glue, decocts and obtains with donkey hide, horse skin, ox-hide, pigskin, tortoise plastron, deer horn respectively.
Epoxy glue sample (containing the donkey-hide gelatin of 5% deer horn glue, the donkey-hide gelatin of the donkey-hide gelatin of 10% deer horn glue, 20% deer horn glue, the donkey-hide gelatin of 40% deer horn glue) by above-mentioned sterling glue, be accurately mixed.(percentage is mass percent)
Reagent: ammonium bicarbonate (analyzing pure), trypsase (sequence is pure, purchased from National Institute for Food and Drugs Control), synthetic polypeptide Gly-Glu-Ala-Gly-Pro-Gln-Gly-Pro-Arg.
2 detection methods
(1) preparation of synthetic polypeptide control sample enzymolysis solution
Get 1.0g sterling donkey-hide gelatin sample in 500mL measuring bottle, add a small amount of 1%NH 4hCO 3solution, the ultrasonic sample that makes dissolves completely, uses 1%NH 4hCO 3solution (pH8.0) is diluted to scale, shakes up, and filtering with microporous membrane, precision measures subsequent filtrate 1mL and adds synthetic polypeptide 0.3 μ g, then adds the trypsin solution 100 μ L of 2mg/mL, 37 ℃ of constant temperature enzymolysis 6h.
(2) preparation of glue sample enzymolysis solution
Get 1.0g glue sample to be measured in 500mL measuring bottle, add a small amount of 1%NH 4hCO 3solution, the ultrasonic sample that makes dissolves completely, uses 1%NH 4hCO 3solution (pH8.0) is diluted to scale, shakes up, and filtering with microporous membrane, precision measures the trypsin solution 100 μ L that subsequent filtrate 1mL adds 2mg/mL, 37 ℃ of constant temperature enzymolysis 6h.
(3) detect
Get 5 μ L enzymolysis solution and put into the detection of LC-MS instrument.Liquid-phase condition: C 18reverse-phase chromatographic column (2.1mm * 100mm, 1.8 μ m), mobile phase A is 0.1% formic acid solution [volume fraction], Mobile phase B is acetonitrile, flow velocity 0.3mL/min; Gradient elution: 0~40min, 2%~50% Mobile phase B (the percentage here represents from 0 minute to the 40th minute, flow B by 2% linear gradient be 50%, mobile phase A fades to 50% by 98%).Mass spectrum condition: electron spray positive ion mode (ESI +) select to carry out many reaction detection, select m/z435.0 → 319.2,554.3,611.3 as detecting ion pair.
The results are shown in Figure Isosorbide-5-Nitrae .42min place and only have in synthetic polypeptide control sample and deer horn glue and detect corresponding quasi-molecular ions, other all do not detect.The percentage composition of deer horn glue of take is horizontal ordinate, and take m/z435.0 → 554.3, to extract chromatographic peak area in ion flow graph be ordinate, drawing standard curve, R 2be more than 0.999, linear good.Visible this method can specific detection deer derived component, comprises that with other various animal derived materials donkey, horse, ox, pig, tortoise etc. distinguish.
Embodiment 2
1 material and reagent
Material: the sterling glue sample of epoxy glue sample in embodiment 1 adds respectively 5% deer horn glue to make, comprise containing the donkey-hide gelatin of 5% deer horn glue, the horse skin glue of 5% deer horn glue, containing the oxhide gelatin of 5% deer horn glue, the pig skin gelatin of 5% deer horn glue, containing the colla carapacis et plastri testudinis of 5% deer horn glue.(percentage is massfraction)
Reagent: ammonium bicarbonate (analyzing pure), trypsase (sequence is pure, purchased from National Institute for Food and Drugs Control), synthetic polypeptide Gly-Glu-Ala-Gly-Pro-Gln-Gly-Pro-Arg.
2 detection methods
(1) preparation of synthetic polypeptide control sample enzymolysis solution
Get 1.0g sterling donkey-hide gelatin to be measured sample in 500mL measuring bottle, add a small amount of 1%NH 4hCO 3solution, the ultrasonic sample that makes dissolves completely, uses 1%NH 4hCO 3solution (pH8.0) is diluted to scale, shakes up, and filtering with microporous membrane, precision measures subsequent filtrate 1mL and adds synthetic polypeptide 0.3 μ g, then adds the trypsin solution 100 μ L of 2mg/mL, 37 ℃ of constant temperature enzymolysis 6h.
(2) preparation of glue sample enzymolysis solution
Get 1.0g testing sample in 500mL measuring bottle, add a small amount of 1%NH 4hCO 3solution, the ultrasonic sample that makes dissolves completely, uses 1%NH 4hCO 3solution (pH8.0) is diluted to scale, shakes up, and filtering with microporous membrane, precision measures the trypsin solution 100 μ L that subsequent filtrate 1mL adds 2mg/mL, 37 ℃ of constant temperature enzymolysis 6h.
(3) detect
Get 5 μ L enzymolysis solution and put into the detection of LC-MS instrument.Liquid-phase condition: C 18reverse-phase chromatographic column (2.1mm * 100mm, 1.8 μ m), mobile phase A is 0.1% formic acid solution [volume fraction], Mobile phase B is acetonitrile, flow velocity 0.3mL/min; Gradient elution: 0~40min, 2%~50%B(percentage here represents from 0 minute to the 40th minute, flow B by 2% linear gradient be 50%, mobile phase A fades to 50% by 98%).Mass spectrum condition: electron spray positive ion mode (ESI +) select to carry out many reaction detection, select m/z435.0 → 319.2,554.3,611.3 as detecting ion pair.
The results are shown in Figure 2,4.42min place synthesizes polypeptide control sample, add in the epoxy glue sample of 5% deer horn glue and all detect corresponding quasi-molecular ions, and all do not detect corresponding quasi-molecular ions in sterling donkey-hide gelatin in case study on implementation 1, horse skin glue, oxhide gelatin, pig skin gelatin, colla carapacis et plastri testudinis.The deer derived component of visible this method in can specific detection glue class Chinese medicine.
Embodiment 3
1 material and reagent
Material: deer horn glue, the celestial oral liquid of tortoise deer two (Dong-E donkey-hide Gelatin Co., Ltd., Shandong Prov.), not containing the celestial oral liquid of tortoise deer two (self-control) of deer horn glue
Reagent: trichloroacetic acid (analyzing pure), ammonium bicarbonate (analyzing pure), trypsase (sequence is pure, purchased from National Institute for Food and Drugs Control), synthetic polypeptide Gly-Glu-Ala-Gly-Pro-Gln-Gly-Pro-Arg.
2 detection methods
(1) preparation of synthetic polypeptide control sample enzymolysis solution
Get 1.0g and containing the celestial oral liquid sample of tortoise deer two of deer horn glue, do not add 1%NH 4hCO 3solution 24mL dissolves, and adds 4g trichloroacetic acid, places 10min for 4 ℃, and the centrifugal 5min of 10000rpm, abandons supernatant, and sediment is proceeded in 100mL measuring bottle, uses 1%NH 4hCO 3solution (pH8.0) dissolves and is settled to scale, shakes up, and filtering with microporous membrane, precision measures subsequent filtrate 1mL and adds synthetic polypeptide 0.3 μ g, then adds the trypsin solution 100 μ L of 2mg/mL, 37 ℃ of constant temperature enzymolysis 6h.
(2) preparation of testing sample enzymolysis solution
Get 1.0g testing sample and add 1%NH 4hCO 3solution 25mL dissolves, and adds 4g trichloroacetic acid, places 10min for 4 ℃, and the centrifugal 5min of 10000rpm, abandons supernatant, and sediment is proceeded in 100mL measuring bottle, uses 1%NH 4hCO 3solution (pH8.0) dissolves and is settled to scale, shakes up, and filtering with microporous membrane, precision measures the trypsin solution 100 μ L that subsequent filtrate 1mL adds 2mg/mL, 37 ℃ of constant temperature enzymolysis 6h.
(3) detect
Get 5 μ L enzymolysis solution and put into the detection of LC-MS instrument.Liquid-phase condition: C 18reverse-phase chromatographic column (2.1mm * 100mm, 1.8 μ m), mobile phase A is 0.1% formic acid solution [volume fraction], Mobile phase B is acetonitrile, flow velocity 0.3mL/min; Gradient elution: 0~40min, 2%~50%B(percentage here represents from 0 minute to the 40th minute, flow B by 2% linear gradient be 50%, mobile phase A fades to 50% by 98%).Mass spectrum condition: electron spray positive ion mode (ESI +) carry out many reaction detection, select m/z435.0 → 319.2,554.3,611.3 as detecting ion pair.
The results are shown in Figure 3, the celestial oral liquid of tortoise deer two that 4.42min place synthesizes polypeptide control sample, deer horn glue and contains deer horn glue detects corresponding quasi-molecular ions, other do not detect, the deer derived component of visible this method in can specific detection deer horn glue goods, and comprise that with other various animal derived materials donkey, horse, ox, pig, tortoise etc. distinguish.

Claims (10)

1. a detection method for deer derived component in glue class Chinese medicine and goods thereof, it utilizes the difference in red deer and sika deer genome with specificity nucleotide sequence or amino acid sequence to detect, and described specific amino acid is as shown in SEQ.ID No1.
2. method according to claim 1, is characterized in that, described glue class Chinese medicine is selected from donkey-hide gelatin, oxhide gelatin, colla carapacis et plastri testudinis or deer horn glue.
3. method according to claim 1, is characterized in that, the described choosing of glue class Chinese herbal product food, health products or medicine that freely corresponding glue class Chinese medicine is made.
4. according to the method described in claim 1~3 any one, it specifically comprises the steps:
(1) choose specific amino acid or amino acid sequence in red deer and sika deer genome, described specific amino acid is as shown in SEQ.ID No1;
(2) glue class Chinese medicine or its goods are dissolved, or extract polypeptide constituents dissolving wherein, add restriction enzyme to carry out enzyme and cut;
(3) put into subsequently LC-MS instrument, take negative sample to be detected as matrix, add step (1) gained amino acid sequence or deer horn glue sterling in contrast, select parent ion m/z435.0 and the daughter ion thereof of this polypeptide to monitor.
5. method according to claim 4, is characterized in that, dissolves reagent used and be 1%, the NH of pH value 8.0 described in step (2) 4hCO 3solution; Described restriction enzyme is trypsase.
6. detect a molecular marked compound for deer derived component in glue class Chinese medicine and goods thereof, the amino acid sequence of this molecular marked compound is as shown in SEQ.ID No1.
Coding albumen claimed in claim 6 gene.
8. the application of molecular marked compound claimed in claim 6 deer derived component in detecting glue class Chinese medicine and goods thereof.
9. a pharmaceutical composition, is characterized in that, the albumen claimed in claim 6 that comprises pharmacy effective dose and one or more pharmaceutically acceptable carriers.
10. the application of pharmaceutical composition claimed in claim 9 deer derived component in detecting glue class Chinese medicine and goods thereof.
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Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104360002A (en) * 2014-09-12 2015-02-18 江西师范大学 Quantitative detection method for cow leather gelatin
CN105301165A (en) * 2015-10-09 2016-02-03 东阿阿胶股份有限公司 Donkey characteristic polypeptide and application thereof to detection on donkey skin derived ingredients
CN106645479A (en) * 2016-12-22 2017-05-10 东阿阿胶股份有限公司 Composition, kit and detection method for detecting content of dog bone source ingredients in glue type traditional Chinese medicines and compound preparations thereof
CN106841496A (en) * 2017-01-04 2017-06-13 东阿阿胶股份有限公司 A kind of composition for detecting dog bone derived components content in glue class Chinese medicine and its compound preparation, kit and its detection method
CN109187783A (en) * 2018-09-03 2019-01-11 贵州广济堂药业有限公司 Deer horn glue feature peptide and identification sample to be tested in whether include deer horn glue method
CN110530999A (en) * 2019-09-18 2019-12-03 山东省食品药品检验研究院 A kind of distinguishing method between true and false of caribou horn
CN111060622A (en) * 2019-12-26 2020-04-24 北京化工大学 Labeled peptide of tuna-derived component and application of labeled peptide in detection of collagen degradation product and product thereof
CN116106554A (en) * 2022-11-30 2023-05-12 山东省食品药品检验研究院 Method for detecting content of deer antler glue by liquid chromatography-mass spectrometry

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1548940A (en) * 2003-05-08 2004-11-24 中国科学院过程工程研究所 Method for distinguishing between donkey-hide gelati and pigskin glue, oxhide glue
CN102749349A (en) * 2012-06-20 2012-10-24 山东大学 Method for testing authenticity of donkey-hide gelatin

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1548940A (en) * 2003-05-08 2004-11-24 中国科学院过程工程研究所 Method for distinguishing between donkey-hide gelati and pigskin glue, oxhide glue
CN102749349A (en) * 2012-06-20 2012-10-24 山东大学 Method for testing authenticity of donkey-hide gelatin

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
ZHANG GUI-FENG ET AL.: "Identification of marker peptides in digested gelatins by high performance liquid chromatography/mass spectrometry", 《CHINESE JOURNAL OF ANALYTICAL CHEMISTRY》 *
张贵锋 等: "高效液相色谱-质谱联用法识别明胶动物来源", 《质谱学报》 *
王前: "明胶制备工艺及溯源方法研究", 《中国优秀硕士学位论文全文数据库》 *
程显隆 等: "UPLC-QTOF-MS结合主成分分析法用于龟甲胶、鹿角胶中添加牛皮源成分的检测研究", 《药物分析杂志》 *

Cited By (13)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104360002A (en) * 2014-09-12 2015-02-18 江西师范大学 Quantitative detection method for cow leather gelatin
CN104360002B (en) * 2014-09-12 2016-08-24 江西师范大学 A kind of quantitative detecting method of cattle hide gelatin
CN105301165A (en) * 2015-10-09 2016-02-03 东阿阿胶股份有限公司 Donkey characteristic polypeptide and application thereof to detection on donkey skin derived ingredients
CN106645479A (en) * 2016-12-22 2017-05-10 东阿阿胶股份有限公司 Composition, kit and detection method for detecting content of dog bone source ingredients in glue type traditional Chinese medicines and compound preparations thereof
CN106645479B (en) * 2016-12-22 2019-08-09 东阿阿胶股份有限公司 It is a kind of for detecting composition, kit and its detection method of dog bone derived components content in glue class Chinese medicine and its compound preparation
CN106841496B (en) * 2017-01-04 2019-02-19 东阿阿胶股份有限公司 It is a kind of for detecting composition, kit and its detection method of dog bone derived components content in glue class Chinese medicine and its compound preparation
CN106841496A (en) * 2017-01-04 2017-06-13 东阿阿胶股份有限公司 A kind of composition for detecting dog bone derived components content in glue class Chinese medicine and its compound preparation, kit and its detection method
CN109187783A (en) * 2018-09-03 2019-01-11 贵州广济堂药业有限公司 Deer horn glue feature peptide and identification sample to be tested in whether include deer horn glue method
CN109187783B (en) * 2018-09-03 2021-06-29 贵州广济堂健康药业有限公司 Deer glue characteristic peptide and method for identifying deer glue in sample to be detected
CN110530999A (en) * 2019-09-18 2019-12-03 山东省食品药品检验研究院 A kind of distinguishing method between true and false of caribou horn
CN111060622A (en) * 2019-12-26 2020-04-24 北京化工大学 Labeled peptide of tuna-derived component and application of labeled peptide in detection of collagen degradation product and product thereof
CN111060622B (en) * 2019-12-26 2021-06-29 北京化工大学 Labeled peptide of tuna-derived component and application of labeled peptide in detection of collagen degradation product and product thereof
CN116106554A (en) * 2022-11-30 2023-05-12 山东省食品药品检验研究院 Method for detecting content of deer antler glue by liquid chromatography-mass spectrometry

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