CN103630634B - The detection thing of a kind of colla carapacis et plastri testudinis, deer horn glue and goods thereof and detection method thereof - Google Patents
The detection thing of a kind of colla carapacis et plastri testudinis, deer horn glue and goods thereof and detection method thereof Download PDFInfo
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Abstract
The invention provides a kind of method identifying colla carapacis et plastri testudinis, deer horn glue and goods thereof, it utilizes colla carapacis et plastri testudinis, deer horn glue and goods thereof and has specific nucleotide sequences in other Counterfeit Item genomes or the different of amino acid sequence are detected, is described specific amino acid as SEQ.ID? shown in No1.Method of the present invention, can detect the adulterant compositions such as the donkey in colla carapacis et plastri testudinis, deer horn glue and goods thereof, horse, ox, pig or sheep fast, thus point evident.The invention still further relates to polypeptide, and the composition be made up of described polypeptide and respective carrier.In addition, described polypeptide and the application of composition in detection colla carapacis et plastri testudinis and goods thereof in donkey, horse, ox, pig or sheep derived material thereof is also comprised.
Description
Technical field
The present invention relates to the detection method of a breeding ass, horse, ox, pig or sheep derived material, specifically can be applicable to the authenticity of colla carapacis et plastri testudinis, deer horn glue and goods thereof, belong to field of traditional Chinese medicine detection.
Background technology
Record according to version Chinese Pharmacopoeia in 2010, colla carapacis et plastri testudinis take tortoise plastron as raw material, and through soak by water, the concentrated solid gum made, property is salty, sweet, cool, returns liver, kidney, the heart channel of Hang-Shaoyin.Can enriching yin, nourish blood, stop blooding, be mainly used in deficiency of Yin hectic fever, hectic fever due to yin night sweat, soreness and weakness of waist and knees, the deficiency of blood be sallow, uterine bleeding band is inferior.Long-term taking colla carapacis et plastri testudinis also helps and strengthens body self-regulation, can promote longevity, also be beneficial to the physical rehabilitation of hyperactivity of yang due to yin deficiency patient.Deer horn glue take deer horn as raw material, and through soak by water, the concentrated solid gum made, property is sweet, salty, warm, returns kidney, Liver Channel.Can warm filling liver kidney, beneficial intensive culture blood, the waist knee acid be mainly used in caused by kidney deficiency and liver is cold, impotence and seminal emission, and consumptive disease is thin thin, metrostaxis, hematuria of having blood in stool, and cloudy subcutaneous ulcer swells and ache.Along with improving constantly of living standard, colla carapacis et plastri testudinis, deer horn glue become one of first-selection that people nourish.
Due to tortoise plastron and deer horn self famousr and precious, medical value is remarkable, and seriously catching and killing in the last few years to various Chelonian and animal in deer family, cause the supply of tortoise plastron, deer horn raw material very limited, and price is high, has directly had influence on the production of colla carapacis et plastri testudinis, deer horn glue.There are some counterfeit and shoddy goods in colla carapacis et plastri testudinis, deer horn glue market, its make raw material be not tortoise plastron, deer horn, but by other animals mix skin, broken bone replace, comprise pigskin, ox-hide, even the skin of animals died of illness, dirty skin, rotten skin etc.Such counterfeit and shoddy goods once be shaped, outward appearance, color and luster, smell all to certified products colla carapacis et plastri testudinis, deer horn glue and similar, true and false difficulty is distinguished.Take these adulterants not only without any curative effect, be also probably harmful to health, consequence is very serious.These adulterants seriously hinder the sound development of whole colla carapacis et plastri testudinis, deer horn glue Market of Chinese Materia Medica.
The glue class Chinese medicine concentrated by the skin of animal, bone, the long-time thermophilic digestion of first-class warp, the very close and production technology of different manufacturers of principal ingredient there are differences, and adopts infrared, the method such as near infrared, X-diffraction to carry out True-false distinguish to glue class Chinese medicine and often exists and judge the situations such as inaccurate.Have and report from the angle of polypeptide and utilize tortoise source property, deer source property, Niu Yuanxing, donkey derived component Mass Spectrometer Method glue class Chinese medicine, result accurately and reliably.But at present also not from the angle of collagen polypeptide, the detection method of adulterant composition in disposable discriminating colla carapacis et plastri testudinis, deer horn glue and goods thereof.
In glue class Chinese medicine, principal ingredient is collagen polypeptide, the collagen, amino acid sequence of different animals there are differences, in different plant species except tortoise, deer, the same polypeptide of the stable existence contained can be used as the characteristic polypeptide of adulterant composition in colla carapacis et plastri testudinis, deer horn glue, by digestion with restriction enzyme, utilize mass spectrum can to trace to the source qualification to glue class Chinese medicine completely.
Summary of the invention
The present invention is directed to the problems referred to above, provide detection thing and the authenticity identification method thereof of a kind of colla carapacis et plastri testudinis, deer horn glue and goods thereof, the method is simple to operate, and characteristic is strong, highly sensitive, can be used for the qualification of tracing to the source of adulterant composition in colla carapacis et plastri testudinis, deer horn glue and goods thereof.
Technical scheme of the present invention is as follows:
First, the invention provides a kind of method identifying colla carapacis et plastri testudinis, deer horn glue and goods thereof, it utilizes colla carapacis et plastri testudinis, deer horn glue and goods thereof and has specific nucleotide sequences in other Counterfeit Item genomes or the different of amino acid sequence are detected, and described specific amino acid is as shown in SEQ.IDNo1.
Preferably, described colla carapacis et plastri testudinis, deer horn glue goods are selected from the one in food, health products or the medicine that colla carapacis et plastri testudinis or deer horn glue make, and not containing donkey, horse, ox, pig, sheep derived material in formula.
Preferably, other Counterfeit Items described refer to one or more in donkey, horse, ox, pig or sheep gelatin substances of boiling.
More specifically, described method comprises the steps:
(1) choose specific amino acid or amino acid sequence in colla carapacis et plastri testudinis, deer horn glue goods and other Counterfeit Item genomes, described specific amino acid is as shown in SEQ.IDNo1;
(2) colla carapacis et plastri testudinis, deer horn glue or its goods are dissolved, or extract the polypeptide constituents of described colla carapacis et plastri testudinis or its goods and dissolve, add restriction enzyme subsequently and carry out enzyme and cut;
(3) solution after being cut by the enzyme that step (2) obtains puts into LC-MS instrument, with colla carapacis et plastri testudinis sterling for matrix, adds step (1) described amino acid sequence in contrast, and selects the parent ion m/z441.8 of this amino acid sequence and daughter ion thereof to monitor.
If the retention time detecting this ion is consistent with reference substance, and its daughter ion is consistent with the daughter ion of reference substance, then contain this adulterant composition in described colla carapacis et plastri testudinis or its goods; If this not consistent with reference substance retention time ion, then not containing this adulterant composition.
Preferably, to dissolve reagent used described in step (2) be mass percent is 1%, the NH of pH value 8.0
4hCO
3solution; Described restriction enzyme is trypsase.
Further, the invention provides a kind of molecular marked compound detecting colla carapacis et plastri testudinis and goods thereof, the amino acid sequence of this molecular marked compound is as shown in SEQ.IDNo1.
Further, the invention provides the gene of the albumen described in coding.
Further, the invention provides described molecular marked compound whether detecting in colla carapacis et plastri testudinis containing the application in donkey, horse, ox, pig or sheep derived material.
Further, the invention provides a kind of pharmaceutical composition, it comprises described albumen and one or more pharmaceutically acceptable carriers of pharmacy effective dose.
Further, the invention provides described pharmaceutical composition whether detecting in colla carapacis et plastri testudinis containing the application in donkey, horse, ox, pig or sheep derived material.
Beneficial effect of the present invention is as follows:
Adopt method of the present invention, the adulterant composition such as donkey, horse, ox, pig, sheep in colla carapacis et plastri testudinis, deer horn glue and goods thereof can be detected fast, thus point evident, although collagen polypeptide has certain hydrolysis and destroys in glue class Chinese medicine, but the otherness polypeptide that the present invention is based in its principal ingredient collagen, whether its content is high, and the degree that is damaged is very little, can identify in colla carapacis et plastri testudinis, deer horn glue and goods thereof containing adulterant compositions such as donkey, horse, ox, pig or sheep.
The invention still further relates to polypeptide, its amino acid sequence is Gly-Val-Val-Gly-Leu-Pro-Gly-Gln-Arg, and the composition be made up of described polypeptide and respective carrier.In addition, described polypeptide and the application of composition in detection colla carapacis et plastri testudinis, deer horn glue and goods thereof in donkey, horse, ox, pig or sheep derived material thereof is also comprised.
Accompanying drawing explanation
Fig. 1 be under improvement on synthesis and sterling glue multiple-reaction monitoring scan pattern Selective ion mode to the mass spectrograms of m/z441.8 → 456.8,627.8 monitorings;
Fig. 2 be under improvement on synthesis and the epoxy glue multiple-reaction monitoring scan pattern adding 5% deer horn glue Selective ion mode to the mass spectrograms of m/z441.8 → 456.8,627.8 monitorings;
Fig. 3 be improvement on synthesis and colla carapacis et plastri testudinis, deer horn glue goods (for the celestial oral liquid of tortoise deer two) under multiple-reaction monitoring scan pattern Selective ion mode to the mass spectrograms of m/z441.8 → 456.8,627.8 monitorings.
Embodiment
Further describe the present invention below in conjunction with specific embodiment, advantage and disadvantage of the present invention will be more clear along with description.But embodiment is only exemplary, does not form any restriction to scope of the present invention.It will be understood by those skilled in the art that and can modify to the details of technical solution of the present invention and form or replace down without departing from the spirit and scope of the present invention, but these amendments 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
1 material and reagent
Material: sterling glue comprises donkey-hide gelatin, horse skin glue, oxhide gelatin, pig skin gelatin, sheepskin glue, colla carapacis et plastri testudinis, deer horn glue, decocts obtain with donkey hide, horse skin, ox-hide, pigskin, sheepskin, tortoise plastron, deer horn respectively.
Reagent: ammonium bicarbonate (analyzing pure), trypsase (sequence is pure, purchased from National Institute for Food and Drugs Control), improvement on synthesis Gly-Val-Val-Gly-Leu-Pro-Gly-Gln-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, filtering with microporous membrane, and precision measures the trypsin solution 100 μ L that subsequent filtrate 1mL adds 2mg/mL, 37 DEG C of constant temperature enzymolysis 6h.
(2) preparation of improvement on synthesis control sample enzymolysis solution
Take a certain amount of improvement on synthesis, add 1%NH
4hCO
3solution (pH8.0), shakes up, and is mixed with concentration and is about 0.3 μ g/mL, filtering with microporous membrane, for subsequent use.
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 μm), mobile phase A is 0.1% formic acid solution (volume fraction), and Mobile phase B is acetonitrile, flow velocity 0.3mL/min; Gradient elution: 0 ~ 40min, 2% ~ 50% Mobile phase B (percentage here represents from 0 minute to the 40th minute, flowing B by 2% linear gradient be 50%, mobile phase A then fades to 50% by 98%).Mass Spectrometry Conditions: electron spray positive ion mode (ESI
+) carry out one-level and entirely sweep, sweep limit m/z400-m/z1000.
Entirely sweep figure from the mass spectrum of respective glue sample and extract ion m/z441.8, find to only have in colla carapacis et plastri testudinis, deer horn glue not this quasi-molecular ions at about 10.84min by contrast, and all contain in donkey-hide gelatin, horse skin glue, oxhide gelatin, pig skin gelatin and sheepskin glue.Show that above-mentioned conclusion is correct by the detection of multiple batches of sample and checking.Illustrate: can, by detecting the ion m/z441.8 in colla carapacis et plastri testudinis, deer horn glue, determine whether to be mixed with donkey, horse, ox, pig or sheep derived material.
(2) the amino acid sequence initial guess of polypeptide
Utilize triple quadrupole bar mass spectrum, daughter ion scanning is carried out to the above-mentioned ion found out, confirm this ion band double charge, by sequence assembly, infer the partial sequence polypeptide.Partial sequence by inference, at NCBI(http: //www.ncbi.nlm.nih.gov/) database carries out sequence retrieval, feature in conjunction with it (does not have in tortoise, deer, and all contain in donkey, horse, ox, pig or sheep, and be about 882.6 by molecular weight after tryptic digestion), find out the sequence Gly-Val-Val-Gly-Leu-Pro-Gly-Gln-Arg that this polypeptide is possible.According to the CID fracture theory of peptide chain, the daughter ion calculating this polypeptide possible is as follows:
| a | b | c″ | Res: | x | y″ | z |
| 30.0 | 58.0 | 77.1 | Gly | - | - | - |
| 129.1 | 157.1 | 176.1 | Val | 849.5 | 825.5 | 806.5 |
| 228.2 | 256.2 | 275.2 | Val | 750.4 | 726.4 | 707.4 |
| 285.2 | 313.2 | 332.2 | Gly | 651.3 | 627.4 | 608.3 |
| 398.3 | 426.3 | 445.3 | Leu | 594.3 | 570.3 | 551.3 |
| 495.3 | 523.3 | 542.4 | Pro | 481.2 | 457.3 | 438.2 |
| 552.4 | 580.3 | 599.4 | Gly | 384.2 | 360.2 | 341.2 |
| 680.4 | 708.4 | 727.4 | Gln | 327.1 | 303.2 | 284.1 |
| - | - | - | Arg | 199.1 | 175.1 | 156.1 |
Coincideing in figure can be entirely swept with the daughter ion of peculiar ion m/z441.8 in donkey-hide gelatin, horse skin glue, oxhide gelatin, pig skin gelatin or sheepskin glue.
(3) sequence is determined
Amino acid sequence by inference, entrusts Peptide systhesis company (the biochemical company limited of gill) to carry out the synthesis of this polypeptide.Then the daughter ion utilizing LC-MS instrument to carry out improvement on synthesis is swept entirely, and in its testing conditions and glue sample, the daughter ion of corresponding polypeptide is swept consistent entirely.Full figure is swept by the ion appearance time, the daughter ion that contrast improvement on synthesis and donkey-hide gelatin, horse skin glue, oxhide gelatin, pig skin gelatin and sheepskin glue, both discoveries fit like a glove, thus determine that this ion m/z441.8 is polypeptide Gly-Val-Val-Gly-Leu-Pro-Gly-Gln-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, sheepskin glue, decocts obtain with donkey hide, horse skin, ox-hide, pigskin, tortoise plastron, deer horn, sheepskin respectively.
Epoxy glue sample (colla carapacis et plastri testudinis, the colla carapacis et plastri testudinis of 10% oxhide gelatin, the colla carapacis et plastri testudinis of 20% oxhide gelatin, the colla carapacis et plastri testudinis of 40% oxhide gelatin containing 5% oxhide gelatin) is accurately mixed (percentage is massfraction) by above-mentioned sterling glue.
Reagent: ammonium bicarbonate (analyzing pure), trypsase (sequence is pure, purchased from National Institute for Food and Drugs Control), improvement on synthesis Gly-Val-Val-Gly-Leu-Pro-Gly-Gln-Arg.
2 detection methods
(1) preparation of improvement on synthesis control sample enzymolysis solution
Get 1.0g sterling colla carapacis et plastri testudinis 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, filtering with microporous membrane, and precision measures subsequent filtrate 1ml and adds improvement on synthesis 0.3 μ g, then adds the trypsin solution 100 μ l of 2mg/ml, 37 DEG C 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, filtering with microporous membrane, and precision measures the trypsin solution 100 μ l that subsequent filtrate 1ml adds 2mg/ml, 37 DEG C 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 μm), mobile phase A is 0.1% formic acid solution (volume fraction), and Mobile phase B is acetonitrile, flow velocity 0.3ml/min; Gradient elution: 0 ~ 40min, 2% ~ 50% Mobile phase B (percentage here represents from 0 minute to the 40th minute, flowing B by 2% linear gradient be 50%, mobile phase A then fades to 50% by 98%).Mass Spectrometry Conditions: electron spray positive ion mode (ESI
+) select to carry out many reaction detection, select m/z441.8 → 456.8,627.8 as detecting ion pair.
The results are shown in Figure 1,10.84min place to only have in colla carapacis et plastri testudinis, deer horn glue sterling and detect corresponding quasi-molecular ions, other all do not detect.The detection of epoxy glue sample, with the percentage composition of oxhide gelatin for horizontal ordinate, with chromatographic peak area in m/z441.8 → 456.8 extraction ion flow graph for ordinate, drawing standard curve, R
2be more than 0.999, linearly well.This method visible can specific detection colla carapacis et plastri testudinis, adulterant composition in deer horn glue, thus carries out authenticity to colla carapacis et plastri testudinis, deer horn glue.
Embodiment 2
1 material and reagent
Material: epoxy glue sample adds 5% adulterant glue respectively by the deer horn glue sterling in embodiment 1 and makes, comprises deer horn glue, the deer horn glue of 5% horse skin glue, the deer horn glue containing 5% oxhide gelatin, the deer horn glue of 5% pig skin gelatin, the deer horn glue (percentage is massfraction) of 5% sheepskin glue containing 5% donkey-hide gelatin.
Reagent: ammonium bicarbonate (analyzing pure), trypsase (sequence is pure, purchased from National Institute for Food and Drugs Control), improvement on synthesis Gly-Val-Val-Gly-Leu-Pro-Gly-Gln-Arg.
2 detection methods
(1) preparation of improvement on synthesis control sample enzymolysis solution
Get 1.0g sterling deer horn glue 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, filtering with microporous membrane, and precision measures subsequent filtrate 1ml and adds improvement on synthesis 0.3 μ g, then adds the trypsin solution 100 μ l of 2mg/ml, 37 DEG C 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, filtering with microporous membrane, and precision measures the trypsin solution 100 μ l that subsequent filtrate 1ml adds 2mg/ml, 37 DEG C 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 μm), mobile phase A is 0.1% formic acid solution (volume fraction), and 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, flowing B by 2% linear gradient be 50%, mobile phase A then fades to 50% by 98%).Mass Spectrometry Conditions: electron spray positive ion mode (ESI
+) select to carry out many reaction detection, select m/z441.8 → 456.8,627.8 as detecting ion pair.
The results are shown in Figure 2,10.84min place improvement on synthesis control sample, add in the epoxy glue sample of 5% adulterant glue and all detect corresponding quasi-molecular ions, and all do not detect corresponding quasi-molecular ions in sterling colla carapacis et plastri testudinis in case study on implementation 1, deer horn glue.This method visible can specific detection colla carapacis et plastri testudinis, adulterant composition in deer horn glue.
Embodiment 3
1 material and reagent
Material: colla carapacis et plastri testudinis, deer horn glue, the celestial oral liquid of tortoise deer two (Dong-E donkey-hide Gelatin Co., Ltd., Shandong Prov.), add the celestial oral liquid of tortoise deer two (self-control) of oxhide gelatin, add the celestial oral liquid of tortoise deer two (self-control) of donkey-hide gelatin
Reagent: trichloroacetic acid (analyzing pure), ammonium bicarbonate (analyzing pure), trypsase (sequence is pure, purchased from National Institute for Food and Drugs Control), improvement on synthesis Gly-Val-Val-Gly-Leu-Pro-Gly-Gln-Arg.
2 detection methods
(1) preparation of improvement on synthesis control sample enzymolysis solution
Get the celestial oral liquid of 1.0g tortoise deer two and add 1%NH
4hCO
3solution 24ml dissolves, and adds 4g trichloroacetic acid, places the centrifugal 5min of 10min, 10000rpm, abandons supernatant, proceeded to by sediment in 100ml measuring bottle, use 1%NH for 4 DEG C
4hCO
3solution (pH8.0) dissolves and is settled to scale, shakes up, filtering with microporous membrane, and precision measures subsequent filtrate 1ml and adds improvement on synthesis 0.3 μ g, then adds the trypsin solution 100 μ l of 2mg/ml, 37 DEG C 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 the centrifugal 5min of 10min, 10000rpm, abandons supernatant, proceeded to by sediment in 100ml measuring bottle, use 1%NH for 4 DEG C
4hCO
3solution (pH8.0) dissolves and is settled to scale, shakes up, filtering with microporous membrane, and precision measures the trypsin solution 100 μ l that subsequent filtrate 1ml adds 2mg/ml, 37 DEG C 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 μm), mobile phase A is 0.1% formic acid solution (volume fraction), and 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, flowing B by 2% linear gradient be 50%, mobile phase A then fades to 50% by 98%).Mass Spectrometry Conditions: electron spray positive ion mode (ESI
+) carry out many reaction detection, select m/z441.8 → 456.8,627.8 as detecting ion pair.
The results are shown in Figure 3,10.84min place improvement on synthesis control sample, the celestial oral liquid of tortoise deer two adding oxhide gelatin, the celestial oral liquid of tortoise deer two adding donkey-hide gelatin all detect corresponding quasi-molecular ions, other do not detect, and this method can specific detection colla carapacis et plastri testudinis, adulterant composition in deer horn glue goods as seen.
Sequence table
<110> Dong-E donkey-hide Gelatin Co., Ltd., Shandong Prov.
The detection thing of <120> a colla carapacis et plastri testudinis, deer horn glue and goods thereof and authentication method thereof
<130>
<160>1
<170>PatentInversion3.3
<210>1
<211>9
<212>PRT
<213> artificial sequence
<400>1
GlyValValGlyLeuProGlyGlnArg
15。
Claims (1)
1. identify a method for colla carapacis et plastri testudinis, deer horn glue and goods thereof, it is characterized in that, it specifically comprises the steps:
(1) choose specific amino acid in colla carapacis et plastri testudinis, deer horn glue goods and other Counterfeit Item genomes, described specific amino acid is as shown in SEQ.IDNo1; Described colla carapacis et plastri testudinis, deer horn glue goods are selected from the one in food, health products or the medicine that colla carapacis et plastri testudinis or deer horn glue make, and do not conform in formula and have donkey, horse, sheep derived material; Other Counterfeit Items described refer to one or more in donkey, horse or sheep gelatin substances of boiling;
(2) colla carapacis et plastri testudinis, deer horn glue or its goods are dissolved, or extract the polypeptide constituents of described colla carapacis et plastri testudinis or its goods and dissolve, add restriction enzyme subsequently and carry out enzyme and cut; To be mass percent be described dissolving reagent used 1%, the NH of pH value 8.0
4hCO
3solution; Described restriction enzyme is trypsase;
(3) solution after being cut by the enzyme that step (2) obtains puts into LC-MS instrument, with colla carapacis et plastri testudinis sterling for matrix, add step (1) described amino acid sequence in contrast, and select the parent ion m/z441.8 of this amino acid sequence and daughter ion thereof to monitor.
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| CN111825744B (en) * | 2016-03-18 | 2022-11-08 | 青岛海关技术中心 | Short peptides for identifying donkey-hide gelatin and bovine-derived components in donkey-hide gelatin product |
| CN106770825B (en) * | 2016-12-22 | 2019-12-03 | 东阿阿胶股份有限公司 | It is a kind of for detecting composition, kit and its detection method of donkey hide derived components content in animal glue |
| CN106841497B (en) * | 2017-01-20 | 2018-08-07 | 东阿阿胶股份有限公司 | Composition, kit and detection method for detecting donkey hide derived components content in colla carapacis et plastri testudinis, deer horn glue, donkey-hide gelatin and its adulterant glue |
| 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 |
| CN109971868A (en) * | 2019-05-07 | 2019-07-05 | 镇江市食品药品监督检验中心 | One group for identifying the specific primer and deer horn authenticity identification method of deer horn |
| CN113880916B (en) * | 2021-08-19 | 2023-06-30 | 青海瑞肽生物科技有限公司 | Yak skin antioxidant polypeptide and preparation method and application thereof |
Family Cites Families (1)
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-
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Non-Patent Citations (8)
| Title |
|---|
| Assessing the Extent of Bone Degradation Using Glutamine Deamidation in Collagen;Julie Wilson et al.;《Analytical Chemistry》;20121231;第84卷;全文 * |
| DNA 分子标记在中药鉴定中的应用及发展趋势分析;李力 等;《时珍国医国药》;20091231;第20卷(第11期);全文 * |
| Identification of five gelatins by ultra performance liquid chromatography/time-of flight mass spectrometry (UPLC/Q-TOF-MS) using principal component analysis;Xian-Long Cheng et al.;《Journal of Pharmaceutical and Biomedical Analysis》;20121231;第62卷;全文 * |
| Mass spectrometric analysis of enzymatic digestion of denatured collagen for identification of collagen type;Guifeng Zhang et al.;《Journal of Chromatography A》;20061231;第1114卷;摘要，第274页，第275页右栏，第276页，表1 * |
| Mass spectrometric detection of marker peptides in tryptic digests of gelatin:A new method to differentiate between bovine and porcine gelatin;Guifeng Zhang et al.;《Food Hydrocolloids》;20091231;第23卷;全文 * |
| 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;程显隆 等;《药物分析杂志》;20121231;第32卷(第6期);摘要，第932页左栏、右栏"2.3样品处理"项，第934页左栏，表1，图3，图4 * |
| 动物胶类药材的鉴别方法研究进展;程显隆 等;《亚太传统医药》;20110331;第7卷(第3期);全文 * |
| 高效液相色谱质谱鉴别龟甲胶与鹿角胶;王前 等;《光谱学与光谱分析》;20081031;第28卷(第10期);全文 * |
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