CN103630646B - The detection method of turtle-derived component in a kind of glue class Chinese medicine and goods thereof - Google Patents
The detection method of turtle-derived component in a kind of glue class Chinese medicine and goods thereof Download PDFInfo
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Abstract
The invention provides the detection method of turtle-derived component in a kind of glue class Chinese medicine and goods thereof, it utilizes the difference in tortoise genome with specific nucleotide sequences or amino acid sequence to detect, is described specific amino acid as SEQ.ID? shown in No1.Adopt method of the present invention, whether can detect fast in glue class Chinese medicine containing turtle-derived component, 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, its content is high, and the degree that is damaged is very little, can identify the turtle-derived component in glue class Chinese medicine and goods thereof.
Description
Technical field
The present invention relates to a kind of detection method of turtle-derived component, be specifically related to the detection method of turtle-derived component in a kind of glue class Chinese medicine and goods thereof, belong to field of traditional Chinese medicine detection.
Background technology
Tortoise plastron, namely the carapace of Testudinidae animal tortoise (Chinemysreevesii) and plastron, belong to traditional rare Chinese medicine.Chinese Pharmacopoeia according to latest edition is recorded, and taking tortoise plastron can be nourishing and suppressing Yang, the strong bone of kidney-nourishing, bushing of nourishing blood.In the last few years, along with the raising of biology level, the correlative study of tortoise plastron also deepens continuously.There are some researches show, tortoise plastron powder and extract thereof can not only strengthen liver function, release the pressure, and also have effect that is anticancer and immunity moderation resistibility.With tortoise plastron be the two series products Guiling Jellies made of raw material and colla carapacis et plastri testudinis on whole Market of Chinese Materia Medica in occupation of important position.
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.Along with improving constantly of living standard, similar with donkey-hide gelatin, colla carapacis et plastri testudinis also becomes one of first-selection that people nourish.
Because tortoise plastron self is famousr and precious, medical value is remarkable, and tortoise meat also usually eats for people for cook various kinds tonic, in the last few years to various Chelonian to catch and kill phenomenon very serious, some Chelonian kind is endangered even.Therefore, the supply of tortoise plastron raw material is very limited, and price is high, has also directly had influence on the production of colla carapacis et plastri testudinis.Colla carapacis et plastri testudinis market exists some counterfeit and shoddy goods, its make raw material be not tortoise plastron, 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 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.
Glue class Chinese medicine forms gelatin substance by the skin of animal, bone, first-class boiling, and comprises donkey-hide gelatin, colla carapacis et plastri testudinis, deer horn glue etc., has the traditional Chinese medicine of typical ethnic characteristic.Glue class Chinese medicine generally concentrates through long-time thermophilic digestion and forms, 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 sterling and often exists and judge the situations such as inaccurate.The discriminating of glue class Chinese medicament turtle-derived component, existing scholar differentiates from DNA angle, but just qualitative, has false negative and false positive phenomenon, and operation is comparatively complicated.Have and report from the angle of polypeptide and utilize Mass Spectrometer Method glue class Chinese medicament turtle-derived component, result accurately and reliably.At present, trace to the source in the goods also not to glue class Chinese medicine qualification method.
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 accurate to glue class Chinese medicine and goods thereof trace to the source qualification and content detection.
Summary of the invention
The present invention is directed to the problems referred to above, provide the detection method of turtle-derived component in a kind of glue class Chinese medicine and goods thereof, the method is simple to operate, and characteristic is strong, highly sensitive, can be used for the qualification of turtle-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 turtle-derived component in a kind of glue class Chinese medicine and goods thereof, it utilizes the difference in tortoise genome with specific nucleotide sequences or amino acid sequence to detect, and described specific amino acid is as shown in SEQ.IDNo1.
Preferably, described glue class Chinese medicine is selected from donkey-hide gelatin, oxhide gelatin, colla carapacis et plastri testudinis or deer horn glue.
Preferably, described glue class Chinese herbal product is selected from food, health products or the medicine be made up of corresponding glue class Chinese medicine.
Particularly, the method comprises the steps:
(1) choose specific amino acid or amino acid sequence in tortoise genome, described specific amino acid is as shown in SEQ.IDNo1;
(2) glue class Chinese medicine or its goods are dissolved, or extract protein and peptide component dissolves wherein, add restriction enzyme and carry out enzyme and cut;
(3) put into LC-MS instrument subsequently, with negative sample to be detected for matrix, add step (1) gained amino acid sequence or colla carapacis et plastri testudinis sterling in contrast, select the parent ion m/z569.1 of this polypeptide 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 described glue class Chinese medicine or its goods contain turtle-derived component; If this not consistent with reference substance retention time ion, then not containing turtle-derived component.
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.
Adopt method of the present invention, whether can detect fast in glue class Chinese medicine containing turtle-derived component, 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, its content is high, and the degree that is damaged is very little, can identify the turtle-derived component in glue class Chinese medicine and goods thereof.
The invention still further relates to polypeptide, its amino acid sequence is Gly-Val-Gln-Gly-Ala-Hyp-Gly-Pro-Gln-Gly-Pro-Arg, and the composition be made up of described polypeptide and respective carrier.In addition, described polypeptide and the application of composition in detection glue class Chinese medicine and goods thereof in turtle-derived component 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 m/z569.1 → 724.4,795.4, the mass spectrograms of 852.4 monitorings;
Fig. 2 be under improvement on synthesis and the epoxy glue multiple-reaction monitoring scan pattern adding 5% colla carapacis et plastri testudinis Selective ion mode to m/z569.1 → 724.4,795.4, the mass spectrograms of 852.4 monitorings;
Fig. 3 be improvement on synthesis and colla carapacis et plastri testudinis goods (for the celestial oral liquid of tortoise deer two) under multiple-reaction monitoring scan pattern Selective ion mode to m/z569.1 → 724.4,795.4, the mass spectrograms of 852.4 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
Material: sterling glue comprises donkey-hide gelatin, horse skin glue, oxhide gelatin, pig skin gelatin, colla carapacis et plastri testudinis, deer horn glue, decocts obtain 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), improvement on synthesis Gly-Val-Gln-Gly-Ala-Hyp-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, 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/z569.1, find to only have containing this quasi-molecular ions in colla carapacis et plastri testudinis at about 4.06min by contrast, and all do not have in donkey-hide gelatin, horse skin glue, oxhide gelatin, pig skin gelatin and deer horn glue.Show that above-mentioned conclusion is correct by the detection of multiple batches of sample and checking.Illustrate: by detecting the ion m/z569.1 in colla carapacis et plastri testudinis, can determine whether containing turtle-derived component.
(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 (has in tortoise, and all do not have in donkey, horse, ox, pig or deer, and be about 1137.2 by molecular weight after tryptic digestion), derive the sequence Gly-Val-Gln-Gly-Ala-Hyp-Gly-Pro-Gln-Gly-Pro-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 | 1103.5 | 1079.6 | 1060.5 |
| 257.2 | 285.2 | 304.2 | Gln | 1004.5 | 980.5 | 961.5 |
| 314.2 | 342.2 | 361.2 | Gly | 876.4 | 852.4 | 833.4 |
| 385.2 | 413.2 | 432.3 | Ala | 819.4 | 795.4 | 776.4 |
| 498.3 | 526.3 | 545.3 | Hyp | 748.3 | 724.4 | 705.3 |
| 555.3 | 583.3 | 602.3 | Gly | 635.3 | 611.3 | 592.3 |
| 652.3 | 680.3 | 699.4 | Pro | 578.3 | 554.3 | 535.3 |
| 780.4 | 808.4 | 827.4 | Gln | 481.2 | 457.3 | 438.2 |
| 837.4 | 865.4 | 884.5 | Gly | 353.2 | 329.2 | 310.2 |
| 934.5 | 962.5 | 981.5 | Pro | 296.1 | 272.2 | 253.1 |
| - | - | - | Arg | 199.1 | 175.1 | 156.1 |
Coincideing in figure can be entirely swept with the daughter ion of peculiar ion m/z569.1 in colla carapacis et plastri testudinis.
(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.Entirely sweep figure by the ion appearance time, the daughter ion that contrast improvement on synthesis and colla carapacis et plastri testudinis, both discoveries fit like a glove, thus determine that this ion m/z569.1 is polypeptide Gly-Val-Gln-Gly-Ala-Hyp-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 obtain with donkey hide, horse skin, ox-hide, pigskin, tortoise plastron, deer horn respectively.
Epoxy glue sample (oxhide gelatin, the oxhide gelatin of 10% colla carapacis et plastri testudinis, the oxhide gelatin of 20% colla carapacis et plastri testudinis, the oxhide gelatin of 40% colla carapacis et plastri testudinis containing 5% colla carapacis et plastri testudinis) 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-Gln-Gly-Ala-Hyp-Gly-Pro-Gln-Gly-Pro-Arg.
2 detection methods
(1) preparation of improvement on synthesis control sample enzymolysis solution
Get 1.0g sterling oxhide 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, 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/z569.1 → 724.4,795.4,852.4 as detecting ion pair.
The results are shown in Figure Isosorbide-5-Nitrae .06min place to only have in improvement on synthesis control sample and colla carapacis et plastri testudinis and detect corresponding quasi-molecular ions, other all do not detect.The detection of epoxy glue sample, with the percentage composition of colla carapacis et plastri testudinis for horizontal ordinate, with chromatographic peak area in m/z569.1 → 724.4 extraction ion flow graph for ordinate, drawing standard curve, R
2be more than 0.999, linearly well.This method visible can specific detection turtle-derived component, comprises donkey, horse, ox, pig, deer etc. distinguish with other various animal derived materials.
Embodiment 2
1 material and reagent
Material: epoxy glue sample adds 5% colla carapacis et plastri testudinis respectively by the sterling glue sample in embodiment 1 and makes, comprises donkey-hide gelatin, the horse skin glue of 5% colla carapacis et plastri testudinis, the oxhide gelatin containing 5% colla carapacis et plastri testudinis, the pig skin gelatin of 5% colla carapacis et plastri testudinis, the deer horn glue containing 5% colla carapacis et plastri testudinis containing 5% colla carapacis et plastri testudinis.
Reagent: ammonium bicarbonate (analyzing pure), trypsase (sequence is pure, purchased from National Institute for Food and Drugs Control), improvement on synthesis Gly-Val-Gln-Gly-Ala-Hyp-Gly-Pro-Gln-Gly-Pro-Arg.
2 detection methods
(1) preparation of improvement on synthesis control sample enzymolysis solution
Get 1.0g sterling oxhide 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, 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/z569.1 → 724.4,795.4,852.4 as detecting ion pair.
The results are shown in Figure 2,4.06min place improvement on synthesis control sample, add 5% colla carapacis et plastri testudinis epoxy glue sample in 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, deer horn glue.This method visible can turtle-derived component in specific detection glue class Chinese medicine.
Embodiment 3
1 material and reagent
Material: colla carapacis et plastri testudinis, 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 colla carapacis et plastri testudinis
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-Gln-Gly-Ala-Hyp-Gly-Pro-Gln-Gly-Pro-Arg.
2 detection methods
(1) preparation of improvement on synthesis control sample enzymolysis solution
Get 1.0g and do not add 1%NH containing the celestial oral liquid sample of tortoise deer two of colla carapacis et plastri testudinis
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/z569.1 → 724.4,795.4,852.4 as detecting ion pair.
The results are shown in Figure 3,4.06min place improvement on synthesis control sample, colla carapacis et plastri testudinis and the celestial oral liquid of tortoise deer two containing colla carapacis et plastri testudinis detect corresponding quasi-molecular ions, other do not detect, this method visible can turtle-derived component in specific detection colla carapacis et plastri testudinis goods, and comprises donkey, horse, ox, pig, deer etc. with other various animal derived materials and distinguish.
Sequence table
<110> Dong-E donkey-hide Gelatin Co., Ltd., Shandong Prov.
The detection method <130> of turtle-derived component in <120> glue class Chinese medicine and goods thereof
<160>1
<170>PatentInversion3.3
<210>1
<211>12
<212>PRT
<213> artificial sequence
<400>1
GlyValGlnGlyAlaHypGlyProGlnGlyProArg
1510。
Claims (9)
1. the detection method of turtle-derived component in glue class Chinese medicine and goods thereof, it utilizes has specific amino acid in tortoise genome and detects, and described specific amino acid is as shown in SEQ.IDNo1; Described detection method is LC-MS detection method.
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, described glue class Chinese herbal product is selected from food, health products or the medicine be made up of glue class Chinese medicine.
4. the method according to claims 1 to 3 any one, it specifically comprises the steps:
(1) choose specific amino acid in tortoise genome, described specific amino acid is as shown in SEQ.IDN01;
(2) glue class Chinese medicine or its goods are dissolved, or extract protein and peptide component dissolves wherein, add restriction enzyme and carry out enzyme and cut;
(3) LC-MS instrument is put into subsequently, with negative sample to be detected for matrix, add the polypeptide with step (1) described amino acid sequence or colla carapacis et plastri testudinis sterling in contrast, select the parent ion m/z569.1 of this contrast and daughter ion thereof to monitor.
5. method according to claim 4, is characterized in that, 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.
6. detect a molecular marked compound for turtle-derived component in glue class Chinese medicine and goods thereof, the amino acid sequence of this molecular marked compound is as shown in SEQ.IDNo1.
7. molecular marked compound according to claim 6 is detecting the application of turtle-derived component in glue class Chinese medicine and goods thereof.
8. a pharmaceutical composition, is characterized in that, comprises molecular marked compound according to claim 6 and one or more pharmaceutically acceptable carriers of pharmacy effective dose.
9. pharmaceutical composition according to claim 8 is detecting the application of turtle-derived component in glue class Chinese medicine and goods thereof.
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| CN102199192A (en) * | 2011-03-29 | 2011-09-28 | 中国科学院过程工程研究所 | Method for separating functional polypeptide from tortoise-shell glue |
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