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Detection method of gelatin medicinal material

A technology of glue-like medicinal materials and detection methods, which is applied in the directions of measuring devices, material separation, and analysis of materials, can solve the problems of difficulty in identifying the source of animal skins, affecting the safety of diet and medication of the general public, and lack of technical support for identification, etc. To achieve the effect of high sensitivity, strong specificity and simple operation

Active Publication Date: 2020-12-18
NANJING UNIVERSITY OF TRADITIONAL CHINESE MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In the existing market, there are counterfeit gelatin medicinal materials such as fake donkey-hide gelatin and fake deerskin glue, and they are sold to the market as inferior ones, seriously affecting the food and drug safety of the broad masses of the people.
At present, unscrupulous companies often use horsehide, cowhide, pigskin, etc. to prepare donkey-hide gelatin, deerskin glue, antler glue, etc. as shoddy products, which leads to one of the reasons why the counterfeiting of precious rubber medicinal materials has been banned repeatedly. The lack of simple, feasible, Supported by accurate identification technology, after the animal skin is boiled, it is difficult to identify the source of the animal skin

Method used

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  • Detection method of gelatin medicinal material
  • Detection method of gelatin medicinal material
  • Detection method of gelatin medicinal material

Examples

Experimental program
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Effect test

Embodiment 1

[0060] Take about 10 mg of deer, donkey, horse, cow and pig respectively, add 5 ml of phosphate buffer solution, dissolve the sample completely by ultrasonication, centrifuge at 12000 rpm for 20 min, take 150 μl of the supernatant, put it in a 2 ml centrifuge tube, and dissolve it with 1 ml of 50 mM PBS Dilute, add 1% trypsin (w / v), shake well, enzymolyze at 37°C for 12 hours, add 60μl of 10% TFA solution after enzymolysis to terminate the reaction, centrifuge at 12000rpm for 20min, and obtain deer, donkey, horse, cow and pig enzymes solution and stored at -20°C.

[0061] The deer, donkey, horse, cattle and pig enzymatic hydrolyzate were injected into the liquid chromatography-mass spectrometer in sequence, and the injection volume was 1 μg. μm×100mm), flow rate 0.3ml / min, mobile phase A (acetonitrile), mobile phase B (0.1% formic acid), 0~5min, 5~25%A linear gradient elution, 5~6min, 25~5%A Linear gradient elution, 6 ~ 7min, 5% A elution. The mass spectrometry conditions de...

Embodiment 2

[0066] Embodiment 2 Deerskin Glue Authenticity Identification and Doping Situation

[0067] Take 4 batches of commercially available buckskin glue samples, about 10 mg for each batch, add 5 ml of phosphate buffer, ultrasonically dissolve the samples completely, centrifuge at 12000 rpm for 20 min, take 150 μl of the supernatant, and put it in a 2 ml centrifuge tube. Dilute with 1ml of 50mMPBS, add 1% trypsin (w / v), shake well, enzymolyze at a constant temperature of 37°C for 12 hours, add 60μl of 10% TFA solution after enzymolysis to stop the reaction, centrifuge at 12000rpm for 20min, and obtain the enzymatic hydrolyzate of buckhide glue , stored at -20°C for later use.

[0068] The deerskin gum enzymatic hydrolysis solution was injected into the LC-MS instrument, the injection volume was 1 μg, and the liquid phase conditions detected by the LC-MS instrument were: the chromatographic column was a 1.7 μm C18 reverse-phase chromatographic column (2.1 μm×100 mm), and the flow rat...

Embodiment 3

[0086] Embodiment 3 donkey-hide gelatin authenticity identification and doping situation

[0087] Take 6 batches of commercially available donkey-hide gelatin samples, about 10 mg for each batch, add 5 ml of phosphate buffer, ultrasonically dissolve the samples completely, centrifuge at 12000 rpm for 20 min, take 150 μl of the supernatant, put it in a 2 ml centrifuge tube, and dilute it with 1 ml Dilute with 50mM PBS, add 1% trypsin (w / v), shake well, ultrasonically hydrolyze for 10min, add 60μl of 10% TFA solution after enzymolysis to terminate the reaction, and centrifuge at 12000rpm for 20min to obtain the enzymatic hydrolyzate of donkey-hide gelatin. Store at 20°C for later use.

[0088] The donkey-hide gelatin enzymatic solution was injected into the LC-MS instrument, and the injection volume was 1 μg. The liquid phase conditions detected by the LC-MS instrument were: the chromatographic column was a 1.7 μm C18 reverse-phase chromatographic column (2.1 μm×100mm), and the ...

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Abstract

The invention discloses a detection method of gelatin medicinal materials. According to the detection method, trypsin is adopted for enzyme digestion of the gelatin medicinal material, so that specific polypeptide is released from collagen; then a liquid chromatography triple quadrupole mass spectrometer is adopted for detection, and if it is detected that the sample is corresponded to the specific polypeptide retentiontime, the parent ions and the daughter ions, determining that the species source of the gelatin medicinal material sample is consistent with the species corresponding to the specific polypeptide; and determining the adulteration proportion of the doped sample according to the peak area ratio of the specific polypeptide. The method has the advantages of strong specificity, high sensitivity, simple operation and the like, and can be used for identification and quality control of the gelatin medicinal materials.

Description

technical field [0001] The invention relates to a detection method for gum medicinal materials, in particular to the authenticity, adulteration sample detection and adulteration ratio of gum medicinal materials. Background technique [0002] Glue medicinal materials include donkey-hide gelatin, buckskin glue, antler glue, yellow gelatin, etc., and more than 80% of the ingredients are different types of collagen, including type I collagen α1 chain (COL1A1), type I collagen α2 chain (COL1A2), type II collagen Collagen α1 chain (COL2A1), type III collagen α1 chain (COL3A1), etc., mainly peptides derived from COL1A2. As a highly conserved protein, COL1A2 widely exists in different animal species, and is one of the important protein components that constitute gum medicinal materials. [0003] Fake donkey-hide gelatin, fake buckskin glue and other glue-like medicines and counterfeit products exist in the existing market, and they are sold as inferior ones to the market, seriously...

Claims

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Application Information

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IPC IPC(8): G01N30/88
CPCG01N30/88G01N2030/8868
Inventor 蔡朔刘睿蒋梦彤赵珂璇段金廒
Owner NANJING UNIVERSITY OF TRADITIONAL CHINESE MEDICINE
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