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Method for detecting sheep-derived ingredients in glue type traditional Chinese medicines and products thereof

A technology of glue-like traditional Chinese medicine and detection method, which is applied in the direction of botany equipment and method, biochemical equipment and method, material separation, etc., to achieve the effect of small damage and high content

Active Publication Date: 2014-03-12
SHAN DONG DONG E E JIAO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But at present, there is no detection method for sheep-derived components in gelatinous traditional Chinese medicine and its products from the perspective of collagen peptides, nor from the perspective of DNA.

Method used

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  • Method for detecting sheep-derived ingredients in glue type traditional Chinese medicines and products thereof
  • Method for detecting sheep-derived ingredients in glue type traditional Chinese medicines and products thereof
  • Method for detecting sheep-derived ingredients in glue type traditional Chinese medicines and products thereof

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0028] (1) Preparation of gel sample enzymatic hydrolysis solution

[0029] Take 1.0g of the gel sample to be tested in a 500mL measuring bottle, add a small amount of 1%NH 4 HCO 3 solution, sonicate to dissolve the sample completely, and use 1% NH 4 HCO 3 Dilute the solution (pH 8.0) to the mark, shake well, filter through a microporous membrane, accurately measure 1 mL of the subsequent filtrate plus 100 μL of 2 mg / mL trypsin solution, and enzymatically hydrolyze at a constant temperature of 37 °C for 6 h.

[0030] (2) Preparation of enzymatic hydrolysis solution of synthetic polypeptide control sample

[0031] Weigh a certain amount of synthetic peptide, add 1% NH 4 HCO 3 Solution (pH8.0), shake well, prepare to a concentration of about 0.3μg / mL, filter through a microporous membrane, and set aside.

[0032] 3 Discovery of peptides

[0033] (1) Discovery of ions

[0034] Take 5 μL of the enzymatic hydrolysis solution and put it into the liquid mass spectrometer for ...

Embodiment 1

[0044] 1 Materials and Reagents

[0045] Materials: Pure glue includes donkey-hide gelatin, horsehide glue, yellow gelatin, pigskin glue, tortoise shell glue, antler glue, and sheepskin glue, which are obtained by decocting donkey hide, horse hide, cowhide, pig skin, tortoise shell, antler, and sheepskin respectively.

[0046] Mixed gum samples (donkey-hide gelatin with 5% sheepskin glue, 10% sheepskin glue, 20% sheepskin glue, and 40% sheepskin glue) are made by mixing the above pure glue .

[0047] Reagents: ammonium bicarbonate (analytical grade), trypsin (sequence grade, purchased from China National Institutes for Food and Drug Control), synthetic peptide Thr-Gly-Glu-Hyp-Gly-Ala-Ala-Gly-Pro-Hyp-Gly- Phe-Val-Gly-Glu-Lys.

[0048] 2 detection method

[0049] (1) Preparation of synthetic peptide control sample enzymatic hydrolysis solution

[0050] Get 1.0g pure donkey-hide gelatin sample in 500mL measuring bottle, add a small amount of 1%NH 4 HCO 3 solution, sonicate ...

Embodiment 2

[0057] 1 Materials and Reagents

[0058] Materials: Mixed glue samples are made by adding 5% sheepskin glue to the pure glue sample in Example 1, including donkey-hide gelatin containing 5% sheepskin glue, horsehide glue with 5% sheepskin glue, and yellow gelatin containing 5% sheepskin glue , Pigskin glue with 5% sheepskin glue, tortoise shell glue with 5% sheepskin glue, and deer antler glue with 5% sheepskin glue (percentages are mass fractions).

[0059] Reagents: ammonium bicarbonate (analytical grade), trypsin (sequence grade, purchased from China National Institutes for Food and Drug Control), synthetic peptide Thr-Gly-Glu-Hyp-Gly-Ala-Ala-Gly-Pro-Hyp-Gly- Phe-Val-Gly-Glu-Lys.

[0060] 2 detection method

[0061] (1) Preparation of synthetic peptide control sample enzymatic hydrolysis solution

[0062] Get 1.0g sample of pure donkey-hide gelatin to be tested in a 500mL measuring bottle, add a small amount of 1%NH 4 HCO 3 solution, sonicate to dissolve the sample com...

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Abstract

The invention provides a method for detecting sheep-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 of a specific nucleotide sequence or amino acid sequence in a genome of a sheep subfamily animal, wherein the specific amino acid sequence is shown in SEQ. ID No1. By adopting the method provided by the invention, whether the glue type traditional Chinese medicines contain the sheep-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 sheep-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 medicine are high in content and little in damaged extent.

Description

technical field [0001] The invention relates to a detection method of goat-derived components, in particular to a detection method of goat-derived components in gelatinous traditional Chinese medicines and products thereof, and belongs to the field of detection of traditional Chinese medicines. Background technique [0002] Glue-like traditional Chinese medicines are boiled from animal skins and bones to make glue-like substances, including donkey-hide gelatin, tortoise shell glue, antler glue, etc. Traditional Chinese medicine with typical ethnic characteristics is widely used in blood, gynecology, obstetrics, respiratory, tumor and other diseases prevention and treatment. [0003] With the continuous development of the glue-type traditional Chinese medicine market, the number of manufacturers producing glue-type traditional Chinese medicine is also increasing, and the production capacity and product quality are uneven. Taking donkey-hide gelatin as an example, because the...

Claims

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Application Information

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IPC IPC(8): G01N30/02C07K7/08C12N15/12
Inventor 郭尚伟秦玉峰周祥山尤金花田守生嵇传良段小波史兆松
Owner SHAN DONG DONG E E JIAO
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