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Donkey characteristic polypeptide and application thereof to detection on donkey skin derived ingredients

A technology of origin ingredients and donkey skin, applied in the field of identification of animal species and animal origin ingredients, can solve the problems of limited differences, limited effective application, difficult to find, etc.

Active Publication Date: 2016-02-03
SHAN DONG DONG E E JIAO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The DNA molecular identification method is a very mature method for identifying animal DNA, but during the boiling process of glue, the DNA is severely damaged and degraded, and the remaining amount is very small, which greatly limits the effective application of this method in the identification of glue
The method for the determination of protein enzymatic peptides by liquid chromatography-mass spectrometry is very mature. As long as the characteristic peptides that are stable in animal-derived components are found, they can be used for detection; Although there are reports on detection methods for ingredients derived from donkey skin, ingredients derived from pig skin, ingredients derived from turtle skin, and ingredients derived from deer, they are actually common ingredients of donkey-hide gelatin and horse skin glue.
At present, there is still no detection method for the detection of donkey skin-derived components by liquid chromatography-mass spectrometry. Recently, their differences in biological essence are also very limited, so it is difficult to find characteristic peptides of donkeys for LC-MS detection of donkey skin-derived components

Method used

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  • Donkey characteristic polypeptide and application thereof to detection on donkey skin derived ingredients
  • Donkey characteristic polypeptide and application thereof to detection on donkey skin derived ingredients
  • Donkey characteristic polypeptide and application thereof to detection on donkey skin derived ingredients

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] The detection of donkey characteristic polypeptide in the various pure product glues of embodiment 1

[0018] 1. Materials and reagents

[0019] Materials: All kinds of pure traditional Chinese medicines, including donkey-hide gelatin, horsehide glue, yellow gelatin, pigskin glue, tortoise shell glue, antler glue, dog bone glue, sea dragon glue, donkey skin, horse hide, cowhide, pig skin, tortoise shell glue , Antlers, Dog Leg Bones, and Sea Dragons are obtained by decocting.

[0020] Reagents: ammonium bicarbonate (analytical pure), trypsin (serial pure, purchased from China Institute for the Control of Pharmaceutical and Biological Products).

[0021] 2. Detection method

[0022] (1) Take 0.05g of the rubber Chinese medicine sample to be tested in a 25ml measuring bottle, add a small amount of 1% (w / w) NH 4 HCO 3 Solution, sonicate to dissolve the sample completely, with 1% (w / w) NH 4 HCO 3 The solution (pH 8.0) was diluted to the mark, shaken well, filtered thr...

Embodiment 2

[0026] Example 2 Detection of Donkey Skin-derived Components in Glue Traditional Chinese Medicines

[0027] 1. Materials and reagents

[0028] Material: the mixed glue sample is made by adding 5% (w / w) donkey-hide gelatin respectively to the pure product glue sample in embodiment 1, including the horsehide glue containing 5% (w / w) donkey-hide gelatin, containing 5% (w / w) donkey-hide gelatin ) yellow gelatin of donkey-hide gelatin, pigskin glue of 5% (w / w) donkey-hide gelatin, tortoise shell glue containing 5% (w / w) donkey-hide gelatin, deer horn glue containing 5% (w / w) donkey-hide gelatin, / w) dog bone glue of donkey-hide gelatin, sea dragon gum containing 5% (w / w) donkey-hide gelatin.

[0029] Reagents: ammonium bicarbonate (analytical pure), trypsin (serial pure, purchased from China Institute for the Control of Pharmaceutical and Biological Products).

[0030] 2. Detection method

[0031] (1) Take 0.05g of the sample to be tested in a 25ml measuring bottle, add a small ...

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Abstract

The invention discloses donkey characteristic polypeptide and application thereof to detection on donkey skin derived ingredients. The amino acid sequence of the donkey characteristic polypeptide provided by the invention is shown as SEQ ID NO.1 and the polypeptide is specifically contained in collagen of a donkey. Moreover, the invention also provides a method for detecting donkey skin derived ingredients in glue traditional Chinese medicine, which comprises: carrying out enzyme digestion on the glue traditional Chinese medicine by adopting trypsin or arginine-C enzyme so as to enable the characteristic polypeptide to be dissociated from the collagen; then carrying out detection by adopting liquid chromatography-tandem mass spectrometry so as to judge whether the donkey skin derived ingredients are included in the glue traditional Chinese medicine. The method has the advantages of strong characteristic performance, high sensitivity, simplicity for operation and the like and can accurately authenticate whether the donkey skin derived ingredients are included in the glue traditional Chinese medicine.

Description

technical field [0001] The invention relates to a method for identifying animal species and animal-derived components, in particular to a detection method for donkey skin-derived components; in addition, the invention also relates to a characteristic polypeptide for detecting donkey skin-derived components. Background technique [0002] Ejiao, as a traditional precious Chinese medicine, has been eaten for thousands of years. It has the functions of nourishing blood, nourishing yin, moistening dryness and stopping bleeding. The Chinese Pharmacopoeia stipulates that donkey-hide gelatin is a solid gelatin made by decocting and concentrating the dried skin or fresh skin of the equine animal donkey Equusasinus L. Pure donkey-hide gelatin must come from 100% donkey skin to ensure the efficacy of the product. From the perspective of raw material taxonomy, donkeys belong to the genus Equus of the order Perissodactyla, including African wild asses, Tibetan wild asses, Central Asian ...

Claims

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Application Information

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IPC IPC(8): G01N30/89
Inventor 秦玉峰周祥山郝向慧郭尚伟段小波嵇传良田守生王静徐云鹏史兆松
Owner SHAN DONG DONG E E JIAO
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