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Common characteristic polypeptide of horse and mule and application thereof

A common characteristic and horse-derived technology, which is applied to the common characteristic peptides of horses and mules and its application fields, can solve the problems of no peptides, no specificity of peptides, and failure to find characteristic peptides, etc., achieving high sensitivity, simple operation, strong characteristic effect

Active Publication Date: 2016-12-07
SHANDONG INST FOR FOOD & DRUG CONTROL
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to incomplete digestion, the detected characteristic peptides may not be completely digested peptides; in addition, due to the limitation of mass spectrometry scanning conditions, it is impossible to scan all the digested peptides
In this way, the characteristic peptides found may not be specific or the true characteristic peptides may not be found

Method used

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  • Common characteristic polypeptide of horse and mule and application thereof
  • Common characteristic polypeptide of horse and mule and application thereof
  • Common characteristic polypeptide of horse and mule and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Screening and determination of characteristic peptides shared by horses and mules:

[0034] Collagen is the main protein in animal dermis, so the present invention selects suitable polypeptides from collagen as characteristic polypeptides of horse skin collagen. Use the PeptideMass function provided by Uniport to obtain the results of trypsin enzymatic hydrolysis of donkey hide, horsehide, cowhide, sheepskin, and pigskin collagen; use the molecular biology software MEGA5. By comparing the sequences of the proteins, theoretically two horse-specific peptides can be obtained with respect to donkeys, cattle, sheep, and pigs. The sequences are GASGPAGVR and GPSGEPGKPGDK, respectively. In addition, because mules have the gene of horse collagen, and mule skin also contains horse collagen, GASGPAGVR and GPSGEPGKPGDK are theoretically the characteristic peptides shared by horse skin collagen and mule skin collagen.

[0035] In order to verify the characteristic peptides GASGPAG...

Embodiment 2

[0041] Multiple Reaction Monitoring (MRM) Detection of Equine and Mule-Derived Components in Leather Samples

[0042] Donkey hide, horse hide, mule hide, cowhide, sheepskin, and pigskin were selected, according to the sample pretreatment method and HPLC-TQ-S MS / MS analysis method described above. The detection results are shown in Table 2, indicating that only the characteristic peptides GASGPAGVR and GPSGEPGKPGDK can be detected in horse hide and mule hide.

[0043] Table 2 Detection results of characteristic peptides in skin samples

[0044] characteristic peptide

[0045] "+" means detected; "-" means not detected

Embodiment 3

[0047] Multiple Reaction Monitoring (MRM) Detection of Horse- and Mule-Derived Components in Pure Gum

[0048] Colloidal traditional Chinese medicine is made by boiling animal skin at high temperature for a long time. In the process of boiling at high temperature, collagen inevitably participates in a series of reactions, which leads to the denaturation and hydrolysis of collagen. In order to verify the present invention To detect the feasibility of horse-derived and mule-derived components in rubber samples, donkey skin, horse hide, mule hide, cowhide, sheepskin, and pig skin were boiled to make glue. According to the gel sample pretreatment method and HPLC-TQ-S MS / MS analysis method described above. The detection results are shown in Table 3, indicating that only the characteristic peptides GASGPAGVR and GPSGEPGKPGDK can be detected in horse hide glue and mule hide glue.

[0049] Table 3 Detection results of characteristic peptides in gel samples

[0050] characte...

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Abstract

The invention discloses a common characteristic polypeptide of horse and mule and application thereof in horse-sourced and mule-sourced ingredient detection of hide type and gum type traditional Chinese medicines. The characteristic polypeptide has an amino acid sequence as shown in GASGPAGVR and GPSGEPGKPGDK, and is specific to horse hide and mule hide collagen. Furthermore, the invention provides a method for identifying horse hide and mule hide. The method comprises the steps of treating the hide with trypsase, and detecting the characteristic polypeptide by adopting liquid chromatography-tandem mass spectrometry, so that whether the hide is horse hide or mule hide can be identified. The invention further provides a method for detecting horse-sourced and mule-sourced ingredients in gum-type traditional Chinese medicines. The method comprises the steps of digesting a gum-type traditional Chinese medicine with trypsase so that the characteristic polypeptide is free from the collagen protein, and detecting by adopting chromatography-tandem mass spectrometry, so that whether the horse-source ingredient or the mule-sourced ingredient is doped in the gum-type traditional Chinese medicine can be determined. The method has the advantages of strong characteristics, high sensitivity, simplicity in operation and the like, and can be used for accurately identifying whether the horse-sourced ingredient or the mule-sourced ingredient is contained in the gum-type traditional Chinese medicine.

Description

(1) Technical field [0001] The invention relates to the field of detection and application, in particular to a characteristic polypeptide shared by horses and mules and its application. (2) Background technology [0002] Donkey-hide gelatin is made from donkey skin, which has multiple functions such as nourishing yin and nourishing blood. It is very popular among consumers. At present, the annual output value of donkey-hide gelatin and related medicines, food, health food and other products has exceeded 30 billion. It is reported that the current production of donkey-hide gelatin in the market far exceeds the amount of donkey hides in stock. Even with the addition of an increasing amount of imported donkey-hide, it cannot meet the production needs of donkey-hide gelatin. Under the lure of the huge profit space of 10 to 20 times the price difference between donkey hide and miscellaneous hide, unscrupulous enterprises took risks and used low-priced horse hide and mule hide raw...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06G01N30/72
CPCG01N30/02G01N30/06G01N30/72
Inventor 石峰巩丽萍李雪杭宝建咸瑞卿迟连利
Owner SHANDONG INST FOR FOOD & DRUG CONTROL
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