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Deer glue characteristic peptide and method for identifying whether to-be-detected sample contains deer glue

A technology for characteristic peptides and deer glue, which is applied in the detection field of high-performance liquid phase-tandem mass spectrometry technology to identify the authenticity of deer glue and its products, can solve the research gap of mass spectrometry identification, the protein sequence library has no deer collagen sequence library, and the identification The method of deer glue is difficult to achieve the effect of expanding the scope of application and high sensitivity

Active Publication Date: 2021-06-22
贵州广济堂健康药业有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006]However, mass spectrometric identification of deer gum medicinal materials and their products is still blank, and there is no complete sequence library of deer collagen in the international protein sequence library, so it is impossible to carry out Complete Library Search
Therefore, it is difficult to establish a method for identifying deer glue based on mass spectrometry

Method used

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  • Deer glue characteristic peptide and method for identifying whether to-be-detected sample contains deer glue
  • Deer glue characteristic peptide and method for identifying whether to-be-detected sample contains deer glue
  • Deer glue characteristic peptide and method for identifying whether to-be-detected sample contains deer glue

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0058] (1') Preparation of deer gum contrast solution: get deer gum as contrast, after adding ammonium solution, through trypsin enzymolysis, C 18 Solid-phase extraction to obtain deer gum reference solution;

[0059] (2') Preparation of the sample solution to be tested: get the sample to be tested and prepare the sample solution to be tested in the same manner as step (1);

[0060] (3') The deer gum reference solution obtained in the step (1') and the sample solution to be tested obtained in (2') are subjected to multiple reaction monitoring by liquid chromatography-tandem mass spectrometry in electrospray negative ion mode , select m / z732.84→875.42, 818.40 and / or m / z675.99→914.46, 746.38 as detection transitions for detection;

[0061] (4') In the liquid chromatography-tandem mass spectrogram of the sample to be tested, if there is a mass spectrum peak consistent with the retention time of the deer glue contrast chromatogram, it is determined that the sample to be tested co...

Embodiment 1

[0067] This embodiment is the screening and identification of the characteristic peptides of deer glue, the specific method is as follows:

[0068] 1. Reagents and instruments

[0069]Sodium dodecyl sulfate (SDS), dithiothreitol (DTT), ammonium bicarbonate, trypsin (Promega mass spectrometry sequencing grade), acetonitrile, methanol, formic acid and other mass spectrometry reagents are mass spectrometry grade (Merck, Germany) .

[0070] Nanoscale liquid chromatography system: Eksigentekspert TM nanoLC liquid phase system, EksigentsoftwareV.3.12 liquid phase workstation. High-resolution mass spectrometry system: TripleTOF5600, equipped with electrospray ionization source (ESI), AnalystTF1.6 chromatographic workstation, Peak View1.2 mass spectrometry analysis software and ProteinPilot4.5 protein analysis software (AB Sciex, USA). Centrifugal concentrator (Labconco, USA); ZipTipC18 Pipette Tips (Millipore, USA).

[0071] 2. Samples of gum medicinal materials

[0072] Collect...

Embodiment 2

[0089] This example is based on the identification of deer glue and other glue-like medicinal materials based on characteristic peptides

[0090] 1. Reagents and instruments

[0091] Sodium dodecyl sulfate (SDS), dithiothreitol (DTT), ammonium bicarbonate, trypsin (Promega mass spectrometry sequencing grade), acetonitrile, methanol, formic acid and other mass spectrometry reagents are mass spectrometry grade (Merck, Germany) .

[0092] High-resolution fast liquid chromatography coupled with triple quadrupole mass spectrometer (AB Company, USA). Centrifugal concentrator (Labconco, USA); ZipTipC 18 Pipette Tips (Millipore, USA).

[0093] 2. Deer gum and other gum medicinal materials

[0094] Collect deer hide glue (deer hide glue), donkey hide glue (donkey-hide gelatin), cowhide glue (yellow gelatin), pig hide glue (new donkey-hide gelatin), horse hide glue, sheepskin glue and other glue-like medicinal materials, see Table 1.

[0095] The deer glue reference medicinal mater...

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Abstract

The invention discloses a deer glue characteristic peptide and a method for identifying whether a to-be-detected sample contains deer glue. When liquid chromatography-tandem mass spectrometry is adopted for multi-reaction detection in an electrospray anion mode, parent ions of the deer glue characteristic peptide are 732.84, and daughter ions of the deer glue characteristic peptide are 875.42 and 818.40; or the parent ions are 675.99, and the daughter ions are 914.46 and 746.38. The deer glue characteristic peptide can identify and identify deer glue medicinal materials, and a high performance liquid chromatography-tandem mass spectrometry rapid detection method (HPLC-MS / MS) with characteristic peptide fragment ion pairs as indexes can be established based on the deer glue characteristic peptide.

Description

[0001] This application is a divisional application with an application date of September 3, 2018, an application number of CN201811021204.8, an invention titled Deer Glue Characteristic Peptide and a method for identifying whether a sample to be tested contains Deer Glue. technical field [0002] The invention belongs to the technical field of medicine and food detection, and in particular relates to a detection technology for identifying the authenticity of deer glue and its products by using high performance liquid phase-tandem mass spectrometry technology. Background technique [0003] Deer glue (also known as deer skin glue) is a special Chinese herbal medicine made from the skin of red deer Cervus elaphus Linnaeus or sika deer Cervusnippon Temminck. "Compendium of Materia Medica" records that "deerskin is non-toxic and can cure all leaks and sores, leucorrhea in women, metrorrhagia, kidney deficiency and slippery essence". Since ancient times, deer gum has been a funct...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/08G01N30/06G01N30/88
CPCC07K7/08G01N30/06G01N30/88
Inventor 黄勇郑云枫徐浩坤刘睿谢宇
Owner 贵州广济堂健康药业有限公司