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LC-MS characteristic chromatogram of leech enzymolysis polypeptide, detection method and application of LC-MS characteristic chromatogram

A technology of LC-MS and feature map, applied in measurement devices, instruments, scientific instruments, etc., can solve the problems of difficulty in guaranteeing the clinical efficacy of leech, quality control discount of leech medicinal materials, etc., to achieve good specificity and efficacy correlation, solving The effect of basic identification and authenticity identification to ensure clinical efficacy

Pending Publication Date: 2022-01-28
车延柠 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These leech fingerprint analysis methods provide a method for evaluating the quality of leech medicinal materials and their preparations from the perspective of material basis. However, none of them involve protein and polypeptide components, which greatly reduces the quality control of leech medicinal materials and makes it difficult to guarantee the clinical efficacy of leeches.

Method used

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  • LC-MS characteristic chromatogram of leech enzymolysis polypeptide, detection method and application of LC-MS characteristic chromatogram
  • LC-MS characteristic chromatogram of leech enzymolysis polypeptide, detection method and application of LC-MS characteristic chromatogram
  • LC-MS characteristic chromatogram of leech enzymolysis polypeptide, detection method and application of LC-MS characteristic chromatogram

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] 1 Experimental materials

[0039] 1.1 Experimental reagents

[0040] Acetonitrile, mass spectrometry grade, American Fisher Company; formic acid, chromatography grade, China Kemel Chemical Reagent Company; methanol, chromatography grade, American Fisher Company; concentrated hydrochloric acid, analytical grade, China Kemel Chemical Reagent Company; pepsin, analytical grade , Solarbio Life Sciences.

[0041] Leech, provided by Harbin No.4 Traditional Chinese Medicine Factory Co., Ltd., is the dry whole body of Hirudo nippocicaWhitman, a leech family animal. The leech, purchased from the medical material market, is the dried whole body of the leeches Whitmania pigra Whitman.

[0042] 1.2 Experimental equipment

[0043] Ultra-high performance liquid chromatography (Acquity TM UPLC, Waters, USA); mass spectrometer (Synapt TM G2-SiHDMS, Waters, USA); electrothermal constant temperature speed-adjustable oscillator (SYC-A, Shanghai Xinmiao Medical Instrument Manufacturing ...

Embodiment 2

[0068] The difference between this embodiment and embodiment 1 is only:

[0069] Take 1.0g of leech fine powder, weigh it accurately, add 20mL of artificial gastric juice, shake and extract in a constant temperature water bath at 37°C for 40min, add 1% pepsin, shake well, continue to shake in a constant temperature water bath at 37°C for 1.5h, and immediately place at 90 Shake in a constant temperature water bath at ℃ for 20 minutes, then cool to room temperature with running water, centrifuge, add an equal volume of methanol to the supernatant, shake well, centrifuge, take the supernatant, and obtain the test solution.

[0070] The preparation method of the test solution of leech fine powder is the same as above.

[0071] In this embodiment, in the chromatographic conditions, the flow rate of the mobile phase is 0.5 mL / min.

Embodiment 3

[0073] The difference between this embodiment and embodiment 1 is only:

[0074] Take 1.0g of 50-mesh leech fine powder, weigh it accurately, add 20mL of artificial gastric juice, shake and extract in a 37°C constant temperature water bath for 30min, add 1% pepsin, shake well, continue to shake in a 30°C constant temperature water bath for 1h, and immediately place Shake in a constant temperature water bath at 80°C for 10 minutes, then cool to room temperature with running water, centrifuge, add an equal volume of methanol to the supernatant, shake well, centrifuge, take the supernatant, and obtain the test solution;

[0075] The preparation method of the test solution of leech fine powder is the same as above.

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Abstract

The invention discloses an LC-MS characteristic chromatogram of leech enzymolysis polypeptide. The LC-MS characteristic chromatogram comprises 52 polypeptide component chromatographic peaks. The invention further discloses a detection method of the LC-MS characteristic chromatogram of the leech enzymolysis polypeptide, application of the detection method in quality control of leech medicinal materials and leech-containing preparations and application of the detection method in leech identification. According to the method, the quality of the medicinal material is controlled through the overall characterization of the leech enzymolysis polypeptide component, so that the method has relatively high accuracy and specificity; according to the method, the characteristic chromatogram for characterizing the leech enzymolysis polypeptide component is established, better specificity and efficacy correlation can be achieved, the clinical curative effect of the leech medicinal material can be guaranteed, and compared with other leech original animals, the characteristic chromatogram can effectively distinguish two leech original animals, therefore, the problems of primitive identification and authenticity identification of the leech medicinal material are effectively solved, and an accurate evaluation method is provided for identification of the leech medicinal material.

Description

technical field [0001] The invention relates to an LC-MS characteristic spectrum of leech enzymatically hydrolyzed polypeptides, a detection method and an application thereof, and belongs to the technical field of quality control of traditional Chinese medicines. Background technique [0002] The traditional Chinese medicine leech was first recorded in "Shen Nong's Materia Medica". The 2020 edition of "Chinese Pharmacopoeia" stipulates that leeches are the dried whole body of leech Whitmania pigra Whitman, Hirudo nipponica Whitman or Willow leech Whitmania acranulata Whitman. The "Xinxiu Materia Medica" of the Tang Dynasty records: "The big one is long-footed, named horse leeches, and called horse beetle, and can suck the blood of cows, horses, and people; today's custom is to take the small ones in the water and use them. It has great effects and does not need to be eaten. A man full of blood." The Song Dynasty "Materia Medica" records: "The leech has a long ruler, and it ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/72
CPCG01N30/02G01N30/72G01N2030/027
Inventor 王喜军宋宏伟闫广利车延柠
Owner 车延柠
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