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HILIC (hydrophilic interaction chromatography)-MRM (multiple-reaction monitoring) MS/MS (tandem mass spectrometry) detection method for basic constitutional units of low-molecular-weight heparin

一种亲水相互作用、低分子肝素的技术，应用在原料检测,药物,原料药领域，能够解决全面表征等问题，达到生产控制和保障药品安全的效果

Active Publication Date: 2016-10-12
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The non-reducing end and reducing end of enoxaparin sodium are the same as the newly generated end after enzymatic digestion, so it cannot be fully characterized using the original strategy

Method used

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  • HILIC (hydrophilic interaction chromatography)-MRM (multiple-reaction monitoring) MS/MS (tandem mass spectrometry) detection method for basic constitutional units of low-molecular-weight heparin
  • HILIC (hydrophilic interaction chromatography)-MRM (multiple-reaction monitoring) MS/MS (tandem mass spectrometry) detection method for basic constitutional units of low-molecular-weight heparin
  • HILIC (hydrophilic interaction chromatography)-MRM (multiple-reaction monitoring) MS/MS (tandem mass spectrometry) detection method for basic constitutional units of low-molecular-weight heparin

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Experimental program
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Effect test

Embodiment 1

[0029] A method for detecting the complete degradation product of low molecular weight heparin combined with hydrophilic interaction chromatography and multiple reaction monitoring secondary mass spectrometry, the steps are as follows:

[0030] (1) ammonium acetate reagent is dissolved in deionized water, and the obtained ammonium acetate concentration is the mobile phase A of 5mM;

[0031] (2) ammonium acetate reagent is dissolved in deionized water, and acetonitrile is added, and mobile phase B is obtained, and in mobile phase B, the concentration of ammonium acetate is 5mM, and the volume percentage of acetonitrile is 95%;

[0032] (3) The enoxaparin sodium standard was degraded with heparanase I, II and III (all purchased from Beijing Adhawk International Technology Co., Ltd.) at 25°C for 48 hours, and the degradation product added with the internal standard was degraded with Millipore 30KDa Vacuum decompression drying after ultrafiltration membrane filtration;

[0033] (...

Embodiment 2

[0048] A method for detecting the complete degradation product of low molecular weight heparin combined with hydrophilic interaction chromatography and multiple reaction monitoring secondary mass spectrometry, the steps are as follows:

[0049] (1) ammonium acetate reagent is dissolved in deionized water, and the obtained ammonium acetate concentration is the mobile phase A of 5mM;

[0050] (2) ammonium acetate reagent is dissolved in deionized water, and acetonitrile is added, and mobile phase B is obtained, and in mobile phase B, the concentration of ammonium acetate is 5mM, and the volume percentage of acetonitrile is 95%;

[0051] (3) The dalteparin sodium standard was degraded with heparanase I, II and III (all purchased from Beijing Adhawk International Technology Co., Ltd.) at 25°C for 48 hours, and the degradation product added with the internal standard was degraded with Millipore 30KDa Vacuum decompression drying after ultrafiltration membrane filtration;

[0052] (...

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Abstract

The invention relates to an HILIC (hydrophilic interaction chromatography)-MRM (multiple-reaction monitoring) MS / MS (tandem mass spectrometry) detection method for complete degradation products of low-molecular-weight heparin. Original reducing end and non-reducing end of enoxaparin sodium are identified by reducing the reducing end of enoxaparin sodium and adopting hydrolysis with hydrogen peroxide. Quantitative analysis is performed on all constitutional units with HILIC-MRM MS / MS, quantification is particularly performed on special structures with lower content, and characterization is performed on the low-molecular-weight heparin.

Description

technical field [0001] The invention relates to a detection method of a complete degradation product of low-molecular heparin combined with hydrophilic interaction chromatography and multiple reaction monitoring secondary mass spectrometry, and belongs to the technical field of medicine, raw material medicine and raw material detection. Background technique [0002] Heparin and its derivatives Low-molecular-weight heparin is a kind of glycosaminoglycan, which has anticoagulant effect and has been used as an anticoagulant drug in clinical practice. In order to reduce the risk of side effects such as hemorrhage, osteoporosis and thrombocytopenia caused by heparin, and improve its bioavailability, low molecular weight heparin is widely used as a new anticoagulant drug. Low-molecular-weight heparin is prepared by enzymatic degradation or chemical degradation. According to different processes, each low-molecular-weight heparin has its own special structure. Enoxaparin sodium is ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/72
CPCG01N30/02G01N30/72G01N2030/027G01N33/6848G01N2400/40G01N2030/8836B01D15/305G01N30/34G01N30/7233H01J49/004H01J49/4215
Inventor 迟连利孙晓君盛安然刘欣悦
Owner SHANDONG UNIV
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