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Method for quick screening angiotemsin invertase inhibitor combined using high effieient liquid chromatograph and mass spectrum

A high-performance liquid chromatography and angiotensin technology, applied in the field of analytical chemistry, can solve problems such as long time, and achieve simple operation, accurate and reliable results

Inactive Publication Date: 2008-11-19
HUNAN NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the high-performance liquid chromatography method developed on its basis, the reaction time of the substrate and angiotensin-converting enzyme water bath is also more than 30 minutes, but because the high-performance liquid chromatography method can completely separate the substrate and the product, the results are generally better than those of the spectroscopic method. The photometric method is accurate, but the HPLC method takes a long time. Generally, the total detection time of a single sample takes 45 to 95 minutes

Method used

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  • Method for quick screening angiotemsin invertase inhibitor combined using high effieient liquid chromatograph and mass spectrum
  • Method for quick screening angiotemsin invertase inhibitor combined using high effieient liquid chromatograph and mass spectrum
  • Method for quick screening angiotemsin invertase inhibitor combined using high effieient liquid chromatograph and mass spectrum

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] (1) Preparation solution: buffer solution A: buffer solution (pH=8.3) of 300 mmol / liter sodium chloride and 100 mmol / liter tris and formic acid;

[0036] Buffer solution B: a buffer solution of 300 mmol / L sodium chloride and 100 mmol / L Tris and formic acid (pH=8.0).

[0037] Use buffer solution B to prepare a 4-8 mmol / L hippuric acid diglyglyptide solution, and use buffer A to prepare a 4-8 mmol / L hippuric acid histidine-leucine solution.

[0038] Prepare purified rabbit pulmonary angiotensin-converting enzyme solutions with buffer solutions A and B, respectively.

[0039] (2) Experiment:

[0040] 2 mmol / L Glypeptide hippurate substrate reacted with blank plasma, the total reaction volume was 100 μl

[0041] Control group: 40 microliters of 4 mmol / L diglyglycerin hippurate+20 microliters (pH=8.0) of buffer solution B+40 microliters of blank plasma

[0042] Test group: 40 microliters of 4 mmol / L hippurate + 20 microliters of 0.8 ng / ml Benazeprilat solution (angiotensi...

Embodiment 2

[0081] 2 mmol / L hippurate substrate and purified rabbit pulmonary angiotensin-converting enzyme (2 mU) were reacted in a total reaction volume of 80 μl

[0082] Control group: 40 microliters of 4 mmol / L hippuric acid diglycyl peptide + 20 microliters of buffer solution B + 20 microliters of angiotensin converting enzyme (2 milliU)

[0083] Experimental group: 40 microliters of 4 mmol / L hippuric acid diglycyl peptide + 20 microliters of 0.8 ng / ml benazeprilat solution + 20 microliters of angiotensin-converting enzyme (2 milliliters U) in a constant temperature water bath at 37°C After reacting in the pot for 8 minutes, add 160 microliters of acetonitrile solution containing internal standard terephthalic acid, vortex and shake for a few seconds, filter with a 0.45 micron water membrane, and analyze according to the steps of Example 1. The result is as figure 2 shown.

Embodiment 3

[0085] 2 mmol / L hippurate histidine leucine substrate reacted with blank plasma, total volume 100 µl

[0086] Control group: 40 μl 4 mmol / L hippuric acid histidine leucine + 20 μl buffer solution B + 40 μl blank plasma

[0087] Test group: 40 μl 4 mmol / L hippurate histidine leucine + 20 μl 0.8 ng / ml Benazeprilat solution + 40 μl blank plasma

[0088] After reacting in a constant temperature water bath at 37° C. for 16 minutes, add 200 microliters of acetonitrile plus internal standard terephthalic acid, centrifuge at 10,000 rpm for 8 minutes, and analyze the supernatant according to the steps in Example 1. The result is as image 3 shown.

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Abstract

A method of using high efficiency chromatograph and mass spectrum unitedly to quickly screen inhibitor of angiotonin converzyme uses hippuric acid bialkyd as substrate and uses purified rabbit lung angiotonin converzyme as source of angiotonin converzyme for carrying out quick analysis on vitality of angiotonin converzyme in human blood plasma.

Description

technical field [0001] The present invention relates to the field of analytical chemistry, in particular to an angiotensin-converting enzyme inhibitor capable of analyzing natural product extracts, monomers and marketed angiotensin-converting enzyme inhibitors in vitro, and human plasma angiotensin High performance liquid chromatography and mass spectrometry analysis method of invertase activity. Background technique [0002] In recent years, angiotensin inhibitors have become one of the drugs of choice for the treatment of hypertension. Many marketed angiotensin inhibitors, such as belazapril, captopril, enalazapril, etc., have good curative effects on hypertensive diseases. However, some of these angiotensin inhibitors can also cause side effects such as coughing, taste disturbance, blotchy skin, and more. In order to find better and safer angiotensin inhibitors, researchers began to turn to natural products, which aroused great interest from colleagues. Existing method...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02C12Q1/00
Inventor 陈波肖晓峰姚守拙
Owner HUNAN NORMAL UNIVERSITY
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