LC-MC method used for determining NNK metabolites in liver microsomes

A technology of liver microsomes and liquid mass spectrometry, applied in the field of biochemical analysis, can solve the problems of limited application, low peak shape quality, radioactive contamination, etc., and achieve high detection sensitivity, simple pretreatment, and fast analysis speed Effect

Active Publication Date: 2012-05-09
ZHENGZHOU TOBACCO RES INST OF CNTC
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  • Summary
  • Abstract
  • Description
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Problems solved by technology

The isotope radioactive labeling test needs to have corresponding safety protection measures and conditions. After each test or phased test, there may be different degrees of radioactive pollution and radioactive waste. It is necessary to face the problems of decontamination treatment and radioactive waste disposal. In recent years, the application has become increasingly limited
Moreover, the established HPLC method takes a long time to analyze, the quality of the peak shape is not high, and the separation effect is not good

Method used

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  • LC-MC method used for determining NNK metabolites in liver microsomes
  • LC-MC method used for determining NNK metabolites in liver microsomes
  • LC-MC method used for determining NNK metabolites in liver microsomes

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Embodiment 1

[0031] 1. Instruments and reagents:

[0032] Ultrapure water (resistivity ≥ 18.2 MΩ.cm); methanol (chromatographically pure, J&T Baker, USA); acetonitrile (chromatographically pure, J&T Baker, USA); formic acid (chromatographically pure, TEDIA, USA); ammonium formate (purity≥ 99%, Alfa Aesar Company); ammonium acetate (chromatographically pure, TEDIA Company, USA); 0.22 μm organic phase syringe filter (Agilent Company, USA).

[0033] 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK); 4-(methylnitrosamino)-1-(3-pyridyl)-1- Butanol (NNAL); 4-(methylnitroso)-1-(3-pyridyl-N-oxide)-1-butanone (NNK-N-Oxide); 4-(methylnitroso) -1-(3-pyridyl-N-oxidation)-1-butanol (NNAL-N-Oxide); 4-hydroxy-1-(3-pyridyl)-1-butanone (HPB); 1-( 3-pyridyl)-1,4-diol (PBD, hydroxy alcohol or diol); 1-(3-pyridyl)-1-butanone-4-carboxylic acid (keto acid); 1-(3- Pyridyl)-1-butanol-4-carboxylic acid (hydroxy acid, ammonium salt); 4-(methylnitrosamine)-1-(3-pyridyl-d 4 )-1-butanone (NNK-d 4 ); 4-Hydroxy-...

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Abstract

The invention relates to an LC-MC method used for determining NNK metabolites in liver microsomes and specifically to an LC-MC method used for analyzing and determining 4-(methyl nitroso)-1-(3-pyridyl)-1-butanone (NNK) metabolites in liver microsomes, which belongs to the field of biochemical analysis. The method provided in the invention mainly comprises the following steps: (1) preparation of an incubation system for liver microsomes; (2) sample treatment; (3) sample analysis and determination of NNK metabolites in the liver microsomes by using the LC-MC method. The invention has the following advantages of simple pre-treatment, a fast analysis speed, good selectivity, high detection sensitivity and capacity of analyzing and comparing the metabolism of NNK in different kinds of liver microsomes, can lay a methodological foundation for further research on in-vivo and in-vitro metabolism of NNK and other nitrosamine compounds and has critical significance.

Description

technical field [0001] The invention relates to the field of biochemical analysis, in particular to a liquid mass spectrometry method for analyzing and measuring 4-(methylnitroso)-1-(3-pyridyl)-1-butanone (NNK) metabolites in liver microsomes use method. Background technique [0002] NNK can induce a variety of cancers in experimental animals, and the International Organization for Research on Cancer (IARC) classifies it as a class of carcinogens (Smokeless tobacco and tobacco-specific nitrosamines. IARC monographs on the evaluation of carcinogenic risks to humans, vol. 89. Lyon (FR): IARC, 2007.). Biological experiments show that NNK is a strong carcinogen of rat lung cancer (Brunnemann K, Hoffmann D. Analytical studies on tobacco-specific N-nitrosamines in tobacco and tobacco smoke[J]. Critical reviews in toxicity, 1991, 21(4): 235 -240.). No matter how NNK is used to treat experimental animals, it can induce the occurrence of lung cancer in experimental animals, and mo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N27/62
Inventor 张建勋王娟卢斌斌王昇赵贝贝宗永立李鹏孙世豪
Owner ZHENGZHOU TOBACCO RES INST OF CNTC
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