High-sensitivity and high-accuracy method for simultaneously determining NNAL and cotinine in urine

A technology of cotinine and accuracy, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of cumbersome and time-consuming processing, reduce matrix effects, improve detection sensitivity and accuracy, and solve the problems of cumbersome pretreatment Effect

Active Publication Date: 2019-10-15
SICHUAN UNIV
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Problems solved by technology

[0004] The technical problem to be solved by the present invention is to provide a method for simultaneous determination of NNAL and cotinine in urine with high sensitivity and high accuracy, which effectively solves th...

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  • High-sensitivity and high-accuracy method for simultaneously determining NNAL and cotinine in urine
  • High-sensitivity and high-accuracy method for simultaneously determining NNAL and cotinine in urine
  • High-sensitivity and high-accuracy method for simultaneously determining NNAL and cotinine in urine

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Embodiment 1

[0051] Randomly collect 20 smokers' urine samples, and use the hydrophilic interaction chromatography tandem mass spectrometry of the present invention to analyze NNAL and cotinine in the urine, including the following steps.

[0052] (1) Select NNAL and cotinine as the characteristic metabolites of tobacco-specific nitrosamines (Tobacco-specific nitrosamines, TSNAs) and tobacco exposure; prepare a single standard stock solution containing the above two characteristic metabolites, containing the above two A single standard stock solution of the isotopic internal standard of the characteristic metabolites, and a mixed standard solution containing the above two characteristic metabolites and their corresponding isotopic internal standards;

[0053] Among them, NNAL standard (purity 95%), 13 C 6 - NNAL standard (purity 98%) was purchased from Toronto Research Chemicals, Canada. Cotinine standard solution (1.0g / L methanol solution) was purchased from U.S. Sigma-Aldrich company; ...

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Abstract

The invention discloses a high-sensitivity and high-accuracy method for simultaneously determining NNAL and cotinine in urine. The method comprises the following steps: 1) preparing a single standardstock solution containing the NNAL, the cotinine, isotope-labeled NNAL and isotope-labeled cotinine; 2) preparing mixed standard sample working solutions having a series of concentrations and containing the NNAL and the cotinine, and making a standard curve; 3) taking a to-be-determined urine sample, adding a phosphate buffer solution containing an isotope-labeled internal standard, and adding 15-25 [mu]L of beta-glucuronidase for enzymolysis; and 4) taking out the sample solution subjected to the enzymolysis, naturally cooling the sample solution into the room temperature, performing solid-phase extraction, performing instrument analysis, and performing calculation according to the standard curve made in the step 2) to obtain the content of the NNAL and the cotinine. The method effectively solves the problems of complex pretreatment and long consumed time of an existing method for simultaneously testing the NNAL and the cotinine in the urine; and meanwhile, the matrix effect in the urine is greatly reduced, so that the detection sensitivity and accuracy are improved.

Description

technical field [0001] The invention belongs to the field of detection of tobacco smoke exposure biomarkers, in particular to a method for simultaneously measuring NNAL and cotinine in urine with high sensitivity and accuracy. Background technique [0002] Tobacco use kills millions of people worldwide each year and poses a serious public health problem. Tobacco smoke contains more than 60 confirmed carcinogens. When evaluating the exposure and harm of these carcinogens, it is often necessary to select appropriate biomarkers for the intake of tobacco smoke, carcinogen intake and health effects. Wait for the evaluation. As an addictive alkaloid present in tobacco, nicotine is metabolized into cotinine in the liver within a short period of time, and finally a part of nicotine and cotinine are excreted with urine. Urinary cotinine is therefore a key biomarker for assessing exposure to environmental tobacco smoke. Tobacco-specific nitrosamines (TSNAs) are a class of nitrosami...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/06G01N30/72
CPCG01N30/02G01N30/06G01N30/72G01N2030/045
Inventor 邹晓莉袁悦王文佳杨明岐王焱郑波曾红燕
Owner SICHUAN UNIV
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