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Method of simultaneously determining contents of 1-OHP ,3-OHB[a]P and 3-OHB[a]A in urine

A 3-OHB, 1-OHP technology, applied in the field of biochemical testing, can solve problems such as less biological matrix, achieve the effect of high accuracy, good sensitivity and repeatability, and eliminate errors

Inactive Publication Date: 2013-04-24
CHINA NAT TOBACCO QUALITY SUPERVISION & TEST CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Using LC / MS with atmospheric pressure chemical ionization (APCI) or electrospray ionization (ESI) can also achieve the determination of various PAH metabolites, but it is rarely used in biological matrices

Method used

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  • Method of simultaneously determining contents of 1-OHP ,3-OHB[a]P and 3-OHB[a]A in urine
  • Method of simultaneously determining contents of 1-OHP ,3-OHB[a]P and 3-OHB[a]A in urine
  • Method of simultaneously determining contents of 1-OHP ,3-OHB[a]P and 3-OHB[a]A in urine

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Experimental program
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Effect test

experiment example 1

[0041] 1. Instruments and reagents: Agilent 1200 ultra-high resolution liquid chromatography (autosampler G1367D, binary solvent pump G1312B, column oven G1316B) (Agilent Technologies, Palo Alto, CA), API5500 triple quadrupole tandem mass spectrometry system (Applied Biosystems, Foster City, CA), data acquisition and processing were implemented on Analyst1.5.1 software (Applied Biosystems).

[0042]1-OHP was purchased from Dr. Ehrenstorfer GmbH (Augsburg, Germany, purity 98.5%), 3-OHB[a]P was purchased from Institute for Reference Materials and Measurements (Belgium, European Commission, purity 99.4%), 3-OHB[a] ]A was purchased from MRIGlobal (Kansas City, Missouri, purity>97%) and internal standard D 9 - 1-OHP was purchased from Toronto Research Chemicals Inc. (Toronto, Canada, chemical purity 97%, isotopic purity 99%). β-glucosidase / arylsulfatase was purchased from Merck, Germany, and HPLC-grade acetonitrile was purchased from TEDIA Company Inc. (Ohio, USA).

[0043] 2. ...

Embodiment 2

[0068] As described in Example 1, urine samples from 20 smokers and 20 non-smokers were selected. The average content levels of 1-OHP, 3-OHBaP and 3-OHBaA in the urine of all volunteers were 212.40ng / 24h, 0.07ng / 24h and 0.64ng / 24h; The average levels of 3-OHBaA and 3-OHBaA were 290.28ng / 24h, 0.08ng / 24h and 0.73ng / 24h, which were significantly higher than those in the urine of non-smokers (1-OHP: 134.53ng / 24h, 3-OHBaP: 134.53ng / 24h, 3-OHBaP: 0.06ng / 24h, 3-OHBaA: 0.55ng / 24h, Table 4).

[0069] Table 4 The levels of 1-OHP, 3-OHBaP and 3-OHBaA in the urine of smokers and non-smokers

[0070]

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Abstract

The invention discloses a method of simultaneously determining contents of 1-OHP ,3-OHB[a]P and 3-OHB[a]A in urine. The method comprises sequentially using beta-glucuronidase for enzymatic hydrolysis of samples, performing solid-phase extraction and purification, concentrating the samples with a vacuum rotary evaporator, and determining by using a liquid chromatography-tandem mass spectrometer, thereby rapidly, accurately and simultaneously detecting contents of 1-hydroxy pyrene (1-OHP), 3-hydroxy benzo [a] pyrene (3-OHB [a] P) and 3-hydroxy benzo [a] anthracene (3-OHB [a] A) in the urine. The method uses deuterated standards as quantitative analysis substances of internal standards and thus can reduce errors in a pretreatment process for the samples; uses a tandem mass spectrometer to relatively improve selectivity and accuracy of the method; and selects a method of preparing standards by matrices, wherein, compared with a method of preparing the standards by pure water, the method of preparing standards by matrices is relatively good in accuracy and can eliminate interference from matrix effects. Through selecting and optimizing chromatographic columns and gradient elution conditions, the method relatively improves a chromatogram separating process and shortens a chromatographic analysis time.

Description

technical field [0001] The invention relates to a method for simultaneously measuring the contents of 1-OHP, 3-OHB[a]P and 3-OHB[a]A in urine, belonging to the technical field of biochemical testing. Background technique [0002] Benzopyrene (B[a]P), benzanthracene (B[a]A) and pyrene are important polycyclic aromatic hydrocarbons (PAH) harmful components in cigarette smoke, and their metabolites produced in the human body— 3-Hydroxybenzopyrene (3-OHB[a]P), 3-hydroxybenzoanthracene (3-OHB[a]A) and 1-hydroxypyrene (1-OHP), available as B[a]P, Exposure biomarkers of B[a]A and other polycyclic aromatic hydrocarbons (PAH) compounds were used to differentiate populations at different exposure doses and predict possible mechanisms of action. [0003] So far, researchers have established methods for detecting PAH metabolites in urine, among which high-performance liquid chromatography-fluorescence detection (HPLC-FL) is the most common method. Due to the high sensitivity of fluore...

Claims

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Application Information

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IPC IPC(8): G01N30/88
Inventor 侯宏卫胡清源熊巍张小涛刘彤陈欢朱风鹏李中皓庞永强边照阳刘楠张洪非唐纲岭
Owner CHINA NAT TOBACCO QUALITY SUPERVISION & TEST CENT
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