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Method for detecting 4-hydroxyl-4-(3-pyridyl) butyric acid in urine

A technology based on pyridyl and hydroxyl, which is applied in the field of physical and chemical testing of tobacco alkaloids, and achieves the effects of good stability, low detection limit, and easy popularization and application

Active Publication Date: 2014-05-07
HONGYUN HONGHE TOBACCO (GRP) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

How to realize the accurate detection of two kinds of 4-hydroxyl-4-(3-pyridyl)butyric acid content in the urine sample simultaneously and carry out in-depth analysis to the ratio of the two, there is not yet a good solution in the prior art

Method used

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  • Method for detecting 4-hydroxyl-4-(3-pyridyl) butyric acid in urine
  • Method for detecting 4-hydroxyl-4-(3-pyridyl) butyric acid in urine
  • Method for detecting 4-hydroxyl-4-(3-pyridyl) butyric acid in urine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032]1. Chemical reagent preparation: (S)-(+)-α-methoxy-α-trifluoromethylphenylacetyl chloride (CAS No. 20445-33-4), (S)-(+)-α-- Methoxy-(trifluoromethyl)phenylacetic anhydride (CAS No. 85541-57-7), (±)-4-hydroxy-4-(3-pyridyl)butanoic acid (CAS No. 15569-97-8) , (±)-4-Hydroxy-4-(3-pyridyl)butanoic acid-d5, triethylamine, diazomethane, dimethyl sulfate.

[0033] 2. Derivatization of (±)-4-hydroxy-4-(3-pyridyl)butanoic acid and establishment of standard curve:

[0034] Using anhydrous acetonitrile as solvent, prepare (±) 4-hydroxy-4-(3-pyridyl) butyric acid standard single standard solution with a concentration of 1 μg / mL, keep four significant figures; use anhydrous acetonitrile in the same way Prepare a single standard solution of (±)-4-hydroxy-4-(3-pyridyl)butanoic acid-d5 with a concentration of 1 μg / mL as the solvent; use anhydrous acetonitrile as the solvent, prepare the solution containing (±) according to Table 2 - Mixed standard solution of 4-hydroxy-4-(3-pyridyl)but...

Embodiment 2

[0044] Repeat Example 1, with the following differences:

[0045] 1. Derivatization of (±)-4-hydroxy-4-(3-pyridyl)butanoic acid and establishment of standard curve

[0046] Using anhydrous acetonitrile as a solvent, prepare a single standard solution of (±)-4-hydroxy-4-(3-pyridyl)butyric acid standard with a concentration of 1 μg / mL, and keep four significant figures; Acetonitrile was used as the solvent to prepare a single standard solution of (±)-4-hydroxy-4-(3-pyridyl)butyric acid-d5 with a concentration of 1 μg / mL; anhydrous acetonitrile was used as the solvent to prepare the solution containing (±) )-4-hydroxy-4-(3-pyridyl)butanoic acid and deuterated (±)-4-hydroxy-4-(3-pyridyl)butyric acid-d5 mixed standard solution; the prepared concentration is 1μg / mL (S)-(+)-α-methoxy-α-trifluoromethylphenylacetic anhydride in anhydrous acetonitrile solution; prepare a solution of triethylamine in anhydrous acetonitrile with a concentration of 1 μg / mL; take 2 mL respectively Mix the...

Embodiment 3

[0050] Repeat Example 1, with the following differences:

[0051] 1. Derivatization of (±)-4-hydroxy-4-(3-pyridyl)butanoic acid and establishment of standard curve

[0052] Using anhydrous acetonitrile as a solvent, prepare a single standard solution of (±)-4-hydroxy-4-(3-pyridyl)butyric acid standard with a concentration of 1 μg / mL, and keep four significant figures; Acetonitrile was used as the solvent to prepare a single standard solution of (±)-4-hydroxy-4-(3-pyridyl)butyric acid-d5 with a concentration of 1 μg / mL; anhydrous acetonitrile was used as the solvent to prepare the solution containing (±) )-4-hydroxy-4-(3-pyridyl)butanoic acid and deuterated (±)-4-hydroxy-4-(3-pyridyl)butyric acid-d5 mixed standard solution; the prepared concentration is 1μg / mL Anhydrous acetonitrile solution of (S)-(+)-α-methoxy-α-trifluoromethylphenylacetyl chloride; prepare anhydrous acetonitrile solution of triethylamine with a concentration of 1 μg / mL; take 0.5 Put the mixed standard solu...

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Abstract

The invention discloses a method for detecting 4-hydroxyl-4-(3-pyridyl) butyric acid in urine. The method comprises the following steps: purifying a urine sample by using an Oasis MCX solid-phase extraction small column; carrying out methyl esterification on carboxyl in the sample and carrying out esterification on alcoholic hydroxyl; then carrying out HPLC-MS-MS (High Performance Liquid Chromatography-Mass Spectrometry-Mass Spectrometry) detection on a derived urine sample so as to calculate the concentrations of (S)-4-hydroxyl-4-(3-pyridyl) butyric acid and (R)-4-hydroxyl-4-(3-pyridyl) butyric acid in the urine sample; and calculating a ratio of the (S)-4-hydroxyl-4-(3-pyridyl) butyric acid to the (R)-4-hydroxyl-4-(3-pyridyl) butyric acid in the urine sample. The method can provide a scientific judgment method for estimating metabolism health conditions to nicotine, nornicotine, NNK (Nicotine-derived Nitrosamine Ketone) and NNAL ((4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol)) by an individual body, and evaluating the exposure degree to substances in smoke, has the advantages of low detection limit, good stability, rapidness and accuracy and the like, and fills the blank of the technical field.

Description

technical field [0001] The invention belongs to the technical field of physical and chemical testing of tobacco alkaloids, in particular to a method for detecting 4-hydroxy-4-(3-pyridyl)butyric acid in urine. Background technique [0002] In recent years, with people's widespread concern about smoking and health issues, the tobacco industry and health agencies at home and abroad urgently need to establish some methods to accurately evaluate the exposure of different groups of people to some chemical components in tobacco, and the biomarker method is a recognized method. and effective methods for monitoring exposure in various populations. At present, there have been some reports on the detection of various specific smoke chemical components in human urine biomarkers at home and abroad, but the number is still relatively small, and the purposes involved are relatively narrow. [0003] The research results show that there are more than 3,000 chemical substances in tobacco and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02
Inventor 蒋举兴者为王文元段焰青夏建军党立志陈兴
Owner HONGYUN HONGHE TOBACCO (GRP) CO LTD
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