Method for simultaneously detecting cotinine, phenyl hydroxyacetic acid and phenylglyoxalic acid in human urine based on derivation method

Abstract

The invention relates to the technical field of biological engineering, in particular to a method for detecting cotinine, mandelic acid and phenylglyoxalic acid in human urine simultaneously on the basis of a derivation method. The method is characterized in that the method comprises: step one, storing collected urine after centrifugal process; step two, taking the urine obtained in step one and adding the urine into internal label benzophenone, diphenylamine, hydrochloric acid and chloroform to carry out extraction through rotating centrifugation, thereby aspirating and recording organic phase as A and taking residual solution for standby; step three, taking the residual solution obtained in step two and adding the residual solution into sodium hydroxide solution and chloroform to carry out secondary extraction through rotating centrifugation, thereby aspirating and recording organic phase as B; step four, mixing the organic phase A extracted in step two and the organic phase B extracted in step three inside a glass tube, and then blowing dry both organic phases; step five, adding acetonitrile into the glass tube adopted in step four for redissolution and adding derivative reagent to carry out derivative reaction; and step six, taking the reaction solution obtained in step five to carry out gas phase chromatography-mass spectrometry combined measurement. The method has the characteristics of simplicity, convenience, quickness, reliable result, high degree of automation and convenient industrialized application.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a method for simultaneously detecting codnine, mandelic acid and phenylglyoxylic acid in human urine based on a derivatization method. Background technique [0002] Styrene acts on the P450 enzyme system, and its main metabolites, mandelic acid and phenylglyoxylic acid, can be used as biomarkers of the dose of styrene in humans. However, styrene is widely used in industry, which pollutes the atmosphere, water and soil. In daily life, people inhale styrene mainly due to the pollution of automobile exhaust. With the development of the polymer industry, the number of workers exposed to styrene is increasing. The chemical structure of styrene is similar to the mutagen and carcinogen vinyl chloride, and it is also a potential mutagen after metabolic activation, and its intermediate metabolite 7,8-epoxystyrene is a strong direct mutagen. Cotinine is the main metabolite of nico...

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Problems solved by technology

[0004] By consulting the literature, we found that there are many methods used by domestic scholars to study the environmental pollutants nicotine and styrene exposure metabolites, including GC, HPLC, GC-MS, thin-layer chromatography, enzyme-linked immunosorbent assay, etc. etc.; the types of instruments used in the method are also different. Taking GC-MS as an example, a combination of Shimadzu GC-14A and American Agilent 6890N-5973MSD are used; there is no special institution for environmental pollutants and The human exposure of its metabolites was tested

Most of the research is dedicated to the detection of single pollutants, and the methods used for the same substance are different, and the focus is on improving the recovery rate of pollutants, ignoring the comprehensiveness and comprehensiveness of the detection types, and the detection cost is high. It is difficult to systematically and comprehensively evaluate the degree of pollutant exposure of the human body, which is not conducive to the promotion of industrialization to the general population

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