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Gas chromatography-mass spectrography combined detection method for eight monohydric polyaromatic hydrocarbon in urine

A polycyclic aromatic hydrocarbon and detection method technology, applied in the field of physical and chemical inspection of biomarkers, can solve the problems of low sensitivity, cumbersome pretreatment, and high detection cost

Inactive Publication Date: 2013-06-26
HONGYUN HONGHE TOBACCO (GRP) CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] The purpose of the present invention is to provide a biomarker gas chromatography- The detection method coupled with mass spectrometry provides a scientific determination method for evaluating the exposure of individuals to polycyclic aromatic hydrocarbons. This method has the advantages of low detection limit, good stability, simple pretreatment, fast and accurate, etc. Detection of Biomarkers of PAH Exposure in Urine

Method used

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  • Gas chromatography-mass spectrography combined detection method for eight monohydric polyaromatic hydrocarbon in urine
  • Gas chromatography-mass spectrography combined detection method for eight monohydric polyaromatic hydrocarbon in urine
  • Gas chromatography-mass spectrography combined detection method for eight monohydric polyaromatic hydrocarbon in urine

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Embodiment 1

[0016] A gas chromatography-mass spectrometry detection method for monohydroxy polycyclic aromatic hydrocarbons in urine, comprising the following steps:

[0017] 1. Chemical reagents

[0018] 1-hydroxyphenanthrene, 2-hydroxyphenanthrene, 3-hydroxyphenanthrene, 4-hydroxyphenanthrene, 9-hydroxyphenanthrene, 1-hydroxypyrene, 3-hydroxybenzo[a]pyrene, 7-hydroxybenzo[a]pyrene, 2-hydroxyphenanthrene-d9, 1-hydroxypyrene-d9, 3-hydroxybenzo[a]pyrene-d11, 25% N,O-bistrimethylsilylacetamide solution in acetonitrile.

[0019] 2. Standard curve establishment

[0020] 8 kinds of standard solutions of 1-hydroxy polycyclic aromatic hydrocarbons: using acetonitrile as solvent, prepare 1-hydroxyphenanthrene, 2-hydroxyphenanthrene, 3-hydroxyphenanthrene, 4-hydroxyphenanthrene, 9-hydroxyphenanthrene, 1-hydroxyphenanthrene at a concentration of 1 μg / mL respectively Hydroxypyrene, 3-hydroxybenzo[a]pyrene, and 7-hydroxybenzo[a]pyrene are single standard solutions of 8 kinds of monohydroxy polycycl...

Embodiment 2

[0035] Actual sample testing:

[0036] Shake the urine sample after thawing at room temperature, accurately pipette 5 mL into a 25 mL test tube, and add 10 μL of 100 ng / mL 2-hydroxyphenanthrene-d9, 1-hydroxypyrene-d9 and 3 -Hydroxybenzo[a]pyrene-d11 three deuterated compound solution (converted to 1000pg / deuterated compound), then add 2 mL of sodium acetate buffer (pH5.5, 1M) and 10 μL of β-glucose Glycocuronidase / arylsulfatase proenzyme (364.25U / mL urine), mixed thoroughly, placed in a microwave-assisted extraction apparatus, and hydrolyzed at a constant temperature of 37 °C for 60 s. Extract the enzymatically hydrolyzed urine sample with 2*5 mL of n-hexane, combine the organic phases, add 10 μL of n-dodecane with a higher boiling point, filter after anhydrous drying, and concentrate to nearly dryness in a nitrogen atmosphere. The liquid is about 10 μL, add 30 μL toluene and shake for 1 minute, transfer it to a gas chromatography vial with a micro syringe, add 20 μL of 25% N...

Embodiment 3

[0038] Actual sample testing:

[0039] Shake the urine sample after thawing at room temperature, accurately pipette 5 mL into a 25 mL test tube, and add 10 μL of 100 ng / mL 2-hydroxyphenanthrene-d9, 1-hydroxypyrene-d9 and 3 -Hydroxybenzo[a]pyrene-d11 three deuterated compound solution (converted to 1000pg / deuterated compound), then add 2 mL of sodium acetate buffer (pH5.5, 1M) and 10 μL of β-glucose Glycocuronidase / arylsulfatase proenzyme (364.25U / mL urine), mixed thoroughly, placed in a microwave-assisted extraction apparatus, and hydrolyzed at a constant temperature of 37 °C for 60 s. Extract the enzymatically hydrolyzed urine sample with 2*5 mL of n-hexane, combine the organic phases, add 10 μL of n-undecane with a higher boiling point, filter after anhydrous drying, and concentrate to near dryness in a helium atmosphere. The remaining liquid is about 10 μL, add 30 μL toluene and shake for 1 minute, transfer it to a gas chromatography vial with a micro syringe, add 30 μL tr...

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Abstract

The invention discloses a gas chromatography-mass spectrography combined detection method for eight monohydric polyaromatic hydrocarbon in urine. The method comprises the following steps of: (1) respectively transforming the monohydric polyaromatic hydrocarbon and the deuterated monohydric polyaromatic hydrocarbon in a mixed standard solution into corresponding trimethylsiloxy polyaromatic hydrocarbon, carrying out GC-MS (gas chromatography-mass spectrometry) analysis on eight derivative mixed standard solutions; (2) shaking up a urine sample unfrozen under room temperature, and carrying out pretreatment and derivatization on the urine sample, sequentially detecting the content of eight monohydric polyaromatic hydrocarbon in urine sample. The method is suitable for detection and research on biomarkers exposed by polyaromatic hydrocarbon in urine of various common samples, can simultaneously detect eight monohydric polyaromatic hydrocarbon in urine, and has the advantages of being low in detection limit, excellent in stability, simple in pretreatment, rapid and accurate.

Description

technical field [0001] The invention belongs to the technical field of biomarker physical and chemical detection of polycyclic aromatic hydrocarbons in urine, and in particular relates to a gas chromatography-mass spectrometry detection method for eight kinds of monohydroxy polycyclic aromatic hydrocarbons in urine. Background technique [0002] Polycyclic aromatic hydrocarbons are a class of carcinogenic compounds that are widely distributed in environmental media such as air, food, water, and even some organisms. Therefore, the content of polycyclic aromatic hydrocarbons in various specific environments has always been valued by the government. In daily life, people may be exposed to PAHs to varying degrees through breathing, eating, drinking, smoking and even skin exposure. Since people are constantly moving between various specific environments, it is difficult to use polycyclic aromatic hydrocarbons in specific environments. Therefore, in recent years, research on bioma...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/06
Inventor 蒋举兴段焰青夏建军王文元杨丽萍刘亚
Owner HONGYUN HONGHE TOBACCO (GRP) CO LTD
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