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A method for measuring cotinine content in urine

A technology of cotinine and urine, which is applied in the field of urine metabolite detection, can solve the problem of poor separation effect, and achieve the effects of simple operation, improved selectivity and sensitivity, and simplified pretreatment process

Active Publication Date: 2017-06-16
SICHUAN BRANCH OF CHINA TOBACCO
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Problems solved by technology

Fan et al established a LC-MS / MS method for direct dilution, filtration, and sample analysis of cotinine in urine. This method simplifies the pretreatment process and selects a conventional liquid chromatography column (column diameter: 4.6mm, Particle size: 5μm), but its separation effect is not as good as that of UPLC columns with small particle size and narrow inner diameter

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  • A method for measuring cotinine content in urine
  • A method for measuring cotinine content in urine
  • A method for measuring cotinine content in urine

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Embodiment Construction

[0032] The following will be described in conjunction with a specific embodiment.

[0033] The determination method of the cotinine content in the urine in this embodiment adopts successively acetonitrile dilution and protein precipitation, deuterated internal standard quantification, vortex oscillation purification, organic phase membrane filtration, ultra high performance liquid chromatography-tandem mass spectrometer determination , can quickly, accurately, and simultaneously detect the content of cotinine in urine.

[0034] The instruments and reagents used in this embodiment are:

[0035] Waters Xevo TQ ultra-high performance liquid chromatography-tandem mass spectrometer (Waters, USA), equipped with an electrospray ionization source (ESI); VtexMixer 230VeU oscillator (Labnet, USA).

[0036] Formic acid is HPLC grade (concentration is 49-51%, German Sigma company); Acetonitrile, methyl alcohol are chromatographically pure (U.S. Thermo-Fisher company); Cotinine, cotinine-...

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Abstract

The invention discloses a method for determining the content of cotinine in urine, comprising the following steps: pipetting the urine into a centrifuge tube, adding a deuterated internal standard, and diluting with acetonitrile; placing the centrifuge tube in a vortex shaker, vortexing Shake and let stand; draw the supernatant and filter it through an organic phase filter membrane, and the filtrate is to be tested; prepare the stock solution and working solution, and use ultra-high performance liquid chromatography-tandem mass spectrometry for determination. Compared with the traditional gas chromatography-mass spectrometry method, the present invention adopts the method of direct dilution with acetonitrile and vortex vibration filtering for sample injection, which simplifies the pretreatment process and improves the analytical sensitivity. The method has the advantages of simple operation, rapidity, accuracy, good sensitivity and repeatability.

Description

technical field [0001] The invention belongs to the technical field of urine metabolite detection and relates to a method for determining cotinine in urine. Background technique [0002] Nicotine, commonly known as nicotine, is an important component of tobacco, accounting for 98% of the total alkaloids in tobacco. Nicotine is a major factor in smoking addiction or tobacco dependence. About 80% of nicotine entering the human body is metabolized into cotinine under the action of cytochrome P4502A6 in the liver. Cotinine enters various parts of the human body with the blood, and is found in body fluids such as blood, urine, saliva, and organs such as the liver and brain. The half-life of cotinine in the body is 18-22 hours, and since nicotine is the only source of cotinine, it is currently recognized as a specific biomarker for evaluating cigarette smoke exposure. [0003] Urine is the most widely used biological matrix in biological exposure risk assessment due to its char...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02G01N30/06
Inventor 熊巍陶晓秋韶济民张海燕庞夙黄玫
Owner SICHUAN BRANCH OF CHINA TOBACCO
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