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Method for determining content of cotinine in urine

A technology for cotinine and urine, applied in the field of urine metabolite detection, can solve the problems of poor separation effect, and achieve the effects of simple operation, improved analytical sensitivity, and improved separation degree.

Active Publication Date: 2015-07-08
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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  • Method for determining content of cotinine in urine
  • Method for determining content of cotinine in urine
  • Method for determining content of cotinine 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 disclose a method for determining the content of cotinine in urine. The method comprises the following steps of: putting urine into a centrifugal tube, adding a deuterated internal standard and diluting with acetonitrile; placing the centrifugal tube in a vortex oscillation instrument, carrying out vortex oscillation and standing; sucking supernatant and filtering by an organic-phase filtering membrane, and obtaining filtrate to be determined; and preparing stock solution and working solution, and carrying out ultra-high-performance liquid chromatography-tandem mass spectrum determination. Compared with the traditional gas chromatography and mass spectrometry, the method has the beneficial effects that a mode of direct dilution with acetonitrile and vortex oscillation filtering sample injection is adopted, so that the pretreatment process is simplified and the analysis sensitivity is improved. The method has the advantages of simple operation, fastness, high accuracy and 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 Applications(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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