Analysis method for dynamically tracing and monitoring nicotine and nicotine metabolites in animal

An analytical method and metabolite technology, applied in the field of nicotine metabolism analysis in animals, can solve the problems of low sensitivity and easy loss of radioactive signals, and achieve the effects of accurate quantitative analysis results, high sensitivity and reliable results

Active Publication Date: 2016-08-24
ZHENGZHOU TOBACCO RES INST OF CNTC
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Problems solved by technology

This method uses a pyridine ring 2 H-labeled nicotine ([ 2 h 4 ]-(±)-Nicotine) to trace the metabolic transformation process of nicotine, combined with high-resolution mass spectrometry, not only has high sensitivity, can obtain accurate molecular mass information of metabolites, but also can use the pyridine ring 2 The pairing of H-labeled isotope peaks and unlabeled nicotine metabolite peaks further confirm

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  • Analysis method for dynamically tracing and monitoring nicotine and nicotine metabolites in animal
  • Analysis method for dynamically tracing and monitoring nicotine and nicotine metabolites in animal
  • Analysis method for dynamically tracing and monitoring nicotine and nicotine metabolites in animal

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Embodiment 1

[0028] Male adult normal SD rats, 10 weeks old, weighing (200 ± 20) g, were raised in independent isolation cages, with free access to water and standard pellets. Rats were anesthetized by intraperitoneal injection of 1% pentobarbital at a dose of 50 mg / kg. With the aid of a stereotaxic instrument, a probe cannula was embedded in the striatum of the rat brain. After 24 h, the microdialysis probe (CMA / 12) was inserted in the rat's conscious and free-moving state. For hemodialysis sample collection, a microdialysis probe (CMA / 20) was implanted in the jugular vein of the right atrium of the rat. Perfuse with filtered Ringer's solution (compound sodium chloride injection) at a rate of 1.5 μL / min, and start collecting samples in microcentrifuge tubes after equilibrating for 120 min (can be used with microdialysis sample collection system and can be used with In the chromatographic sample bottle), a tube of brain and hemodialysate were collected synchronously every 20 min at 4°C. ...

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Abstract

An analysis method for dynamically tracing and monitoring nicotine and nicotine metabolites in an animal is characterized in that the animal is exposed after pyridine ring 2H labeled nicotine ([2H4]-(+/-)-Nicotine) and unlabeled nicotine are mixed in equal proportion, a micro-dialysis system is utilized to respectively collect blood and brain dialysate samples in a micro-centrifuge tube, the samples are directly put on an ultra high performance liquid chromatography-orbit trap to conduct high-resolution mass spectrometry detection on the nicotine metabolites and pyridine ring 2H labelers of the nicotine metabolites in the micro-dialysis samples after an analyzing interior label is added for short-term low-speed centrifugation, the nicotine metabolites are identified according to test results, and a corresponding time-concentration curve is drawn for dynamic monitoring and analysis. The analysis method has the advantages that a variety of nicotine metabolites in animals can be accurately traced and identified, a sample pretreatment step is greatly simplified, accurate analysis and synchronous comparison of nicotine metabolite changes in different systems in animals are achieved, and a novel method for researching the nicotine metabolites in animals is created.

Description

technical field [0001] The invention relates to a method for analyzing nicotine metabolism in animals. In this method, stable isotopes are used to trace the metabolic process of nicotine, rat brain tissue and blood samples are collected synchronously through microdialysis technology, and nicotine metabolites in the samples are traced and detected by ultra-high performance liquid chromatography-Orbitrap high-resolution mass spectrometry Analysis, so as to comprehensively reveal the metabolic characteristics of nicotine in animals. Background technique [0002] Nicotine (Nic) is an important characteristic component of tobacco, and the occurrence of nicotine dependence is closely related to the individual's ability to metabolize nicotine. Although the nicotine ingested in the body will quickly pass through the blood-brain barrier and enter the brain tissue, most of the body's metabolism of nicotine is completed in the liver. The conversion and metabolism process of nicotine ...

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Application Information

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IPC IPC(8): G01N30/02G01N27/62
CPCG01N27/626G01N30/02G01N2030/027
Inventor 毛健李鹏卢斌斌孙世豪刘俊辉王丁众张文娟柴国璧张启东曾世通张建勋
Owner ZHENGZHOU TOBACCO RES INST OF CNTC
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