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Synchronous analysis method for trace nicotine in blood-brain samples of animal and main metabolites thereof

A simultaneous analysis and animal blood technology, applied to the analysis of materials, measuring devices, material separation, etc., can solve the problems of cumbersome operation, affecting the accuracy of quantitative analysis, and less research on the characteristics of nicotine metabolism, so as to achieve simple and fast operation and reliable results , the effect of high sensitivity

Inactive Publication Date: 2014-09-03
ZHENGZHOU TOBACCO RES INST OF CNTC
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Problems solved by technology

However, these studies also have certain deficiencies, such as only focusing on collecting samples from a single site in the peripheral system for analysis and determination, and less research on the metabolic characteristics of nicotine in its target organ—brain tissue; moreover, the collected biological samples All belong to complex matrix samples, and pretreatment steps such as protein precipitation, centrifugation, and purification are often required for analysis and detection, which not only is cumbersome to operate, but also affects the accuracy of quantitative analysis

Method used

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  • Synchronous analysis method for trace nicotine in blood-brain samples of animal and main metabolites thereof
  • Synchronous analysis method for trace nicotine in blood-brain samples of animal and main metabolites thereof
  • Synchronous analysis method for trace nicotine in blood-brain samples of animal and main metabolites thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0021] 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. Compound sodium chloride injection was used to perfuse at a rate of 2.0 μL / min, and samples were collected after 120 minutes of equilibration, and a tube of brain and hemodialysate were collected synchronously every 15 minutes at 4°C. Start timing at the end of the collection of the fourth tube, inject nicotine solution (0.2 mg / mL, prepared with compound sodium chloride injection) intra...

Embodiment 2

[0041] This example is basically the same as Example 1, except that the dose to rats is different, which is 0.8 mg / kg nicotine. The results are shown in Table 5.

[0042]

[0043] Table 5 Contents of NIC and COT in rats at different time points after single-dose nicotine injection

[0044]

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Abstract

The invention discloses a synchronous analysis method for trace nicotine in blood-brain samples of an animal and main metabolites thereof. The synchronous analysis method is characterized by comprising the following steps of: utilizing a microdialysis system to synchronously collect blood dialysate and brain tissue dialysate samples of an animal in lined pipes of different chromatographic sampling bottles, respectively adding NIC-d3 and COT-d3 to be mixed with internal labeled solution, and after mixing uniformly, directly analyzing and detecting the content of the nicotine and cotinine in a microdialysis sample by using UPLC-MS / MS. The synchronous analysis method disclosed by the invention has the advantages that the pretreatment steps of protein precipitation and purification for the samples are not needed, the synchronous and accurate analysis of nicotine and metabolites in an animal body is realized; and compared with the prior art, the synchronous analysis method has the characteristics of simple operation, high sensitivity and reliable result and the like, and a new method for researching the metabolism of the nicotine in the animal body is provided.

Description

technical field [0001] The invention relates to a method for synchronous analysis of trace nicotine and its main metabolites in animal blood and brain samples. The method is aimed at animal microdialysate samples, uses stable isotope markers as internal standards, and utilizes ultra-high performance liquid chromatography-tandem Mass spectrometry (UPLC-MS / MS) detects trace amounts of nicotine and cotinine in the sample, revealing the metabolism of nicotine in animals. Background technique [0002] Nicotine (NIC) is an important characteristic component of tobacco. Nicotine dependence caused by smoking is the deep-seated reason for continuous smoking and endangering health. The occurrence of nicotine dependence is closely related to the individual's metabolism of nicotine. Studying the metabolic characteristics of nicotine in vivo, especially in the central nervous system, can provide a basis for the formation mechanism of nicotine dependence. [0003] In most cases, ni...

Claims

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Application Information

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IPC IPC(8): G01N30/88
Inventor 毛健张太平卢斌斌孙世豪李鹏张建勋宗永立曾世通柴国璧
Owner ZHENGZHOU TOBACCO RES INST OF CNTC
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