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Analysis method for trace tobacco specific N-nitrosamine (TSNAs) in animal blood sample

A blood sample and analysis method technology, applied in the analysis field of tobacco-specific N-nitrosamines (TSNAs), can solve the problems of deviation in measurement results, discrimination between analytes and internal standards, etc.

Active Publication Date: 2011-04-13
ZHENGZHOU TOBACCO RES INST OF CNTC
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the HPLC-MS method also has certain defects. For example, in the process of quantitative analysis, due to the inconsistency between the internal standard and the analyte, there is discrimination between the analyte and the internal standard in the sample pretreatment, resulting in deviations in the measurement results. The use of isotope labels as internal standards can effectively eliminate this discrimination and improve the accuracy of quantitative analysis

Method used

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  • Analysis method for trace tobacco specific N-nitrosamine (TSNAs) in animal blood sample
  • Analysis method for trace tobacco specific N-nitrosamine (TSNAs) in animal blood sample
  • Analysis method for trace tobacco specific N-nitrosamine (TSNAs) in animal blood sample

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Purchase 5 male Japanese white rabbits with big ears, weighing 3-4 kg, healthy and disease-free, fasting for 12 hours before intragastric administration, one dose: NNK 1.5 mg, NNN 1.02 mg, NAT 1.05 mg, NAB 1.26 mg. After the administration, the timing was started, and the blood sampling method of the ear vein was adopted. 1 mL blood sample was collected every 10 minutes, and the blood was collected 10 times continuously. The blood sample was put into 0.1 mL saturated aqueous solution of EDTA-k2 Centrifuge at 10000 rpm in a 1.5 mL centrifuge tube, take 0.5 mL of the upper plasma and add 0.5 mL of 1 ng / mL acetonitrile solution to the internal standard. After standing for 30 minutes, filter and analyze and detect by LC-MS / MS.

[0035] Chromatographic conditions

[0036] Column: Agilent Extend-C18 column (100×4.6mm i.d., 1.8 μm, USA).

[0037] Column temperature: 50 ℃; mobile phase: A: water (containing 0.1% ammonium formate, mass fraction), B: acetonitrile (containing 0.1% form...

Embodiment 2

[0048] This example is basically the same as Example 1, except that the dosage to animals is different, which are NNK 1.05 mg, NNN 0.5 mg, NAT 0.52 mg, and NAB 0.56. The results are shown in Table 5.

[0049]

Embodiment 3

[0051] This example is basically the same as Example 1, except that the dosage to animals is different, which are NNK 2.05 mg, NNN 1.51 mg, NAT 1.45 mg, and NAB 1.58. The results are shown in Table 6.

[0052]

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Abstract

An analysis method for a trace tobacco specific N-nitrosamine (TSNAs) in an animal blood sample is characterized by comprising the following steps of: placing the animal blood sample into a centrifuge tube with ethylene diamine tetraacetic acid EDTA-k2 saturated aqueous solution, performing the centrifugation at 10,000 rpm; adding the upper blood plasma into the mixed internal standard solution of NNN-d4, NNK-d4, NAT-d4, NAB-d4 and NNAL-d3; after standing for 30min to precipitate protein, and filtering upper solution; and detecting the content of the tobacco specific N-nitrosamine in the sample by adopting LC-MS / MS. The method has the advantages of exactly determining the content of the tobacco specific N-nitrosamine (TSNAs) and the content of metabolites thereof in the complex matrix sample of the animal blood. Compared with the prior art, the invention has the characteristics of simplicity in operation, high sensitivity and reliability in result, and creates the method of determining the content of the tobacco specific N-nitrosamine (TSNAs) and the content of the metabolites thereof in the complex matrix sample of the animal blood.

Description

technical field [0001] The invention relates to an analysis method for tobacco-specific N-nitrosamines (TSNAs) in complex matrices of animal blood. This method uses isotope labels as internal standards, and high-performance liquid chromatography-mass spectrometry (HPLC-MS / MS) is used to detect trace amounts of TSNAs and their metabolites in animal blood, revealing the metabolism of TSNAs in animals. Background technique [0002] Smoking and health issues have aroused great concern. Some harmful components in tobacco leaves and cigarette smoke, especially tobacco-specific N-nitrosamines (TSNAs), which are highly carcinogenic, have attracted great attention from scientists from all over the world.[ Liu Wanfeng, Wang Yingyuan. Research progress of tobacco-specific nitrosamines in tobacco. China Tobacco Science, 2002, (2): 11-15]. To study the metabolites and metabolic process of TSNAs in animals, and to carry out the research on the metabolic process of TSNAs can provide the ...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/84
Inventor 卢斌斌王兆宇王昇张建勋刘惠民张晓兵宗永立
Owner ZHENGZHOU TOBACCO RES INST OF CNTC
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