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System and method for measuring NNK and metabolites thereof by liquid-phase-liquid-phase two-dimensional chromatography based on solid-phase extraction

A chromatographic system and two-dimensional chromatographic technology, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of large health hazards for experimental operators, cumbersome operations, and environmental pollution, so as to avoid broadening of chromatographic peaks and shorten analysis time , The effect of improving the analytical sensitivity

Active Publication Date: 2020-02-21
CHINA TOBACCO YUNNAN IND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, the commonly used sample pretreatment solid-phase extraction or column chromatography separation and purification for the analysis of TSNAs and their metabolites, this kind of method needs to concentrate and transfer the sample many times, and the operation is cumbersome; and it will also use a large amount of organic solvents in an open environment. The pollution of the environment is a great hazard to the health of the experimental operators.

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  • System and method for measuring NNK and metabolites thereof by liquid-phase-liquid-phase two-dimensional chromatography based on solid-phase extraction
  • System and method for measuring NNK and metabolites thereof by liquid-phase-liquid-phase two-dimensional chromatography based on solid-phase extraction
  • System and method for measuring NNK and metabolites thereof by liquid-phase-liquid-phase two-dimensional chromatography based on solid-phase extraction

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0078] Such as figure 1 As shown, the system for determining NNK and its metabolites based on liquid phase-liquid two-dimensional chromatography of the present embodiment includes a primary chromatography system 1 and a secondary chromatography system 5, a primary chromatography system 1 and a secondary chromatography system The chromatographic system 5 is connected to the solid phase extraction column 3 through the six-way valve 2; the gel purification effluent fraction dilution pump 4 is connected to the three-way valve 1.5 of the gel purification system.

[0079] The primary chromatographic system 1 includes a sampler 1.2, one end of the sampler 1.2 is connected to the first pump 1.1, the other end is connected to the purification column 1.3, one end of the purification column 1.3 is connected to the detector 1.4, and one end of the detector 1.4 is connected to the three-way valve 1.5 Connect, this end of three-way valve 1.5 is also connected with the second pump, the seco...

Embodiment 2

[0091] The device of this embodiment is the same as that of Embodiment 1.

[0092] NNK and NNAL determination method in the smoker's blood and urine of the present embodiment, carry out as follows:

[0093] The samples measured in this embodiment are urine samples from heavy smokers of mixed cigarettes.

[0094] After sample collection, accurately pipette 5.0 mL, add 100 µL of internal standard working solution, then dilute to 10 mL with methanol and shake well to keep the proportion of methanol in the actual sample to be tested at 50%, and the concentration of internal standard at 5 ng / L . The samples were quickly brought back to the laboratory and centrifuged at a high speed of 20000 r / min for 10 min; the supernatant was filtered for sample analysis. The sample injection volume was 1.0 mL, 30% acetonitrile (containing 0.01% ammonium acetate) was used as the mobile phase, the Agilent Zorbax SB-C18 (9.4×150 mm, 5.0 µm) column was used as the stationary phase, and the flow ra...

Embodiment 3

[0098] The device of this embodiment is the same as that of Embodiment 1.

[0099] NNK and NNAL determination method in the smoker's blood and urine of the present embodiment, carry out as follows:

[0100] The samples tested in this embodiment are urine samples from moderate smokers of mixed cigarettes.

[0101] After sample collection, accurately pipette 5.0 mL, add 100 µL of internal standard working solution, dilute to 10 mL with methanol and shake well to keep the proportion of methanol in the actual sample to be tested at 50%, and the concentration of internal standard at 5 ng / L .

[0102] The samples were quickly brought back to the laboratory and centrifuged at a high speed of 20000 r / min for 10 min; the supernatant was filtered for sample analysis. The sample injection volume was 2.0 mL, 30% acetonitrile (containing 0.01% ammonium acetate) was used as the mobile phase, the Agilent Zorbax SB-C18 (9.4×150 mm, 5.0 µm) column was used as the stationary phase, and the fl...

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Abstract

The invention relates to a system and a method for measuring NNK and metabolites thereof by liquid-phase-liquid-phase two-dimensional chromatography based on solid-phase extraction. The method comprises the following steps: after a prepared sample is introduced, carrying out high-efficiency purification by primary chromatography; cutting part of fractions flowing out of NNK and NNAL in the primarychromatography; diluting the cut fractions by a dilution pump to reduce the solvent strength of the fractions; mixing by a mixer; enriching by a solid-phase extraction column; after the solid-phase extraction and enrichment are completed, switching by a valve; conveying a mobile phase by an analysis pump of secondary chromatography; eluting the NNK and NNAL enriched on the solid-phase extractioncolumn from the flow direction opposite to the sample enrichment; and separating by an ultra-high performance liquid chromatographic column and detecting by tandem mass spectrometry. The method can simultaneously realize high-efficiency purification and high-multiple enrichment of the sample, greatly improves the analysis sensitivity of the method, greatly simplifies the pretreatment of the analysis sample, can accurately measure the NNK and NNAL of ng / L magnitude order in blood and urine samples, and provides a sensitive, accurate and reliable new method for the research of the correlation between smoking and health.

Description

technical field [0001] The present invention is a system for measuring NNK and its metabolites, especially a system for measuring NNK and its metabolites by liquid phase-liquid two-dimensional chromatography based on solid phase extraction, and also relates to the determination of NNK and its metabolites in the blood and urine of smokers. A method for metabolites, belonging to the field of analytical instruments. Background technique [0002] Tobacco-specific nitrosamines (TSNAs) are an important class of harmful components in cigarette smoke. There are currently 8 reported TSNAs, including N-nitrosonornicotine (NNN), 4-(methylnitrosamine In-depth research on 4 kinds of -1-(3-pyridyl)-1-butanone (NNK), N-nitrosoanatabine (NAT) and N-nitrosoanabatin (NAB). In particular, the State Tobacco Monopoly Administration of my country considers carbon monoxide, hydrogen cyanide, NNK, ammonia, B[a]P, phenol and crotonaldehyde to be the most harmful indicators of cigarette smoke, and c...

Claims

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Application Information

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IPC IPC(8): G01N30/08G01N30/88
CPCG01N30/08G01N30/88
Inventor 李晶杨叶昆刘欣孔维松黄海涛许永王晋米其利高茜向海英曾婉利杨光宇李雪梅汪伟光
Owner CHINA TOBACCO YUNNAN IND
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