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
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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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