Sample pretreatment method for detecting special nitrosamine in tobacco and tobacco products by LC-MS/MS (liquid chromatogram-mass spectrum/mass spectrum)
A technology for sample pretreatment and tobacco products, which is applied in the field of sample pretreatment for LC-MS/MS detection of specific nitrosamines in tobacco and tobacco products, which can solve the problems of large reagent consumption and long time consumption, and achieve reagent consumption Small size, short extraction time, safe and convenient operation
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example 1
[0035] Weigh 45-50 mg of freeze-dried fresh Burley tobacco leaf samples into a glass test tube, add 1.5 mL of 100 mM ammonium acetate buffer solution into the glass tube (4 kinds of deuterated TSNAs internal standards were added to the ammonium acetate buffer solution in advance , with a concentration of 5 ng mL -1 NNN-d4, 4 ng mL -1 NNK-d4, 20 ng mL -1 NAT-d4, 0.24 ng mL -1 NAB-d4), capped and sealed, placed in an ultrasonic cleaner, ultrasonically extracted at room temperature for 30 min, ultrasonic power was 100 HZ, and then centrifuged at 8000 rpm for 5 min, 0.8 mL supernatant was taken, and placed in Add 0.8 mL of ethyl acetate to the supernatant, cover and seal, shake at room temperature for 5 min, then centrifuge at 8000 rpm for 5 min, take 0.4 mL of the supernatant and dry it with nitrogen, wash with 0.4 mL of deionized water Reconstitute and transfer to a chromatographic vial for LC-MS / MS analysis.
[0036] By LC-MS / MS analysis, the resulting total ion current...
example 2
[0038] Weigh 20-25 mg flue-cured tobacco shred samples into a glass test tube, add 0.75 mL of 100 mM ammonium acetate buffer solution into the glass tube (4 kinds of deuterated TSNAs internal standards were added to the ammonium acetate buffer solution in advance, the concentrations were 5 ng mL -1 NNN-d4, 4 ng mL -1 NNK-d4, 20 ng mL -1 NAT-d4, 0.24 ng mL -1 NAB-d4), capped and sealed, placed in an ultrasonic cleaning machine, ultrasonically extracted at room temperature for 30 min, ultrasonic power was 100 HZ, and then centrifuged at 8000 rpm for 5 min, 0.4 mL supernatant was taken, and placed in Add 0.4 mL of ethyl acetate to the supernatant, cover and seal, shake at room temperature for 5 min, then centrifuge at 8000 rpm for 5 min, take 0.2 mL of the supernatant and dry it with nitrogen, wash with 0.2 mL of deionized water Reconstitute and transfer to a chromatographic vial for LC-MS / MS analysis.
[0039] By LC-MS / MS analysis, the resulting total ion current TIC and ...
example 3
[0041] Weigh 90-100 mg of freeze-dried oriental tobacco into a glass test tube, add 3.0 mL of 100 mM ammonium acetate buffer solution into the glass tube (4 kinds of deuterated TSNAs internal standards are added to the ammonium acetate buffer solution in advance, the concentrations are respectively 5ngmL -1 NNN-d4, 4 ng mL -1 NNK-d4, 20 ng mL -1 NAT-d4, 0.24 ng mL -1NAB-d4), capped and sealed, placed in an ultrasonic cleaner, ultrasonically extracted at room temperature for 30 min, with an ultrasonic power of 100 HZ, and then centrifuged at 8000 rpm for 5 min, and 1.6 mL of the supernatant was taken in Add 1.6 mL of ethyl acetate to the supernatant, cover and seal, shake at room temperature for 5 min, then centrifuge at 8000 rpm for 5 min, take 0.8 mL of the supernatant and dry it with nitrogen, wash with 0.8 mL of deionized water Reconstitute and transfer to a chromatographic vial for LC-MS / MS analysis.
[0042] By LC-MS / MS analysis, the resulting total ion current TIC...
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