Determination of NNK and its 7 Metabolites in Rabbit Blood with HILIC-MS/MS

A method of determining NNK and its seven metabolites in rabbit blood with HILIC-MS/MS (hydrophilic interaction liquid chromatography tandem mass spectrometry) was established. Using NNK-d₄, NNAL-d₃ and HPB-d₄ as internal standards, immediately after protein precipitating with acetonitrile, the plasma samples were separated by a HILIC column, then determined by mass spectrometry under multi-reaction monitoring (MRM) mode. The results showed that the limits of detection of NNK and its metabolites ranged from 0.016 to 0.082 ng/mL with r of 0.9991 to 1.0000, and the recoveries ranged from 76.9% to 116.3% with the intra-day precision of 0.53% to 6.52%. The method is suitable for the simultaneous analysis of NNK and its seven metabolites in blood.