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HIGH PRESSURE LIQUID CHROMATOGRAPHIC METHOD FOR THE DETERMINATION OF NICOTINE METABOLITES AND THEIR IN VITRO METABOLISM BY RAT BRAIN

Nicotine is the major alkaloid in commercial tobacco and it is metabolised both in vivo [1] and in vitro [2,3] by human and many mammalians. Since nicotine has a very important role in neuropharmacological effects in smokers [4], the brain tissue was chosen as an enzyme source. These effects could be produced by not only nicotine but also its metabolites. Therefore, it is important to understand the further biotransformation of these metabolites. In the present work, the metabolism of nornicotine, cotinine, norcotinine and some N-oxides being nicotine derivatives ie cotinine-N-oxide and 3’-hydroxycotinine-N-oxide were studied in rat to detect whether they could be metabolised into corresponding potential metabolites ie norcotinine, trans-3’-hydroxycotinine, 5’-hydroxycotinine, 4-(3-pyridyl)-4-oxo-butyramide, nicotinamide and cotinine. A reverse phase high performance liquid chromatographic system was also developed for the determination of these nicotine analogues using a diode-array UV detector.