Metabolism of (-)-(S)-nicotine by guinea pig and rat brain: identification of cotinine

Abstract

Since the brain is the major site of pharmacological activity of nicotine, it was of interest to investigate the metabolism of nicotine by this organ. We now report our findings using guinea pig and rat brain as the enzyme source. Whole brains were removed and washed with isotonic KCl, blotted dry and cut into small pieces. The tissue was weighed and homogenized in pH 7.4 Tris-KCl buffer, 2 ml/g tissue. Incubations were carried out using 0.5 mi of brain homogenate and 0.1-1 mu mol of nicotine at 37 degrees C. The reactions were terminated by freezing at -80 degrees C. The samples were extracted and analyzed by capillary GC with nitrogen-phosphorus detection. Cotinine was detected as the major metabolite and its identity confirmed by GC-MS. Cotinine formation may contribute to the detoxication pathway of nicotine and may be important in controlling nicotine levels in the brain. Furthermore, the conversion of nicotine to cotinine involves the intermediacy of nicotine-Delta(1'(5'))-iminium ion, which is an alkylating agent. This finding supports the concept that reactive intermediates may play a role in the pharmacology and toxicology of nicotine. [GRAPHICS] S]