DETERMINATION OF AMITRIPTYLINE AND NORTRIPTYLINE IN HUMAN LIVER-MICROSOMES WITH REVERSED-PHASE HPLC IN-VITRO

AIM: To develop a method for simultaneous determinations of amitriptyl ine (Ami) and its metabolite nortriptyline (Nor) in human liver micros omes. METHODS: An incubation buffer containing microsomes, NADPH-gener ating system, and Ami, after termination of enzyme reaction and desipr amine (Des) as internal standard (IS), was extracted with diethy ether and separated on a reversed-phase ODS column. Detection was achieved at 242 nm by ultraviolet detector. RESULTS: No potential interfering p eaks were found. Ami and Nor gave rapid elution and baseline resolutio n. The linear curves of both analyses ranged 0.02 - 10 nmol and the li mit of detection was 0.01 nmol. The recovery (94 % - 101 %) had good p recision with relative s of < 8.3 %. CONCLUSION: This:method is rapid, sensitive, and simple for studying the metabolism of Ami and Nor.