Simultaneous Assay for Tramadol Hydrochloride and Metabolite M 1 in Human Plasma by HPLC with Fluorecence Detection

Objective:To develop a simultaneous assay for tramadol hydrochloride (TMD) and metabolite M 1 in human plasma by HPLC.Method:Expriments were performed on Waters 208 HPLC system with a Shimadzu RF_530 fluorecence detector(λ Ex = 275 nm,λ Em =304 nm) and a Inertsil C 18 column(4 6 mm×200 mm,5 μ m) The mobile phase was consisted of borax buffer ( 0. 03 mol·L -1 ,containing 0 5% triethylamine,pH 4 0) - methanol( 68∶32) . The column temperature was 50 ℃. Atenolol was used as internal standard.Results:The calibration was linear in the range within 4 5～446 ng·mL -1 for TMD and 3 1～312 ng·mL -1 for M 1, and detection limit was 2 ng·mL -1 and 1 5 ng·mL -1 respectively The recovery of the two substance was both within 95%～100 3%.Conclusion:The method is sensitive,simple and easy to do, with accurate and dependable results.