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Development and Validation of a new HPLC Method for Tramadol Determination in Human Plasma using L/L Extraction

Domenii publicaţii > Ştiinţe medicale + Tipuri publicaţii > Articol în volumul unei conferinţe

Autori: Lavinia G. Hinescu, M. C. Ranetti, M. Ionescu, M.E. Hinescu

Editorial: 9th Congress of Balkan Military Medical Committee, Varna - Bulgaria, 2005.


Tramadol is an effective and well tolerated agent to reduce pain resulting from trauma, renal or biliary colic and labor, and also for the management of chronic pain of malignant or nonmalignant origin, particularly neuropathic pain. It appears to produce less dependence than equianalgesic doses of strong opioids.
Tramadol, a centrally acting analgesic structurally related to codeine and morphine, consists of two enantiomers, both of which contribute to analgesic activity via different mechanisms. The complementary and synergistic actions of the two enantiomers improve the analgesic efficacy and tolerability profile of the racemate.
Aim of the study. This paper describes a new bioanalytical method involving a simple liquid–liquid extraction for the simultaneous HPLC determination of the tramadol (TR) and its active metabolite O-desmethyltramadol (M1) in biological samples.
Material and method. A reversed-phase mechanism with FLD detection and liquid/liquid extraction from basic plasma samples was chosen for determination of tramadol.
Results. The ratio between phases was 1:3; after vigorously mix for 30 minutes, the organic layer was evaporated to dryness under N2 stream at 40°C. The recovery for O-desmethyltramadol (95-101%) and Tramadol (80-84.81%) is satisfactory to allow the determination of the lowest plasma level concentration of the drug. An aliquot of 100l sample was automatically injected into Inertsil ODS-2 chromatographic column. We choose an isocratic elution at 40°C using a timetable for FLD detector. The variation of analytical wavelengths Ex/Em was between 276/310 and 228/310nm. The internal standard (IS) used was Metoprolol.
The calibration graphic is linear between 10 and 600 ng/mL (r=0.9924, n=8) for tramadol 5 to 300ng/mL (r=0.9957, n=8) for metabolite.

Cuvinte cheie: Tramadol, HPLC, human plasma