Development and Validation of Spectrofluorimetric Methods for Estimation of Tramadol Hydro chloride in Human Plasma

Abstract

This paper describes validated Spectrofluorimetric methods for the estimation of Tramadol hydrochloride in human plasma using liquid-liquid extraction method. Tramadol hydrochloride measured at an excitation wavelength of 276nm and emission wavelength of 301nm. Drug extraction form plasma was achieved using a mixture of ethyl acetate and hexane in a ratio of 1:4. The method is validated as per USFDA bioanalytical guideline and linearity was found between 10-1000 ng/ml with correlation coefficient of 0.997. The percent recovery was found more than 95% and precision was found between 1.79% and 7.68%. Stability studies were performed under different conditions like, freeze and thaw, solution stability and bench top. Stability was performed at LQC and HQC level and % RSD was found between 3.2 to 5.6%. Selectivity and matrix effect was performed at lower limit of quantification that was 10ng/ml and found well within the acceptance criteria. This indicates there is no plasma interference for estimation of drug hence method is specific. Limit of quantification is 10ng/ml for developed spectrofluorimetric method which is as low as reported HPLC method.