Bio-Analytical Method Development and Validation for The Estimation of Tapentadol Hydrochloride in Human Plasma By RP-HPLC Method

A rapid, sensitive and selective HPLC method for the determination of tapentadol hydrochloride in human plasma was developed and validated. Sample preparation was carried out using solid-phase extraction (SPE) method. Separation was done on a Phenomenex C18 reverse phase column with a mobile phase of methanol and acetonitrile (50:50) – phosphate buffer (pH 6.8) in the ratio of 55:45 % v/v and detection at 275 nm. The retention time of 6.4 and 8.6 mins for tapentadol hydrochloride and ibuprofen (IS) is highly advantageous. The standard curve is linear (r > 0.999) over the concentration range of 200–1000 ng/ml. Validated analytical method was based on the parameters: selectivity, accuracy, precision, repeatability, linearity, LOD, LOQ, and the suitability of the system. Validation analysis showed selectivity test Rs>1.5, Based on the area ratio of peak height and a segment of the chromatogram obtained LOD values 59.33 ng/ml, LLOQ value of 177.9 ng/ml, percent accuracy from 99.4 to 100.4% for interaday studies and 99.7 to 101.7% for interday studies and precision CV (%) 3.44 to 6.94 for interaday studies and 6.65 to 7.88 for interday studies. System suitability test results on the retention time, area ratio, and high ratios of peak chromatogram shows the CV (%) <10%. Thus it is concluded that the analytical methods used have validity in accordance with the requirements.