Bioanalytical Method Development and Validation of Lamivudine by Rp-hplc Method

A simple, accurate, precise and sensitive HPLC method with UV detection was developed and validated to separate and detect lamivudine in human plasma using Stavudine as an internal standard.Lamivudine (3-TC) and Stavudine (internal standard) were extracted from human plasma using methanol protein precipitation and were chromatographed on a Phenomenex C18 (250X4.6mm.,5mμ particle size) column using 20μl injection volume and detection at 270nm.An isocratic mobile phase consisting of Methanol: Water (85:15%v/v) was used to separate these drugs. The retention times of lamivudine and I.S were 4.6 and 6.2 respectively. The method was validated over the range of 406.104020.05 ng/ml. The limit of detection was 200 ng/ml and the limit of quantification was 400 ng/ml for 3TC. Within and betweenday precisions are less than 6.5% for all quality control samples. The absolute recoveries of 3-TC was greater than 90% were achieved. The described method can be readily utilized for analysis of pharmaceutical products.