Application of high-performance thin-layer chromatographic method for the simultaneous determination of lamivudine and tenofovir disoproxil fumarate in pharmaceutical dosage form

Abstract

A simple, precise and accurate high performance thin layer chromatographic method has been developed for the simultaneous determination of lamivudine (LAM) and tenofovir disoproxil fumarate (TDF) in pharmaceutical dosage form. The separation was carried out on Merck HPTLC aluminum plates of silica gel 60 F254, (20 × 10 cm) with 250 μm thickness using chloroform: methanol: toluene (8: 2: 2, v/v/v) as mobile phase. HPTLC separation of the two drugs followed by densitometric measurement was carried out in the absorbance mode at 265 nm. The drugs were satisfactorily resolved with Rf values of 0.27 ± 0.01 and 0.51 ± 0.01 for LAM and TDF, respectively. The linear regression analysis data for the calibration plots showed good linear relationship with r2=0.9999 and 0.9996 for LAM and TDF, respectively in the concentration range of 60-210 ng spot-1 for each drug. The method was validated for precision, robustness, specificity and accuracy. The limit of detection and quantitation were 20 and 40 ng spot-1, respectively for LAM and 30 and 60 ng spot-1, respectively for TDF. The proposed developed HPTLC method can be applied for identification and quantitative determination of LAM and TDF in bulk drug and pharmaceutical dosage form.