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A sensitive and selective LC-MS/MS method to quantify Lamivudine and Nevirapine in K3EDTA human plasma over the concentration range of 25.0240 to 3997.1740 ng/mL and 37.5080 to 5991.0460 ng/mL, respectively were developed and validated. Lamivudine and Nevirapine and its internal standard (Lamivudine 13C 15N2 and Nevirapine D5) were selectively extracted from 300 µL plasma by direct elution using solid phase extraction technique. Elution was achieved by reverse phase liquid chromatography on Hypurity C18 (100 mm × 4.6 mm, 5.0 µm) column was done. LLOQ of 25.0240 ng/mL and 37.5080 ng/mL were resolute for this technique for Lamivudine and Nevirapine respectively. Accuracy and precision values for Lamivudine were 99.55 & 1.50 and for Nevirapine the accuracy and precision values are 99.33 & 1.03%, respectively.
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