Background: A fast, reliable, and sensitive RP-HPLC method was developed for the simultaneous quantification of Lamivudine (3TC), Tenofovir (TFV), and Efavirenz (EFV) in pure form and their combined

pharmaceutical formulations.

Methods: The analysis was performed on an Inertsil ODS-3V column (250 × 4.6 mm, 5 μm) using an isocratic mobile phase composed of Acetonitrile and 1% Isopropyl Alcohol (IPA) in the ratio 85:15, flowing at 1 mL/min. Detection was carried out at 256 nm with a PDA detector.

Results: Retention times for 3TC, TFV, and EFV were observed at 2.4, 2.8, and 4.5 minutes, respectively (± 0.5 min). The method showed excellent linearity for all three drugs with correlation coefficients (R²) of 0.999. Recovery studies ranged between 98% and 102%, and precision (%RSD) values were below 2%. The method’s sensitivity was confirmed by determining the limits of detection (LOD) and quantification (LOQ), which were 0.06, 0.09, 0.17 μg/mL and 0.18, 0.27, 0.53 μg/mL for 3TC, TFV, and EFV, respectively.

Conclusion: This RP-HPLC method is rapid, robust, and highly reproducible, making it suitable for routine quality control and high-throughput analysis of these antiretroviral drugs in combined dosage forms. Its simplicity and accuracy ensure reliable application in pharmaceutical laboratories.