Title:Development and Validation of RP-HPLC Method for the Estimation of
Tenofovir and Emtricitabine in Bulk and Pharmaceutical Dosage Form
Volume: 16
Issue: 1
Author(s): Biswa Mohan Sahoo*, Pallepogu Venkateswara Rao and Naidu Srinivasa Rao
Affiliation:
- Department of Pharmaceutical Chemistry, Roland Institute of Pharmaceutical Sciences, Berhampur- 760010, Odisha, India
Keywords:
Emtricitabine, tenofovir, RP-HPLC, mobile phase, flow rate, linearity, validation.
Abstract:
Background: A novel, simple, efficient, rapid, and precise reverse-phase highperformance
liquid chromatography (RP-HPLC) method was developed for the estimation of
Tenofovir and Emtricitabine in the bulk and pharmaceutical dosage form. The currently developed
method was subsequently validated according to ICH guidelines in terms of linearity, accuracy,
precision, the limit of detection, the limit of quantification, robustness, etc.
Methods: The separation of the selected drugs was optimized after several trials including change
of mobile phase and its composition, stationary phase, flow rate, column temperature, etc. The
separation was performed by using an Inertsil ODS C18 column (250 mm x 4.6 mm, 5μ) and UV
absorption was measured at 231 nm. Methanol: Acetonitrile: Water was selected as the mobile
phase in the ratio of 50:20:30 (V/V/V) at a flow rate of 1 mL/min. As per International Conference
on Harmonization (ICH) Q2 R1 guidelines, several validation parameters were evaluated
which include specificity, linearity, precision, accuracy, the limit of detection (LOD), and the limit
of quantitation (LOQ).
Results: The acceptable degree of linearity range was found to be 40-100 μg/mL. The standard solution
exhibited retention times of 3.06 minutes and 5.07 minutes for Tenofovir and Emtricitabine respectively.
The LOD and LOQ obtained were 0.05 μg/ml and 0.02 μg/mL, 15 μg/mL, and 0.08 μg/mL for
Tenofovir and Emtricitabine respectively. The percent recovery was found to be 98 to 102%.
Conclusion: Hence, the proposed method is simple, selective, and specifically meets the requirements
of ICH guidelines for the validation of the analytical method.
