Analytical Method Development and Validation of Efavirenz by Using RP- HPLC

Background

Efavirenz is a widely used antiretroviral medication for managing HIV infection, known for its role as a Non-Nucleoside Reverse Transcriptase Inhibitor (NNRTI). By targeting and inhibiting the reverse transcriptase enzyme, it effectively hinders the replication of HIV, making it a crucial component of combination therapy for the virus.

Materials and Methods

The purpose of this research is to develop an accurate, selective and sensitive HPLC method for the regular quantification of efavirenz in laboratory settings. In this research, tert butyl methyl ether is used as a diluent and methanol is used as a mobile phase while the assay test involved analyzing the Active Pharmaceutical Ingredient (API) and market formulations of efavirenz. EFZ was extracted from market formulations, diluted in tert-butyl methyl ether and analyzed using the developed HPLC method. The results indicated that the method accurately quantified EFZ in both the API and market formulations.

Results

The Separation process utilized using a (X-Bridge® C8 5 μm 4.6×250 mm), with mobile phase consisting of Methanol. The Flow rate was maintained at 0.8 mL/min and EFZ detection occurred at a wavelength of 252 nm, using a manual injection volume of10 μL and retention time was found to be 3.7 min. Linearity was examined and proven at different concentrations in the range of EFZ (2-64 μg/mL), respectively and correlation coefficient R² was found to be 0.9998. The method achieved very good validation parameters LOQ about 0.03 mg/mL and LOD about 0.01 mg/mL for EFZ. The high value of recoveries obtained for EFZ indicates that the proposed method was found to be accurate.

Conclusion

The developed HPLC method for quantifying efavirenz in pharmaceutical formulations is straightforward, accurate and reproducible. It satisfies all validation criteria, making it suitable for routine quality control and ensuring the consistency and safety of efavirenz products.