Int. J. Pharm. Investigation, 2024; 14(4):1-11. https://www.jpionline.org Original Article International Journal of Pharmaceutical Investigation, Vol 14, Issue 4, Oct-Dec, 2024 1 DOI: ["DOI"].11.2.1 Copyright Information : Copyright Author (s) 2024 Distributed under Creative Commons CC-BY 4.0 Publishing Partner : EManuscript Tech. [www.emanuscript.in] Analytical Method Development and Validation of Efavirenz by Using RP- HPLC Ishu Tyagi 1 , Kandasamy Nagarajan 2, * , Snigdha Bhardwaj 3 1 Department of Pharmaceutical Quality Assurance, KIET School of Pharmacy, KIET Group of Institutions, Ghaziabad, Delhi-NCR, INDIA. 2 Department of Pharmaceutical Chemistry, KIET School of Pharmacy, KIET Group of Institutions, Ghaziabad, Delhi-NCR, INDIA. 3 Department of Pharmaceutics, KIET School of Pharmacy, KIET Group of Institutions, Ghaziabad, Delhi-NCR, INDIA. ABSTRACT 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 of 10 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 R 2 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. Keywords: Efavirenz, HIV/AIDS, RP-HPLC, LOD, LOQ, Rt. INTRODUCTION Efavirenz, an antiretroviral medication primarily used in the treatment of HIV/AIDS. It belongs to the class of drugs known as Non-Nucleoside Reverse Transcriptase Inhibitors (NNRTIs). It works by inhibiting the activity of the enzyme reverse transcriptase, which is essential for the replication of HIV. By blocking reverse transcriptase, Efavirenz prevents HIV from converting its RNA to DNA, thereby halting the virus’s ability to replicate and reducing the viral load in the body. Efavirenz is typically administered orally in the form of tablets or capsules. e recommended dosage for adults is usually 600 mg once daily, preferably taken on an empty stomach. Some formulations of efavirenz may be taken with food to reduce the risk of gastrointestinal side effects. Efavirenz can interact with a wide range of medications, including certain antibiotics, antifungals, antidepressants and anticonvulsants. It’s crucial for patients to inform their healthcare provider about all the medications, supplements and herbal remedies they are taking to avoid potential interactions that could affect the efficacy or safety of Efavirenz. Antiretroviral drugs from different classes, Efavirenz helps to control the progression of HIV infection and improve immune function. 1,2 Efavirenz may include dizziness, drowsiness, trouble sleeping, vivid dreams and rash. Some individuals may experience psychiatric symptoms such as mood changes, depression, anxiety, or hallucinations, particularly during the initial weeks of treatment. Most side effects tend to be temporary and diminish over time as the body adjusts to the medication. However, it's essential for patients to report any severe or persistent side effects to their healthcare provider promptly. 3-5 Efavirenz is not recommended for use during pregnancy, particularly during the first trimester, due to concerns about potential adverse effects on fatal development. Pregnant individuals living with HIV should discuss the risks and benefits Received: x-x-x; Revised: x-x-x; Accepted: x-x-x. Correspondence: Dr. Kandasamy Nagarajan Principal and Professor, Department of Pharmaceutical Chemistry, KIET School of Pharmacy, KIET Group of Institutions, Ghaziabad, Delhi-NCR, INDIA. Email: rknagaraaja@gmail.com