UFLC Method Development and Validation for Anti-retroviral Drugs, Investigation of Greenness Assessment using Complex GAPI, AGREE and AMGS SPREADSHEET

Introduction: The study’s proposal was to establish a method of development and validation by green chromatography technique for antiviral drugs, as well as to assess the proposed method’s greenness using various tools, which is one of the emerging developments in the analytical field. The chromatographic separation is achieved by using Eclipse plus C18 (250 x 4.6, 5 μm) column by applying isocratic elution using the mobile phase containing methanol and isopropyl acetate in the ratio of (60:40% v/v) with 1-mL/min flow rate. The separation of drugs is achieved by using a greener mobile phase. Results: The retention time of ritonavir and ombitasvir was found to be 1.854 and 8.09 minutes, respectively. The regression coefficient (R2) is 0.997 for both drugs. Accuracy and precision is evaluated for the method and found be within the limit and the results were reproducible. Assessment of the method was carried out using the three different tools. The developed method is anticipated to be eco-friendly, an alternative to the developed method HPLC method in regard to safe solvent, less toxic and less run time. The developed method was found suitable for the simultaneous estimation in their combined dosage form.