"A QBD-ORIENTED RP-HPLC ANALYTICAL METHOD FOR CONCURRENT ESTIMATION OF LAMIVUDINE AND ZIDOVUDINE IN TABLET DOSAGE FORM"
DOI:
https://doi.org/10.22159/ijap.2026v18i4.59178Keywords:
Lamivudine, Zidovudine, Quality by design, Box-behnken design, Anti-retroviral agents, HPLC, Simultaneous estimation, Method development, Validation, Design expert® 13, 3TC, AZTAbstract
Objective: This article aimed to develop a method for the simultaneous estimation of Lamivudine (3TC) and Zidovudine (AZT) by incorporating a quality-by-design (QbD) approach, which is used to develop the most accurate and precise analytical method for tablet formulation in comparison with traditional method development.
Methods: At the initial phase, researchers conducted reverse phase high performance liquid chromatography (RP-HPLC) trials as per the traditional method development protocol and the factors like mobile phase, flow rate and wavelength are taken into consideration for finalizing most suitable trial. Analytical quality by design (AQbD) approach is then implemented as per ICH Q14 guidelines for further steps in method development. The analytical method was systematically optimized by implementing QbD principles, utilizing a design of experiments (DoE) approach specifically through Box-Behnken design (BBD) by using Design Expert® 13 software. The most suitable optimized solution is used for validation. Method was validated in accordance with ICH Q2(R2) guidelines.
Results: Based on traditional method development, Methanol (MeOH): Buffer (Potassium dihydrogen phosphate buffer with sodium salt of pentane sulphonic acid) selected as a mobile phase in 40: 60 (v/v) ratio, the flow rate was selected as 1.5 ml/min and wavelength was selected at 268 nm. On the basis of these factors 17 RP-HPLC runs performed as per BBD protocol. Method was optimized by QbD and the outcomes were 50:50 (v/v) same mobile phase, 1.7 ml/min flow rate and 268 nm wavelength. Optimized parameters were validated. The accuracy for Lamivudine and Zidovudine was found to be 99.43% and 99.84% respectively. The linearity was calculated from range of 5-45 µg/ml for Lamivudine and 10-90 µg/ml for Zidovudine, correlation coefficient (R2) was found to be 0.9998 and 0.9984 for Lamivudine and Zidovudine respectively. Recovery data lies in between 98-102%.
Conclusion: Through systematic risk assessment and Box-Behnken design, a stable method operable design region (MODR) was established. The QbD optimized method achieved a ~39.5% reduction in analysis time (from 5.953 to 3.603 min), which resulted in a ~31.5% net saving in mobile phase volume per injection (from 8.93 to 6.12 ml) despite a slight increase in flow rate. This proves the cost effectiveness of the developed method. The MODR ensures method reliability and consistency across different pharmaceutical scaffolds, providing a cost-effective, high-throughput tool for routine industrial quality control and laboratory testing.
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