Method Development and Validation of Elabasvir and Grazoprevir by using RP-HPLC

Sri Mounika, M. Shymala*
Department of Pharmaceutical Analysis, Joginapally B.R. Pharmacy College, Yenkapally, Hyderabad, Telangana–500075

A B S T R A C T
A new method was established for simultaneous estimation of Elbasvir and Grazopravir by RP-HPLC method.  The chromatographic conditions were successfully developed for the separation of Elbasvir and Grazopravir by using Symmetry (4.6 x 250mm, 5mm), flow rate was 1.0ml/min, detection wave length was 264nm. The instrument used was WATERS HPLC Auto Sampler, Separation module 2695, PDA Detector, Empower-software version-2. The retention times were found to be 2.8 mins and 4.3 mins. The assay of Elbasvir and Grazopravir was performed with tablets and the % assay was found to be 100.18 and 100.00 which shows that the method is useful for routine analysis. The linearity of Elbosvir and Grazopravir was found to be linear with a correlation coefficient of 0.999 and 0.999, which shows that the method is capable of producing good sensitivity. The acceptance criteria of precision is RSD should be not more than 2.0% and the method show  precision 0.5 and 0.1 for Elbasvir and Grazopravir which shows that the method is precise. The acceptance criteria of intermediate precision is RSD should be not more than 2.0% and the method show  precision 0.6 and 0.2 for Elbasvir and Grazopravir which shows that the method is repeatable when performed in different days also. The accuracy limit is the percentage recovery should be in the range of 97.0% – 103.0%.  The total recovery was found to be 100.11% and 100.38% for Elbasvir and Grazopravir. The validation of developed method shows that the accuracy is well within the limit, which shows that the method is capable of showing good accuracy and reproducibility. The acceptance criteria for LOD and LOQ is 3 and 10.The LOD and LOQ for Elbosvir was found to be 2.97 and 9.91 and LOD and LOQ for Grazopravir was found to be 3.09 and 10.07. The robustness limit for mobile phase variation and flow rate variation are well within the limit, which shows that the method is having good system suitability and precision under given set of conditions. 
Keywords: Elbasvir, Grazopravir, HPLC