Stability indicating novel analytical method development by RP-UPLC for the estimation of Gitingensine in bulk

Abstract

A new method was proposed using RP-UPLC for the determination of Gitingensine in bulk, which exhibits its power of stability output. Gitingensine is a natural product found in Kibatalia laurifolia belonging to Apocynaceae which is a steroid having the activities such as anti-inflammatory, anti-spasmodic and anti-cancer activity. Cevadine is used as an internal standard for chromatographic analysis. The elution was performed on BEH C18 (2.1 × 50 mm, 1.7 µm) column at 30 °C with a mobile phase distribution Acetonitrile: 0.1% orthophosphoric acid (60: 40) respectively. The flow rate was well- kept at 0.3 mL min-1. Retention times for Gitingensine and Cevadine were found to be 2.005 and 1. 395 min, respectively. The regression equation was found to be linear in the range of 12.5 – 75 µg/mL with a high correlation coefficient (0.999). Recovery of Gitingensine was obtained as 100.04%. Validation was done as claimed by ICH guidelines with respect to accuracy, sensitivity, robustness, and precision studies. From the insignificant variations in the analysis by changing the mobile phase, temperature, and flow rate, the robustness was studied. All the validation parameters were found to be within the specifications. Forced degradation studies revealed that when the influence of acid, alkali, peroxide, thermal, photolytic, and hydrolytic conditions were applied on the drug, it was stable. Hence, it can be concluded that the developed RP-UPLC method is economical, precise, and robust and can be adopted in regular Quality control analysis.