Development and Validation of a Rapid and Sensitive Stability-indicating RP-HPLC Method for Resveratrol Quantification in Pharmaceutical Formulation

Abstract:

Background: Trans-resveratrol (RVT) is a phytoconstituent belonging to the family of polyphenols, extensively studied for its numerous health benefits. However, its susceptibility for photolytic, oxidative, and hydrolytic degradation, hinders the quantification of RVT present in different formulations. In the present study, a novel, precise, simple, reproducible, accurate, and stability indicating reversed-phase high performance liquid chromatography (RP-HPLC) method was developed and validated using quality by design for the quantification of RVT in pharmaceutical formulation. Force degradation studies were carried out, and the intrinsic stability of RVT was assessed using the proposed method. Materials and Methods: Chromatographic separation of RVT was obtained on Phenomenex Kinetex C18 (250×4.6 mm, 5 μ, 100 Å) stationary phase with isocratic elution using 0.2% v/v trifluoroacetic acid (TFA) in HPLC grade water and acetonitrile (ACN) in the ratio 75:25% v/v. A flow rate of 1.0 ml/min was maintained for pumping the mobile phase, and the eluent was detected at a wavelength of 306 nm. Results: RVT was found to elute at a retention time (tR) of 6.96 ± 0.08 min. The method was linear in the range of 0.1 μg/ml to 2.0 μg/ml concentration with correlation coefficient 0.9998. The % recovery was found to be 100.16 to 101.73, % RSD of six replicate standards was found to be 0.58. The excipients present in the formulation did not show any interference in the analysis of RVT, indicating the specificity of the developed method. Conclusion: The proposed method can be applied for the quantification of RVT in a pharmaceutical formulation, dietary supplements, and bulk drug.

Key words: Trans-resveratrol, RP-HPLC, Method validation, Force degradation, Specificity