Development and Validation of UV Spectrophotometric Method for the Estimation of Kaempferol in Kaempferol: Hydrogenated Soy PhosphatidylCholine (HSPC) Complex

Abstract

Introduction: Kaempferol (3,5,7-trihydroxy-2-(4-hydroxyphenyl)-4H-1-benzopyran-4-one) is a natural flavonoid belongs to subcategory of flavonol family. The Kaempferol – Hydrogenated Soy Phosphatidylcholine (HSPC) Complex was obtained by refluxing and freeze drying method. UV – Visible Spectrophotometric method has been developed for determination of Kaempferol in Kaempferol – HSPC Complex. Objective: A validated UV – Visible spectrophotometric method for determination of Kaempferol in Kaempferol – HSPC complex. Methods: The Kaempferol – HSPC Complex (Phytosomes) were prepared by dissolving both Kaempferol and Hydrogenated Soy Phosphatidylcholine (HSPC) in 1, 4 – dioxane for refluxing up to 2h and freeze dried. The spectrophotometric detection of kaempferol was done at absorption maximum (λmax) of 365 nm and 265 nm using methanol as solvent. The developed method was validated as per ICH guidelines. Result: The Kaempferol content in Kaempferol – HSPC Complex was found to be 79.32% and 79.19% at 365 nm and 265 nm. Kaempferol demonstrated good linearity in concentration range of 2 – 12 µg/ml (r2>0.99) at 365 nm and 2 – 14 µg/ml (r 2 >0.99) at 265nm. Precision and mean recoveries were found to be in the range of (% RSD 0.0957 & 0.0580) and (% RSD 0.1461 & 0.0959) and 99.70 % & 91.85 % at 365 nm and at 265 nm. LOD and LOQ were found to be (0.015µg/ml & 0.0191µg/ml) and (0.0457µg/ml & 0.0579µg/ml) respectively. Conclusion: The developed method was found to be simple, specific, economic, reliable, accurate, precise, reproducible and used as a quality control tool for analysis of Kaempferol. Key Words: Freeze drying, HSPC, Kaempferol, Method validation, UV – Visible spectrophotometer.