Method Development and Validation of Kaempferol by using High Performance Liquid Chromatography

Pharmacy

Kaempferol, a natural flavonoid compound found in various plant sources, possesses numerous pharmacological properties, including antioxidant, anti-inflammatory, and anticancer activities. In this study, a novel analytical method for the quantitative determination of kaempferol using Reverse- phase high-performance liquid chromatography (RP-HPLC) was developed and validated. The chromatographic separation was achieved on a C18 column using a mobile phase consisting of Acetonitrile: Water with 0.1 % formic acid 50:50 with isocratic elution. The detection was performed at a wavelength of 265 nm. The developed method demonstrated excellent linearity over the concentration range of 10-60 μg/mL with a correlation coefficient (R2) of 0.9989. The method exhibited satisfactory precision with intra-day and inter-day relative standard deviations (RSD) below 2%. The accuracy of the method was confirmed by recovery studies, with mean recovery values ranging from 99.96% to 100.17%. Furthermore, the robustness and stability of the method were evaluated under various experimental conditions, demonstrating its reliability and reproducibility. The developed HPLC method provides a rapid, sensitive, and reliable means for the quantitative analysis of kaempferol in plant extracts, dietary supplements, and pharmaceutical formulations. This validated method can be effectively employed for routine quality control and pharmacokinetic studies of kaempferol-containing products.

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