34 Pharmaceutical Methods Vol 5 ● Issue 1 ● Jan-Jun 2014 Development and Validation of UV Spectrophotometric Method for the Estimation of Kaempferol in Kaempferol: Hydrogenated Soy PhosphatidylCholine (HSPC) Complex Darshan R. Telange 1 *, Arun T. Patil 1 , Amol Tatode 1 , Bhushan Bhoyar 1 1 Department of Pharmaceutics, University Department of Pharmaceutical Sciences, Rashtrasant Tukadoji Maharaj Nagpur University, Nagpur, Maharashtra, India ABSTRACT Introduction: Kaempferol (3, 5, 7-trihydroxy-2-(4-hydroxyphenyl)-4H-1-benzopyran-4-one) is a natural flavonoid belongs to a subcategory of flavonol family. The kaempferol – hydrogenated soy phosphatidylcholine (HSPC) complex was obtained by refluxing and freeze drying method. Ultra violet (UV) – visible spectrophotometric method has been developed for the determination of kaempferol in kaempferol – HSPC complex. Objective: A validated UV – visible spectrophotometric method for determination of kaempferol in Kaempferol – HSPC complex. Materials and Methods: The Kaempferol – HSPC complex (Phytosomes) were prepared by dissolving both kaempferol and HSPC in 1, 4 – dioxane for refluxing up to 2 h and freeze dried. The spectrophotometric detection of kaempferol was done at absorption maximum (λ max ) 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 (r 2 > 0.99) at 365 nm and 2-14 μg/ml (r 2 > 0.99) at 265 nm. Precision and mean recoveries were found to be in the range of (%RSD 0.0957 and 0.0580) and (% RSD 0.1461 and 0.0959) and 99.70% and 91.85% at 365 nm and at 265 nm. Limit of detection and limit of quantification were found to be (0.015 μg/ml and 0.0191 μg/ml) and (0.0457 μg/ml and 0.0579 μg/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. Keywords: Freeze drying, hydrogenated soy phosphatidylcholine, Kaempferol, method validation, UV – visible spectrophotometer *Corresponding address: Dr. Darshan R. Telange, Department of Pharmaceutics, University Department of Pharmaceutical Sciences, Rashtrasant Tukadoji Maharaj Nagpur University, Nagpur, Maharashtra - 440 033, India. E-mail: telange.darshan@gmail.com DOI: 10.5530/phm.2014.1.6 INTRODUCTION Kaempferol (3, 5, 7-trihydroxy-2-(4-hydroxyphenyl)-4H- 1-benzopyran-4-one), a yellow compound with a low molecular weight (MW: 286.2 g/mol), is a common natural flavonoid which representative of the subcategory of flavonol. This flavonoid is abundant in many plant-derived foods and traditional medicine. 1 It is widely distributed in the plant kingdom such as onion, kale, endive and tea along with formed as secondary metabolites through the phenylpropenoid biosynthetic pathway. 2 Although it has broad spectrum importance, researcher have been isolated it from different plants like Asclepias cyriaca L., 3 Crocus sativus L., 4 Cassia alata L., 5 Capsella bursa-patoris L., 6 Leptadenia pyrotechnica L., 7 and broccoli 8 and also reported its various pharmacotherapeutic effects like anticancer, 7 antioxidant, 9,10 anti-inflammatory 11 and hepatoprotective 12 etc. kaempferol revealed low to moderate absorption, which results poor bioavailability ~2%. 13 It is hydrophobic in nature and freely soluble in methanol, 1, 4 – dioxane, Ethanol and dimethylformamide. 14,15 In a wide range of HPLC and HPTLC techniques were reported for estimation of kaempferol in extracts, 16-20 gingko biloba tablets, 21,22 phytosome formulation 23 (Kaempferol – phospholipids complex) at around 360 nm. Literature survey revealed that no simple UV – visible method has Original Article