Spectrofluorimetric and HPLC Determination of Morin in Human Serum

Abstract

Morin is a flavonol antioxidant. In ethanol-water mixtures (70 wt% of ethanol) it reacts with Al3+ to give Al(Morin)(2) in the pH range 3-6. The conditional stability constant of this complex at 298 K was found to be log beta(2) = 16.96 +/- 0.02 at pH 4.40. The complex shows strong fluorescence emission at 500 nm upon excitation at 410 nm. The fluorescence intensity is pH dependent with maximum emission at pH 4.40. Since the complexation reaction enhances the fluorescence of morin, this property was used for the determination of morin in human serum. A linear dependence of the intensity of fluorescence of the complex on the concentration of morin was obtained in morin concentration range from 1.5-30.5 ng mL(-1), relative standard error of measurements was 1.4%. The LOD was 0.02 ng mL(-1) while LOQ was 1.0 ng mL(-1). Serum concentration of morin was also determined using HPLC as a reference method. A C-18 Hypersil Gold AQ column was used with acetonitrile-0.1% v/v phosphoric acid (30:70% v/v) as the mobile phase at 1.0 mL min(-1) flow rate and UV detection at 250 nm. Acceptable relative standard errors (less than 5%) between determinations obtained by the two methods indicate that the fluorescence method is reliable.