Effects of flavonoids on plasma coagulation tested in vitro by "screening tests"

The effects of four flavonoids: rutin, quercetin, morin and hesperidin on plasma coagulation has been examined by measuring aPTT, TT and PT. It is found that water solution of these flavonoids shows no effects in the measured quantities. This is due to their low solubility in water, and therefore, their low concentrations. Ethanolic solutions of these studied flavonoids, however, show significant prolongation of all measured coagulation times, but compared to ethanol alone, their anticoagulant effects are smaller. It is suggested that the observed effects can be attributed to the interaction of flavonoids with human plasma proteins

Development and validation of spectrofluorimetric and LC–MS/MS methods for the determination of hesperidin in human plasma and pharmaceutical forms

The spectrofluorometric method, based on fluorescence properties of aluminium (III)–hesperidin complex, for the determination of hesperidin in human plasma and pharmaceutical forms has been developed and validated. The complex shows strong emission in the presence of surfactant betain sulfonate SB 12 at 476 nm with excitation at 390 nm. Linearity range in pharmaceutical forms of hesperidin was 0.06 – 24.4 μg mL-1 with LOD 0.016 μg mL-1 and LOQ 0.049 μg mL-1. Recovery values in the range 99.3 – 99.7% indicate good accuracy of the method. A linear dependence of the intensity of fluorescence of the complex on the concentration of hesperidin in plasma was obtained in concentration range from 0.1 – 12.2 μg mL-1. The LOD was 0.032 μg mL-1 while LOQ was 0.096 μg mL-1. Recovery values were in the range 98.4 – 99.8%. The reliability of the method was checked by LC-MS/MS method for plasma samples and HPLC/UV method for tablets with direct determination of hesperidin after separation. Linearity range in determination of hesperidin in pharmaceutical forms was obtained in the range from 0.05 to 10.00 μg mL-1. The LOD was 0.01 μg mL-1 and LOQ was 0.03 μg mL-1. Linearity range in plasma determination of hesperidin was 0.02 – 10.00 μg mL-1 with LOD 0.005 μg mL-1 and LOQ 0.015 μg mL-1. Good agreement between two methods indicate the usability of the proposed spectroflurometric method for the simple, precise and accurate determination of hesperidin in clinical and quality control laboratories