Determination of rutin in human plasma by high-performance liquid chromatography utilizing solid-phase extraction and ultraviolet detection.
An HPLC method for determining a flavonol glycoside, rutin, in human plasma is presented for application to the pharmacokinetic study. Isocratic reversed-phase HPLC was employed for the quantitative analysis by using kaempferol-3-rutinoside as an internal standard. Solid-phase extraction was performed on an Oasis MAX cartridge possessing reversed-phase and anion-exchange functions (recovery, approximately 80%). The HPLC assay was carried out using a Luna ODS-2 column (150 x 2.1 mm I.D., 5 microm particle size). The mobile phase was acetonitrile-10 mM ammonium acetate solution containing 0.3 mM EDTA-glacial acetic acid (16.5:82.5:1, v/v, pH 3.8). The flow-rate was 0.3 ml/min. The detection wavelength was set at 370 nm. Calibration of the overall analytical procedure gave a linear signal (r>0.9999) over a concentration range of 3-1,000 ng/ml of rutin in plasma. The lower limit of quantification was ca. 5 ng/ml of rutin in plasma. The detection limit (defined as signal-to-noise ratio of about 3) was approximately 0.75 ng/ml. A preliminary experiment to investigate the plasma concentration of rutin after oral administration of 500 mg of rutin to a healthy volunteer demonstrated that the present method was suitable for determining rutin in human plasma.
Ishii
K
K
Kyorin University, School of Health Sciences, Hachioji, Tokyo, Japan. ishiikaz@kyorin-u.ac.jp
Furuta
T
T
Kasuya
Y
Y
D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002852:Chromatography, Ion Exchange; D006801:Humans; D012015:Reference Standards; D012431:Rutin / Q000097:blood*; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization; D013056:Spectrophotometry, Ultraviolet
D012431:Rutin