Determination of (+)-catechin in plasma by high-performance liquid chromatography using fluorescence detection.
A high-performance liquid chromatographic method, using fluorescence detection, was developed for the determination of (+)-catechin in rabbit plasma. The procedure involved the precipitation of plasma protein using acetonitrile, followed by solid-phase adsorption onto alumina. After washing with water and methanol, the residue was vortex-mixed with perchloric acid solution to release the adsorbed (+)-catechin. Separation was performed on a reversed-phase column using an eluent consisting of phosphoric acid solution with 12% acetonitrile. The excitation and emission wavelengths were set at 280 and 310 nm, respectively. The retention times for (+)-catechin and the internal standard (deoxyhigenamine) were 6.87 and 8.47 min respectively, without any interference. Validations of accuracy and precision were satisfactory in both within- and between-run assays. All coefficients of variance were less than 6% and mean relative errors were within +/- 3.75%. The average recovery was 73.77%. The limit of detection and quantitation were 1 ng and 0.02 micrograms/ml, respectively. Application of this method was successfully assessed by intravenous administration of a 15 mg/kg dose of (+)-catechin in rabbits. This new method provides a simple, specific and sensitive determination for (+)-catechin in rabbit plasma and is suitable for pharmacokinetic studies.
Ho
Y
Y
Department of Pharmaceutics, School of Pharmacy, Taipei Medical College, Taiwan.
Lee
Y L
YL
Hsu
K Y
KY
D000097:Acetonitriles; D000327:Adsorption; D000537:Aluminum Oxide; D000818:Animals; D002392:Catechin / Q000097:blood* / Q000493:pharmacokinetics; D011232:Chemical Precipitation; D002851:Chromatography, High Pressure Liquid / Q000379:methods* / Q000706:statistics & numerical data; D004355:Drug Stability; D008297:Male; D011817:Rabbits; D012680:Sensitivity and Specificity; D013053:Spectrophotometry
D000097:Acetonitriles; D002392:Catechin; D000537:Aluminum Oxide; C032159:acetonitrile