Journal
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
Volume 823, Issue 2, Pages 108-114Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jchromb.2005.06.016
Keywords
puerarin; Kudzu; human plasma; reversed-phase HPLC
Funding
- NIAAA NIH HHS [AA10536] Funding Source: Medline
- NIDA NIH HHS [DA00345] Funding Source: Medline
- PHS HHS [N44A02006] Funding Source: Medline
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Puerarin, an isoflavone C-glycoside, has been identified as the major active component isolated from Pueraria lobata (Kudzu) responsible for suppression of alcohol drinking. In order to conduct clinical studies of Kudzu's efficacy, a method for measuring its bioavailability and pharmacokinetic profile is needed. We have developed a gradient reversed-phase HPLC system for pharmacokinetic study of puerarin in human plasma. Solid-phase extraction was performed on an abselut Nexus cartridge (60 mg/3 ml) possessing adsorbent function with a recovery of > 97% and 4-hydroxybenzoic acid was used as an internal standard. The HPLC assay was performed on a YMC ODS-A column (150 mm x 4.6 mm i.d., 5 mu m particle size). The HPLC mobilephase consisted of methanol/0.5% acetic acid with 20-35% methanol gradient at a flow-rate of 0.8 ml/min. The UV wavelength was set at 254 nm. Calibration of the overall analytical procedure gave a linear signal (r > 0.999) over a puerarin concentration range of 5-500 ng/ml in human plasma. The lower limit of quantification was ca. at 8 ng/ml of puerarin in plasma. The detection limit (defined as signal-to-noise ratio of about 3) was approximately 3 ng/ml. The preliminary pharmacokinetic study after oral administration of the Kudzu capsules containing 400 mg of puerarin to a healthy volunteer confirmed that the present method was suitable for determining puerarin in human plasma. (c) 2005 Elsevier B.V. All rights reserved.
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