Determination of free and glucuronidated kaempferol in rat plasma by LC-MS/MS: application to pharmacokinetic study.
Flavanoid kaempferol is mainly present as glucuronides and sulfates in rat plasma, and small amounts of the intact aglycone are also detected. In the this study, a rapid, specific and sensitive liquid chromatography-electrospray ionization-tandem mass spectrometry method (HPLC-MS/MS) was developed and validated for determination of kaempferol and its major metabolite glucuronidated kaempferol in rat plasma. A liquid-liquid extraction with acetic ether was involved for the extraction of kaempferol and internal standard. Analytes were separated on a C18 column (150 mm x 2.1 mm, 4.5 microm, Waters Corp.) with isocratic elution at a flow-rate of 0.3 ml min(-1). The mobile phase was consisted of 0.5% formic acid and acetonitrile (50:50, v/v). The Quattro Premier HPLC-MS/MS was operated under the multiple reaction-monitoring mode (MRM) using the electrospray ionization technique. The method was validated according to the FDA guidelines for validation of bioanalytical method. The validated method was successfully applied to the study of the pharmacokinetics in rats after oral administration of kaempferol with different doses.
Zhang
Wei-Dong
WD
Department of Pharmacy, School of Stomatology, Fourth Military Medical University, 145 West Changle Road, Xi'an 710032, PR China.
Wang
Xiao-Juan
XJ
Zhou
Si-Yuan
SY
Gu
Yi
Y
Wang
Rong
R
Zhang
Tao-Li
TL
Gan
Hong-Quan
HQ
D000818:Animals; D002853:Chromatography, Liquid; D004333:Drug Administration Routes; D020719:Glucuronides / Q000008:administration & dosage / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics*; D044949:Kaempferols / Q000008:administration & dosage / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics*; D013058:Mass Spectrometry / Q000379:methods*; D051381:Rats; D012015:Reference Standards; D013997:Time Factors
D020719:Glucuronides; D044949:Kaempferols; C006552:kaempferol