Simultaneous determination of imperatorin and its 2 metabolites in dog plasma by using liquid chromatography-tandem mass spectrometry.
In this study, 2 metabolites of imperatorin, imperatorin hydroxylate (IMH) and imperatorin epoxide (IME), were identified for the first time in dog plasma. A sensitive, specific, and accurate high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was then developed for the simultaneous quantification of imperatorin and its 2 metabolites in dog plasma. Separation was achieved on an Agilent ZORBAX Extend-C(18) column (2.1 mm × 50 mm, 3.5 μm) at 30 °C. The mobile phase consisted of 0.02% ammonium acetate solution-methanol with a gradient program at a flow rate of 0.3 mL/min. Detection was performed using an electrospray ionization source operating in positive ion multiple reaction monitoring mode and by monitoring the ion transitions from 271 to 203 m/z for imperatorin, 309.4-224.1 m/z for IMH, 287-203 m/z for IME, and 441.3-325.2 m/z for simvastatin (the internal standard). Good linearity was shown over the concentration range of 1-500 ng/mL for imperatorin, and 0.2-500 ng/mL for IMH and IME. The validated method was successfully applied to a pharmacokinetic study of imperatorin in beagle dogs. The pharmacokinetic profiles of imperatorin and its 2 metabolites showed sex differences after the i.v. administration of imperatorin at a dose of 5 mg/kg.
College of Pharmaceutical Sciences, Zhejiang University, 886 Yuhangtang Road, Hangzhou 310058, China.
Lu
L
Wang
Wen
W
Lu
Qi
Q
Shen
Shengjia
S
Wang
Hui
H
Zhou
Lushan
L
Yu
Sicen
S
Wang
Huidi
H
Jiang
Langchong
L
He
Su
S
Zeng
D000085:Acetates / Q000737:chemistry; D000284:Administration, Oral; D000818:Animals; D001711:Biotransformation; D002138:Calibration; D002853:Chromatography, Liquid / Q000592:standards; D004285:Dogs; D004365:Drugs, Chinese Herbal / Q000008:administration & dosage / Q000737:chemistry / Q000378:metabolism* / Q000493:pharmacokinetics*; D004852:Epoxy Compounds / Q000097:blood; D005243:Feces / Q000737:chemistry; D005260:Female; D011564:Furocoumarins / Q000008:administration & dosage / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics*; D006900:Hydroxylation; D007275:Injections, Intravenous; D016014:Linear Models; D009682:Magnetic Resonance Spectroscopy; D008297:Male; D000432:Methanol / Q000737:chemistry; D015394:Molecular Structure; D051381:Rats; D017207:Rats, Sprague-Dawley; D012015:Reference Standards; D012680:Sensitivity and Specificity; D012737:Sex Factors; D021241:Spectrometry, Mass, Electrospray Ionization; D053719:Tandem Mass Spectrometry / Q000592:standards
D000085:Acetates; D004365:Drugs, Chinese Herbal; D004852:Epoxy Compounds; D011564:Furocoumarins; C031534:imperatorin; C018824:ammonium acetate; D000432:Methanol