Novel LC-MS/MS method for analyzing imperialine in rat plasma: development, validation, and application to pharmacokinetics.
A sensitive, selective, rapid liquid chromatography-electrospray ionization-tandem mass spectrometric method was developed and validated in rat plasma to quantify imperialine, a major active constituent extracted from Bulbus Fritillariae Cirrhosae. Before analysis, plasma samples were pre-treated using cost-effective protein precipitation in order to extract imperialine and the internal standard, carbamzepine. The two analytes were then separated on a Diamonsil ODS chromatography column using gradient elution with a mobile phase of 0.1% aqueous formic acid and acetonitrile. Mass spectrometry was carried out in multiple reaction monitoring mode using a positive electrospray ionization interface. The calibration curve was linear (r(2)=0.9998) over the concentration range 2-1000ng/mL, while the validated limit of determination (LOD) was 0.5ng/mL. Precision varied from 0.1% to 7.1%, and the accuracy varied within ±2%. The method proved robust to sample freezing and thawing, as well as short- and long-term sample storage. The developed method was successfully applied to the pharmacokinetic study of imperialine in rats. Different amounts of imperialine were administered in single doses orally or through the caudal vena cava, and pharmacokinetic parameters were evaluated. Oral bioavailability with a dose of 1mg/kg was 31.2%; 5mg/kg, 53.6%; and 10mg/kg, 47.4%.
Key Laboratory of Drug Targeting and Drug Delivery Systems, Ministry of Education, West China School of Pharmacy, Sichuan University, Chengdu 610041, PR China.
Qing
Q
Lin
Quan
Q
Zhang
Xu
X
Song
Tao
T
Gong
Xun
X
Sun
Zhirong
Z
Zhang
D000284:Administration, Oral; D000818:Animals; D001682:Biological Availability; D002595:Cevanes / Q000008:administration & dosage / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics*; D002853:Chromatography, Liquid / Q000379:methods*; D004355:Drug Stability; D057230:Limit of Detection; D016014:Linear Models; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods*
D002595:Cevanes; C047331:peiminine
Imperialine; LC–MS/MS; Pharmacokinetic study; Rat plasma; Validation