Simple, sensitive and rapid HPLC-MS/MS method for the determination of cepharanthine in human plasma.
A rapid, sensitive and specific method for the determination of cepharanthine in human plasma using high performance liquid chromatography coupled with tandem mass spectrometry (HPLC-MS/MS) was described. Cepharanthine and the internal standard (I.S.), telmisartan, were extracted from human plasma by methanol to precipitate the protein. A centrifuged upper layer was then evaporated and reconstituted with 100μL methanol. Chromatographic separation was performed on an AGILENT XDB-C(8) column (150mm×2.1mm, 5.0μm, Agilent, USA) using a gradient mobile phase with 1mmol/L ammonium acetate in water with 0.05% formic acid and methanol. Detection and quantitation was performed by MS/MS using electrospray ionization (ESI) and multiple reaction monitoring (MRM) in the positive ion mode. The most intense [M+H](+) MRM transition of cepharanthine at m/z 607.3→365.3 was used for quantitation and the transition at m/z 515.5→276.4 was used to monitor telmisartan. The calibration curve was linear within the concentration range of 0.5-200.0ng/mL (r=0.9994). The limit of quantification (LOQ) was 0.5ng/mL. The extraction recovery was above 81.1%. The accuracy was higher than 92.3%. The intra- and inter-day precisions were less than 9.66%. The method was accurate, sensitive and simple and was successfully applied to a pharmacokinetic study after single intravenous administration of 50mg cepharanthine in 12 healthy Chinese volunteers.
Hao
Guangtao
G
Department of Pharmacology, Affiliated Hospital of the Academy of Military Medical Sciences, No. 8, East Street, Fengtai District, Beijing 100071, PR China.
Liang
Haixia
H
Li
Yuanyuan
Y
Li
Haiyan
H
Gao
Hongzhi
H
Liu
Guang
G
Liu
Zeyuan
Z
D044182:Benzylisoquinolines / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004355:Drug Stability; D006801:Humans; D016014:Linear Models; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods*; D052117:Benzodioxoles
D044182:Benzylisoquinolines; C006947:cepharanthine; D052117:Benzodioxoles