title: Quantitation of clevidipine in dog blood by liquid chromatography tandem mass spectrometry: application to a pharmacokinetic study.
abstract: Clevidipine, a vascular selective calcium channel antagonist of the dihydropyridine class, is rapidly metabolized by ester hydrolysis because of incorporation of an ester linkage into the drug molecule. To characterize its pharmacokinetic profiles in dogs, a simple, rapid and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for quantitation of clevidipine in dog blood. After one-step protein precipitation with methanol, the chromatographic separation was carried out on an Ecosil C18 column (150mm×4.6mm, 5μm) with a gradient mobile phase consisting of methanol and 5mM ammonium formate at a flow rate of 0.5mL/min. The quantitation analysis was performed using multiple reaction monitoring (MRM) at the specific ion transitions of m/z 454.1 [M-H](-)→m/z 234.1 for clevidipine and m/z 256.1 [M-H](-)→m/z 227.1 for elofesalamide (internal standard) in the negative ion mode with electrospray ionization (ESI) source. This validated LC-MS/MS method showed good linearity over the range 0.5-100ng/mL with the lower limit of quantitation (LLOQ) of 0.5ng/mL together with the satisfied intra- and inter-day precision, accuracy, extraction recovery and matrix effect. Stability testing indicated that clevidipine in dog blood with the addition of denaturant methanol was stable on workbench for 1h, at -80°C for up to 30 days, and after three freeze-thaw cycles. Extracted samples were also observed to be stable over 24h in an auto-sampler at 4°C. The validated method has been successfully applied to a pharmacokinetic study of clevidipine injection to 8 healthy Beagle dogs following intravenous infusion at a flow rate of 5mg/h for 0.5h.
authors.affiliation: Hubei Province Key Laboratory of Biotechnology of Chinese Traditional Medicine, Hubei Collaborative Innovation Center for Green Transformation of Bio-resources, Hubei University, Wuhan 430062, China; State Key Laboratory of Drug Delivery Technology and Pharmacokinetics, Tianjin Institute of Pharmaceutical Research, Tianjin 300193, China.
authors.forename: Huihui
authors.initials: H
authors.lastname: Wei
authors.affiliation: State Key Laboratory of Drug Delivery Technology and Pharmacokinetics, Tianjin Institute of Pharmaceutical Research, Tianjin 300193, China.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Gu
authors.affiliation: State Key Laboratory of Drug Delivery Technology and Pharmacokinetics, Tianjin Institute of Pharmaceutical Research, Tianjin 300193, China.
authors.forename: Yanping
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Hubei Province Key Laboratory of Biotechnology of Chinese Traditional Medicine, Hubei Collaborative Innovation Center for Green Transformation of Bio-resources, Hubei University, Wuhan 430062, China. Electronic address: cy101610@qq.com.
authors.forename: Yong
authors.initials: Y
authors.lastname: Chen
authors.affiliation: State Key Laboratory of Drug Delivery Technology and Pharmacokinetics, Tianjin Institute of Pharmaceutical Research, Tianjin 300193, China.
authors.forename: Changxiao
authors.initials: C
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Drug Delivery Technology and Pharmacokinetics, Tianjin Institute of Pharmaceutical Research, Tianjin 300193, China. Electronic address: tjpks@163.com.
authors.forename: Duanyun
authors.initials: D
authors.lastname: Si
mesh_terms: D000818:Animals; D001798:Blood Proteins / Q000737:chemistry; D002121:Calcium Channel Blockers; D011232:Chemical Precipitation; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004285:Dogs; D011725:Pyridines / Q000097:blood* / Q000493:pharmacokinetics*; D011786:Quality Control; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D001798:Blood Proteins; D002121:Calcium Channel Blockers; D011725:Pyridines; C118563:clevidipine
keywords: Clevidipine; Dog blood; LC–MS/MS; Pharmacokinetics