title: Validation of a HPLC-ESI MS/MS method for the determination of clonidine in human plasma and its application in a bioequivalence study in Chinese healthy volunteers.
abstract: A rapid and sensitive liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method to determine clonidine in human plasma was developed and fully validated. Sample preparation was involved an one-step extraction with diethyl ether. Donepezil was employed as the internal standard (IS). Chromatographic separation was performed on a Hypersil BDS C18 column (i.d. 2.1 × 50 mm, particle size 3μm) with a mobile phase of methanol-water (containing 0.1% formic acid; 60:40, v/v) at a flow rate of 200 μL/min. The peaks were detected by mass spectrometry using the electrospray ion source in selected reaction monitoring mode. The extraction recovery was 72.53-85.25%. The method was found to be linear in a concentration range of 0.02-6.00 ng/mL and the lower limit of quantification was 0.02 ng/mL. The within- and between-batch precisions at three concentrations were 4.33-16.47 and 7.24-17.24% with accuracies of -2.47-10.91 and 1.86-10.19%, respectively. This validated method was successfully used for a bioequivalence study of two clonidine transdermal patches on healthy volunteers. The results suggested that the test formulation of clonidine patch met the regulatory criterion for bioequivalence to the reference formulation, but a larger sample size should be needed for the estimation of bioequivalence.
authors.affiliation: Laboratory of Drug Metabolism and Pharmacokinetics, School of Pharmaceutical Sciences, Sun Yat-Sen University, Guang Zhou, 510006, China.
authors.forename: Jialang
authors.identifier: http://orcid.org/0000-0002-9548-3767
authors.initials: J
authors.lastname: Zhuang
authors.affiliation: Laboratory of Drug Metabolism and Pharmacokinetics, School of Pharmaceutical Sciences, Sun Yat-Sen University, Guang Zhou, 510006, China.
authors.forename: Jiangying
authors.initials: J
authors.lastname: Chen
authors.affiliation: Laboratory of Drug Metabolism and Pharmacokinetics, School of Pharmaceutical Sciences, Sun Yat-Sen University, Guang Zhou, 510006, China.
authors.forename: Xueding
authors.initials: X
authors.lastname: Wang
authors.affiliation: Sun Yat-sen University Cancer Center, Guang Zhou, 510060, China.
authors.forename: Yin
authors.initials: Y
authors.lastname: Pang
authors.affiliation: Laboratory of Drug Metabolism and Pharmacokinetics, School of Pharmaceutical Sciences, Sun Yat-Sen University, Guang Zhou, 510006, China.
authors.forename: Huichang
authors.initials: H
authors.lastname: Bi
authors.affiliation: Laboratory of Drug Metabolism and Pharmacokinetics, School of Pharmaceutical Sciences, Sun Yat-Sen University, Guang Zhou, 510006, China.
authors.forename: Lihui
authors.initials: L
authors.lastname: Huang
authors.affiliation: Laboratory of Drug Metabolism and Pharmacokinetics, School of Pharmaceutical Sciences, Sun Yat-Sen University, Guang Zhou, 510006, China.
authors.forename: Guixiong
authors.initials: G
authors.lastname: Zeng
authors.affiliation: The First Affiliated Hospital, Sun Yat-Sen University, Guang Zhou, 510080, China.
authors.forename: Xiaoxing
authors.initials: X
authors.lastname: Liao
authors.affiliation: The First Affiliated Hospital, Sun Yat-Sen University, Guang Zhou, 510080, China.
authors.forename: Zhongfu
authors.initials: Z
authors.lastname: Ma
authors.affiliation: The First Affiliated Hospital, Sun Yat-Sen University, Guang Zhou, 510080, China.
authors.forename: Xiao
authors.initials: X
authors.lastname: Chen
authors.affiliation: Laboratory of Drug Metabolism and Pharmacokinetics, School of Pharmaceutical Sciences, Sun Yat-Sen University, Guang Zhou, 510006, China.
authors.forename: Guoping
authors.initials: G
authors.lastname: Zhong
authors.affiliation: Laboratory of Drug Metabolism and Pharmacokinetics, School of Pharmaceutical Sciences, Sun Yat-Sen University, Guang Zhou, 510006, China.
authors.forename: Min
authors.initials: M
authors.lastname: Huang
authors.affiliation: Laboratory of Drug Metabolism and Pharmacokinetics, School of Pharmaceutical Sciences, Sun Yat-Sen University, Guang Zhou, 510006, China.
authors.forename: Xianglan
authors.initials: X
authors.lastname: Zhao
mesh_terms: D000328:Adult; D019540:Area Under Curve; D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003000:Clonidine / Q000008:administration & dosage / Q000097:blood* / Q000493:pharmacokinetics*; D064368:Healthy Volunteers; D006801:Humans; D008297:Male; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000379:methods; D013810:Therapeutic Equivalency; D057968:Transdermal Patch
chemical_list: D003000:Clonidine
keywords: Clonidine; HPLC-MS/MS; bioequivalence; human plasma