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.

Validation of a HPLC/MS method for simultaneous quantification of clonidine, morphine and its metabolites in human plasma

“…Solid-phase extraction was adopted for effective clean-up and enrichment and its simplicity over liquid-liquid extraction, which was adopted by most of the published assays for clonidine (Nirogi et al, 2008;Qin et al, 2015;Zhuang et al, 2015). Both morphine and clonidine are both weakly basic with similar pK a values (8.2 and 8.05, respectively) (Veigure et al, 2017).…”

“…A number of high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) methods that quantified morphine, M3G and M6G and those that quantified clonidine in human plasma were reviewed. The lowest limits of quantification (LLOQ) were 0.25-3.5 ng/mL for morphine, 1-3.5 ng/mL for M3G/M6G (Clavijo et al, 2011;Eckart et al, 2015;Ghassabian et al, 2012) and 0.01-0.25 ng/mL for clonidine (Danafar & Hamidi, 2016;Muller et al, 2007;Nirogi, Kandikere, Mudigonda, & Komarneni, 2008;Parekh, Pudage, Joshi, Vaidya, & Gomes, 2008;Qin et al, 2015;Zhuang et al, 2015), and these methods required plasma volumes ranging from 100 μL up to 1 mL. There was one published HPLC-MS/MS method that simultaneously quantified clonidine, morphine and its two major metabolites that was highly sensitive and only required 100 μL of plasma (Veigure et al, 2017).…”

Review on analytical methods for quantification of ADHD drugs in human biological samples

“…The m/z was 230 for the analyte and 194 and 237 for the IS, respectively [162]. Other LC-MS/MS methods for the quantification of CLN in human plasma samples [163][164][165][166] and their LC-MS chromatographic conditions are reported in Table 2.…”

Development and Validation of an HPLC-UV Detection Assay for the Determination of Clonidine in Mouse Plasma and Its Application to a Pharmacokinetic Study

“…Because of the reported variability in exposure to clonidine, the development of a sensitive and selective procedure for the determination of clonidine level in plasma is required. Various analytical techniques have been reported for clonidine determination by using spectrophotometry [ 6 , 7 , 8 ], potentiometry [ 9 , 10 ], capillary electrophoresis [ 11 ], liquid chromatography-mass spectrometry (LC-MS) [ 12 , 13 , 14 , 15 , 16 ], and gas chromatography-mass spectrometry (GC-MS) [ 17 , 18 ].…”