Liquid chromatography tandem mass spectrometry method for the quantification of clonidine with LLOQ of 10 pg/mL in human plasma

Abstract

A sensitive high-performance liquid chromatography-positive ion electrospray tandem mass spectrometry method was developed and validated for the quantification of clonidine in human plasma. Following liquid-liquid extraction, the analytes were separated using an isocratic mobile phase on a reverse-phase column and analyzed by MS/MS in the multiple reaction monitoring mode using the respective [M + H](+) ions, m/z 230 to 44 for clonidine and m/z 254 to 44 for the internal standard. The assay exhibited a linear dynamic range of 10-2000 pg/mL for clonidine in human plasma. The lower limit of quantification was 10 pg/mL with a relative standard deviation of less than 6.8%. Acceptable precision and accuracy were obtained for concentrations over the standard curve range. A run time of 2.5 min for each sample made it possible to analyze more than 250 human plasma samples per day. The validated method was successfully used to analyze human plasma samples for application in pharmacokinetic studies.

A highly sensitive method for the simultaneous UHPLC–MS/MS analysis of clonidine, morphine, midazolam and their metabolites in blood plasma using HFIP as the eluent additive

“…iFunnel voltage in the high pressure region was 210 V and at low pressure it was 220 V. Optimized collision energies for each analyte and the internal standard transitions are listed in Table 1. Fragmentation patterns for these analytes have been documented previously [13,16,20] and support the chosen m/z values for the method. Table 1 goes here***…”

“…In total, 10 liquid chromatography-mass spectrometry (LC-MS) methods were reviewed. The lowest limits of quantification (LLOQ) ranging from 2 -9 ng/mL for morphine and its two major metabolites [7,10,11], 0.01 -100 ng/mL for clonidinebut clonidine was the only single compound analyzed in these assays [3,4,12,13], 0.025 -5 ng/mL for midazolam [10,11,14,15] and 0.1 -2.5 ng/mL for MiOH [10,15]. The exception was the LLOQ of 50 fg/mL for midazolam and 0.25 pg/mL for MiOH [16].…”

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).…”

Avaliações farmacometabolômicas e de ancestralidade genética em pacientes hipertensos de um estudo clínico randomizado

“…Para isso, foi utilizada uma coluna de RACNTs que provou ser robusta e eficiente em cerca de 300 ciclos analíticos, exibindo maior ou igual vida útil em comparação com outros estudos que empregam o mesmo tipo de material 62,70,71,82 . A vida útil dos RACNTs também pode ser confirmada quando comparada a outros materiais de RAM, sendo até 15 vezes maior do que alguns deles [83][84][85][86][87] . Microextração por sorvente embalado.…”