Determination of Levocetirizine in Human Plasma by LC–MS-MS: Validation and Application in a Pharmacokinetic Study

Abstract

A fast and simple sample cleanup approach for levocetirizine in human was developed using protein precipitation coupled with LC-MS-MS. Samples were treated with 6% trichloroacetic acid in water prior to LC-MS-MS analysis. Chromatographic separation was performed on a reverse phase column with an isocratic mobile phase of acetonitrile and 10 mM ammonium formate pH 3.5 (80:20, v/v) at a flow rate of 1.0 mL/min. The run time was 3.5 min. Mass parameters were optimized to monitor transitions at m/z [M+H] + 389.0→201.0 for levocetirizine and m/z [M+H] + 375.3→201.0 for hydroxyzine as internal standard. The lower limit of quantification and the dynamic range were 1.00 and 1.00-500 ng/mL, respectively. Linearity was good for intraday and interday validations (r 2 ≥ 0.995). The mean recoveries were 59 and 69% for levocetirizine and hydroxyzine, respectively. Matrix effect was acceptable with %CV < 15. Hemolytic effect was negligible. Levocetirizine was stable in human plasma for 27 h at room temperature (25°C), for 16 weeks frozen at −70°C, 4 weeks frozen at −20°C, for 24 h in an autosampler at 15°C and for three freeze/thaw cycles. The validated method was applied in a pharmacokinetic study to determine the concentration of levocetirizine in plasma samples. The study provides a fast and simple bioanalytical method for routine analysis and may be particularly useful for bioequivalence studies.

Bioanalysis of antihistamines for clinical or forensic purposes

“…The published chromatographic methods for the determination of antihistamines include mainly liquid–liquid extraction (LLE) and solid‐phase extraction (SPE) procedures or direct injection into the chromatographic system (Tables ). Protein precipitation of the samples with methanol (Ming et al , ), acetonitrile (Naidong et al , ; Gupta et al , ; Vlase et al , ), HClO 4 25% (Moncrieff, ) or trichloroacetic acid 5–6% (Lange et al , ; Shakya et al , ; Wichitnithad et al , ) have been used prior to LC analysis.…”

“…The published chromatographic methods for the determination of antihistamines include mainly liquid-liquid extraction (LLE) and solid-phase extraction (SPE) procedures or direct injection into the chromatographic system (Tables 2-5). Protein precipitation of the samples with methanol (Ming et al, 2007), acetonitrile (Naidong et al, 2002;Gupta et al, 2005;Vlase et al, 2007), HClO 4 25% (Moncrieff, 1992) or trichloroacetic acid 5-6% (Lange et al, 1968;Shakya et al, 2009;Wichitnithad et al, 2015) have been used prior to LC analysis. Solans et al (2007) subjected plasma samples to enzymatic hydrolysis with β-glucuronidase/arylsulfatase from Helix pomatia in order to quantify free and glucuronoconjugated 3-hydroxydesloratadine, a common metabolite of desloratadine and rupatadine.…”

Continuous synthesis of BaFe2O4 and BaFe12O19 nanoparticles in a droplet microreactor for efficient detection of antihistamine drugs in oral fluid using surface-assisted laser desorption/ionization mass spectrometry

“…These observations have been reported in previous studies. 58,59 The further investigation involved the analysis of levocetirizine tablet-spiked OF (Fig. 6c and d).…”