title: 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.
authors.affiliation: Department of Analytical Development, Pharma Nueva Co., Ltd, Bangkok 10900, Thailand Department of Food and Pharmaceutical Chemistry, Faculty of Pharmaceutical Sciences, Chulalongkorn University, 254 Phayathai Road, Patumwan, Bangkok 10330, Thailand.
authors.forename: Wisut
authors.initials: W
authors.lastname: Wichitnithad
authors.affiliation: Department of Clinical Development, Pharma Nueva Co., Ltd, Bangkok 10900, Thailand.
authors.forename: Ponsiree
authors.initials: P
authors.lastname: Jithavech
authors.affiliation: Department of Clinical Development, Pharma Nueva Co., Ltd, Bangkok 10900, Thailand.
authors.forename: Kingkan
authors.initials: K
authors.lastname: Sanphanya
authors.affiliation: Department of Clinical Development, Pharma Nueva Co., Ltd, Bangkok 10900, Thailand.
authors.forename: Petploy
authors.initials: P
authors.lastname: Vicheantawatchai
authors.affiliation: Department of Food and Pharmaceutical Chemistry, Faculty of Pharmaceutical Sciences, Chulalongkorn University, 254 Phayathai Road, Patumwan, Bangkok 10330, Thailand pornchai.r@chula.ac.th.
authors.forename: Pornchai
authors.initials: P
authors.lastname: Rojsitthisak
mesh_terms: D002138:Calibration; D017332:Cetirizine / Q000097:blood* / Q000493:pharmacokinetics; D002853:Chromatography, Liquid / Q000379:methods*; D039563:Histamine H1 Antagonists, Non-Sedating / Q000097:blood* / Q000493:pharmacokinetics; D006801:Humans; D012015:Reference Standards; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D039563:Histamine H1 Antagonists, Non-Sedating; C472067:levocetirizine; D017332:Cetirizine