title: Sensitive and rapid liquid chromatography/tandem mass spectrometric assay for the quantification of chloroquine in dog plasma.
abstract: A simple, sensitive and rapid liquid chromatography/tandem mass spectrometric (LC-MS/MS) method was developed and validated for quantification of chloroquine, an antimalarial drug, in plasma using its structural analogue, piperazine bis chloroquinoline as internal standard (IS). The method is based on simple protein precipitation with methanol followed by a rapid isocratic elution with 10 mM ammonium acetate buffer/methanol (25/75, v/v, pH 4.6) on Chromolith SpeedROD RP-18e reversed phase chromatographic column and subsequent analysis by mass spectrometry in the multiple reaction monitoring mode (MRM). The precursor to product ion transitions of m/z 320.3-->247.2 and m/z 409.1-->205.2 were used to measure the analyte and the IS, respectively. The assay exhibited a linear dynamic range of 2.0-489.1 ng/mL for chloroquine in dog plasma. The limit of detection (LOD) and lower limit of quantification (LLOQ) were 0.4 and 2.0 ng/mL, respectively in 0.05 mL plasma. Acceptable precision and accuracy were obtained for concentrations over the standard curve range of 2.0-489.1 ng/mL. A run time of 2.0 min for a sample made it possible to achieve a throughput of more than 400 plasma samples analyzed per day. The validated method was successfully used to analyze samples of dog plasma during non-clinical study of chloroquine.
authors.affiliation: Metabolism and Pharmacokinetic Department, Ranbaxy Research Laboratories, Plot # 20, Sector-18, Udhyog Vihar Industrial Area, Gurgaon, Haryana 122015, India.
authors.forename: Puran
authors.initials: P
authors.lastname: Singhal
authors.forename: Ashwani
authors.initials: A
authors.lastname: Gaur
authors.forename: Vanita
authors.initials: V
authors.lastname: Behl
authors.forename: Anirudh
authors.initials: A
authors.lastname: Gautam
authors.forename: Brijesh
authors.initials: B
authors.lastname: Varshney
authors.forename: Jyoti
authors.initials: J
authors.lastname: Paliwal
authors.forename: Vijay
authors.initials: V
authors.lastname: Batra
mesh_terms: D000818:Animals; D000962:Antimalarials / Q000097:blood* / Q000493:pharmacokinetics; D002738:Chloroquine / Q000097:blood* / Q000493:pharmacokinetics; D002853:Chromatography, Liquid / Q000379:methods*; D004285:Dogs; D012015:Reference Standards; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D000962:Antimalarials; D002738:Chloroquine