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.

Simultaneous Determination of Chloroquine and Its Metabolite Desethyl Chloroquine in Human Plasma Using Liquid Chromatography Tandem Mass Spectrometry

“…The precursor ion and product ion mass spectrums are illustrated in Figure 1. The MRM transition of CQ was in accordance with the reported result (Singhal 2007). The declustering potential, collision energy, collision cell entrance, and exit potentials were optimized sequentially with MRM mode.…”

“…In recent decades, LC-MS=MS has become a popular and highly efficient technique for the trace analysis in bioassay especially for pharmacokinetic and toxicological studies. Recently, Puran Singhal et al (2007) reported a method for determining CQ in human and dog plasma using LC-MS=MS. An, Hodel et al (2009) developed an LC-MS=MS for the simultaneous determination of 14 antimalarials including CQ in human plasma.…”

A modified serial blood sampling technique and utility of dried-blood spot technique in estimation of blood concentration: application in mouse pharmacokinetics

“…Also, the run time of the method was only 2.4 min with the use of UPLC, which further increased the throughput. Various methods are reported in literature for the quantitation of amodiaquine (Stepniewska et al 2009), chloroquine (Hodel et al 2009;Singhal et al 2007) and chlorthalidone (Khuroo et al 2008;Salvadori et al 2009) in plasma. However, all the reported methods use time and resource-intensive extraction methods or have a long run time.…”

Development and Validation of an Hilic–MS/MS Method by One-Step Precipitation for Chloroquine in Miniature Pig Plasma

“…The C18 column was the most frequently used columns in previously published research [7][8][9][10][12][13][14] and we have investigated the peak shape and response of several C18 columns in the optimization process. With additives of 5 mM ammonium formate and formic acid (0.015∼0.1% formic acid) in mobile phase system consisting of acetonitrile and water, chloroquine did not reserve on an Eclipse Plus C18 column (5 μm, 100 × 4.6 mm, Agilent, USA) and the retention time remained almost unchanged (∼1.8 min) with different proportion of acetonitrile ranged between 13 and 70%.…”

“…Singhal [14] Zuluaga-Idarraga [13] Cheomung [10] Internal standard Chloroquine-d4 Piperazine bis chloroquinoline [14] published an LC-MS/MS method for the detection of chloroquine in dog plasma for the first time. A structural analog instead of deuterated isotope internal standard (IS) and a simple protein precipitation by methanol were employed in this study.…”