Enantioselective analyses of chloroquine and hydroxychloroquine in rat liver microsomes through chiral liquid chromatography–tandem mass spectrometry

Abstract

An efficient, sensitive and selective liquid chromatography-tandem mass spectrometry (LC-MS/MS) chiral analysis method was established for determination of chloroquine and hydroxychloroquine enantiomers in rat liver microsomes. Effects of polysaccharide chiral stationary phases and basic additives on chiral separations of two analytes were discussed in detail. Amylose tris(3, 5-dimethylphenylcarbamate)-coated chiral stationary phase showed the best separation performance for them with acetonitrile-diethylamine-ethanoldiethylamine mixture (90:0.1:10:0.1, v/v/v/v) among four chiral stationary phases. Then, multiple reaction monitoring mode was selected as the data acquisition for determination of two pairs of enantiomers. The proposed LC-MS/MS chiral analysis method was validated in terms of linearity, accuracy, precision, and specificity. Good linearity with correlation coefficient over 0.998 was obtained in the concentration range of 0.05-5 μM. Limits of quantification for chloroquine and hydroxychloroquine enantiomers were 5.0 and 1.0 nM, respectively. The recoveries ranged from 81.14% to 111.09%. The intra-day and inter-day relative standard deviation were less than 6.5%. Moreover, concentrations of chloroquine and hydroxychloroquine enantiomers in rat liver microsomes were determined through the proposed LC-MS/MS analysis method. After incubated with rat liver microsomes for 10 min, the enantiomeric factor of hydroxychloroquine decreased from 0.50 to 0.45 (p < 0.001). In brief, our developed determination method for chloroquine and hydroxychloroquine enantiomers through LC-MS/MS spectrometry showed the characteristics of high-efficiency, fast speed, and very low detection limit, and would be greatly beneficial for screening and quantitation of them in biological matrices.

Copper Nanoparticles and Reduced Graphene Oxide as an Electrode Modifier for the Development of an Electrochemical Sensing Platform for Chloroquine Phosphate Determination

“…Certain analytical methods have been proposed for the analysis of chloroquine and its derivatives, such as ultraviolet spectrophotometry [10][11][12][13], immunoassay [14], chemiluminescence [15], nuclear magnetic resonance [16], spectrofluorimetry [17], and capillary electrophoresis [18], and most studies' methodologies are based on chromatography [19][20][21][22][23][24][25][26][27][28], including high-performance liquid chromatography (HPLC) [22][23][24][25][26][27] and gas chromatography [28]. However, some of these techniques require solid-phase extraction, liquid-liquid extraction, high-performance solvents (chromatography grade), and highly trained personnel [20,29], which makes it difficult to perform these techniques in some cases.…”

Stereoselective quantitative analysis of ranolazine in plasma and tissue samples: application in pharmacokinetics and tissue distribution studies

“…It is prominent that for most chiral compounds, only one enantiomer is pharmacologically active but the other may be inactive or exert completely diverse activity secondary to the stereoselective binding effects of each enantiomer with biological macromolecules (such as enzymes, proteins, and nucleic acids). [10][11][12] For ranolazine, the S-enantiomer displayed stronger anti-anginal and anti-ischemic activities than the R-enantiomer or rac-ranolazine whereas the R-form improved the glucose levels in diabetic rats by preserving the b-cells and enhancing the secretion of insulin. 13,14 Aside from different biological activities to target objects, the absorption and distribution processes of chiral drugs in organisms are typically stereoselective.…”