Development and bioanalytical validation of a liquid chromatographic-tandem mass spectrometric (LC-MS/MS) method for the quantification of the CCR5 antagonist maraviroc in human plasma

Abstract

Background Maraviroc is a CCR5 antagonist that has been utilized as a viral entry inhibitor in the management of HIV-1. Current clinical trials are pursuing maraviroc drug efficacy in both oral and topical formulations. Therefore, in order to fully understand drug pharmacokinetics, a sensitive method is required to quantify plasma drug concentrations. Methods Maraviroc-spiked plasma was combined with acetonitrile containing an isotopically-labeled internal standard, and following protein precipitation, samples were evaporated to dryness and reconstituted for liquid chromatographic-tandem mass spectrometric (LC-MS/MS) analysis. Chromatographic separation was achieved on a Waters BEH C8, 50 × 2.1 mm UPLC column, with a 1.7 μm particle size and the eluent was analyzed using an API 4000 mass analyzer in selected reaction monitoring mode. The method was validated as per FDA Bioanalytical Method Validation guidelines. Results The analytical measuring range of the LC-MS/MS method is 0.5-1000 ng/ml. Calibration curves were generated using weighted 1/x2 quadratic regression. Inter-and intra-assay precision was ≤ 5.38% and ≤ 5.98%, respectively; inter-and intra-assay accuracy (%DEV) was ≤ 10.2% and ≤ 8.44%, respectively. Additional studies illustrated similar matrix effects between maraviroc and its internal standard, and that maraviroc is stable under a variety of conditions. Method comparison studies with a reference LC-MS/MS method show a slope of 0.948 with a Spearman coefficient of 0.98. Conclusions Based on the validation metrics, we have generated a sensitive and automated LC-MS/MS method for maraviroc quantification in human plasma.

Dual quantification of dapivirine and maraviroc in cervicovaginal secretions from ophthalmic tear strips and polyester-based swabs via liquid chromatographic–tandem mass spectrometric (LC–MS/MS) analysis

“…Similar to previously reported observations describing the selectivity of dapivirine and maraviroc in plasma, no interfering peaks were observed in drug-free cervicovaginal fluid at the retention times at which dapivirine, maraviroc and the respective internal standards, elute (Figure 4) [29, 30]. Further, to assess the potential influence of endogenous compounds on analyte suppression or enhancement, quantitative matrix effects studies were performed following the guidelines of Matuszewskiand colleagues [32].…”

“…For dapivirine and maraviroc quantification in cervicovaginal fluid using ophthalmic tear strips or polyester-based swabs, the same chromatographic and mass spectrometric conditions were implemented as previously described for drug quantification in plasma [29, 30]. However, significant development was required for the preparation of a specimen for LC-MS/MS analysis, as the matrix may be acquired using a variety of collection devices.…”

“…Chromatographic separation of dapivirine and maraviroc was performed using a Waters BEH C8, 50 × 2.1 mm UPLC column with a 1.7 µm particle size, on a Waters Acquity UPLC system as previously described [29, 30]. A mobile phase system consisting of water containing 0.1% formic acid (mobile phase A) and acetonitrile containing 0.1% formic acid (mobile phase B) was used for separation.…”

“…Further, a method for the dual quantification of both the NNRTI dapivirine and the CCR-5 antagonist maraviroc has not been previously reported. Our group has previously developed and validated liquid chromatographic-tandem mass spectrometric (LC-MS/MS) assays for the quantification of dapivirine and maraviroc in plasma, with lower limits of quantification of 20 pg/ml and 500 pg/ml, respectively [29, 30]. However, a method for quantification in cervicovaginal fluid must be amended for a wide analytical measuring range, with extensive linearity to accommodate previously reported localized drug concentrations.…”

CYP3A5 Genotype Impacts Maraviroc Concentrations in Healthy Volunteers

“…1. Plasma maraviroc concentrations were determined using a validated ultra-performance liquid chromatography-tandem mass spectrometry method as previously reported (Emory et al, 2014), with inter-and intra-assay precision of #5.4 and #6.0%, and inter-and intra-assay accuracy within 610.2 and 68.4%, respectively. The validation results exceeded standards recommended by the FDA (2013).…”