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

Abstract

Background Topical microbicidal agents are being actively pursued as a modality to prevent HIV viral transmission during sexual intercourse. Quantification of antiretroviral agents in specimen sources where antiviral activity is elicited is critical, and drug measurements in cervicovaginal fluid can provide key information on local drug concentrations. Two antiretroviral drugs, dapivirine and maraviroc, have gained interest as vaginal microbicidal agents, and rugged methods are required for their quantification in cervicovaginal secretions. Methods Cervicovaginal fluid spiked with dapivirine and maraviroc were applied to ophthalmic tear strips or polyester-based swabs to mimic collection procedures used in clinical studies. Following sample extraction and the addition of isotopically-labeled internal standards, samples were subjected to liquid chromatographic-tandem mass spectrometric (LC-MS/MS) analysis using a Waters BEH C8, 50 × 2.1 mm, 1.7 µm particle size column, on an API 4000 mass analyzer operated in selective reaction monitoring mode. The method was validated according to FDA Bioanalytical Method Validation guidelines. Results Due to the disparate saturation capacity of the tested collection devices, the analytical measuring ranges for dapivirine and maravirocin cervicovaginal fluid on the ophthalmic tear strip were 0.05 to 25 ng/tear strip, and 0.025 to 25 ng/tear strip, respectively. As for the polyester-based swab, the analytical measuring ranges were 0.25 to 125 ng/swab for dapivirine and 0.125 to 125 ng/swab for maraviroc. Dilutional studies were performed for both analytes to extended ranges of 25,000 ng/tear strip and 11,250 ng/swab. Standard curves were generated via weighted (1/x2) linear or quadratic regression of calibrators. Precision, accuracy, stability and matrix effects studies were all performed and deemed acceptable according to the recommendations of the FDA Bioanalytical Method Validation guidelines. Conclusions A rugged LC-MS/MS method for the dual quantification of dapivirine and maraviroc in cervicovaginal fluid using two unique collection devices has been developed and validated. The described method meets the criteria to support large research trials.

Phase I trial of pod-intravaginal rings delivering antiretroviral agents for HIV-1 prevention: Rectal drug exposure from vaginal dosing with tenofovir disoproxil fumarate, emtricitabine, and maraviroc

“…Concentrations of TDF, TFV, TFV-DP, FTC, and MVC in fluids/tissue were determined via previously described liquid chromatographic-tandem mass spectrometric (LC-MS/MS) assays [ 11 , 28 – 30 ]. All assays were developed and validated following the Food and Drug Administration Guidance for Industry, Bioanalytical Method Validation recommendations and met all acceptability criteria [ 31 ].…”

A Phase 1 Trial to Assess the Safety, Acceptability, Pharmacokinetics, and Pharmacodynamics of a Novel Dapivirine Vaginal Film

“…23,24 The lower limits of quantification (LLOQ) for dapivirine in plasma and CVL were 0.02 ng/mL and 2 ng/mL, respectively. One set of tissue biopsies intended for drug concentration was weighed, snap frozen, and stored at -80°C prior to drug quantification.…”

Combination Pod-Intravaginal Ring Delivers Antiretroviral Agents for HIV Prophylaxis: Pharmacokinetic Evaluation in an Ovine Model

“…An Agilent Zorbax Eclipse XDB-C18 Rapid Resolution column Concentrations of TFV, TFV diphosphate (TFV-DP), and MVC in VT homogenate were measured by LC-MS/MS at Johns Hopkins University using established methods (31)(32)(33). The lower limits of quantitation (LLQs) for these analytes in the sample matrices described above are presented (see Table 3).…”