Simultaneous measurement of etravirine, maraviroc and raltegravir in pigtail macaque plasma, vaginal secretions and vaginal tissue using a LC–MS/MS assay

Abstract

Etravirine (ETR), maraviroc (MVC) and raltegravir (RAL) are promising antiretroviral drugs being used in HIV treatment and may be interesting for prevention applications such as oral or topical pre-exposure prophylaxis. Here we describe a sensitive and accurate method for the simultaneous detection of ETR, MVC and RAL from pigtail macaque plasma, vaginal secretions, and vaginal tissue. This method is characterized by a straightforward precipitation extraction method, a limit of quantification <0.5 ng mL−1 for all three antiretrovirals bolstered by a corresponding internal standard for each drug analyte, and short run time. Quantification is performed using positive ion electrospray triple quadrupole mass spectrometry. This method was validated over clinically relevant ranges for the three ARV drugs in all three matrices: 0.1–100 ng mL−1 for ETR, 0.05–100 ng mL−1 for MVC and 1–100 ng mL−1 for RAL. Our method is accurate and precise, with measured mean inter-assay precision (%CV) and accuracy (% bias) of 5.08% and 1.96%, respectively, while the mean intra-assay precision and accuracy were 3.44% and 1.08 %. The overall post-extraction recovery for ETR, MVC and RAL was >94% in all cases. We also show that extracted biological samples are stable after storage at room temperature or 4 °C and after three freeze/thaw cycles. This is the first analytical method capable of quantifying ETR, MVC and RAL in biological matrices relevant for pre-clinical testing of oral or topical HIV prevention methods in pigtailed macaques.

Lipid Vesicles Loaded with an HIV-1 Fusion Inhibitor Peptide as a Potential Microbicide

“…Therefore, a peptide recovery of 24 ± 3% was obtained. This percentage is lower than those obtained by other authors studying drug recovery from vaginal tissue, but it allowed us to achieve a suitable selectivity avoiding interferences caused by proteins, tissue compounds and other peptides [ 58 ].…”

Glycan targeted polymeric antibiotic prodrugs for alveolar macrophage infections

“…The supernatant was collected and stored on ice until analyzed for ciprofloxacin via LC-MS/MS. [40], and previously employed by our group for evaluation of ciprofloxacin concentration in mouse tissues [34]. Ciprofloxacin was separated on an Agilent 1290 Infinity LC system (Agilent Technologies, Santa Clara, CA, USA) under gradient elution using a Hypersil GOLD PFP column (100mm × 2.1 mm internal diameter, 1.9μm particle size) (Thermo Scientific, Waltham, MA, USA).…”

Chromatographic methods in HIV medicine: Application to therapeutic drug monitoring

“…Under these conditions, the authors achieve baseline separation of four out of five target drugs (TNF/FTC, EVG, RPV and DTG) in <5 min (Prathipati, Mandal, & Destache, ). Vaginal tissues and secretions have also been sampled in an effort to quantify drug levels following oral or local administration pre‐prophylaxis ARV drugs (Barreiros et al, ; Blakney, Jiang, Whittington, & Woodrow, ). Instances of suboptimal drug concentrations during and following delivery have been detected in vaginal secretions (Yeh et al, ).…”

“…However, combining strategies such as isotopically labeled internal standards, high‐resolution chromatography and MS/MS detection can compensate for the limited selectivity of PP sample preparation. Blakney and colleagues demonstrated the minimization of matrix effects, coupled with high recovery (>94%) for ETR, MVC and RAL when using matched isotopically labeled internal standards with a PP sample preparation (Blakney et al, ). (Likewise, the incorporation of five deuterium labeled internal standards (APV‐d 4 , TFV‐d 6 , EFV‐d 4 , NVP‐d 5 , LPV‐d 8 ) with UHPLC and MS/MS detection made PP sufficient for sample preparation of human plasma for quantification of 16 ARV drugs; Djerada et al, .)…”

“…Finally, the most sensitive assay published relies on ESI‐MS/MS for quantification of ETR, MVC and RAL in vaginal tissues and secretions. Demonstration of sub‐ng/mL quantification limits involved coupling isotopically labeled internal standards (ETR‐13C 6 , MVC‐d 6 , and RAL‐d 6 ) with multiple reaction monitoring using a triple quadrupole mass spectrometer (Blakney et al, ).…”