Simultaneous determination of intracellular concentrations of tenofovir, emtricitabine, and dolutegravir in human brain microvascular endothelial cells using liquid chromatography-tandem mass spectrometry (LC-MS/MS)

Abstract

Combination antiretroviral therapy (cART) regimens are recommended for HIV patients to better achieve and maintain plasma viral suppression. Despite adequate plasma viral suppression, HIV persists inside the brain, which is, in part thought to result from poor brain penetration of antiretroviral drugs. In this study, a simple and ultra-sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for simultaneous determination of tenofovir, emtricitabine, and dolutegravir in cell lysates of an immortalized human brain microvascular endothelial cell line (hCMEC/D3) was developed and validated. Analytes were separated on a reverse phase C 18 column using water and 0.1% formic acid in acetonitrile as mobile phases. The analytes were detected using positive electrospray ionization mode with multiple reaction monitoring (MRM). The assay was linear in the concentration range of 0.1-100 ng mL −1 for all analytes. Intra-and inter-assay precision and accuracy were within ± 13.33% and ± 10.53%, respectively. This approach described herein was used to determine the intracellular accumulation of tenofovir, emtricitabine, dolutegravir simultaneously in hCMEC/D3 cells samples.

Rapid quantification of tenofovir in umbilical cord plasma and amniotic fluid in hepatitis B mono‐infected pregnant women during labor by ultra‐performance liquid chromatography/tandem mass spectrometry

“…In positive ion mode, [M + H] + peaks were observed at m/z 288.2 and 278.2 for TFV and the IS, respectively. The most abundant and representative product ions were found at m/z 176.2 for TFV and 152.2 for the IS (Figure ), which were consistent with previous studies . The representative chromatograms of blank matrix, blank matrix spiked with IS and TFV at LLOQ, and umbilical cord plasma and amniotic fluid sample of pregnant women during labor are presented in Figures and .…”

Determination of intracellular anlotinib, osimertinib, afatinib and gefitinib accumulations in human brain microvascular endothelial cells by liquid chromatography/tandem mass spectrometry

“…As anlotinib has exhibited a good overall response rate in clinical evaporation and reconstitution. 18 In our study, a simpler and more cost-effective sample preparation procedure was developed.…”

“…34 We optimized the procedure and extended the range to 1-200 ng mL −1 , which was more applicable for the determination of afatinib in cell extracts. Two studies for the determination of osimertinib in human plasma used LLE or protein precipitation followed by evaporation to obtain a wide linear range of 5-1000 ng mL −1 and 1-1000 ng mL −1 , 14,18 respectively, compared with our range of 1-500 ng mL −1 in the cell extract. However, in contrast to our method, there was a slight ion suppression in the middle QC sample, and the precision of the absolute matrix effect in the low QC sample exceeded 15%.…”

Validation of a UHPLC-MS/MS Method to Quantify Twelve Antiretroviral Drugs within Peripheral Blood Mononuclear Cells from People Living with HIV

“…Therefore, thoroughly validated methods for the quantification of ARVs in PBMCs are needed in order to estimate their penetration in viral reservoirs. Several methods have been reported for the quantification of ARVs in PBMCs [ 25 , 27 , 31 , 32 ] and in other cell types [ 33 ], but these included only a few currently used ARVs; moreover, these methods mainly focused on NRTIs [ 21 ]. Furthermore, there are no reported methods for the simultaneous intracellular quantification of dolutegravir (DTG), elvitegravir (ELV), and rilpivirine (RPV), together with other ARVs of non-nucleos(t)ide nature.…”