High‐performance liquid chromatography–tandem mass spectrometry for simultaneous determination of raltegravir, dolutegravir and elvitegravir concentrations in human plasma and cerebrospinal fluid samples

Abstract

A simple sample treatment procedure and sensitive liquid chromatography-tandem mass spectrometry method were developed for the simultaneous quantification of the concentrations of human immunodeficiency virus-1 integrase strand transfer inhibitors - raltegravir, dolutegravir and elvitegravir - in human plasma and cerebrospinal fluid (CSF). Plasma and CSF samples (20 μL each) were deproteinized with acetonitrile. Raltegravir-d was used as the internal standard. Chromatographic separation was achieved on an XBridge C column (50 × 2.1 mm i.d., particle size 3.5 μm) using acetonitrile-water (7:3, v/v) containing 0.1% formic acid as the mobile phase at a flow rate of 0.2 mL/min. The run time was 5 min. Calibration curves for all three drugs were linear in the range 5-1500 ng/mL for plasma and 1-200 ng/mL for CSF. The intra- and inter-day precision and accuracy of all three drugs in plasma were coefficient of variation (CV) <12.9% and 100.0 ± 12.2%, respectively, while those in CSF were CV <12.3% and 100.0 ± 7.9%, respectively. Successful validation under the same LC-MS/MS conditions for both plasma and CSF indicates this analytical method is useful for monitoring the levels of these integrase strand transfer inhibitors in the management of treatment of HIV-1 carriers.

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)

“…No significant ion suppression or enhancement were observed for the studied analytes using normalized internal standard results; however, low extraction recoveries (22.1-63.1%) were reported [36]. An LC-MS/MS method for simultaneous determination of dolutegravir, raltegravir, and elvitegravir levels has been reported [39] using protein precipitation by acetonitrile in human plasma and CSF. The method was developed on Xbridge C18 Column ( 50 × 2.0 mm, 3.5 µm) using isocratic mobile phase of acetonitrile:water (70:30) containing 0.1% formic acid.…”

“…Tenofovir and its phosphorylated metabolites were individually estimated in hepatocytes infected with hepatitis B virus using protein precipitation by methanol and a gradient elution consisting of water with 5 mM ammonium acetate and acetonitrile-methanol, 50:50, v/v mobile phases [38]. Tsuchiya et al simultaneously determined dolutegravir and other INSTI concentrations in human plasma and CSF using protein precipitation by acetonitrile with an isocratic mobile phase consisting of acetonitrile:water (70:30, v/v) with 0.1% formic acid [39]. To date, this particular three-drug combination has been quantified concurrently in mouse plasma and tissue by SPE using isocratic mobile phase consisting of 0.1% formic acid in water and 0.1% formic acid in acetonitrile mobile phases [36].…”

Development and validation of an HPLC‐UV method for quantification of elvitegravir and two other new antiretrovirals, dolutegravir and rilpivirine, in the plasma of HIV‐positive patients

“…Djerada et al, 2013;Grégoire et al, 2014;Penchala et al, 2016;Simiele et al, 2017;Tsuchiya et al, 2018). Djerada et al, 2013;Grégoire et al, 2014;Penchala et al, 2016;Simiele et al, 2017;Tsuchiya et al, 2018).…”

Development and validation of a UHPLC–MS/MS assay for elvitegravir measurement in human plasma and cerebrospinal fluid

“…Previously, EVG has been quantified in human biological matrixes such as plasma , seminal plasma , and peripheral blood mononuclear cells , as well as mouse plasma and tissue using LC–MS. A recent report has described the measurement of the integrase inhibitors raltegravir, dolutegravir, and EVG in human plasma and CSF . The assay had a calibration range of 5–1500 ng/mL in plasma and 1–200 ng/mL in CSF, which is not ideal considering the average trough concentrations of EVG in plasma at steady state is typically 409 ng/mL while the Cmax concentrations are 2500 ng/mL .…”

“…Furthermore, Tsuchiya et al. reported plasma EVG concentrations of 3200 ng/mL (3 h post dose) and 168 ng/mL (20 h post dose) which suggested the requirement of multiple dilutions from samples taken at early time points due to the narrow plasma range of 5–1500 ng/mL. Lastly, Tsuchiya et al.…”