Development of an assay method for the detection and quantification of protease and non-nucleoside reverse transcriptase inhibitors in plasma and in peripherical blood mononuclear cells by liquid chromatography coupled with ultraviolet or tandem mass spectrometry detection

Simultaneous determination of HIV protease inhibitors amprenavir, atazanavir, indinavir, lopinavir, nelfinavir, ritonavir and saquinavir in human plasma by high-performance liquid chromatography-tandem mass spectrometry

“…Early work only needed to determine one or a small number of PIs [16][17][18][19] but due to an increase in the number of available PIs and the introduction of combination PI therapy bioanalytical assays capable of measuring multiple PIs were necessary. Recently, techniques to determine intracellular concentrations of PIs have also been described [20][21][22]. Several methods have been reported to concurrently determine a number HIV PIs in human plasma by liquid chromatography coupled to mass spectrometry [23][24][25][26][27][28][29].…”

Monitoring of lopinavir and ritonavir in peripheral blood mononuclear cells, plasma, and ultrafiltrate using a selective and highly sensitive LC/MS/MS assay

“…For this purpose monitor- ing of intracellular concentrations will be a useful tool to ensure effective drug levels at the site of virus replication. Meanwhile different FDA conform validated methods have been established to analyse intracellular drug amounts [15][16][17][18][19] and they always depend on LC/MS or LC/MS/MS to meet the selectivity and sensitivity, which is a prerequisite to support respective studies in vivo. The differences between the methods can be found e.g.…”

“…Whereas plasma concentrations are high enough to analyse the drugs using UV detection, determinations in other matrices (e.g. blood cells, cerebrospinal fluid, plasma ultrafiltrate, sanctuary sites) were mainly processed using HPLC coupled to mass spectrometry (LC/MS) or tandem mass spectrometry (LC/MS/MS) [15][16][17][18][19]. These methods are highly specific and sensitive and allow quantifications of HIV drugs below 1 ng/mL in fluids, respectively below 1 ng/cell pellet.…”

“…These methods are highly specific and sensitive and allow quantifications of HIV drugs below 1 ng/mL in fluids, respectively below 1 ng/cell pellet. Beside the LC/MS determination cell isolation procedures have to be developed mainly depending on ficoll density gradient centrifugation [15][16][17][18][19].…”

Quantification of 8 HIV-Protease Inhibitors and 2 Nonnucleoside Reverse Transcriptase Inhibitors by Ultra-Performance Liquid Chromatography with Diode Array Detection

“…Furthermore, UV detection remains an available option for many laboratories. Unfortunately, analysis times are usually very long with conventional HPLC columns (up to 70 min) (2)(3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17) because UV detection requires complete resolution of all chromatographic peaks to avoid interferences. Faster and higher-resolution separation can be achieved with ultra-performance liquid chromatography (UPLC) technology.…”