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.
We present a simple chromatographic method to detect and quantify protease inhibitors (PI), metabolites and non-nucleoside reverse transcriptase inhibitors (NNRTIs) in human plasma of HIV-1 infected patients and in peripheral blood mononuclear cells (PBMCs) using either liquid chromatography coupled with ultraviolet (LC-UV) or liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). A solid-liquid extraction was carried out on 500 microl of plasma as pre-treatment. Calibration curve ranges were from 50 (100) to 5000 ng/ml (indinavir). PBMC pellets from 7 ml of blood were lysed with methanol/tris with a calibration curve ranging from 0.25 to 250 ng/pellet. Simple modifications in the mobile phase composition (slight increase of ammonium acetate concentration and addition of methanol for LC-UV) easily linked the two analytical systems.
Pèlerin
Hélène
H
CEA, Service de Pharmacologie et d'Immunologie, DSV/DRM, 91191 Gif sur Yvette Cedex, France.
Compain
Séverine
S
Duval
Xavier
X
Gimenez
François
F
Bénech
Henri
H
Mabondzo
Aloïse
A
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D015658:HIV Infections / Q000097:blood; D015497:HIV-1; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000737:chemistry*; D013058:Mass Spectrometry / Q000379:methods*; D011480:Protease Inhibitors / Q000097:blood*; D015203:Reproducibility of Results; D018894:Reverse Transcriptase Inhibitors / Q000097:blood*; D012680:Sensitivity and Specificity; D013056:Spectrophotometry, Ultraviolet
D011480:Protease Inhibitors; D018894:Reverse Transcriptase Inhibitors