Quantification of seven nucleoside/nucleotide reverse transcriptase inhibitors in human plasma by high-performance liquid chromatography with tandem mass-spectrometry.
A simple analytical method was developed in 100 microL of plasma for the simultaneous assay of the 7 nucleoside/nucleotide reverse transcriptase inhibitors (abacavir, didanosine, emtricitabine, lamivudine, stavudine, tenofovir, and zidovudine) currently used for the treatment of HIV-infected patients. After adding the internal standard, 6-beta-hydroxy-theophyline, plasma samples were precipitated with 500 microL acetonitrile and the supernatants were evaporated to dryness. The residues were reconstituted with 500 microL of water and 10 microL of the extracts were injected in the chromatographic system. The chromatographic separation was performed with a C-18 column and a gradient mobile phase consisting of a mixture of water and acetonitrile, both containing 0.05% formic acid. Analytes quantification was performed by electrospray ionisation triple quadrupole mass-spectrometry in the positive mode using selected reaction monitoring (SRM). Intra- and inter-assay precision and accuracy were lower than 20% for the limit of quantification, and 15% for higher concentrations. The method has been implemented to assess plasma concentrations of patients infected by HIV and was found suitable for therapeutic drug monitoring.
Le Saux
Thomas
T
Université Paris-Descartes, Faculté de Médecine, Service de Pharmacologie Clinique, Groupe Hospitalier Cochin - Saint-Vincent de Paul, Assistance Publique, Hôpitaux de Paris, France.
Chhun
Stéphanie
S
Rey
Elisabeth
E
Launay
Odile
O
Weiss
Laurence
L
Viard
Jean-Paul
JP
Pons
Gérard
G
Jullien
Vincent
V
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D015203:Reproducibility of Results; D018894:Reverse Transcriptase Inhibitors / Q000097:blood*; D053719:Tandem Mass Spectrometry / Q000379:methods*
D018894:Reverse Transcriptase Inhibitors