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

Elens, Laure;Veriter, Sophie;Di Fazio, Vincent;Vanbinst, Roger;Haufroid, Vincent;et.al.
(2008) Clinical Chemistry : international journal of molecular diagnostics and laboratory medicine — Vol. 55, n° 1, p. 170-174 (2009)

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Authors
  • Elens, LaureUCLouvain
    Author
  • Veriter, SophieUCLouvain
    Author
  • Di Fazio, VincentUCLouvain
    Author
  • Vanbinst, RogerUCLouvain
    Author
  • Boesmans, DanielUCLouvain
    Author
  • Wallemacq, PierreUCLouvain
    Author
  • Haufroid, Vincentorcid-logoUCLouvain
    Author
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Abstract
BACKGROUND: Most HPLC-UV methods for therapeutic drug monitoring of anti-HIV drugs have long run times, which reduce their applicability for high-throughput analysis. We developed an ultra-performance liquid chromatography (UPLC)-diode array detection method for the simultaneous quantification of the HIV-protease inhibitors (PIs) amprenavir, atazanavir, indinavir, lopinavir, nelfinavir, ritonavir, saquinavir, and tipranavir (TPV), and the nonnucleoside reverse transcriptase inhibitors (NNRTIs) efavirenz and nevirapine. METHODS: Solid-phase extraction of 1 mL plasma was performed with Waters HLB cartridges. After 3 wash steps, we eluted the drugs with methanol, evaporated the alcohol, and reconstituted the residue with 50 microL methanol. We injected a 4-microL volume into the UPLC system (Waters ACQUITY UPLC BEH C8 column maintained at 60 degrees C) and used a linear gradient of 50 mmol/L ammonium acetate and 50 mmol/L formic acid in water versus acetonitrile to achieve chromatographic separation of the drugs and internal standard (A-86093). Three wavelengths (215, 240, and 260 nm) were monitored. RESULTS: All drugs were eluted within 15 min. Calibration curves with concentrations of 0.025-10 mg/L (1.875-75 mg/L for TPV) showed coefficients of determination (r(2)) between 0.993 and 0.999. The lower limits of quantification were well below the trough concentrations reported in the literature. Inter- and intraassay CVs and the deviations between the nominal and measured concentrations were <15%. The method was validated by successful participation in an international interlaboratory QC program. CONCLUSIONS: This method allows fast and simultaneous quantification of all commercially available PIs and NNRTIs for therapeutic drug monitoring.
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Elens, L., Veriter, S., Di Fazio, V., Vanbinst, R., Boesmans, D., Wallemacq, P., & Haufroid, V. (2008). Quantification of 8 HIV-Protease Inhibitors and 2 Nonnucleoside Reverse Transcriptase Inhibitors by Ultra-Performance Liquid Chromatography with Diode Array Detection. Clinical Chemistry : international journal of molecular diagnostics and laboratory medicine, 55(1), 170-174. https://doi.org/10.1373/clinchem.2008.108647 (Original work published 2009)