Quantification of 8 HIV-protease inhibitors and 2 nonnucleoside reverse transcriptase inhibitors by ultra-performance liquid chromatography with diode array detection.

Clin Chem 2009 Jan 21;55(1):170-4. Epub 2008 Nov 21.

Industrial Toxicology and Occupational Medicine Unit, Université Catholique de Louvain, Brussels, Belgium.

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.

Download full-text PDF

Source
http://dx.doi.org/10.1373/clinchem.2008.108647DOI Listing
January 2009

Publication Analysis

Top Keywords

nonnucleoside reverse
8
reverse transcriptase
8
transcriptase inhibitors
8
array detection
8
ultra-performance liquid
8
quantification hiv-protease
8
liquid chromatography
8
hiv-protease inhibitors
8
maintained degrees
4
degrees linear
4
column maintained
4
beh column
4
system waters
4
waters acquity
4
acquity uplc
4
uplc beh
4
linear gradient
4
ammonium acetate
4
acid water
4
water versus
4

Similar Publications