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New assays in development:

• darunavir assay, which will include atazanavir, efavirenz, lopinavir, saquinavir, and ritonavir
• nevirapine, lamivudine and stavudine

Efavirenz

The concentration of DMP-266 is measured in plasma using a method developed at Dupont and validated at the Laboratory for Antiviral Research at the State University of New York at Buffalo. DMP-266 is analyzed by extraction from 100 μL of plasma (EDTA) via a liquid-liquid extraction with ethylene dichloride, followed by centrifugation, aspiration of aqueous layer and evaporation under a nitrogen stream. Samples are reconstituted with mobile phase and injected into the chromatography system. UV detection is used at a wavelength of 250 nm.

The chromatographic system consists of a Waters Model 501 pump, a Waters Model 717plus autosampler, a Waters column heater and temperature control module, and a Waters Model 490 detector. The chromatographic system is controlled by Waters Millennium 2010 software, version 2.15 which also collects all chromatographic data generated during analysis. This program was utilized to assess: (1) all chromatographic data for the analysis, (2) generate calibration standard curves and their corresponding statistics, and (3) calculate specimen concentrations.

Characteristics of the method are as follows: