35. Simultaneous Quantitative Analysis of 13 Anti-Retroviral Drugs in Human Serum using HPLC Triple Quadrupole Mass Spectrometry
Poster: Tue 2:00-3:00PM
Susan C. Leonard
Applied Biosystems
Susan C. Leonard1, Keith Goodman1, Lisa M. Sapp1, Jeffrey D. Miller1,

1Applied Biosystems, Framingham, Massachusetts., U.S.A.
Anti Retroviral drug therapy has been used for approximately 20 years and each year new drugs are introduced to address various parts of the viral particle replication cycle in treatment of Human Immunodeficiency Virus type 1 (HIV -1) infections. The current therapy involves administering combinations of drugs with maintenance of high circulating concentrations so multiple analytes need to be monitored both effectively and efficiently.
This work describes a sensitive method for the simultaneous determination of 13 antiretroviral drugs over a wide concentration range including non-nucleoside analog reverse transcriptase inhibitors, protease inhibitors, an integrase inhibitor, an entry inhibitor and a nucleotide analog reverse transcriptase inhibitor in human serum. This method employs high-performance liquid chromatography–tandem mass spectrometry with electrospray ionization. The analytes were monitored in multiple reaction monitoring mode in positive polarity mode to detect all compounds after a single injection. A protein precipitation sample preparation was used to extract all compounds.

The compounds in this panel are Saquinavir, Ritonavir, Indinavir, Nelfinavir, Amprenavir, Lopinavir, Nevirapine, Maraviroc, Tipranavir, Efavirenz, Delavirdine, Raltegravir and Darunavir. The linear dynamic range for the assay reported to be from 20 ng/mL up to 2000ng/mL with 10 uL injection volume to the LCMSMS system. Within- run precision and between-day precision results were <15% for the various concentrations analyzed in the linear range. This method may be very effective for quantifying and screening antiretroviral drugs in clinical samples with limited sample volumes and may be applied for therapeutic drug monitoring and clinical toxicology laboratory studies.

1. Jung, B.Y., Reich , N.L., Bridges, A.S., Corbett, A.H., Kashia, D.M., “Simultaneous Determination of antiretroviral drugs in human plasma for quantitative analysis with liquid chromatography-tandem mass spectrometry”, Biomedical Chromatography, volume 21, issue 10, 2007.
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