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Antimicrobial Agents and Chemotherapy, November 2000, p. 3097-3100, Vol. 44, No. 11
0066-4804/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Simultaneous Quantitation of Intracellular Zidovudine and Lamivudine Triphosphates in Human Immunodeficiency Virus-Infected Individuals

Jose F. Rodriguez,1,2,* Jorge L. Rodriguez,1 Jorge Santana,3 Hermes García,4 and Osvaldo Rosario2

Department of Biochemistry1 and Department of Medicine,3 School of Medicine, Medical Sciences Campus, University of Puerto Rico, and Puerto Rico Health Department (CLETS),4 San Juan, and Department of Chemistry, Rio Piedras Campus, University of Puerto Rico, Rio Piedras,2 Puerto Rico

Received 11 February 2000/Returned for modification 28 May 2000/Accepted 21 August 2000

Highly active antiretroviral therapy (HAART) is the standard treatment for infection with human immunodeficiency virus (HIV). The most common HAART regimen consists of the combination of at least one protease inhibitor (PI) with two nucleoside reverse transcriptase inhibitors (NRTIs). Contrary to PIs, NRTIs require intracellular activation from the parent compound of their triphosphate moiety to suppress HIV replication. Simultaneous intracellular determination of two NRTI triphosphates is difficult to accomplish due to their relatively small concentrations in peripheral blood mononuclear cells (PBMCs), requiring large amounts of blood from HIV-positive patients. Recently, we described a method to determine intracellular zidovudine triphosphate (ZDV-TP) concentrations in HIV-infected patients by using solid-phase extraction and tandem mass spectrometry. The limit of quantitation (LOQ) for ZDV-TP was 0.10 pmol, and the method was successfully used for the determination of ZDV-TP in HIV-positive patients. In this study, we enhanced the aforementioned method by the simultaneous quantitation of ZDV-TP and lamivudine triphosphate (3TC-TP) in PBMCs from HIV-infected patients. The LOQ for 3TC-TP was 4.0 pmol, with an interassay coefficient of variation and an accuracy of 7 and 12%, respectively. This method was successfully applied to the simultaneous in vivo determination of the ZDV-TP and 3TC-TP pharmacokinetic profiles from HIV-infected patients receiving HAART.

* Corresponding author. Mailing address: Department of Biochemistry, P.O. Box 365067, School of Medicine, Medical Sciences Campus, University of Puerto Rico, San Juan Puerto Rico 00936-5067. Phone and Fax: (787) 754-4929. E-mail: j_rodriguez{at}rcmaxp.upr.clu.edu.

Antimicrobial Agents and Chemotherapy, November 2000, p. 3097-3100, Vol. 44, No. 11
0066-4804/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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