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Antimicrobial Agents and Chemotherapy, December 1999, p. 2964-2968, Vol. 43, No. 12
0066-4804/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Determination of Zidovudine Triphosphate Intracellular Concentrations in Peripheral Blood Mononuclear Cells from Human Immunodeficiency Virus-Infected Individuals by Tandem Mass Spectrometry

Eva Font,1 Osvaldo Rosario,1 Jorge Santana,2 Hermes García,3 Jean-Pierre Sommadossi,4 and Jose F. Rodriguez5,*

Department of Chemistry, Rio Piedras Campus,1 and Departments of Medicine2 and Biochemistry,5 School of Medicine, Medical Sciences Campus, University of Puerto Rico, and CLETS Center, Puerto Rican Health Department,3 San Juan, Puerto Rico, and Department of Clinical Pharmacology, School of Medicine, University of Alabama at Birmingham, Birmingham, Alabama4

Received 15 March 1999/Returned for modification 10 September 1999/Accepted 28 September 1999

Nucleoside reverse transcriptase inhibitors (NRTIs) used against the human immunodeficiency virus (HIV) need to be activated intracellularly to their triphosphate moiety to inhibit HIV replication. Intracellular concentrations of these NRTI triphosphates, especially zidovudine triphosphate (ZDV-TP), are relatively low (low numbers of femtomoles per 106 cells) in HIV-infected patient peripheral blood mononuclear cells. Recently, several methods have used either high-performance liquid chromatography (HPLC) or solid-phase extraction (SPE) coupled with radioimmunoassay to obtain in vivo measurements of ZDV-TP. The limit of detection (LOD) by these methods ranged from 20 to 200 fmol/106 cells. In this report, we describe the development of a method to determine intracellular ZDV-TP concentrations in HIV-infected patients using SPE and HPLC with tandem mass spectrometry for analysis. The LOD by this method is 4.0 fmol/106 cells with a linear concentration range of at least 4 orders of magnitude from 4.0 to 10,000 fmol/106 cells. In hispanic HIV-infected patients, ZDV-TP was detectable even when the sampling time after drug administration was 15 h. Intracellular ZDV-TP concentrations in these patients ranged from 41 to 193 fmol/106 cells. The low LOD obtained with this method will provide the opportunity for further in vivo pharmacokinetic studies of intracellular ZDV-TP in different HIV-infected populations. Furthermore, this methodology could be used to perform simultaneous detection of two or more NRTIs, such as ZDV-TP and lamivudine triphosphate.

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

Antimicrobial Agents and Chemotherapy, December 1999, p. 2964-2968, Vol. 43, No. 12
0066-4804/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Benech, H., Becher, F., Pruvost, A., Grassi, J. J., Melendez, M., Rodriguez, J. F. (2006). Is Stavudine triphosphate a natural metabolite of Zidovudine?. Antimicrob. Agents Chemother. 50: 2899-2901 [Full Text]  
  • Melendez, M., Blanco, R., Delgado, W., Garcia, R., Santana, J., Garcia, H., Rosario, O., Rodriguez, J. F. (2006). Lack of Evidence for In Vivo Transformation of Zidovudine Triphosphate to Stavudine Triphosphate in Human Immunodeficiency Virus-Infected Patients. Antimicrob. Agents Chemother. 50: 835-840 [Abstract] [Full Text]  
  • Rodriguez, J. F., Rodriguez, J. L., Santana, J., García, H., Rosario, O. (2000). Simultaneous Quantitation of Intracellular Zidovudine and Lamivudine Triphosphates in Human Immunodeficiency Virus-Infected Individuals. Antimicrob. Agents Chemother. 44: 3097-3100 [Abstract] [Full Text]  
  • Rigourd, M., Lanchy, J.-M., Le Grice, S. F. J., Ehresmann, B., Ehresmann, C., Marquet, R. (2000). Inhibition of the Initiation of HIV-1 Reverse Transcription by 3'-Azido-3'-deoxythymidine. COMPARISON WITH ELONGATION. J. Biol. Chem. 275: 26944-26951 [Abstract] [Full Text]  


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