Simultaneous measurement of intracellular triphosphate metabolites of zidovudine, lamivudine and abacavir (carbovir) in human peripheral blood mononuclear cells by combined anion exchange solid phase extraction and LC-MS/MS

Brian L. Robbins, Philip A. Poston, Erin F. Neal, Clive Slaughter, John H. Rodman

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

All nucleoside reverse transcriptase inhibitors (NRTI) must first be metabolized to their triphosphate forms in order to be active against HIV. Zidovudine (ZDV), abacavir (ABC) and lamivudine (3TC) have proven to be an efficacious combination. In order simultaneously to measure intracellular levels of the triphosphates (-TP) of ZDV, ABC (carbovir, CBV) and 3TC, either together or individually, we have developed a cartridge-LC-MS/MS method. The quantitation range was 2.5-250 pg/μl for 3TC-TP, 0.1-10.0 pg/μl for ZDV-TP and 0.05-5.00 pg/μl for CBV-TP. This corresponds to 0.1-11.0 pmol 3TC-TP per million cells, 4-375 fmol ZDV-TP per million cells and 2-200 fmol CBV-TP per million cells, extracted from 10 million cells. Patient samples demonstrated measured levels in the middle regions of our standard curves both at pre-dose and 4 h post-dose times.

Original languageEnglish (US)
Pages (from-to)310-317
Number of pages8
JournalJournal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
Volume850
Issue number1-2
DOIs
StatePublished - May 1 2007
Externally publishedYes

Fingerprint

carbovir
Zidovudine
Solid Phase Extraction
Metabolites
Anions
Blood Cells
Ion exchange
Blood
Lamivudine
Reverse Transcriptase Inhibitors
Nucleosides
HIV
lamivudine drug combination abacavir
triphosphoric acid
3'-azido-3'-deoxythymidine 5'-triphosphate

Keywords

  • Antiviral drug metabolism
  • Carbovir triphosphate
  • Human immunodeficiency virus
  • Lamivudine triphosphate
  • Zidovudine triphosphate

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Clinical Biochemistry
  • Cell Biology

Cite this

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title = "Simultaneous measurement of intracellular triphosphate metabolites of zidovudine, lamivudine and abacavir (carbovir) in human peripheral blood mononuclear cells by combined anion exchange solid phase extraction and LC-MS/MS",
abstract = "All nucleoside reverse transcriptase inhibitors (NRTI) must first be metabolized to their triphosphate forms in order to be active against HIV. Zidovudine (ZDV), abacavir (ABC) and lamivudine (3TC) have proven to be an efficacious combination. In order simultaneously to measure intracellular levels of the triphosphates (-TP) of ZDV, ABC (carbovir, CBV) and 3TC, either together or individually, we have developed a cartridge-LC-MS/MS method. The quantitation range was 2.5-250 pg/μl for 3TC-TP, 0.1-10.0 pg/μl for ZDV-TP and 0.05-5.00 pg/μl for CBV-TP. This corresponds to 0.1-11.0 pmol 3TC-TP per million cells, 4-375 fmol ZDV-TP per million cells and 2-200 fmol CBV-TP per million cells, extracted from 10 million cells. Patient samples demonstrated measured levels in the middle regions of our standard curves both at pre-dose and 4 h post-dose times.",
keywords = "Antiviral drug metabolism, Carbovir triphosphate, Human immunodeficiency virus, Lamivudine triphosphate, Zidovudine triphosphate",
author = "Robbins, {Brian L.} and Poston, {Philip A.} and Neal, {Erin F.} and Clive Slaughter and Rodman, {John H.}",
year = "2007",
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T1 - Simultaneous measurement of intracellular triphosphate metabolites of zidovudine, lamivudine and abacavir (carbovir) in human peripheral blood mononuclear cells by combined anion exchange solid phase extraction and LC-MS/MS

AU - Robbins, Brian L.

AU - Poston, Philip A.

AU - Neal, Erin F.

AU - Slaughter, Clive

AU - Rodman, John H.

PY - 2007/5/1

Y1 - 2007/5/1

N2 - All nucleoside reverse transcriptase inhibitors (NRTI) must first be metabolized to their triphosphate forms in order to be active against HIV. Zidovudine (ZDV), abacavir (ABC) and lamivudine (3TC) have proven to be an efficacious combination. In order simultaneously to measure intracellular levels of the triphosphates (-TP) of ZDV, ABC (carbovir, CBV) and 3TC, either together or individually, we have developed a cartridge-LC-MS/MS method. The quantitation range was 2.5-250 pg/μl for 3TC-TP, 0.1-10.0 pg/μl for ZDV-TP and 0.05-5.00 pg/μl for CBV-TP. This corresponds to 0.1-11.0 pmol 3TC-TP per million cells, 4-375 fmol ZDV-TP per million cells and 2-200 fmol CBV-TP per million cells, extracted from 10 million cells. Patient samples demonstrated measured levels in the middle regions of our standard curves both at pre-dose and 4 h post-dose times.

AB - All nucleoside reverse transcriptase inhibitors (NRTI) must first be metabolized to their triphosphate forms in order to be active against HIV. Zidovudine (ZDV), abacavir (ABC) and lamivudine (3TC) have proven to be an efficacious combination. In order simultaneously to measure intracellular levels of the triphosphates (-TP) of ZDV, ABC (carbovir, CBV) and 3TC, either together or individually, we have developed a cartridge-LC-MS/MS method. The quantitation range was 2.5-250 pg/μl for 3TC-TP, 0.1-10.0 pg/μl for ZDV-TP and 0.05-5.00 pg/μl for CBV-TP. This corresponds to 0.1-11.0 pmol 3TC-TP per million cells, 4-375 fmol ZDV-TP per million cells and 2-200 fmol CBV-TP per million cells, extracted from 10 million cells. Patient samples demonstrated measured levels in the middle regions of our standard curves both at pre-dose and 4 h post-dose times.

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