1. On-line coupling of anion exchange and ion-pair chromatography for measurement of intracellular triphosphate metabolites of reverse transcriptase inhibitors
- Author
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Kuklenyik, Zsuzsanna, Martin, Amy, Pau, Chou-Pong, Holder, Angela, Youngpairoj, Ae S., Zheng, Qi, Cong, Mian-Er, Garcia-Lerma, J. Gerardo, Heneine, Walid, Pirkle, James L., and Barr, John R.
- Subjects
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SOLID phase extraction , *ION exchange (Chemistry) , *HIGH performance liquid chromatography , *ELECTROSPRAY ionization mass spectrometry , *TANDEM mass spectrometry , *METABOLITES , *REVERSE transcriptase , *ENZYME inhibitors , *SEPARATION (Technology) - Abstract
Abstract: We developed an automated on-line weak anion exchange (WAX) solid-phase extraction (SPE) method coupled with ion-pair (IP) chromatography–tandem mass spectrometry (MS/MS) detection for quantitatively measuring triphosphorylated metabolites of three reverse transcriptase inhibitors (RTI). The administered pro-drugs were Tenofovir disoproxil fumarate (TDF), Emtricitabine (FTC) and Lamivudine (3TC). Their intracellular metabolites Tenofovir-diphosphate (TFV-DP), Emtricitabine-triphosphate (FTC-TP), and Lamivudine-triphosphate (3TC-TP) were measured in peripheral blood mononuclear cells (PBMC). We coupled the WAX and IP chromatography systems using a combination of 6-port and 10-port switching valves, and we mixed the WAX elute with 1,5-dimethyl-hexyl-amine before IP chromatography separation. Multiple waste outlets allowed for eliminating potential matrix components interfering with MS/MS detection. Limits of detection were 9, 200 and 75pg per sample for TFV-DP (448/176 m/z), FTC-TP (488/130 m/z) and 3TC-TP (468/119 m/z), respectively. [Copyright &y& Elsevier]
- Published
- 2009
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