Abstract |
HIV-1 integrase strand transfer inhibitors are an important class of compounds targeted for the treatment of HIV-1 infection. Microdosing has emerged as an attractive tool to assist in drug candidate screening for clinical development, but necessitates extremely sensitive bioanalytical assays, typically in the pg/mL concentration range. Currently, accelerator mass spectrometry is the predominant tool for microdosing support, which requires a specialized facility and synthesis of radiolabeled compounds. There have been few studies attempted to comprehensively assess a liquid chromatography-tandem mass spectrometry (LC-MS/MS) approach in the context of microdosing applications. Herein, we describe the development of automated LC-MS/MS methods to quantify five integrase inhibitors in plasma with the limits of quantification at 1 pg/mL for raltegravir and 2 pg/mL for four proprietary compounds. The assays involved double extractions followed by UPLC coupled with negative ion electrospray MS/MS analysis. All methods were fully validated to the rigor of regulated bioanalysis requirements, with intraday precision between 1.20 and 14.1% and accuracy between 93.8 and 107% at the standard curve concentration range. These methods were successfully applied to a human microdose study and demonstrated to be accurate, reproducible, and cost-effective. Results of the study indicate that raltegravir displayed linear pharmacokinetics between a microdose and a pharmacologically active dose.
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Authors | Li Sun, Hankun Li, Kenneth Willson, Sheila Breidinger, Matthew L Rizk, Larissa Wenning, Eric J Woolf |
Journal | Analytical chemistry
(Anal Chem)
Vol. 84
Issue 20
Pg. 8614-21
(Oct 16 2012)
ISSN: 1520-6882 [Electronic] United States |
PMID | 23030780
(Publication Type: Clinical Trial, Journal Article, Research Support, Non-U.S. Gov't)
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Chemical References |
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Topics |
- Adolescent
- Adult
- Chromatography, Liquid
(methods)
- Drug Dosage Calculations
- HIV Integrase Inhibitors
(administration & dosage, blood)
- Humans
- Limit of Detection
- Male
- Middle Aged
- Tandem Mass Spectrometry
(methods)
- Young Adult
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