Development and validation of a dried blood spot assay for the quantification of ribavirin using liquid chromatography coupled to mass spectrometry

作者：Jimmerson Leah C; Zheng Jia Hua; Bushman Lane R; MacBrayne Christine E; Anderson Peter L; Kiser Jennifer J^*

来源：Journal of Chromatography B-Analytical Technologies in the Biomedical and Life Sciences, 2014, 944: 18-24.

DOI：10.1016/j.jchromb.2013.10.035

• Ribavirin
• Dried blood spot
• Analytical method
• Nucleoside analog
• Hepatitis C
• LC-MS/MS

Efficient, inexpensive and sensitive assays for the measurement of drugs are of interest for pharmacokinetic and pharmacodynamics (PK-PD) analysis. Dried blood spots (DBS) are a unique bioanaltyical matrix with the potential to fulfill this interest for the measurement of numerous analytes. Here we describe the development and validation of a reversed-phase high performance liquid chromatographic (LC), tandem mass spectrometry (MS/MS) assay for the determination of ribavirin (RBV) in DBS. A 3 mm punch from spotted and dried whole blood was extracted in methanol utilizing isotopically labeled internal standard for LC-MS/MS analysis. Validation was performed over a range of 0.05 mu g/mL to 10.0 mu g/mL and the method was shown to be precise (coefficient of variation <= 15%) and accurate (within +/- 15% of control). These acceptance criteria were met for hematocrit ranges of 20-54%, for center versus edge punches and for spot volumes from 10 to 60 mu L. RBV was stable for up to 140 days at room temperature and -20 degrees C as well as for three freeze/thaw cycles. Correlation of RBV in DBS versus in plasma yielded r(2) >= 0.98 demonstrating that DBS can be used as an alternative to plasma for PK-PD studies in human subjects.

• 出版日期2014-1-1