Journal
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
Volume 944, Issue -, Pages 18-24Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jchromb.2013.10.035
Keywords
Ribavirin; Dried blood spot; Analytical method; Nucleoside analog; Hepatitis C; LC-MS/MS
Funding
- NIH NIDDK [K23 DK082621, R03 DK096121]
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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. (C) 2013 Elsevier B.V. All rights reserved.
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