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Journal Article
Research Support, N.I.H., Extramural
Determination of cidofovir in human plasma after low dose drug administration using high-performance liquid chromatography-tandem mass spectrometry.
Journal of Pharmaceutical and Biomedical Analysis 2010 December 2
A sensitive and specific method for the determination of cidofovir (CDV) in human plasma using high-performance liquid chromatography with tandem mass spectrometry (LC-MS/MS) was developed and validated. Plasma samples were processed by a solid phase extraction (SPE) procedure using Varian SAX extraction cartridges prior to chromatography. The internal standard was (13)C5-Folic acid ((13)C5-FA). Chromatography was performed using a Luna C8(2) analytical column, 5 microm, 150 mm x 3.0 mm, using an isocratic elution with a mobile phase consisting of 43% methanol in water containing 12 mM ammonium acetate, at a flow rate of 0.3 mL/min. The retention times of CDV and (13)C5-FA were 2.1 min and 1.9 min, respectively, with a total run time of 5 min. The analytes were detected by a Micromass Quattro Micro triple quadrupole mass spectrometer in positive electron spray ionization (ESI) mode using multiple reaction monitoring (MRM). The extracted ions monitored following MRM transitions were m/z 280.0-->262.1 for CDV and m/z 447.0-->294.8 for (13)C5-FA (IS). The assay was linear over the range 20-1000 ng/mL. Accuracy (101.6-105.7%), intra-assay precision (4.1-5.4%), and inter-assay precision (5.6-6.8%) were within FDA limits. No significant variation in the concentration of CDV was observed with different sample storage conditions. This method is simple, adaptable to routine application, and allows easy and accurate measurement of CDV in human plasma.
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