Simultaneous determination of ribavirin and ribavirin base in monkey plasma by high performance liquid chromatography with tandem mass spectrometry

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Analytical technologies in the biomedical and life sciences, 2007-02, Vol.846 (1), p.57-68
• Main Authors: Li, Wenkui, Luo, Suyi, Li, Shaoyong, Athill, Lawrence, Wu, Amy, Ray, Tapan, Zhou, Wei, Ke, June, Smith, Harold T., Tse, Francis L.S.
• Format: Article
• Language: English
• Subjects: Analysis; Analytical, structural and metabolic biochemistry; Animals; Antiviral Agents - blood; Biological and medical sciences; Chromatography, High Pressure Liquid - methods; Fundamental and applied biological sciences. Psychology; General pharmacology; Haplorhini; LC–MS/MS; Medical sciences; Pharmacology. Drug treatments; Reproducibility of Results; Ribavirin; Ribavirin - blood; Ribavirin base; Sensitivity and Specificity; Simultaneous analysis; Tandem Mass Spectrometry - methods
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/j.jchromb.2006.08.014

For the first time, a liquid chromatographic method with tandem mass spectrometric detection (LC–MS/MS) for the simultaneous determination of ribavirin and rabavirin base was developed and validated over the concentration range of 10–5000 ng/ml, respectively, using a 0.025 ml monkey plasma sample. Ribavirin, ribavirin base, and the internal standards were extracted from monkey plasma via protein precipitation. After evaporation of the supernatant, the extract was reconstituted with 5% methanol (containing 0.1% formic acid) and injected onto the LC–MS/MS system. Optimum chromatographic separation was achieved on a Waters Atlantis dc18 (150 mm × 2.1 mm, 5 μm) column with mobile phase run in gradient with 100% water containing 0.5% formic acid ( A) and 90% acetonitrile (containing 0.5% formic acid ( B). The flow rate was 0.4–0.6 ml/min with total cycle time of approximately 7.0 min. Post-column addition of acetonitrile (containing 0.1% formic acid) at 0.3 ml/min was used to increase the ionization efficiency in the MS source. The method was validated for sensitivity, linearity, reproducibility, stability and recovery. Lack of adverse matrix effect and carry-over was also demonstrated. The intra-day and inter-day precision and accuracy of the quality control (QC) samples were