Simultaneous determination of ribavirin and ribavirin base in monkey plasma by high performance liquid chromatography with tandem mass spectrometry.
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 10-5,000 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 x 2.1mm, 5 microm) 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 <9.0% relative standard deviation (R.S.D.) and 10.8% bias for ribavirin, and 10.3% R.S.D. and 11.3% bias for ribavirin base. The current specific, accurate and precise assay is useful in support of the toxicokinetic and pharmacokinetic studies of these compounds.
Li
Wenkui
W
Department of Drug Metabolism and Pharmacokinetics, Novartis Pharmaceuticals Corporation, One Health Plaza, East Hanover, NJ 07936, USA. wenkui.li@novartis.com
Luo
Suyi
S
Li
Shaoyong
S
Athill
Lawrence
L
Wu
Amy
A
Ray
Tapan
T
Zhou
Wei
W
Ke
June
J
Smith
Harold T
HT
Tse
Francis L S
FL
D000818:Animals; D000998:Antiviral Agents / Q000097:blood*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D000882:Haplorhini; D015203:Reproducibility of Results; D012254:Ribavirin / Q000097:blood*; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods*
D000998:Antiviral Agents; D012254:Ribavirin