Highly sensitive determination of HCV protease inhibitors boceprevir (SCH 503034) and telaprevir (VX 950) in human plasma by LC-MS/MS.
The purpose of this study was to develop a specific and highly sensitive method based on fast sample preparation and LC-MS/MS techniques for the determination of the HCV protease inhibitors boceprevir (SCH 503034) and telaprevir (VX 950) in human plasma. Boceprevir, telaprevir and the internal standard dimethylcelecoxib were separated on a Luna C18 column (150 mm x 2.0 mm I.D., 5 microm particle size) under gradient conditions with a mobile phase A consisting of water/ammonia solution (25%) (100:0.05, v/v) and mobile phase B consisting of methanol/ammonia solution (25%) (100:0.05, v/v) and a chromatographic run time of 11 min. The lower limit of quantification (LLOQ) of boceprevir and telaprevir is 0.25 pg on column (25 pg/mL at injection volume of 10 microL). The method possesses a reliable calibration range of 0.025-2.5 ng/mL. Due to the dilution of real life plasma samples by a factor of 10 during the precipitation process the method is suitable to quantify boceprevir and telaprevir at a concentration range of 0.25-25 ng/mL. Variations in accuracy and intraday and interday precision (n=6 for each concentration) were <15% over the whole range of calibration. For the first time, a rapid, specific, sensitive, accurate and reproducible LC-MS/MS method in human plasma has been developed and validated. It is suitable to quantify the concentrations of the hepatitis C virus protease inhibitors boceprevir and telaprevir in human plasma.
Pharmazentrum Frankfurt/ZAFES, Institut für Klinische Pharmakologie, Klinikum der Johann Wolfgang Goethe-Universität Frankfurt, Theodor-Stern-Kai 7, Haus 74, 60590 Frankfurt am Main, Germany.
Harald
H
Farnik
Juliane
J
El-Duweik
Christoph
C
Welsch
Christoph
C
Sarrazin
Jörn
J
Lötsch
Stefan
S
Zeuzem
Gerd
G
Geisslinger
Helmut
H
Schmidt
D000998:Antiviral Agents / Q000097:blood; D002853:Chromatography, Liquid / Q000379:methods*; D016174:Hepacivirus / Q000201:enzymology*; D006801:Humans; D009842:Oligopeptides / Q000097:blood*; D010447:Peptide Hydrolases / Q000378:metabolism*; D011392:Proline / Q000031:analogs & derivatives* / Q000097:blood; D011480:Protease Inhibitors / Q000097:blood*; D012015:Reference Standards; D053719:Tandem Mass Spectrometry / Q000379:methods*
D000998:Antiviral Agents; D009842:Oligopeptides; D011480:Protease Inhibitors; C486464:telaprevir; C512204:N-(3-amino-1-(cyclobutylmethyl)-2,3-dioxopropyl)-3-(2-((((1,1-dimethylethyl)amino)carbonyl)amino)-3,3-dimethyl-1-oxobutyl)-6,6-dimethyl-3-azabicyclo(3.1.0)hexan-2-carboxamide; D011392:Proline; D010447:Peptide Hydrolases