A versatile system of high-flow high performance liquid chromatography with tandem mass spectrometry for rapid direct-injection analysis of plasma samples for quantitation of a β-lactam drug candidate and its open-ring biotransformation product

✍ Scribed by Mohammed Jemal; Zheng Ouyang; Yuan-Qing Xia; Mark L Powell

• Publisher: John Wiley and Sons
• Year: 1999
• Tongue: English
• Weight: 144 KB
• Volume: 13
• Category: Article
• ISSN: 0951-4198
• DOI: 10.1002/(sici)1097-0231(19990730)13:14<1462::aid-rcm668>3.0.co;2-z

✦ Synopsis

A bioanalytical method has been developed and validated for quantitation of a b-lactam drug candidate and its open-ring biotransformation product utilizing high-flow high-performance liquid chromatography (HPLC) for on-line purification of plasma samples and electrospray tandem mass spectrometry for detection and quantitation. The HPLC system used two columns: an Oasis column (1 Â 50 mm, 30 mm) as the on-line extraction column and a conventional C18 column (2 Â 50 mm, 5 mm) as the analytical column. Each plasma standard or quality control (QC) sample (50 mL) was mixed with 50 mL of a working solution of the internal standard in aqueous 0.5 M ammonium acetate (pH 4.0). Portions (10 mL) of these samples were then injected into an Oasis column with a mobile phase consisting of 100% aqueous 1 mM formic acid at a high flow rate (4.0 mL/min), with the effluent from the Oasis column directed to waste and not to the mass spectrometer. After the purification step, the Oasis column effluent was directed to the analytical column and the mass spectrometer and the analytes were eluted with methanol/aqueous 1 mM formic acid (70:30) at a flow rate of 1.0 mL/min. The total analysis time was 1.6 min per sample. The standard curve range was 0.980 to 250 ng/mL. The accuracy, inter-day precision and intra-day precision were within 10% for both compounds.