Simultaneous quantification of plasma levels of 12 antimicrobial agents including carbapenem, anti-methicillin-resistant Staphylococcus aureus agent, quinolone and azole used in intensive care unit using UHPLC-MS/MS method

• Published in: Clinical biochemistry 2021-04, Vol.90, p.40-49
• Main Authors: Kai, Makoto, Tanaka, Ryota, Suzuki, Yosuke, Goto, Koji, Ohchi, Yoshifumi, Yasuda, Norihisa, Tatsuta, Ryosuke, Kitano, Takaaki, Itoh, Hiroki
• Format: Article
• Language: English
• Subjects: Antimicrobial; Cmax and Ctrough; Intensive care unit; Pharmacokinetics; Tandem mass spectrometry; Ultra-high performance liquid chromatography
• ISSN: 0009-9120; 1873-2933
• DOI: 10.1016/j.clinbiochem.2021.01.012

•A sensitive and selective method is developed using UHPLC-MS/MS for quantitative measurement of 12 antimicrobial agents commonly used in ICU.•Calibration curves have sufficiently wide ranges to measure both Cmax and Ctrough in ICU patients. Critically ill patients in intensive care unit (ICU) are susceptible to infectious diseases, thus empirical therapy is recommended. However, the therapeutic effect in ICU patients is difficult to predict due to fluctuation in pharmacokinetics because of various factors. This problem can be solved by developing personalized medicine through therapeutic drug monitoring. However, when different measurement systems are used for various drugs, measurements are complicated and time consuming in clinical practice. In this study, we aimed to develop an assay using ultra-high performance liquid chromatography coupled with tandem mass spectrometry for simultaneous quantification of 12 antimicrobial agents commonly used in ICU: doripenem, meropenem, linezolid, tedizolid, daptomycin, ciprofloxacin, levofloxacin, pazufloxacin, fluconazole, voriconazole, voriconazole N-oxide which is a major metabolite of voriconazole, and posaconazole. Plasma protein was precipitated by adding acetonitrile and 50% MeOH containing standard and labeled IS. The analytes were separated with an ACQUITY UHPLC CSH C18 column, under a gradient mobile phase consisting of water and acetonitrile containing 0.1% formic acid and 2 mM ammonium formate. The method fulfilled the criteria of US Food and Drug Administration for assay validation. The recovery rate was more than 84.8%. Matrix effect ranged from 79.1% to 119.3%. All the calibration curves showed good linearity (back calculation of calibrators: relative error ≤ 15%) over wide concentration ranges, which allowed determination of Cmax and Ctrough. Clinical applicability of the novel method was confirmed. We have developed an assay for simultaneous quantification of 12 antimicrobial agents using a small sample volume of 50 μL with a short assay time of 7 min. Our novel method may contribute to simultaneous calculation of pharmacokinetic and pharmacodynamic parameters.