Abstract
Background: Recent studies have demonstrated highly variable blood concentrations of piperacillin, tazobactam, cefepime, meropenem, ciprofloxacin and linezolid in critically ill patients with a high incidence of sub-therapeutic levels. Consequently, therapeutic drug monitoring (TDM) of these antibiotics has to be considered, requiring robust and reliable routine analytical methods. The aim of the present work was to develop and validate a multi-analyte ultra high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) method for the simultaneous quantification of the above mentioned antibiotics.
Methods: Sample preparation included a manual protein precipitation step followed by two-dimensional ultra high performance liquid chromatography (2D-UHPLC). Corresponding stable isotope-labeled substances were used as internal standards for all of the analytes, with the exception of tazobactam. The injected sample volume was 7 μL. The run time was 5.0 min.
Results: Inaccuracy was ≤8% and imprecision coefficient of variation (CV) was <9% for all analytes. Only minor matrix effects and negligible carry-over was observed. The method was found to be robust during the validation period.
Conclusions: We were able to develop a reliable 2D-UHPLC-MS/MS method addressing analytes with highly heterogeneous physico-chemical properties. The novel assay may be an efficient tool for an optimized process workflow in clinical laboratories for important antibiotics in regards to TDM.
Acknowledgments
This study was supported by a Mérieux Research Grant (Institut Mérieux, Lyon, France).
Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.
Financial support: None declared.
Employment or leadership: None declared.
Honorarium: None declared.
Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.
References
1. Karlsson S, Varpula M, Ruokonen E, Pettila V, Parviainen I, Ala-Kokko TI, et al. Incidence, treatment, and outcome of severe sepsis in ICU-treated adults in Finland: the Finnsepsis study. Intensive Care Med 2007;33:435–43.10.1007/s00134-006-0504-zSearch in Google Scholar PubMed
2. Martin CM, Priestap F, Fisher H, Fowler RA, Heyland DK, Keenan SP, et al. A prospective, observational registry of patients with severe sepsis: the Canadian Sepsis Treatment and Response Registry. Crit Care Med 2009;37:81–8.10.1097/CCM.0b013e31819285f0Search in Google Scholar PubMed
3. Quenot JP, Binquet C, Kara F, Martinet O, Ganster F, Navellou JC, et al. The epidemiology of septic shock in French intensive care units: the prospective multicenter cohort EPISS study. Crit Care 2013;17:R65.10.1186/cc12598Search in Google Scholar PubMed PubMed Central
4. Kollef MH, Sherman G, Ward S, Fraser VJ. Inadequate antimicrobial treatment of infections: a risk factor for hospital mortality among critically ill patients. Chest 1999;115:462–74.10.1378/chest.115.2.462Search in Google Scholar PubMed
5. Kumar A, Roberts D, Wood KE, Light B, Parrillo JE, Sharma S, et al. Duration of hypotension before initiation of effective antimicrobial therapy is the critical determinant of survival in human septic shock. Crit Care Med 2006;34:1589–96.10.1097/01.CCM.0000217961.75225.E9Search in Google Scholar PubMed
6. Pea F, Viale P, Furlanut M. Antimicrobial therapy in critically ill patients: a review of pathophysiological conditions responsible for altered disposition and pharmacokinetic variability. Clin Pharmacokinet 2005;44:1009–34.10.2165/00003088-200544100-00002Search in Google Scholar PubMed
7. Reinhart K, Brunkhorst FM, Bone HG, Bardutzky J, Dempfle CE, Forst H, et al. Prevention, diagnosis, therapy and follow-up care of sepsis: 1st revision of S-2k guidelines of the German Sepsis Society (Deutsche Sepsis-Gesellschaft e.V. (DSG)) and the German Interdisciplinary Association of Intensive Care and Emergency Medicine (Deutsche Interdisziplinäre Vereinigung für Intensiv- und Notfallmedizin (DIVI)). Ger Med Sci 2010;8:Doc14.Search in Google Scholar
8. Bodmann Klaus-Friedrich GB. Empfehlungen zur kalkulierten parenteralen intitialtherapie bakterieller erkrankungen bei erwachsenen update. Paul-Ehrlich-Gesellschaft 2010; Chemother J 2010;19:179–255.Search in Google Scholar
9. Chapuis TM, Giannoni E, Majcherczyk PA, Chioléro R, Schaller MD, Berger MM, et al. Prospective monitoring of cefepime in intensive care unit adult patients. Crit Care 2010;14:R51.10.1186/cc8941Search in Google Scholar PubMed PubMed Central
10. Forrest A, Nix DE, Ballow CH, Goss TF, Birmingham MC, Schentag JJ. Pharmacodynamics of intravenous ciprofloxacin in seriously ill patients. Antimicrob Agents Chemother 1993;37:1073–81.10.1128/AAC.37.5.1073Search in Google Scholar PubMed PubMed Central
11. Gonçalves-Pereira J, Póvoa P. Antibiotics in critically ill patients: a systematic review of the pharmacokinetics of β-lactams. Crit Care 2011;15:R206.10.1186/cc10441Search in Google Scholar PubMed PubMed Central
12. Rayner CR, Forrest A, Meagher AK, Birmingham MC, Schentag JJ. Clinical pharmacodynamics of linezolid in seriously ill patients treated in a compassionate use programme. Clin Pharmacokinet 2003;42:1411–23.10.2165/00003088-200342150-00007Search in Google Scholar PubMed
13. Taccone FS, Laterre PF, Dugernier T, Spapen H, Delattre I, Wittebole X, et al. Insufficient β-lactam concentrations in the early phase of severe sepsis and septic shock. Crit Care 2010;14:R126.10.1186/cc9091Search in Google Scholar PubMed PubMed Central
14. Ariano RE, Nyhlén A, Donnelly JP, Sitar DS, Harding GK, Zelenitsky SA. Pharmacokinetics and pharmacodynamics of meropenem in febrile neutropenic patients with bacteremia. Ann Pharmacother 2005;39:32–8.10.1345/aph.1E271Search in Google Scholar PubMed
15. Craig WA. Pharmacokinetic/pharmacodynamic parameters: rationale for antibacterial dosing of mice and men. Clin Infect Dis 1998;26:1–10.10.1086/516284Search in Google Scholar PubMed
16. McKinnon PS, Paladino JA, Schentag JJ. Evaluation of area under the inhibitory curve (AUIC) and time above the minimum inhibitory concentration (T>MIC) as predictors of outcome for cefepime and ceftazidime in serious bacterial infections. Int J Antimicrob Agents 2008;31:345–51.10.1016/j.ijantimicag.2007.12.009Search in Google Scholar PubMed
17. Preston SL, Drusano GL, Berman AL, Fowler CL, Chow AT, Dornseif B, et al. Pharmacodynamics of levofloxacin: a new paradigm for early clinical trials. J Am Med Assoc 1998;279:125–9.10.1001/jama.279.2.125Search in Google Scholar PubMed
18. Dellinger RP, Levy MM, Rhodes A, Annane D, Gerlach H, Opal SM, et al. Surviving sepsis campaign: international guidelines for management of severe sepsis and septic shock: 2012. Crit Care Med 2013;41:580–637.10.1097/CCM.0b013e31827e83afSearch in Google Scholar PubMed
19. Ahsman MJ, Wildschut ED, Tibboel D, Mathot RA. Microanalysis of beta-lactam antibiotics and vancomycin in plasma for pharmacokinetic studies in neonates. Antimicrob Agents Chemother 2009;53:75–80.10.1128/AAC.00636-08Search in Google Scholar PubMed PubMed Central
20. Bu W, Sexton H, Fan X, Torres P, Houston P, Heyman I, et al. The novel sensitive and high throughput determination of cefepime in mouse plasma by SCX-LC/MS/MS method following off-line microElution 96-well solid-phase extraction to support systemic antibiotic programs. J Chromatogr B Analyt Technol Biomed Life Sci 2010;878:1623–8.10.1016/j.jchromb.2010.03.045Search in Google Scholar PubMed
21. Carlier M, Stove V, Roberts JA, Van de Velde E, De Waele JJ, Verstraete AG. Quantification of seven β-lactam antibiotics and two β-lactamase inhibitors in human plasma using a validated UPLC-MS/MS method. Int J Antimicrob Agents 2012;40:416–22.10.1016/j.ijantimicag.2012.06.022Search in Google Scholar PubMed
22. Cazorla-Reyes R, Romero-González R, Frenich AG, Rodríguez Maresca MA, Martínez Vidal JL. Simultaneous analysis of antibiotics in biological samples by ultra high performance liquid chromatography-tandem mass spectrometry. J Pharm Biomed Anal 2014;89:203–12.10.1016/j.jpba.2013.11.004Search in Google Scholar PubMed
23. Cohen-Wolkowiez M, White NR, Bridges A, Benjamin DK Jr, Kashuba AD. Development of a liquid chromatography-tandem mass spectrometry assay of six antimicrobials in plasma for pharmacokinetic studies in premature infants. J Chromatogr B Analyt Technol Biomed Life Sci 2011;879:3497–506.10.1016/j.jchromb.2011.09.031Search in Google Scholar PubMed PubMed Central
24. Colin P, De Bock L, T’jollyn H, Boussery K, Van Bocxlaer J. Development and validation of a fast and uniform approach to quantify β-lactam antibiotics in human plasma by solid phase extraction-liquid chromatography-electrospray-tandem mass spectrometry. Talanta 2013;103:285–93.10.1016/j.talanta.2012.10.046Search in Google Scholar PubMed
25. Jourdil JF, Tonini J, Stanke-Labesque F. Simultaneous quantitation of azole antifungals, antibiotics, imatinib, and raltegravir in human plasma by two-dimensional high-performance liquid chromatography-tandem mass spectrometry. J Chromatogr B Analyt Technol Biomed Life Sci 2013;919–920:1–9.10.1016/j.jchromb.2012.12.028Search in Google Scholar PubMed
26. Ohmori T, Suzuki A, Niwa T, Ushikoshi H, Shirai K, Yoshida S, et al. Simultaneous determination of eight β-lactam antibiotics in human serum by liquid chromatography-tandem mass spectrometry. J Chromatogr B Analyt Technol Biomed Life Sci 2011;879:1038–42.10.1016/j.jchromb.2011.03.001Search in Google Scholar PubMed
27. Szultka M, Krzeminski R, Jackowski M, Buszewski B. Simultaneous determination of selected chemotherapeutics in human whole blood by molecularly imprinted polymers coated solid phase microextraction fibers and liquid chromatography-tandem mass spectrometry. J Chromatogr B Analyt Technol Biomed Life Sci 2013;940:66–76.10.1016/j.jchromb.2013.09.009Search in Google Scholar PubMed
28. Zander J, Maier B, Zoller M, Teupser D, Vogeser M. Quantification of linezolid in serum by LC-MS/MS using semi-automated sample preparation and isotope dilution internal standardization. Clin Chem Lab Med 2014;52:381–9.10.1515/cclm-2013-0594Search in Google Scholar PubMed
29. Klee GG. Establishment of outcome-related analytic performance goals. Clin Chem 2010;56:714–22.10.1373/clinchem.2009.133660Search in Google Scholar PubMed
30. Richtlinie der Bundesaerztekammer zur Qualitätssicherung laboratoriums-medizinischer Untersuchungen. Dt Aerzteblatt 2008;105:C301–13.Search in Google Scholar
31. Clinical and Laboratory Standards Institute. Liquid chromatography-mass spectrometry methods; draft guideline C62 (Draft 2). Wayne, PA: CLSI, 2013.Search in Google Scholar
32. European Medicines Agency. Guideline on bioanalytical method validation. London, UK: Committee for Medicinal Product for Human Use (CHMP), 2011.Search in Google Scholar
33. Matuszewski BK, Constanzer ML, Chavez-Eng CM. Strategies for the assessment of matrix effect in quantitative bioanalytical methods based on HPLC-MS/MS. Anal Chem 2003;75:3019–30.10.1021/ac020361sSearch in Google Scholar PubMed
34. Harmelink IM, Alffenaar JC, Wessels AM, Greijdanus B, Uges DR. A rapid and simple liquid chromatography-tandem mass spectrometry method for the determination of linezolid in human serum. EJHP Science 2008;14:3–7.Search in Google Scholar
35. Phillips OA, Abdel-Hamid ME, al-Hassawi NA. Determination of linezolid in human plasma by LC-MS-MS. Analyst 2001;126:609–14.10.1039/b100076oSearch in Google Scholar PubMed
36. Meyer E, Gastmeier P, Deja M, Schwab F. Antibiotic consumption and resistance: data from Europe and Germany. Int J Med Microbiol 2013;303:388–95.10.1016/j.ijmm.2013.04.004Search in Google Scholar PubMed
37. Website: surveillance der antibiotika-anwendung und der bakteriellen resistenzen auf intensivstationen. SARI information for antibiotic consumption over the time of all intensive care units participating at SARI. Available from: http://sari.eu-burden.info/auswertung/down/AD-ZEIT.pdf. Accessed 5 June, 2014.Search in Google Scholar
38. Zoller M, Barbara M, Hornuss C, Neugebauer C, Döbbeler G, Nagel D, et al. Variability of linezolid concentrations after standard dosing in critically ill patients: a prospective observational study. Crit Care 2014;18:R148.10.1186/cc13984Search in Google Scholar PubMed PubMed Central
39. Reed MD, Goldfarb J, Yamashita TS, Lemon E, Blumer JL. Single-dose pharmacokinetics of piperacillin and tazobactam in infants and children. Antimicrob Agents Chemother 1994;38:2817–26.10.1128/AAC.38.12.2817Search in Google Scholar PubMed PubMed Central
40. Vogeser M, Seger C. Pitfalls associated with the use of liquid chromatography-tandem mass spectrometry in the clinical laboratory. Clin Chem 2010;56:1234–44.10.1373/clinchem.2009.138602Search in Google Scholar PubMed
41. Roberts JA, Norris R, Paterson DL, Martin JH. Therapeutic drug monitoring of antimicrobials. Br J Clin Pharmacol 2012;73:27–36.10.1111/j.1365-2125.2011.04080.xSearch in Google Scholar PubMed PubMed Central
42. Turnidge JD. The pharmacodynamics of beta-lactams. Clin Infect Dis 1998,27:10–22.10.1086/514622Search in Google Scholar PubMed
43. EUCAST Website: European Committee on Antimicrobial Susceptibility Testing. Information on clinical breakpoint tables. Available from: http://www.eucast.org/clinical_breakpoints/. Accessed 2 July, 2014.Search in Google Scholar
44. Shorr AF. Review of studies of the impact on Gram-negative bacterial resistance on outcomes in the intensive care unit. Crit Care Med 2009;37:1463–9.10.1097/CCM.0b013e31819ced02Search in Google Scholar PubMed
45. Choi G, Gomersall CD, Tian Q, Joynt GM, Li AM, Lipman J. Principles of antibacterial dosing in continuous renal replacement therapy. Blood Purif 2010;30:195–212.10.1159/000321488Search in Google Scholar PubMed
46. Stalker DJ, Jungbluth GL, Hopkins NK, Batts DH. Pharmacokinetics and tolerance of single- and multiple-dose oral or intravenous linezolid, an oxazolidinone antibiotic, in healthy volunteers. J Antimicrob Chemother 2003;51:1239–46.10.1093/jac/dkg180Search in Google Scholar PubMed
Supplemental Material
The online version of this article (DOI: 10.1515/cclm-2014-0746) offers supplementary material, available to authorized users.
©2015 by De Gruyter
