Novel treatment options for some carbapenem-resistant Gram-negative pathogens have been identified by the World Health Organization as being of the highest priority. Ceftolozane–tazobactam is a novel cephalosporin–beta-lactamase inhibitor combination antibiotic with potent bactericidal activity against the most difficult-to-treat multi-drug resistant and extensively drug resistant Gram-negative pathogens. This study aimed to develop and validate a liquid chromatography – tandem mass spectrometry method for the simultaneous quantification of ceftolozane and tazobactam in plasma (total and unbound), renal replacement therapy effluent (RRTE), cerebrospinal fluid (CSF) and urine.
Analytes were separated using mixed-mode chromatography with an intrinsically base-deactivated C18 column and a gradient mobile phase consisting of 0.1% formic acid, 10 mM ammonium formate and acetonitrile. The analytes and internal standards were detected using rapid ionisation switching between positive and negative modes with simultaneous selected reaction monitoring.
A quadratic calibration was obtained for plasma (total and unbound), RRTE and CSF over the concentration range of 1–200 mg/L for ceftolozane and 0.5–100 mg/L for tazobactam, and for urine the concentration range of 10–2,000 mg/L for ceftolozane and 5–1,000 mg/L for tazobactam. For both ceftolozane and tazobactam, validation testing for matrix effects, precision and accuracy, specificity and stability were all within the acceptance criteria of ±15%.
This methodology was successfully applied to one pilot pharmacokinetic study in infected critically ill patients, including patients receiving renal replacement therapy, and one case study of a patient with ventriculitis, where all patients received ceftolozane–tazobactam.
Funding source: Merck Sharp and Dohme
Research funding: This study was funded in part by an investigator-initiated grant from MSD. SLP is a recipient of a National Health and Medical Research Council-funded Fellowship (APP1142757), JAR is a recipient of a National Health and Medical Research Council-funded Centre for Research Excellence Research Excellence (APP1044941), Project Grant (1062040) and Fellowship (APP1048652).
Author contributions: SLP: method design, data analysis, interpretation of results and writing of manuscript; SP: method design, data analysis, writing of manuscript; FS: study protocol design and writing of manuscript; JL: study protocol design and writing of manuscript; JAR: study protocol design and interpretation of results and writing of manuscript; SCW: method design, data interpretation and writing of manuscript.
Competing interests: JAR has provided consultancy and has received grant funding from MSD. All other authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
Ethical approval: The study was performed in accordance with the ethical standards, with ethical approval obtained for the use of drug-free human blood from the Royal Brisbane and Women’s Hospital Human Research Ethics Committee HREC/16/QRBW/211and HREC/17/QRBW/117.
1. World Health Organization. WHO publishes list of bacteria for which new antibiotics are urgently needed. Geneva: World Health Organization; 2017.Search in Google Scholar
2. Koulenti, D, Song, A, Ellingboe, A, Abdul-Aziz, MH, Harris, P, Gavey, E, et al.. Infections by multidrug-resistant Gram-negative Bacteria: what’s new in our arsenal and what’s in the pipeline? Int J Antimicrob Agents 2019;53:211–24. https://doi.org/10.1016/j.ijantimicag.2018.10.011.Search in Google Scholar
3. Nelson, RE, Slayton, RB, Stevens, VW, Jones, MM, Khader, K, Rubin, MA, et al.. Attributable mortality of healthcare-associated infections due to multidrug-resistant Gram-negative bacteria and methicillin-resistant Staphylococcus aureus. Infect Control Hosp Epidemiol 2017;38:848–56. https://doi.org/10.1017/ice.2017.83.Search in Google Scholar
4. Viala, B, Zaidi, FZ, Bastide, M, Dumont, Y, Le Moing, V, Jean-Pierre, H, et al.. Assessment of the in vitro activities of ceftolozane/tazobactam and ceftazidime/avibactam in a collection of beta-lactam-resistant Enterobacteriaceae and Pseudomonas aeruginosa clinical isolates at Montpellier University Hospital, France. Microb Drug Resist 2019;5:1325–9.10.1089/mdr.2018.0439Search in Google Scholar
5. Zhanel, GG, Chung, P, Adam, H, Zelenitsky, S, Denisuik, A, Schweizer, F, et al.. Ceftolozane/tazobactam: a novel cephalosporin/beta-lactamase inhibitor combination with activity against multidrug-resistant Gram-negative bacilli. Drugs 2014;74:31–51. https://doi.org/10.1007/s40265-013-0168-2.Search in Google Scholar
6. Farrell, DJ, Flamm, RK, Sader, HS, Jones, RN. Antimicrobial activity of ceftolozane–tazobactam tested against Enterobacteriaceae and Pseudomonas aeruginosa with various resistance patterns isolated in U.S. Hospitals (2011-2012). Antimicrob Agents Chemother 2013;57:6305–10. https://doi.org/10.1128/aac.01802-13.Search in Google Scholar
7. U.S. Food and Drug Administration. FDA approves new antibacterial drug Zerbaxa; Silver Spring: U.S. Food and Drug Administration; 2014.Search in Google Scholar
8. Solomkin, J, Hershberger, E, Miller, B, Popejoy, M, Friedland, I, Steenbergen, J, et al.. Ceftolozane/tazobactam plus metronidazole for complicated intra-abdominal infections in an era of multidrug resistance: results from a randomized, double-blind, phase 3 trial (ASPECT-cIAI). Clin Infect Dis 2015;60:1462–71. https://doi.org/10.1093/cid/civ097.Search in Google Scholar
9. Wagenlehner, FM, Umeh, O, Steenbergen, J, Yuan, GJ, Darouiche, RO. Ceftolozane-tazobactam compared with levofloxacin in the treatment of complicated urinary-tract infections, including pyelonephritis: a randomised, double-blind, phase 3 trial (ASPECT-cUTI). Lancet 2015;385:1949–56. https://doi.org/10.1016/s0140-6736(14)62220-0.Search in Google Scholar
10. Sader, HS, Farrell, DJ, Flamm, RK, Jones, RN. Ceftolozane/tazobactam activity tested against aerobic Gram-negative organisms isolated from intra-abdominal and urinary tract infections in European and United States hospitals (2012). J Infect 2014;69:266–77. https://doi.org/10.1016/j.jinf.2014.04.004.Search in Google Scholar PubMed
11. Lerma, FA, Bermudez, RM, Grau, S, Arnillas, MPG, Sorli, L, Recasens, L, et al.. Ceftolozane-tazobactam for the treatment of ventilator-associated infections by colistin-resistant Pseudomonas aeruginosa. Rev Esp Quimioter 2017;30:224–8.Search in Google Scholar
12. U.S. Food and Drug Administration. FDA approves new treatment for hospital-acquired and ventilator-associated bacterial pneumonia; Silver Spring: U.S. Food and Drug Administration; 2019.Search in Google Scholar
13. de Kraker, MEA, Davey, PG, Grundmann, H, Grp, BS. Mortality and hospital stay associated with resistant Staphylococcus aureus and Escherichia coli bacteremia: estimating the burden of antibiotic resistance in Europe. PLoS Med 2011;8:8. https://doi.org/10.1371/journal.pmed.1001104.Search in Google Scholar PubMed PubMed Central
14. Chandorkar, G, Xiao, A, Mouksassi, MS, Hershberger, E, Krishna, G. Population pharmacokinetics of ceftolozane/tazobactam in healthy volunteers, subjects with varying degrees of renal function and patients with bacterial infections. J Clin Pharmacol 2015;55:230–9. https://doi.org/10.1002/jcph.395.Search in Google Scholar PubMed PubMed Central
15. Miller, B, Hershberger, E, Benziger, D, Trinh, M, Friedland, I. Pharmacokinetics and safety of intravenous ceftolozane-tazobactam in healthy adult subjects following single and multiple ascending doses. Antimicrob Agents Chemother 2012;56:3086–91. https://doi.org/10.1128/aac.06349-11.Search in Google Scholar PubMed PubMed Central
16. Sime, FB, Lassig-Smith, M, Starr, T, Stuart, J, Pandey, S, Parker, SL, et al.. Population pharmacokinetics of unbound ceftolozane and tazobactam in critically ill patients without renal dysfunction. Antimicrob Agents Chemother 2019;63:e01265–19. https://doi.org/10.1128/aac.01265-19.Search in Google Scholar
17. Roger, C, Cotta, MO, Muller, L, Wallis, SC, Lipman, J, Lefrant, JY, et al.. Impact of renal replacement modalities on the clearance of piperacillin-tazobactam administered via continuous infusion in critically ill patients. Int J Antimicrob Agents 2017;50:227–31. https://doi.org/10.1016/j.ijantimicag.2017.03.018.Search in Google Scholar PubMed
18. Carlier, M, Taccone, FS, Beumier, M, Seyler, L, Cotton, F, Jacobs, F, et al.. Population pharmacokinetics and dosing simulations of cefepime in septic shock patients receiving continuous renal replacement therapy. Int J Antimicrob Agents 2015;46:413–9. https://doi.org/10.1016/j.ijantimicag.2015.05.020.Search in Google Scholar PubMed
19. Gao, CL, Tong, J, Yu, KJ, Sun, ZD, An, R, Du, ZM. Pharmacokinetics of cefoperazone/sulbactam in critically ill patients receiving continuous venovenous hemofiltration. Eur J Clin Pharmacol 2016;72:823–30. https://doi.org/10.1007/s00228-016-2045-x.Search in Google Scholar PubMed
20. Mariat, C, Venet, C, Jehl, F, Mwewa, S, Lazarevic, V, Diconne, E, et al.. Continuous infusion of ceftazidime in critically ill patients undergoing continuous venovenous haemodiafiltration: pharmacokinetic evaluation and dose recommendation. Crit Care 2006;10:7. https://doi.org/10.1186/cc3993.Search in Google Scholar PubMed PubMed Central
21. Seyler, L, Cotton, F, Taccone, FS, De Backer, D, Macours, P, Vincent, JL, et al.. Recommended beta-lactam regimens are inadequate in septic patients treated with continuous renal replacement therapy. Crit Care 2011;15:9. https://doi.org/10.1186/cc10257.Search in Google Scholar PubMed PubMed Central
22. Rawlins, M, Cheng, V, Raby, E, Dyer, J, Regli, A, Ingram, P, et al.. Pharmacokinetics of ceftolozane-tazobactam during prolonged intermittent renal replacement therapy. Chemotherapy 2018;63:203–6. https://doi.org/10.1159/000493196.Search in Google Scholar PubMed
23. Sime, FB, Lassig-Smith, M, Starr, T, Stuart, J, Pandey, S, Parker, SL, et al.. A population pharmacokinetic model-guided evaluation of ceftolozane-tazobactam dosing in critically ill patients undergoing continuous venovenous hemodiafiltration. Antimicrob Agents Chemother 2020;64:e01655. https://doi.org/10.1128/aac.01655-19.Search in Google Scholar PubMed PubMed Central
24. Heffernan, AJ, Germano, A, Sime, FB, Roberts, JA, Kimura, E. Vancomycin population pharmacokinetics for adult patients with sepsis or septic shock: are current dosing regimens sufficient? Eur J Clin Pharmacol 2019;75:1219–26. https://doi.org/10.1007/s00228-019-02694-1.Search in Google Scholar PubMed
25. Sime, FB, Byrne, CJ, Parker, S, Stuart, J, Butler, J, Starr, T, et al.. Population pharmacokinetics of total and unbound concentrations of intravenous posaconazole in adult critically ill patients. Crit Care 2019;23:205. https://doi.org/10.1186/s13054-019-2483-9.Search in Google Scholar
26. Ezquer-Garin, C, Ferriols-Lisart, R, Alos-Alminana, M, Aguilar-Aguilar, G, Belda-Nacher, JF, Carbonell, JA. Validated HPLG-UV detection method for the simultaneous determination of ceftolozane and tazobactam in human plasma. Bioanalysis 2018;10:461–73. https://doi.org/10.4155/bio-2017-0257.Search in Google Scholar
27. Rigo-Bonnin, R, Gomez-Junyent, J, Garcia-Tejada, L, Benavent, E, Soldevila, L, Tubau, F, et al.. Measurement of ceftolozane and tazobactam concentrations in plasma by UHPLC-MS/MS. Clinical application in the management of difficult-to-treat osteoarticular infections. Clin Chim Acta 2019;488:50–60. https://doi.org/10.1016/j.cca.2018.10.034.Search in Google Scholar
28. Sutherland, CA, Ozbal, C, Nicolau, DP. Development of an HPLC-MS/MS method for the determination of ceftolozane/tazobactam in bronchoalveolar lavage fluid. Future Sci 2019;5:FSO352. https://doi.org/10.4155/fsoa-2018-0079.Search in Google Scholar
29. U.S. Food and Drug Administration. Bioanalytical method validation: guidance for industry. Rockville, MD: U.S. Department of Health and Human Services; 2018.Search in Google Scholar
30. 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. https://doi.org/10.1021/ac020361s.Search in Google Scholar
31. Sofronescu, AG. Cerebrospinal fluid analysis; 2015. Available from: https://emedicine.medscape.com/article/2093316-overview [Accessed 5 Oct 2019].Search in Google Scholar
32. Venkatesh, B, Scott, P, Ziegenfuss, M. Cerebrospinal fluid in critical illness. Crit Care Resuscitation 2000:42–54.Search in Google Scholar
34. Liao, J, Sheng, HM, Sauri, J, Xiang, R, Martin, G. Structural elucidation of a dimeric impurity in the process development of ceftolozane using LC/HRMS and 2D-NMR. J Pharmaceut Biomed Anal 2019;174:242–7. https://doi.org/10.1016/j.jpba.2019.05.057.Search in Google Scholar
35. Naicker, S, Valero, YCG, Meija, JLO, Lipman, J, Roberts, JA, Wallis, SC, et al.. A UHPLC-MS/MS method for the simultaneous determination of piperacillin and tazobactam in plasma (total and unbound), urine and renal replacement therapy-effluent. J Pharmaceut Biomed Anal 2018;148:324–33. https://doi.org/10.1016/j.jpba.2017.10.023.Search in Google Scholar
36. Rabbolini, S, Verardo, E, Da Col, M, Gioacchini, AM, Traldi, P. Negative ion electrospray ionization tandem mass spectrometry in the structural characterization of penicillins. Rapid Commun Mass Spectrom 1998;12:1820–6. https://doi.org/10.1002/(sici)1097-0231(19981130)12:22<1820::aid-rcm387>3.0.co;2-c.10.1002/(SICI)1097-0231(19981130)12:22<1820::AID-RCM387>3.0.CO;2-CSearch in Google Scholar
37. Sime, FB, Lassig-Smith, M, Starr, T, Stuart, J, Pandey, SL, Parker, SL, et al.. A population pharmacokinetic model-guided evaluation of ceftolozane/tazobactam dosing in critically ill patients undergoing continuous venovenous hemodiafiltration. Antimicrob Agents Chemother 2019;64:e01655–19.10.1128/AAC.01655-19Search in Google Scholar
38. Sime, FB, Lassig-Smith, M, Starr, T, Stuart, J, Pandey, S, Parker, SL, et al.. Cerebrospinal fluid penetration of ceftolozane/tazobactam in critically ill patients with an indwelling external ventricular drain. Antimicrob Agents Chemother 2020. https://doi.org/10.1128/aac.01698-20.Search in Google Scholar
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