UQ Centre for Clinical Research, Faculty of Medicine, The University of Queensland, Brisbane, Australia.
Centre for Translational Anti-Infective Pharmacodynamics, School of Pharmacy, The University of Queensland, Brisbane, Australia.
Clin Chem Lab Med. 2020 Nov 25;59(5):921-933. doi: 10.1515/cclm-2020-1196. Print 2021 Apr 27.
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.
世界卫生组织确定了一些碳青霉烯类耐药革兰氏阴性病原体的新型治疗选择方案，这些方案被列为最高优先级。头孢洛扎他唑巴坦是一种新型头孢菌素-内酰胺酶抑制剂复合抗生素，对最难治疗的多药耐药和广泛耐药革兰氏阴性病原体具有强大的杀菌活性。本研究旨在开发和验证一种同时定量测定血浆（总蛋白和未结合蛋白）、肾替代治疗流出液（RRTE）、脑脊液（CSF）和尿液中头孢洛扎他唑巴坦的液相色谱-串联质谱法。
采用混合模式色谱法，使用内源性碱去活的 C18 柱和由 0.1%甲酸、10 mM 甲酸铵和乙腈组成的梯度流动相分离分析物。通过正、负模式之间快速离子化切换，同时进行选择反应监测，检测分析物和内标。
头孢洛扎他唑巴坦在血浆（总蛋白和未结合蛋白）、RRTE 和 CSF 中的浓度范围为 1-200mg/L，tazobactam 的浓度范围为 0.5-100mg/L，在尿液中的浓度范围为 10-2000mg/L，头孢洛扎他唑巴坦和 tazobactam 均获得了二次校准。对于基质效应、精密度和准确度、特异性和稳定性的验证测试，头孢洛扎他唑巴坦和 tazobactam 的所有结果均在±15%的可接受范围内。
该方法学成功应用于一项感染性危重症患者的药代动力学研究，包括接受肾替代治疗的患者，以及一例脑室炎患者的病例研究，所有患者均接受了头孢洛扎他唑巴坦治疗。
