Boccardi Davide, Marini Valeria, Baiardi Giammarco, Cameran Caviglia Michela, Sacco Fabio, Piras Fabio, Del Puente Filippo, Boni Silvia, Pontali Emanuele, Mattioli Francesca
Pharmacology and Toxicology Unit, Department of Internal Medicine, University of Genoa, Genoa, Italy; Clinical Pharmacology Unit, Ente Ospedaliero Ospedali Galliera, Genoa, Italy.
Pharmacology and Toxicology Unit, Department of Internal Medicine, University of Genoa, Genoa, Italy; Clinical Pharmacology Unit, Ente Ospedaliero Ospedali Galliera, Genoa, Italy.
Clin Chim Acta. 2024 Jul 15;561:119806. doi: 10.1016/j.cca.2024.119806. Epub 2024 Jun 7.
Ceftobiprole is a recent 5 generation parenteral cephalosporin with antibacterial activity against a large range Gram+ and Gram- bacteria. Therapeutic drug monitoring (TDM) is an essential tool for maintaining plasma concentrations of antibiotics above the MIC by the end of the dosing interval, thus preventing the resistant strain diffusion. TDM is already recommended for other cephalosporins, and it is a reasonable tool contributing to the safety and efficacy of these drugs. During the treatment of patients in real-life, a number of pharmacokinetic (PK) changes not normally seen in healthy volunteers can occur which can impair the pharmacokinetic/pharmacodynamic target attainment. We aimed to develop simple and rapid HPLC-UV method for determination of ceftobiprole in human serum to implement TDM in clinical practice and support PKs and pharmacokinetic/pharmacodynamic (PK/PD) studies.
Samples preparation of calibration standards, QC, and anonymous patients serum samples was performed by protein precipitation by adding 0.01 ml of sulphosalicylic acid at 30 % to 0.1 ml of each sample. Then samples were vortexed and the centrifuged at 12,000 rpm for 10 min at 4 °C. Fifty microlitres of clear supernatant were diluted 1:1 with mobile phase and transferred into HPLC autosampler held at 8 °C. Chromatographic separation was carried out in a gradient mode at 35 °C on an ultra-Biphenyl column using a Thermo Scientific chromatographic system with a Diode array. Data management was performed with Chromeleon 7.4 software.
The HPLC-UV method proved to be linear over wide concentration ranges (0.5-50.0 mg/L) and was accurate and reproducible in the absence of matrix effects, allowing for robust, specific, and rapid quantification of ceftobiprole from a low amount of serum (0.1 mL). The mean steady state C and C values measured in the anonymous patients' samples were 6.26 ± 3.81 mg/L and 22.56 ± 15.69 mg/L, respectively.
We report a broadened simple and fast HPLC with UV detection method for quantification of ceftobiprole in human serum to implement ceftobiprole TDM as clinical routine, and support future (PK/PD) studies in special patients' population.
头孢比普是一种新型的五代胃肠外头孢菌素，对多种革兰氏阳性菌和革兰氏阴性菌具有抗菌活性。治疗药物监测（TDM）是一种重要工具，可在给药间隔结束时将抗生素血浆浓度维持在最低抑菌浓度（MIC）以上，从而防止耐药菌株扩散。TDM已被推荐用于其他头孢菌素，是有助于这些药物安全性和有效性的合理工具。在现实生活中治疗患者时，可能会出现一些健康志愿者中通常未见的药代动力学（PK）变化，这可能会损害药代动力学/药效学目标的实现。我们旨在开发一种简单快速的HPLC-UV方法，用于测定人血清中的头孢比普，以便在临床实践中实施TDM，并支持PK和药代动力学/药效学（PK/PD）研究。
校准标准品、质量控制品和匿名患者血清样本的样品制备采用蛋白质沉淀法，向每个样本的0.1 ml中加入0.01 ml 30%的磺基水杨酸。然后将样品涡旋，在4℃下以12000 rpm离心10分钟。取50微升清亮上清液，用流动相1:1稀释，转移至8℃的HPLC自动进样器中。使用带二极管阵列的Thermo Scientific色谱系统，在35℃下于超联苯柱上进行梯度模式的色谱分离。使用Chromeleon 7.4软件进行数据管理。
HPLC-UV方法在宽浓度范围（0.5-50.0 mg/L）内呈线性，在无基质效应的情况下准确且可重复，能够从少量血清（0.1 mL）中对头孢比普进行可靠、特异且快速的定量。在匿名患者样本中测得的平均稳态Cmax和AUC值分别为6.26±3.81 mg/L和22.56±15.69 mg/L。
我们报告了一种扩展的简单快速的带紫外检测的HPLC方法，用于定量人血清中的头孢比普，以便将头孢比普TDM作为临床常规实施，并支持未来在特殊患者群体中的（PK/PD）研究。
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