Laboratoire de Pharmacocinétique et Toxicologie, IFB, Hôpital Purpan, 330 Avenue de Grande-Bretagne, 31059 Toulouse Cedex 9, France; Laboratoire Départemental 31, Eau - Vétérinaire - Air, 76 chemin de Boudou, CS 50013, 31140 Launaguet, France.
Laboratoire de Pharmacocinétique et Toxicologie, IFB, Hôpital Purpan, 330 Avenue de Grande-Bretagne, 31059 Toulouse Cedex 9, France.
J Pharm Biomed Anal. 2022 Sep 20;219:114900. doi: 10.1016/j.jpba.2022.114900. Epub 2022 Jun 19.
Antibiotic (ATB) prescription in an intensive care unit (ICU) requires continuous monitoring of serum dosages due to the patient's pathophysiological condition. Dosing adjustment is necessary to achieve effective targeted concentrations. Since ICUs routinely use a large number of ATBs, global monitoring needs to be developed. In the present study, we developed a global analytical method for extracting, separating and quantifying the most widely used ATBs in ICUs: amoxicillin, piperacillin, cefazolin, cefepime, cefotaxime, ceftazidime, ceftolozane, ceftriaxone, ertapenem, meropenem, ciprofloxacin, moxifloxacin, levofloxacin, daptomycin, dalbavancin, linezolid and a beta-lactamase inhibitor: tazobactam. To guarantee the robustness of the quantification, we differentiated the 16 ATBs and the beta lactamase inhibitor into 4 pools (ATB1 to ATB4), taking into account prescription frequency in the ICU, the physicochemical properties and the calibration ranges of the ATBs selected. The whole ATB was then separated with two LC columns in reversed phase: Kinetex Polar-C18 100 Å and Polar-RP-80 synergy, in less than 6.5 min. Detection was carried out by electrospray in positive ion mode, by tandem mass spectrometry (LC-MS/MS. The four quantification methods were validated according to the European guidelines on bioanalytical method validation (EMEA guide), after determining the extraction yields, matrix effects, recovery, precision, accuracy, within-run precision and between-run precision. For all analyses, bias is < 15% and is comparable to the literature and LOQs vary from 0.05 mg.L for ciprofloxacin to 1.00 mg.L for ceftriaxone and dalbavancin. The stability time of cefepime and piperacillin is 3 hrs and for the other ATBs 6 hrs in serum at room temperature. For long-term stability, freezing at - 80 °C guarantees 3 months of stability for ceftriaxone and dalbavancin and more than 6 months for the other ATBs.
在重症监护病房（ICU）中，由于患者的病理生理状况，抗生素（ATB）的处方需要持续监测血清剂量。为了达到有效的靶向浓度，需要进行剂量调整。由于 ICU 通常使用大量的 ATB，因此需要开发全球监测。在本研究中，我们开发了一种提取、分离和定量 ICU 中使用最广泛的 ATB 的全球分析方法：阿莫西林、哌拉西林、头孢唑林、头孢吡肟、头孢噻肟、头孢他啶、头孢唑肟、头孢曲松、厄他培南、美罗培南、环丙沙星、莫西沙星、左氧氟沙星、达托霉素、达巴万星、利奈唑胺和一种β-内酰胺酶抑制剂：他唑巴坦。为了保证定量的稳健性，我们将 16 种 ATB 和β-内酰胺酶抑制剂分为 4 个池（ATB1 至 ATB4），考虑到 ICU 中的处方频率、所选 ATB 的物理化学性质和校准范围。然后，整个 ATB 用两种反相 LC 柱分离：Kinetex Polar-C18 100Å 和 Polar-RP-80 协同作用，不到 6.5 分钟。检测采用电喷雾正离子模式，通过串联质谱（LC-MS/MS）进行。根据欧洲生物分析方法验证指南（EMEA 指南），对四种定量方法进行了验证，确定了提取率、基质效应、回收率、精密度、准确度、日内精密度和日间精密度。对于所有分析，偏差均<15%，与文献相比可比较，LOQ 从环丙沙星的 0.05mg.L 到头孢曲松和达巴万星的 1.00mg.L 不等。头孢吡肟和哌拉西林的稳定性时间为 3 小时，其他 ATB 的稳定性时间为 6 小时，室温下血清中。对于长期稳定性，-80°C 冷冻可保证头孢曲松和达巴万星 3 个月的稳定性，其他 ATB 超过 6 个月的稳定性。
