KU Leuven, Department of Pharmaceutical and Pharmacological Sciences, Pharmaceutical Analysis, Herestraat 49, O&N2, PB 923, 3000 Leuven, Belgium.
Hospital Pharmacy Department, University Hospitals Leuven, Herestraat 49, 3000 Leuven, Belgium.
J Pharm Biomed Anal. 2023 May 10;228:115319. doi: 10.1016/j.jpba.2023.115319. Epub 2023 Feb 26.
A sensitive, accurate and precise liquid chromatography (LC) method for the simultaneous determination of ceftazidime and pyridine in human plasma has been developed and validated. Acetonitrile (ACN) was employed to precipitate the proteins in the plasma samples. Chromatographic separation was performed with a Kinetex® C18 (150 mm × 3 mm, 2.6 µm) column with gradient elution. Ammonium formate 20 mM and ACN were mixed in a ratio of 98:2 (v/v) for mobile phase A and 85:15 (v/v) for mobile phase B. Both were adjusted to pH 4.5 with formic acid. The flow rate was 0.4 mL/min. UV detection was performed at 254 nm. Calibration curves were linear in the range from 0.3 to 225 μg/mL for ceftazidime and from 0.2 to 10 μg/mL for pyridine with correlation coefficients ≥ 0.999. Within- and between-run precision and accuracy were satisfactory with coefficients of variation (CV) ≤ 8.0% and deviations ≤ 7.0%, respectively. The method fulfilled all validation criteria prescribed by the European Medicines Agency guidelines. Next, it has been used successfully to analyze plasma samples of patients who received ceftazidime under intermittent and continuous administration. With intermittent administration, the concentration of the antibiotics reached a peak and then dropped quickly, which may be below the minimal inhibitory concentration (MIC). With continuous administration, the concentration of the antibiotics remained stable over 24 h, certainly above the MIC. Although the same tendency in ceftazidime concentration changes over time was observed, a difference in concentration amongst the patients was noticeable. The concentration of pyridine in plasma was negligible.
建立并验证了一种灵敏、准确、精密的同时测定人血浆中头孢他啶和吡啶的液相色谱法。采用乙腈沉淀血浆样品中的蛋白质。色谱分离采用 Kinetex® C18（150mm×3mm，2.6μm）柱，以梯度洗脱方式进行。流动相 A 为 20mM 甲酸铵和乙腈（98:2，v/v），流动相 B 为 85:15（v/v），均用甲酸调至 pH 4.5。流速为 0.4mL/min。紫外检测波长为 254nm。头孢他啶在 0.3-225μg/mL 范围内、吡啶在 0.2-10μg/mL 范围内线性良好，相关系数均≥0.999。日内和日间精密度和准确度良好，变异系数（CV）≤8.0%，偏差≤7.0%。该方法符合欧洲药品管理局指南规定的所有验证标准。随后，成功地用于分析接受间歇性和连续性头孢他啶给药的患者的血浆样本。间歇性给药时，抗生素浓度迅速达到峰值，然后迅速下降，可能低于最低抑菌浓度（MIC）。连续性给药时，抗生素浓度在 24 小时内保持稳定，肯定高于 MIC。尽管观察到抗生素浓度随时间的变化趋势相同，但患者间的浓度存在差异。血浆中吡啶的浓度可以忽略不计。
