Ferrone Vincenzo, Carlucci Maura, Cotellese Roberto, Raimondi Paolo, Cichella Annadomenica, Di Marco Lorenzo, Genovese Salvatore, Carlucci Giuseppe
Department of Pharmacy, Università degli Studi, G. d'Annunzio' Chieti-Pescara, via dei Vestini, 66100, Chieti, Italy.
Department of Oral Health Sciences and Biotechnology, Università degli Studi, G. d'Annunzio' Chieti-Pescara, via dei Vestini, 66100, Chieti, Italy.
Drug Test Anal. 2017 Oct;9(10):1611-1619. doi: 10.1002/dta.2195. Epub 2017 May 12.
This study developed a high performance liquid chromatography (HPLC) method involving dried blood spotting (DBS) as a sampling method for the therapeutic drug monitoring of antimicrobial combination therapy with linezolid and ciprofloxacin. DBS for standards, quality control samples, and patient samples was excised and then extracted using a mixture of methanol/water/formic acid 80:20:0.1 (v/v/v), respectively. Linezolid (LZD) and ciprofloxacin (CPR) were measured by HPLC using a Kinetex EVO C (100 × 4.6 mm I.D. 2.6 μm particle size). Mobile phase consisted of 10 mM ammonium acetate (A) and a mixture of acetonitrile and methanol (B), both containing 0.1% triethylamine, in gradient elution. Detection was carried out at 251 nm for linezolid (LZD) and 277 for ciprofloxacin (CPR). Ulifloxacin was used as an internal standard. The internal standard, LZD, and CPR were eluted in 7.90, 7.18, and 8.89 min, respectively. Total run-time was 20 min. Calibration curves were constructed in the range of 0.05-30 μg/mL for LZD and 0.02-10 μg/mL for CPR, respectively. The intra- and inter-day precision (RSD values) did not exceed 9.43%, the intra- and inter-day accuracy (accuracy %) ranged between 96.2 and 106.2%. Haematocrit (Hct) effects were investigated to obtain a linear correlation between haematocrit values and volume of blood. Copyright © 2017 John Wiley & Sons, Ltd.
本研究开发了一种高效液相色谱（HPLC）方法，该方法采用干血斑（DBS）作为采样方法，用于监测利奈唑胺和环丙沙星联合抗菌治疗的治疗药物。分别将标准品、质量控制样品和患者样品的干血斑剪下，然后用甲醇/水/甲酸80:20:0.1（v/v/v）的混合物进行提取。利奈唑胺（LZD）和环丙沙星（CPR）采用HPLC法测定，使用Kinetex EVO C（内径100×4.6 mm，粒径2.6μm）色谱柱。流动相由10 mM醋酸铵（A）和乙腈与甲醇的混合物（B）组成，两者均含有0.1%三乙胺，采用梯度洗脱。利奈唑胺（LZD）在251 nm波长处检测，环丙沙星（CPR）在277 nm波长处检测。乌利沙星用作内标。内标、LZD和CPR的洗脱时间分别为7.90、7.18和8.89分钟。总运行时间为20分钟。分别在0.05 - 30μg/mL范围内构建LZD的校准曲线，在0.02 - 10μg/mL范围内构建CPR的校准曲线。日内和日间精密度（RSD值）不超过9.43%，日内和日间准确度（准确度%）在96.2%至106.2%之间。研究了血细胞比容（Hct）的影响，以获得血细胞比容值与血量之间的线性相关性。版权所有© 2017约翰威立父子有限公司。
