*Unit of Infectious Diseases, Department of Medical Sciences, University of Turin, Amedeo di Savoia Hospital and †Pharmacy Department, Amedeo di Savoia Hospital, Turin, Italy.
Ther Drug Monit. 2013 Dec;35(6):853-8. doi: 10.1097/FTD.0b013e31829403b1.
: A simple ultra performance liquid chromatography with photodiode array method for the quantification of human plasma concentrations of tigecycline was developed and validated. Quinaxoline, used as an internal standard, was added to 500 μL of plasma before adding 1 mL of protein precipitation solution. The extracts were dried in a vacuum centrifuge system at 60°C and reconstituted with 60 μL of water and acetonitrile (95:5, vol/vol), and 5 μL was injected onto an ACQUITY UPLC H-Class system. Chromatographic separation was performed on a C18 ACQUITY UPLC HSS T3 column using a gradient of potassium phosphate buffer (pH 3.2) and acetonitrile. Detection was performed using a photodiode array detector at 350 nm. Relative error at 3 quality control concentrations ranged from -2.49% to -8.74%. Intraday and interday (percent relative standard error) precision ranged from 3.93% to 12.27% and from 9.53% to 13.32%, respectively. Limit of quantification and limit of detection were 0.024 and 0.006 μg/mL, respectively. Mean recovery was 95%. The calibration curve was linear up to 6 μg/mL. This concentration range proved to be adequate to measure tigecycline concentrations in patients treated with the drug, therefore this method would be suitable for therapeutic drug monitoring.
开发并验证了一种简单的超高效液相色谱-光电二极管阵列法，用于定量人血浆中替加环素的浓度。喹诺酮啉作为内标，在加入 1 mL 蛋白沉淀溶液前加入 500 μL 血浆。提取物在 60°C 下的真空离心系统中干燥，并用 60 μL 水和乙腈（95:5，体积/体积）重新溶解，然后取 5 μL 注入 ACQUITY UPLC H-Class 系统。色谱分离在 C18 ACQUITY UPLC HSS T3 柱上进行，使用磷酸钾缓冲液（pH 3.2）和乙腈的梯度洗脱。使用光电二极管阵列检测器在 350nm 处进行检测。在 3 个质控浓度下的相对误差范围为-2.49%至-8.74%。日内和日间（相对标准误差的百分比）精密度分别为 3.93%至 12.27%和 9.53%至 13.32%。定量下限和检测下限分别为 0.024 和 0.006μg/mL。平均回收率为 95%。校准曲线在 6μg/mL 内呈线性。该浓度范围足以测量接受替加环素治疗的患者的替加环素浓度，因此该方法适用于治疗药物监测。
