Utilizing tigecycline-d9 as an internal standard (IS), we establish and validate a simple, effective, and rapid liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the quantitative measurement of tigecycline (TGC) in patient plasma. Acetonitrile was used as a precipitant to process plasma samples by a protein precipitation method. The analyte and IS were separated on an HSS T3 (2.1 × 100 mm, 3.5 μm) chromatographic column using isocratic program with a mobile phase comprising of 80% solvent A (water containing 0.1% formic acid (v/v) with 5 mM ammonium acetate) and 20% solvent B (acetonitrile) with a flow rate of 0.3 mL/min. The mass spectrometer, scanning in multireaction monitoring (MRM) mode and using an electrospray ion source (ESI), operated in the positive-ion mode. The ion pairs used for quantitative analysis were m/z 586.4 ⟶ 513.3 and m/z 595.5 ⟶ 514.3 for TGC and the IS, respectively. The range of the linear calibration curve obtained with this approach was 50–5000 ng/ml. Intra- and interbatch precision for TGC quantitation were less than 7.2%, and the accuracy ranged from 93.4 to 101.8%. The IS-normalized matrix effect was 87 to 104%. Due to its high precision and accuracy, this novel method allows for fast quantitation of TGC with a total analysis time of 2 min. This approach was effectively applied to study the pharmacokinetics of TGC in critically ill adult patients.

中文翻译：

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液相色谱-串联质谱法测定重症患者替加环素的建立与验证

我们以tigecycline-d9作为内标（IS），建立并验证了一种简单，有效，快速的液相色谱-串联质谱（LC-MS / MS）方法，用于定量测量患者血浆中的tigecycline（TGC）。乙腈用作沉淀剂，可通过蛋白质沉淀法处理血浆样品。分析物和IS被上的HSS T3分离（2.1×100mm的，3.5  μm）使用等度程序的色谱柱，其流动相包含80％的溶剂A（含0.1％甲酸（v / v）和5 mM乙酸铵的水）和20％的溶剂B（乙腈），流速为0.3 mL /分钟。质谱仪以正离子模式运行，在多反应监测（MRM）模式下扫描并使用电喷雾离子源（ESI）。用于定量分析的离子对为m / z 586.4⟶513.3和m / z595.5⟶514.3分别用于TGC和IS。用这种方法获得的线性校准曲线的范围是50-5000 ng / ml。TGC定量的批内和批间准确度均小于7.2％，准确度范围为93.4至101.8％。IS归一化矩阵效应为87％至104％。由于其高精度和准确性，这种新颖的方法可以快速定量TGC，总分析时间为2分钟。该方法有效地用于研究重症成年患者中TGC的药代动力学。