全自动二维液相色谱法测定腹腔感染患者血清中替加环素的浓度

目的：建立全自动二维色谱法（2D-LC-UV）测定血清中替加环素浓度，用于腹腔感染患者替加环素药物浓度的监测。方法：150μL样品进样，在一维柱ASTON C8（100 mm×4.6 mm，5μL）上初步分离，通过中间柱ASTON SCX（20 mm×4.6 mm，5μm）截取保留并在二维柱Agilent Extent-C18（150 mm×4.6 mm，5μm）上进一步分离，直至最终测定。一维柱流动相为30 mmol·L-1磷酸二氢钠-乙腈-甲醇（40∶30∶30，pH=5.4）；中间柱流动相为纯水；二维柱流动相为30 mmol·L-1磷酸二氢钠-乙腈（75∶25，pH=3.5），流速均为1.0 mL·min-1，柱温40℃，检测波长244 nm。采集40名腹腔感染患者静滴替加环素（首剂量100 mg，维持剂量50 mg q12h）血清样本，测定峰浓度、中浓度、谷浓度，并估算AUC值。结果：本色谱条件下，替加环素与各杂质分离良好，线性范围0.03~30μg·mL-1，最低检测限10 ng·mL-1。40名腹腔感染患者平均血清峰浓度（1.60±0.77）μg·mL-1，中浓度（0.84±0.36）μg·mL-1，谷浓度（0.57±0.25）μg·mL-1，估算平均AUC值（22.66±9.77）μg·h·mL-1。结论：此法操作简单，准确、精密度好，适用于替加环素浓度-药效研究及临床监测。

Establishment of a 2D-LC-UV Method for the Determination of tigecycline in Human serum and its Clinical Application

Objective: In this study, tigecycline, a new glycylcycline antimicrobial agent was quantified by a two-dimensional liquid chromatography coupled with ultraviolet detection (2D-LC-UV). Methods: In this setup, two reversed phase liquid chromatography columns, ASTON C8 (100 mm×4.6 mm, 5μm) and Agi-lent Extent-C18 (150 mm×4.6 mm, 5μm), were used as the first and second separation dimensions, respectively. An ASTON SCX (20 mm×4.6 mm, 5μm) column was used as the connector to transfer the eluent from the first dimensional column to the second one for further separation. The mobile phases of the two dimensions consisted of 30 mmol·L-1-acetonitrile-methanol NaH2PO4 (40∶30∶30, v/v, pH=5.4) and acetonitrile-30 mmol·L-1 NaH2PO4 (75∶25, v/v, pH=3.5), respectively. The flow rates were both 1 mL·min-1 and the detection wave-length was 244 nm. Serum peak, middle and trough concentration samples from 40 patients with intra-ab-dominal infections administrated with tigecycline (50 mg q12h) were analyzed and values of area under curve (AUC) were estimated. Results: The calibration curves were linear over the range of 0.03-30μg·mL-1. The average serum peak, middle and trough concentrations were (1.60±0.77)μg·mL-1, (0.84±0.36)μg·mL-1 and (0.57±0.25)μg·mL-1, respectively. And the estimated AUC values were (22.66±9.77)μg·h·mL-1. Con-clusions: This method can be used effectively in clinic and research fields.