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

Establishment of a 2D-LC-UV Method for the Determination of Tigecycline in Human Serum and Its Clinical Application

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,p H=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,p H=3.5),流速均为1.0 m L·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。

Objective： In this study, tigecycline, a new glycylcycline antimicrobial agent was quantifiedby a two-dimensional liquid chromatography coupled with ultraviolet detection （2D- LC -UV）. Methods： Inthis setup, two reversed phase liquid chromatography columns, ASTON C8 （100mm×4.6mm, 5 pm） and AgilentExtent-Ct8 （150mm×4.6mm, 5μm）, were used as the first and second separation dimensions, respectively.An ASTON SCX （20 mm×4.6 mm, 5 pm） column was used as the connector to transfer the eluent from thefirst dimensional column to the second one for further separation. The mobile phases of the two dimensionsconsisted of 3 0 mmol.L-1-acetonitrile-methanol NaH〗PO4 （40：30：30, v/v , pH=5.4） and acetonitrile- 3 0 mmol.L-1NaH2PO4 （75：25, v/v , pH=3.5）, respectively. The flow rates were both 1 mL-min-1 and the detection wavelengthwas 244 nm. Serum peak, middle and trough concentration samples from 40 patients with intra-abdominalinfections administrated with tigecycline （50 mg q12h） were analyzed and values of area undercurve （AUC） were estimated. Results： The calibration curves were linear over the range of 0.03-30 μg·mL_1.The average serum peak, middle and trough concentrations were （1.60±0.77） μg·m L-1, （0.84±0.36） μg·mL-1and （0.57±0.25）μg·mL-1, respectively. And the estimated AUC values were （22.66±9.77）μg·h -mL-1. Conclusions：This method can be used effectively in clinic and research fields.