反相高效液相色谱法测定隐球菌性脑膜炎患者脑脊液中两性霉素B的含量

目的:建立反相高效液相色谱法测定人脑脊液中两性霉素B,并用于隐球菌性脑膜炎患者脑脊液样品的测定。方法:采用Agilent C18(5μm,4.6 mm×250 mm)色谱柱,流动相为0.005 mol.L-1乙二胺四乙酸二钠-甲醇(21∶79),流速1.0 mL.min-1,检测波长380 nm,室温下进样20μL。结果:由于患者脑脊液中药物的含量测定范围较大,故采用2个浓度线性范围:1.25～30μg.mL-1(r=0.9999),高、中、低3种浓度的平均回收率为94.9%,日内RSD≤1.38%,日间RSD≤2.67%;0.11～5μg.mL-1(r=0.9999),高、中、低3种浓度的平均回收率为93.1%,日内RSD≤1.73%,日间RSD≤4.09%。结论:本法灵敏度高,重现性好,专属性强,可用于椎管内持续给予两性霉素B的临床药代动力学研究及脑脊液中该药物浓度的临床检测。

RP-HPLC determination of amphotericin B in cerebrospinal fluid of patients with cryptococcal meningitis

Objective: To establish an RP-HPLC method for the determination of amphotericin B in human cerebrospinal fluid(CSF) and apply the method to determination of CSF samples of patients with cryptococcal meningitis.Methods: The chromatographic separation was performed on Agilent C18(5 μm,4.6 mm × 250 mm) column.0.005 mol.L-1 EDTA-2Na-methanol(21∶ 79,v / v) was taken as the mobile phase with the flow rate at 1.0 mL.min-1,and the detection wavelength was set at 380 nm.The sample injection volume was 20 μL under room temperature.Results: Because of a wide measurement range of the drug content in patients’CSF,double linear concentration ranges were used: one was 1.25-30 μg.mL-1;Its average recovery rate for the 3(high,middle and low) concentrations was 94.9%,and the intra-day RSD and the inter-day RSD were separately less than 1.38% and 2.67%.The other was 0.11-5 μg.mL-1;Its average recovery rate for the 3(high,middle and low) concentrations was 93.1%,and the intra-day RSD and the inter-day RSD were separately less than 1.73% and 4.09%.Conclusions: The method is sensitive,reproducible and specific,which can be used for clinical pharmacokinetic study of continuous infusion of amphotericin B into the spinal canal and the clinical monitoring of the concentration of amphotericin B in CSF.