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(5 μm,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20 μL。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:1.25~30 μg·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%;0.11~5 μg·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。

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.

1 CHEN Jiang-han(陈江汉),WEN Hai(温海),WU Jian-hua(吴建华),et al.Adverse effects of amphotericin B during treatment of cryptococcal meningitis and their management(两性霉素B治疗隐球菌性脑膜脑炎的不良反应及其处理).Acad J Second Mil Med Univ(第二军医大学学报),2005, 26(4):456
2 CHEN Ming-quan(陈明泉),SHEN Guang-feng(施光峰),QIN Gang(秦刚),et al.Clinical analysis of 146 patients with cryptococcal meningitis(隐球菌脑膜炎146例临床分析).Chin J Infect Dis(中华传染病杂志),2007,25(4):199
3 LV Tian-ming(吕田明),YIN Jia(尹恝),LIU Xiao-jia(刘晓加),et al.Intrathecal continuous infusion of amphotericin B treatment of cryptococcal meningitis (with the first case report).J South Med Univ(南方医科大学学报),2007,27(9):1461,1464
4 De Socio GV,Fiorio M,Stagni G.AmBisome administration for Candida albicans shunt infections.J Antimicrob Chemother,2003,52(6):1048
5 DENG Bin(邓斌),REN Bin(任斌),LI Shu-xia(黎曙霞),et al.Determination of amphotericin B in human plasma by HPLC(高效液相色谱法测定人血浆中两性霉素B浓度).Chin J Mod Appl Pharm(中国现代应用药学),2003,20(6):481
6 CHEN Hua-hong(陈画虹),LUO Ke(骆科).Determination of amphotericin B in human plasma by RP-HPLC(反相高效液相色谱法测定人血浆中两性霉素B浓度).China Pharm(中国药房),2005,16(11):842