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和伊曲康唑含量(英文)

A sensitive HPLC method for determination of amphotericin B and itraconazole in cerebrospinal fluid and its application to patients with cryptococcal meningitis

目的建立人脑脊液(cerebrospinal fluid,CSF)中两性霉素B(amphotericin B,AmB)和伊曲康唑(itraconazole,ITZ)含量测定的高效液相色谱方法,并用于隐球菌性脑膜炎患者脑脊液样品的测定。方法脑脊液样品经固相萃取后直接进样。AmB测定采用Gemini 5w C18柱,流动相为5 mmol/L乙二胺四乙酸二钠(醋酸调pH至4.5)/乙腈(62/38,V/V),流速0.3 mL/min,DAD检测器:检测波长410 nm。ITZ测定采用YMC C18柱,流动相为0.2%三氟乙酸/乙腈(45/55,V/V),流速1.1 mL/min,荧光检测器:激发波长260 nm,发射波长365 nm。结果AmB和ITZ的线性范围均为5 ng/mL^100 ng/mL,线性相关系数大于0.999,最低定量限为5 ng/mL,低、中、高浓度质控样品在日内、日间精密度均小于7.6%。结论该方法灵敏、快速、简便,适用于隐球菌性脑膜炎患者治疗药物监测。

Objective To establish a sensitive, rapid and simple high-performance liquid chromatography （HPLC） method for the determination of amphotericin B （AmB） and itraconazole （ITZ） in human cerebrospinal fluid （CSF）. Methods Solid-phase extraction （SPE） was involved in sample preparation in this method, followed by reverse-phase separation under two different chromatographic systems. Results The linearity for both Arab and ITZ ranged from 5 ng/mL to 100 ng/mL and the inter-day and intra-day RSD were both below 7.6% at the concentrations of 20,50 and 100 ng/mL. Conclusions The method we established has been applied to the therapeutic drug monitoring on cryptococcal-meningitis-infected patients who were given comedication of these two drugs.