以对羟基苯甲酸甲酯为内标用HPLC.docVIP

以对羟基苯甲酸甲酯为内标用HPLC 作者:孟玲，王永庆，李娜，贾楠，陈玲 【摘要】 目的 建立用内标法测定人血浆中氟康唑浓度的HPLC-UV方法。方法 以对羟基苯甲酸甲酯为内标，血浆用氨水碱化后，用乙酸乙酯提取，测定血浆中氟康唑的浓度。结果 血浆中杂质不干扰样品峰和内标峰，、、μg/ml的回收率分别为%，%及%；批内精密度均小于8%，批间精密度均小于10%，最低检测限为μg/ml，线性范围为～μg/ml；血浆中氟康唑冻融是稳定的；精密度也符合要求。结论 该方法稳定且精密度好，准确可靠，适用于氟康唑血浆浓度测定。 【关键词】 氟康唑；对羟基苯甲酸甲酯；高效液相色谱-紫外；内标 Determination the human plasma content of fluconazole using the methyl parahy-droxybenzoate as a internal standard by HPLC-UV method 【Abstract】 Objective To establish the determination method of fluconazole in healthy volunteers plasma. Methods The plasma was alkalified by ammonium hydroxide， and fluconazole was extracted by acetic ether； and methyl parahydroxybenzoate was used as internal standard. Plasma concentrations of fluconazole and internal standard were determined by HPLC -UV. Results The chromatographic peak of fluconazole and internal standard was not interfered by foreign matter. Recovery rate of 、、μg/ml was %，% and %， respectively. Precision in or between batch was less than % and %， respectively. Lowest detectable limit was μg/ml and linear range was ～μg/ml. The freeze thawing of plasma was stable. Conclusion The method was reliable and stable， it was suitable for determination the concentration of fl{http://以对羟基苯甲酸甲酯为内标用HPLC-UV法测定人血浆中氟康唑的含量欢迎您访问范,文,家}uconazole in plasma. 【Key words】 fluconazole； methyl parahydroxybenzoate； HPLC-UV； internal standard 氟康唑是新型抗真菌药物，由于其穿透力强，分布容积大，血清消除半衰期长，故具有长效特点，可用于多种组织器官深部真菌感染的治疗，具有一定的临床价值。文献报道的检测方法有微生物法、GC法和高效液相法。高效液相法是常用的血浆氟康唑的检测方法，有外标法和内标法，常用内标有非那西丁［1］、美托洛尔［2］、氨苯蝶啶［3］和MHD［4］。本文采用HPLC-UV法，以对羟基苯甲酸甲酯为内标，测定氟康唑血药浓度。 1 实验材料 试验药品 氟康唑标准品： 中国药品生物制品检定所提供，用超纯水配制成相当于/ml 贮备液。临用时稀释至相应的浓度。对羟基苯甲酸甲酯江苏省药品检验所提供，用甲醇配制成相当于μg/ml 贮备液。 试剂 乙腈， Fisher Company， Inc，USA； 其余试剂均为市售分析纯。 仪器 导津高效液相色谱仪；Milli-Q Gradient A10超纯水器。 2 血浆中氟康唑的HPLC法测定 色谱条件 流动相由/L KH2PO4和乙腈组成，流速 ml/min；色谱柱为Alltima C18 μm 250mm× ；柱温箱温度 30℃；检测波长261 nm。 血浆样品的提取与分离 精密吸取血浆置15 ml玻璃离心管中，加入10μl对羟基苯甲