高效液相色谱法测定人血浆中霉酚酸及其代谢产物的浓度  

HPLC determination of mycophenolic acid and its metabolite in human plasma

作　　者：谢志红[1] 叶丽卡[1] 韩珂[1] 

机构地区：[1]广州医科大学附属第二医院,广州510260

出　　处：《中南药学》2015年第4期354-357,共4页Central South Pharmacy

基　　金：广东省科技计划项目(No.2010B031600295);国家自然科学基金资助(No.81202600)

摘　　要：目的建立高效液相色谱法建立测定人血浆中霉酚酸（MPA）、霉酚酸葡萄糖苷（MPAG）、霉酚酸酰基葡萄糖苷（Ac MPAG）的浓度。方法血浆处理采用乙腈蛋白沉淀法。色谱柱为Gemini C18（250mm×4.6 mm,5μm）,柱温为40℃。Ac MPAG、MPA采用荧光检测,流动相：甲醇-10 mmol·L^-1KH2PO4（43：57,p H=8.0）,荧光激发波长343 nm,发射波长425 nm。MPAG采用紫外检测,流动相：甲醇-10 mmol·L^-1 KH2PO4（45：55,p H=3.0）,检测波长254 nm。结果 MPA、MPAG、Ac MPAG血药浓度分别在0.2-20μg·m L^-1、4.75-475μg·m L^-1、0.02-2μg·m L^-1内与峰面积线性关系良好。MPA、MPAG、Ac MPAG的相对回收率分别为88.1%-104.3%、95.7%-99.6%、94.2%-96.0%。日内及日间RSD均〈10%。结论该方法测定人血浆中MPA、MPAG、Ac MPAG的浓度,操作简便,方法灵敏度高,适合用于MPA及其代谢产物的药动学研究及血药浓度监测。Objective To establish a method for the determination of mycophenolic acid（MPA）, mycophenolic acid glucuronide（MPAG） and mycophenolic acid acyl glucuronide（Ac MPAG） concentration in human plasma with HPLC. Methods The samples were precipitated with acetonitrile before injection. Gemini C18（250 mm×4.6 mm,5 μm） column was used and the flow rate was 1 m L·min^- 1. The column temperature was 40 ℃. MPA and Ac MPAG were determined by fluorescence detection with λex 343 nm and λem 425 nm. The mobile phase consisted of a mixture of methanol-10 mmol·L^- 1 KH2PO4（43 ：57,p H = 8.0）. MPAG was determined by UV detection and the detection wavelength was 254 nm. The mobile phase consisted of a mixture of methanol-10 mmol·L^- 1 KH2PO4（45 ：55,p H = 3.0）. Results The assay was linear within 0.2- 20 μg·m L- 1 for MPA, 4.75- 475 μg·m L^- 1 for MPAG, and 0.02- 2 μg·m L^- 1 for Ac MPAG. The average recoveries of MPA, MPAG and Ac MPAG were 88.1%- 104.3%, 95.7%- 99.6% and 94.2%- 96.0%,respectively. The intra-day and inter-day were both lower than 10%. Conclusion The method is simple and sensitive,and is suitable for the pharmacokinetic study and therapeutic drug monitoring of MPA,MPAG and Ac MPAG.

关 键 词：霉酚酸 霉酚酸葡萄糖苷 霉酚酸酰基葡萄糖苷 高效液相色谱法 血药浓度 

分 类 号：R969.1[医药卫生—药理学]