HPLC法同时测定人血浆中霉酚酸与霉酚酸葡糖苷酸的浓度

目的:建立以高效液相色谱法同时测定人血浆中霉酚酸(MPA)与霉酚酸葡糖苷酸(MPAG)浓度的方法。方法:色谱柱为LunaC18,流动相为(20mmol·L-1磷酸二氢钾+40mmol·L-1四丁基溴化铵(pH5·5))-乙腈=73:27,流速为1·6mL·min-1,MPA与MPAG采用二极管阵列检测器和荧光检测器串联检测,MPAG检测波长为250nm,MPA荧光检测波长为316nm(激发波长)、430nm(发射波长),柱温为38℃,其中血样加乙腈沉淀蛋白后进样10μL。结果:MPA、MPAG检测浓度分别在0·2～20(r=0·9993)、1～100(r=0·9999)μg·mL-1范围内与峰面积积分值呈良好线性关系,最低检测限分别为0·05、0·1μg·mL-1;平均回收率分别为95·54%、100·22%;MPA日内、日间RSD均<6%,MPAG日内、日间RSD均<4%。结论:本方法快捷、简便、灵敏,可用于测定人血浆中MPA与MPAG的浓度。

Determination of Mycophenolic Acid and Glucuronide in Human Plasma by HPLC

OBJECTIVE:To determine the contents of mycophenolic acid(MPA) and its glucuronide(MPAG) in human plasma by HPLC.METHODS:Chromatographic separation was performed on Luna C18 column.The mobile phase consisted of(20mmol· L-1 KH2PO4+ 40mmol· L-1 TBA,pH5.5)-acetonitrile=73:27 at a flow rate of 1.6mL· min-1.MPA acid and MPAG were detected by photodiode array detector and fluorescence detector in tandem.The detection wavelength of MPAG was 250nm;the excitation wavelength and emission wavelength of MPA were 316nm and 430nm,respectively.The column tem-perature was 38℃ and the sample size was 10μ L.The plasma sample was added with acetonitrile for protein precipitation.RE-SULTS:The linear ranges of MPA and MPAG were 0.2～ 20μg· mL-1(r=0.999 3) and 1～ 100μg· mL-1(r=0.999 9),with lowest detectable limit at 0.05μg· mL-1 and 0.1μg· mL-1,respectively.The average recoveries were 95.54% and 100.22%,respectively.The intra-day and inter-day RSD of MPA were less than 6%,and those of MPAG were less than 4%.CONCL-USION:The method is rapid,simple and sensitive,and suitable for the determination of MPA and MPAG in human plasma.