HPLC-荧光法测定肾移植患者血浆中霉酚酸的浓度

Determination of Plasma Concentration of Mycophenolic Acid in Renal Transplantation Patients by HPLC-Fluoremetry

目的:建立以高效液相色谱-荧光法测定肾移植患者血浆中霉酚酸浓度的方法。方法:样品用5%硫酸锌的甲醇饱和溶液沉淀蛋白后,取上清液进样20μL进行测定,色谱柱为Zorbax Eclipse XDBC18,流动相为乙腈-甲醇-0.2mol·L-1甘氨酸缓冲液(pH9.0)=18∶2∶80,柱温为25℃,流速为1.0mL·min-1,激发波长(EX)为342nm,发射波长(EM)为425nm。结果:霉酚酸血药浓度在0.5～40mg·L-1范围内线性关系良好,最低定量限为0.5mg·L-1;方法回收率为98.23%～101.00%,萃取回收率为91.56%～94.46%,日内、日间RSD分别为0.64%～3.22%和5.12%～6.10%。结论:本方法灵敏、快速、准确、方便,可用于霉酚酸的血药浓度测定。

OBJECTIVE： To determine the plasma concentration of mycophenolic acid （MPA） in renal transplantation patients by HPLC- Fluoremetry. METHODS： The sample was subjected to precipitation of proteins using 5% Zinc Sulfate methanol saturated solution, and the supernatant （20μL） was taken for sample injection and determination on Zorbax Eclipse XDB C18 with mobile phase consisted of acetonitrile - methanol - 0.2 mol·L^-1glycine buffer（18 ： 2 ： 80, pH = 9.0） at a flow rate of 1.0 mL·min^-1. The column temperature was of 25℃ ; the excitation wavelength（Ex） was 342 nm and the emission wavelength（EM） was 425 nm. RESULTS： The linear range of MPA was 0.5-40 mg·L^-1, with its lowest limit of quantita- tion at 0.5 mg·L^-1. The methodology recovery was 98.23% --101.00% ; the extraction recovery of MPA was 91.56%- 94.46%; the intraday RSD was 0.64%- 3.22% and the inter- day RSD was 5.12%--6.10%. CONCLUSION： The method is sensitive, rapid, accurate, convenient, and applicable for the quantitative determination of plasma concentration of MPA in renal transplantation patients.