人血浆中霉酚酸浓度测定方法研究及药物监测应用

 目的建立用高效液相色谱快速测定人血浆中霉酚酸浓度的方法,对我院肾移植病人霉酚酸血药浓度进行监测。方法血浆样品加20μLH₃PO₄酸化,乙醚提取,萘啶酸为内标,采用Kromasil 100-5C₈(4.6 mm×150 mm,5μm)色谱柱,以甲醇-乙腈-0.05 mol·L^-1 KH₂PO₄(用磷酸调pH2.8)(12∶38∶50)为流动相,流速0.5 mL·min^-1,柱温25℃,检测波长254 nm。结果霉酚酸在0.191～24.45 mg·L^-1内线性关系良好(r=0.999 8),最低定量限为0.191 mg·L^-1(S/N=10)。高、中、低3个质控样品的平均方法学回收率为89.7%～101.5%,提取回收率大于85%,日内、日间精密度的RSD均小于10%。监测肾移植患者环孢霉素或他克莫司的谷浓度同时,测定其MPA血药浓度,82.3%的患者MPA浓度小于2 mg·L^-1,低于有效治疗谷浓度。结论本法简便、灵敏、重现性好,可用于肾移植病人服用霉酚酸酯后霉酚酸血药浓度监测和霉酚酸酯药动学研究。

Determination of Mycophenolic Acid Concentration in Human Plasma and Its Application to Therapeutic Drug Monitoring in Renal Transplantation Patient

OBJECTIVE To establish a HPLC method for the determination of mycophenolic acid in human plasma and monitoring mycophenolic acid concentration in renal transplantation patients.METHODS The plasma was mixed with 20 μL H₃PO₄.Mycophenolic acid was extracted from plasma with ether and nalidixic acid as internal standard.The content of mycophenolic acid was detected by ultraviolet detector at 254 nm with Kromasil 100-5C₈(4.6 mm×150 mm,5 μm) column.The mobile phase consisted of methol-acetonitrile-0.05 mol·L^-1 KH₂PO₄(adjusted to pH 2.8 with H₃PO₄)(12∶38∶50)at a flow rate of 0.5 mL·min^-1.The column temperature was 25 ℃.RESULTS The linear range was 0.191～24.45 mg·L^-1(r=0.999 8).The lowest determination concentration was 0.191 mg·L^-1(S/N=10).The average method recovery was 89.7%～101.5%,and extract recovery was over 85% for the low,middle,high control samples.The relative standard deviation obtained for inter-and intra-assay precisions were less than 10%.The MPA plasma concentration in 83.2% renal transplantation patient was below 2 mg·L^-1.CONCLUSION This method is convenient,sensitive and reproducible,suitable for the determination of mycophenolic acid in human plasma for pharmacokinetics and monitoring mycophenolic acid concentration in renal transplantation patient.