高效液相色谱测定儿童体内霉酚酸血药浓度  

Determination of mycophenolic acid in children’s plasma by high performance liquid chromatography

作　　者：许泽君[1] 孙芳[1] 景霞[1] XU Zejun;SUN Fang;JING Xia(Pharmaceutical Department,Children’s Hospital of Nanjing Medical University,Nanjing,Jiangsu 210008,China)

机构地区：[1]南京医科大学附属儿童医院药学部

出　　处：《安徽医药》2020年第3期610-613,共4页Anhui Medical and Pharmaceutical Journal

摘　　要：目的建立儿童体内霉酚酸血药浓度高效液相色谱测定方法,为临床调整用药提供参考。方法内标(萘啶酸)法,血样先经甲醇沉淀,后经磷酸酸化,进样。色谱柱为Agilent Eclipse Plus–C18(4.6 mm×50 mm,5μm)色谱柱;流动相为0.1%三乙胺溶液(磷酸调节PH为3.5±0.1)⁃乙腈=60∶40;流速为1.0 mL/min;柱温25 oC;进样体积为40μL。按有限采样法模型,取南京医科大学附属儿童医院病儿清晨服药前(0 h),服药后0.5 h和2 h 3个时间点血样,从2017年11月至2018年4月6个月内共检测177例。结果霉酚酸在0.20~50.00μg/mL浓度范围内色谱峰面积与其相应的内标的峰面积比值呈良好线性关系(r=0.9996);低中高三种浓度(0.39,25.00,50.00μg/mL)日内相对标准偏差(RSD)(n=5)为2.00%~3.41%,日间RSD(n=3)为3.40%~4.38%;方法学回收率为97.44%~104.80%。按经验公式0~12 h药-时曲线下面积(AUC0~12 h)=7.75+6.49×C0+0.76×C0.5+2.43×C2计算其中63例小于30 mg·L^-1·h^-1,占35.6%,102例在30~60 mg·L^-1·h^-1之间,占57.6%,12例大于60 mg·L^-1·h^-1,占6.8%。结论该方法专属性、重复性、准确性较好,可用于儿童血浆中霉酚酸的含量检测。Objective To establish a HPLC method for the determination of mycophenolic acid in plasma for children and provide a reference for clinical adjustment of medication.Methods With internal standard(nalidixic acid)method,the plasma was precipitated by methanol,then acidified by phosphoric acid and injected.The chromatographic column was Agilent Eclipse Plus–C18(4.6 mm×50 mm,5μm).The mobile phase consisted of acetonitrile⁃water(contain 0.1%triethylamine pH=3.5±0.1)=40∶60,with a flow rate of 1.0 mL/min.Column temperature was 25℃;The volume of injection was 40μL.With the limited sampling strategy,the blood samples were collected immediately before and 0.5,2h after the morning dose of mycophenolate mofetil in Children’s Hospital of Nanjing Medical University,and a total of 177 samples were detected in the six months from November 2017 to April 2018.Results The peak area of mycophenolic acid showed a good linear relationship with the corresponding peak area ratio of internal markers in the range of 0.20~50.00 g/mL(r=0.9996).The inter⁃run RSDs for the low,middle and high concentrations(0.39,25.00,50.00μg/mL)were 2.00%to 3.41%(n=5)and intra⁃run RSDs were 3.40%to 4.38%(n=3),method recoveries were 97.44%to 104.80%.As calculated by the regression equation(AUC0⁃12 h=7.75+6.49×C0+0.76×C0.5+2.43×C2),AUC of 63 cases was under 30 mg·L^-1·h^-1,accounting for 35.6%;AUC of 102 cases was between 30 and 60 mg·L^-1·h^-1,accounting for 57.6%;AUC of 12 cases was over 60 mg·L^-1·h^-1,accounting for 6.8%.Conclusion The method is specific,repeatable and accurate,and can be used for determination of mycophenolic acid in children’s plasma.

关 键 词：色谱法 高压液相 霉酚酸 内标法 血药浓度 儿童 

分 类 号：R969.1[医药卫生—药理学]