反相高效液相色谱法测定人血清中替考拉宁血药浓度

Determination of Teicoplanin in Human Plasma by RP-HPLC

目的建立测定人血清中替考拉宁浓度的反相高效液相色谱法。方法采用高效液相色谱法,以哌拉西林钠为内标进行检测,分析柱采用InertSustain C_(18)色谱柱(150 mm×4.6 mm,5μm);流动相:0.01 mol·L^(-1) NaH_(2)PO_(4)(pH=3.3)-乙腈(75∶25,H_(3)PO_(4)调pH值为3.2)等度洗脱;流速:1 mL·min^(-1);检测波长:240 nm;柱温:40℃;血浆样品经乙腈沉淀,二氯甲烷萃取后进样,进样量为20μL;二极管阵列检测。结果替考拉宁血药浓度在3.197~102.3μg·mL^(-1)内线性关系良好(R 2=0.9998,n=6),峰型对称与血浆中其他组分分离分离良好。替考拉宁低、中、高3个浓度(3.79、11.37、34.10μg·mL^(-1))的平均方法回收率为94.3%,同样条件下测得平均提取回收率为94.8%。日内精密度RSD小于4.97%,日间精密度RSD小于4.89%。结论本方法准确度高,灵敏度好,操作简便,适用于临床替考拉宁血药浓度监测和药代动力学研究。

OBJECTIVE To establish the method for determination of teicoplanin in human plasma by RP-HPLC.METHODS We used RP-HPLC to determine and regarded Piperacillin Sodium as the internal standard.The liquid chromatography was performed on InertSustain C_(18)(150×4.6 mm,2.6μm),and 0.01 mol·L^(-1) NaH_(2)PO_(4)-acetonitrile(75∶25,pH=3.2)was used as the mobile phase to equivalent elute.The flow rate was 1.0 mL·min^(-1),the column temperature was 40℃,and the UV detector wave length was 240 nm.RESULTS The calibration curve was linear through the range of 3.197^(-1)02.3μg·mL^(-1)(R 2=0.9998,n=6).The average extraction recovery rate and average method recovery rate were 94.8%and 94.3%,respectively.The relative standard deviations(RSDs)of within-day below 4.97%and between-day below 4.89%.This method shows to be sensitive and accurate for monitoring the serum con centration of teicoplanin in clinical practiceand and pharmacokinetic study.