高效液相色谱法测定替考拉宁血浆总浓度和游离浓度

Determination the Total and Free Concentrations of Teicoplanin in Plasma by HPLC

目的建立高效液相色谱法测定血浆中替考拉宁(TEIC)总浓度(Ct)和游离浓度(Cf)的方法。方法游离浓度血浆样本经超滤离心管处理后进行HPLC分析。采用C1g色谱柱,以乙腈0.01·mL^-1磷酸二氢钠(25:75,磷酸调pH值至3.3)为流动相,检测波长240 nm,柱温35 C,流速1.0 mL·mL^-1,考察该方法的专属性、标准曲线、定量下限、精密度、回收率和稳定性。结果游离浓度在0.5~50μg·mL^-1内线性关系良好(y=12801x-2574.1,r^2=0.9998),定量下限为0.5μg·mL^-1,方法回收率为94.63%~103.72%,批内、批间RSD均<4.00%;总质量浓度在1.5625~100μg·mL^-1内线性关系良好(y=10576:x+9950.1,r^2=0.9998),定量下限为1.5625μg·mL^-1,方法回收率为94.55%~99.59%,批内、批间RSD均<4.00%。在室温放置10h、冻融循环3次和4℃冷藏72 h稳定性良好。结论本方法简便、准确、灵敏,适用于临床替考拉宁血药总浓度和游离浓度的检测及药动学的研究。

OBJECTIVE To establish an HPLC method for the determination of total(Ct)and free(Cf)concentrations of teico-planin(TEIC)in plasma.METHODS For determing the free concentration,the plasma samples were prepared by ulrafiltration.A Cx colunn was used,with acetonitrile-0.01·mL^-1sodium dihydrogen phosphate(25:75,pH adjusted to 3.3 by phosphoric acid)as the mobile phase,the detection wavelength was set at 240 nm,the column temperature was maintained at 35 C,and the flow rate was 1.0·mL^-1.The speificity,linearity,the lower limit of quantitation,precession.recovery and stability of the developed meth-od were validated.RESULTS The free concentration was linear with in 0.5-50μg·mL^-1,the lower limit of quantification was 0.5μg·mL^-1,and the method recovery rate was 94.63%-103.72%。The intra-day and inter-day precision(RSD)were all less than 4.00%;the linearity was good with in the total concentration of 1.5625-100μg·mL^-1,the lower limit of quantification was 1.5625μg·mL^-1,the method reeovery rale was 94.55%-99.59%.The intra-day and inter day precision(RSD)were all less than 4.00%。The TEIC was all stability at the conditions of maintaining at room temperature for 10 h,after freeze-thaw cycle and keeping at 4℃ for 72 h.CONCLUSION The method is simple,accurate,and sensitive,and is suitable for the clinical determination of total and free concentrations of teicoplanin and the studly of pharmacokinetis.