万古霉素血药浓度监测方法的建立及稳定性考察

Establisnment of Method of Vancomycin in TDM and Study of Stablility

目的:建立一快速、准确的RP-HPLC方法测定人血清中万古霉素血药浓度,并考察方法的稳定性。方法:以甲硝唑为内标,采用Agilent 1100系列高效液相色谱仪,以Kromasil C18柱(4.6 mm×250 mm,5μm)为分析柱;流动相为乙腈-0.01mol.L-1 KH2 PO4溶液(7∶93);流速1.0 mL.min-1;检测波长236 nm;柱温30℃。血浆样品用30%的硫酸锌溶液沉淀蛋白后,取上清液20μL进样分析,对血样处理前后及标准溶液保存不同时间后的稳定性进行了考察。结果:色谱峰分离良好,无干扰。万古霉素最低检测浓度为0.252 4 mg.L-1,线性范围0.504 8～126.2 mg.L-1,线性方程为Y=-1.94×10-2 X+0.653(r=0.999 3),相对回收率大于95%,高、中、低3个浓度的样品萃取回收率均大于78%,血样未经处理前4℃条件下能在1周内保持较好稳定性,血样处理后48 h内样品能保持较好稳定性,而标准溶液有效保存期为6个月。结论:本方法是一种可靠、快速灵敏的检测方法,适用于万古霉素的血药浓度监测。

Objective： To develop a HPLC method to determine the concentration of vancomycin（VAN） in human plasma and study the stability of vancomycin in different condition.Method： The metonidazole was used as inter standard（I.S.） The analytical column was a Kromasil C18 column connected with the Agilent 1100 HPLC system.The isocratic mobile phase was the acetonitrile and buffer of KH2PO4（0.01 mol·L-1） with the rate of（7∶93）.The detective UV wavelength was 236 nm and the column temperature was 30 ℃.The plasma samples,added metonidazole as internal standard,were deposited down the protein with 30% ZnSO4 and 20 μL of the upper phase was injected into the sampler.The stability of the samples in different conditon were studied.Result： The chromatography was good and not interfered by the components of the plasma.The linear equation was Y=-1.94×10-2X＋0.653（r=0.999 3）.The recovery of vancomycin in different concention were all 78%.The samples were stability in 4 ℃ for a week,the standard solution of vancomycin were stablity for a half year.Conclusion： This is a accurate,sensitive and conventient method for the dertermination of VAN in the therapeutic drug monitoring.