目的利用单因素实验法建立和优化临床万古霉素HPLC分析方法,为临床监测万古霉素血药浓度、调整万古霉素临床给药剂量和个体化差异给药提供定量分析方法。方法比较国内外已报道的万古霉素HPLC测定方法,利用本院WATERS液相系统进行方法验证,以40%的硫酸锌沉淀处理血液标本,最终选择Waters ODS2(250 mm×4.6 mm,5μm)色谱柱为分析柱。实验设计通过梯度改变柱温、pH、流动相的组成比例,验证每种条件对实验结果的容量因子(K),拖尾因子(T),分离度(R)的影响,最终得到最佳色谱条件为流动相0.05 mol·L-1KH2PO4缓冲溶液(用磷酸调pH至2.85)-乙腈-甲醇(91.5∶8∶0.5),紫外检测波长为200 nm,流速为1 mL·min-1,柱温30℃,进样量为20μL。结果万古霉素的最低检测质量浓度为0.05 mg·L-1,在0.1~64.0 mg·L-1时线性较好(r=0.999 2),相对回收率为97.5%~100.1%,日内、日间RSD均<2%,R为2.1。结论该方法灵敏,快速,简便,特异性且稳定可靠,适用于临床万古霉素血药浓度的监测及临床药学的研究。 Objective To establish and optimize the method of clinical analysis of vancomycin by single factor experiment and provide a quantitative analysis method for clinical monitoring of vancomycin blood drug concentration and adjustment of vancomycin clinical administration dose and individualized difference administration. Methods The method of HPLC determination of vancomycin was reported at home and abroad. The method was validated by the WATERS liquid-phase system in our hospital. The blood samples were precipitated with 40% zinc sulfate, and finally the Waters ODS2 (250 mm × 4.6 mm, 5 μm) Column for the analytical column. Experimental design The column temperature, pH, mobile phase composition ratio were changed by gradient to verify the effect of each condition on the capacity factor (K), tailing factor (T) and resolution (R) of the experimental results, and finally the best chromatogram The mobile phase consisted of 0.05 mol·L-1 KH2PO4 buffer solution (pH adjusted to 2.85 with phosphoric acid) -acetonitrile-methanol (91.5: 8:0.5), the UV detection wavelength was 200 nm and the flow rate was 1 mL · min-1. 30 ℃, injection volume of 20μL. Results The minimum detectable concentration of vancomycin was 0.05 mg · L-1, the linearity was good at 0.1 ~ 64.0 mg · L-1 (r = 0.999 2) and the relative recoveries were 97.5% ~ 100.1% Between 2% RSD, R is 2.1. Conclusion The method is sensitive, rapid, simple, specific and stable. It is suitable for the monitoring of clinical vancomycin and clinical pharmacy.