HPLC法测定人血清中去甲万古霉素浓度

目的建立人血清中去甲万古霉素浓度的高效液相色谱测定方法及临床应用。方法以万古霉素为内标,采用10%高氯酸溶液直接沉淀蛋白后进样。色谱柱为SHIMADZUVP-ODS(150 mm×4.6 mm,5μm),流动相为磷酸二氢钾缓冲液-乙腈(92∶8),流速1 mL.min-1,柱温40℃,检测波长236 nm。结果去甲万古霉素血清浓度Y与内标峰面积比X进行线性回归,回归方程为Y=32.544 56X-3.147 39,r=0.999 3;线性范围2.5-80.0μg.mL-1;日内RSD小于3.0%;日间RSD小于5.7%;去甲万古霉素低、中、高浓度的平均方法回收率分别为97.36%、99.51%、101.17%;平均提取回收率分别为86.82%、89.84%、92.68%。结论方法操作简便快速,回收率稳定,适于临床血药浓度监测及人体药动学研究,尤其适于ICU或NICU患者接受去甲万古霉素治疗时的血药浓度监测。

Determination the concentration of norvancomycin in human serum by HPLC

Objective To establish an HPLC method for the determination of norvancomycin in human serum. Methods The vancomycin was used as internal standard （ IS ）. The pretreatment of the blood samples was sedimentation with 10% HClO4. Then the sample was separated on SHIMADZU VP - ODS Colunmn（ 150 mm ×4.6mm,5μm） with potassium dihydrogen phosphate buffer solution - acetonitrile （ 92：8 ） as the mobile phase. The detection wavelength was 236 am. The flow rate was 1.0 mL · min^-1. The temperature of the column was 40 ℃. Results Norvancomycin serum concentrations Y and the internal standard peak area X were doing linear regression The regression equation was Y = 32. 544 56X - 3. 147 39 with linear range of 2.5 to 80 mg · L^-1 （ r = 0. 999 3 ）. The values for intra-day and inter-day RSD were less than 3.0% and 5.7% respectively. The average recoveries of high,medium and low concentrations of norvancomycin were 97.36% ,99.51% and 101.17% ,the average extrac- tion recoveries were 86.82%, 89.84%, 92.68%. Conclusion The method was rapid, simple and recovery was stable. It was suitable to monitor the concentration of norvancomycin in serum and study human pharmacokinetic. It was particularly suitable for the blood concentration monitoring of norvancomycin for ICU or NICU patients.