反相高效液相色谱法测定血液中去甲万古霉素浓度的研究  

Determination of Norvancomycin in plasma by RP-HPLC

作　　者：李顺炜[1] 袁孔现[1] 朱虹[1] 詹三华[1] 

机构地区：[1]铜陵市人民医院药剂科,安徽铜陵244000

出　　处：《安徽医药》2015年第8期1470-1473,共4页Anhui Medical and Pharmaceutical Journal

摘　　要：目的：建立去甲万古霉素血药浓度的反相高效液相色谱（RP －HPLC）测定法。方法以替硝唑或甲硝唑为内标，血液样品处理采用15％ HClO4沉淀、高速离心，NH4 Ac 溶液稀释后进样。色谱分析使用 Kromasil C18柱（150 mm×4．6 mm，5μm），流动相组成为甲醇—乙腈—40 mmol·L －1 KH2 PO4缓冲液（含0．1％磷酸）＝15∶5∶80（v／v），进样量为20μL，流速为1．0 mL· min －1，柱温35℃，紫外检测波长236 nm。灵敏度 AUFs ＝0．02，以峰面积、内标法定量。结果该法在去甲万古霉素浓度（X）为5．0～160 mg·L －1间呈良好线性关系，相关系数 r ＝0．9985（n ＝6），血清最低检测浓度为2．5 mg·L －1（S ／N≥3），日内、日间 RSD ＜10％，一个月内、反复冻融后的稳定性考察 RSD ＜10％。结论此法快速、简便、灵敏、可靠，适用于去甲万古霉素血药浓度监测。Objective To establish reversed phase high -performance liquid chromatography（RP -HPLC）method for determination of Norvancomycin plasma level.Methods Tinidazole or Metronidazole were used as internal standard,and the Norvancomycin in plasma was extracted by the sedimentation with 15% HClO4 and high -speed centrifugation,and diluted with NH4 Ac solution.Liquid chroma-tography was performed on Kromasil C18 column（150 mm ×4.6 mm,5 μm）The mobile phase was composed of CH3 OH -CH3 CN -40 mmol·L -1 KH2 PO4 （contains 0.1%H3 PO4 ）（15∶5∶80）.The injection volume was 20 μL,the flow rate was 1.0 mL·min -1 ,the col-umn temperature was 35℃ and the detection wavelength was 236 nm.The sensitivity was 0.02 AUFs.The experiment data were calcu-lated by peak area and internal standard method.Results A linearity was observed within the range of Norvancomycin concentrations from 5 to 160 mg·L -1 （r =0.998 5,n =6）,and the inspection limitation was 2.5 mg·L -1 （S /N≥3）.And the intra -day and inter-day RSD were less than 10%,the stable inspection RSD was less than 10% after one month and freeze -melt repeatedly.Conclu-sions This method is simple,rapid,sensitive,reliable and suitable for the determination of concentrations of Norvancomycin in plasma.

关 键 词：去甲万古霉素 药物浓度监测 高效液相色谱法 

分 类 号：R96[医药卫生—药理学]