HPLC柱前衍生化测定组织液中妥布霉素的含量  

HPLC derivatization method established for the content determination of tobramycin in tissue fluid

作　　者：程仕虎[1] 刘又宁[1] 王睿[2] 李照霞[1] 崔俊昌[1] 

机构地区：[1]解放军总医院呼吸科,北京100853 [2]解放军总医院药理研究室,北京100853

出　　处：《中国药物应用与监测》2007年第3期26-28,共3页Chinese Journal of Drug Application and Monitoring

基　　金：国家自然科学基金资助项目(30672505)

摘　　要：目的:建立HPLC柱前衍生化法测定组织液中妥布霉素的含量。方法:使用含异硫氰酸-1-奈酯(NITC)的吡啶溶液在70℃衍生化,然后使用甲胺/乙腈溶液消除过度衍生化。HPLC使用AgilentHypersilODS色谱柱,流动相为乙腈:水(60:40),流速0.9mL/min,检测波长:230nm。结果:较好的检测出了组织液中妥布霉素的含量,检测范围为0.9μg/mL～20.0μg/mL,检测限为0.75μg/mL(信:噪=3,注射10μL),日内变异率小于2.0%(n=6),日间变异率小于5.3%,相对回收率大于99%。结论:NITC柱前衍生化HPLC分离测定注射用硫酸妥布霉素的含量方法简单,衍生化稳定可靠,使用高效液相可进行大规模样本测试。Objective：High-performance liquid chromatography （HPLC） method is established for the content determination of tobramycin in rabbit tissue fluid by derivatization.Methods：The method is based on the chemical derivatization of aminoglycoside antibiotic, tobramycin in human plasma, with NITC in pyridine at 70℃. After derivatization reaction, a methylamine/acetonitrile solution was added to the reaction mixture to eliminate the excess derivatizing agent. The derivative result was separated using a Agilent Hypersil ODS column and a water-acetonitrile （60：40） mobile phase （detection at 230nm）.Results：Optimization conditions for the derivatization of tobramycin were investigated by HPLC. The linear range for the quantitation of tobramycin in tissue fluid was over 0.9μg/mL-20.0μg/mL; the detection limit （signal-to-noise ratio≡3; injection volume：10μL） was about 0.75μg/mL.The relative standard deviation was less than 2.0% for intra-day assay （n=6） and 5.3% for inter-day assay （n=6） and relative recoveries were found greater than 99%.Conclusion：The HPLC method can be used to determine the tobramycin in rabbit tissue fluid.

关 键 词：妥布霉素 高效液相 衍生化 

分 类 号：R96[医药卫生—药理学]