Shim Jae Han, Lee Mi Hyun, Kim Mi Ra, Lee Chang Joo, Kim In Seon
Division of Applied Bioscience and Biotechnology, Institute of Agricultural Science and Technology, College of Agriculture and Life Science, Chonnam National University, Gwangju 500-757, South Korea.
Biosci Biotechnol Biochem. 2003 Jun;67(6):1342-8. doi: 10.1271/bbb.67.1342.
Simultaneous detection of the fluoroquinolone antibiotics ciprofloxacin, enrofloxacin, ofloxacin, and norfloxacin in eggs by a combination of supercritical fluid extraction (SFE) and high pressure liquid chromatography (HPLC) was studied. Lipid matrices that have been considered to result in poor extraction and isolation of fluoroquinolones in eggs were removed first by SFE with supercritical CO(2) alone, and then the fluoroquinolones were extracted by SFE with supercritical CO(2) containing 20% (v/v) methanol for HPLC analysis. A time-course study of the extraction of lipid matrices of eggs suggested that the SFE method successfully removed the matrices within 20 min. When the fluoroquinolones added to control eggs were extracted by SFE, the extraction efficiency was similar to that by the solvent extraction method, giving the recovery percentages from 83 to 96% in a 40 min-extraction time. The fluoroquinolones extracted from eggs by SFE were analyzed simultaneously by HPLC equipped with a fluorescence detector with detection sensitivity at about 10 ppb for the detection limit. The standard calibration profiles of fluoroquinolones showed linear responses to HPLC, showing more than 0.995 for the mean r(2) value. This is the first report of the simultaneous measurement of fluoroquinolones in eggs by a combination of SFE and HPLC. Using the SFE method allowed us to avoid extensive sample preparation such as solvent extraction and chromatographic cleanup that are basically required in extraction of fluoroquinolones.
研究了通过超临界流体萃取(SFE)和高压液相色谱(HPLC)联用同时检测鸡蛋中氟喹诺酮类抗生素环丙沙星、恩诺沙星、氧氟沙星和诺氟沙星的方法。首先单独使用超临界CO₂通过SFE去除被认为会导致鸡蛋中氟喹诺酮类提取和分离效果不佳的脂质基质,然后使用含有20%(v/v)甲醇的超临界CO₂通过SFE提取氟喹诺酮类用于HPLC分析。对鸡蛋脂质基质提取的时间进程研究表明,SFE方法在20分钟内成功去除了基质。当通过SFE提取添加到对照鸡蛋中的氟喹诺酮类时,提取效率与溶剂萃取法相似,在40分钟的提取时间内回收率为83%至96%。通过配备荧光检测器的HPLC同时分析通过SFE从鸡蛋中提取的氟喹诺酮类,检测限的检测灵敏度约为10 ppb。氟喹诺酮类的标准校准曲线对HPLC呈线性响应,平均r²值大于0.995。这是首次报道通过SFE和HPLC联用同时测量鸡蛋中的氟喹诺酮类。使用SFE方法使我们能够避免在氟喹诺酮类提取中基本需要的广泛样品制备,如溶剂萃取和色谱净化。