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中空纤维膜微萃取/超高效液相色谱法检测大肠杆菌中氨基酸的研究 预览

Determination of Amino Acids in Escherichia Coli. by Ultrahigh Performance Liquid Chromatography Combined with Hollow Fiber Membrane Microextraction
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摘要 建立了三相中空纤维膜微萃取/超高效液相色谱(HF-LPME/UHPLC)方法,用于大肠杆菌中8种氨基酸的测定。设计了三相中空纤维膜微萃取系统,优化了HF-LPME最佳萃取条件:液膜为正辛醇,接收相Na OH的浓度为0.30 mol/L,给出相样品溶液中盐酸浓度为5.0 mmol/L,Na Cl质量浓度为200 g/L,萃取温度为25℃,搅拌速度为500 r/min,萃取时间为4 h。8种氨基酸经衍生并萃取后进行色谱分析,在5 min内达基线分离,其峰面积与浓度在0.20-4.9×10^-3μmol/L范围内呈良好的线性关系,相关系数r2均大于0.999,富集倍数为110-290倍,检出限为0.08-0.35μmol/L。在10μmol/L加标水平下,大肠杆菌样品的平均回收率为88.7%-103.1%,相对标准偏差(n=5)为3.2%-4.3%。该方法灵敏度高、重复性好,可用于细菌中氨基酸的检测。 A three-phase hollow fiber membrane microextraction/ultrahigh performance liquid chromatography(HF-LPME/UPLC) was developed for the determination of eight amino acids in Escherichia coli.. The three-phase hollow fiber membrane micro-extraction system was designed. The extraction conditions for HF-LPME were optimized as follows: the supported liquid membrane: n-octanol,the concentration for Na OH in receiving phase: 0. 30 mol/L,the concentration for hydrochloric acid in sample solution: 5. 0 mmol/L,the concentration for Na Cl: 200 g/L,the extraction temperature: 25 ℃,the stirring speed: 500 r/min and the extraction time: 4 hours. Eight kinds of amino acids were extracted after derivatization,and finally analyzed by UPLC. A baseline separation was achieved within 5 min. The calibration curves for the analytes were linear in range of 0. 20-4. 9 ×10杜一平-3μmol/L with their correlation coefficients(r2) more than 0. 999,the enrichment factors of 110-290 and the detection limits of 0. 0 l/L. At a spiked concentration of 10 μmol/L,the recoveries for eight amino acids in Escherichia coli. were 88. 7%-103. 1% with the relative standard deviations of3. 2%-4. 3%(n = 5). The proposed method was highly sensitive and reproducible,and could be used for the detection of amino acids in bacteria.
作者 李仪 赵双丽 张炎 何品刚 王清江 LI Yi,ZHAO Shuang-li,ZHANG Yan,HE Pin-gang,WANG Qing-jiang (School of Chemistry and Molecular Engineering,East China Normal University,Shanghai 200241,China )
出处 《分析测试学报》 CSCD 北大核心 2018年第7期790-795,共6页 Journal of Instrumental Analysis
基金 国家自然科学基金项目(21575042)
关键词 大肠杆菌 氨基酸 超高效液相色谱 中空纤维膜微萃取 Escherichia coli. amino acid;ultra high performance liquid chromatography (UHPLC ) hollow fiber membrane phase microextraction
作者简介 通讯作者:王清江,教授,研究方向:微纳分离分析,Tel:021-54340100,E-mail:qjwang@chem.ecnu.cn
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