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反相离子对高效液相色谱-电感耦合等离子体质谱法测定富硒酵母中硒

Ion-Pair RP-HPLC-ICP-MS Determination of Selenium in Selenium-Enriched Yeast

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【作者】 高愈希蒲云霞彭晓敏吴刚

【Author】 GAO Yu-xi1,PU Yun-xia1,2,PENG Xiao-min1,WU Gang2(1.Key Lab.of Nuclear Analytical Techniques,Research Institute of High Energy Physics, Chinese Academy of Sciences,Beijing 100049,China; 2.School of Basic Medicine,Baotou Medical College,Baotou 014040,China)

【机构】 中国科学院高能物理研究所核分析技术重点实验室包头医学院基础医学部

【摘要】 富硒酵母样品用pH 7.5的Tris-盐酸缓冲溶液经超声破碎提取30min,于提取液中加链霉蛋白酶E在37℃搅拌18h,高速离心分离。取上清液经Millipore Corp超滤膜过滤,滤液中无机硒(Ⅵ及Ⅳ)和有机硒,即硒代半胱氨酸、硒代尿素和硒代蛋氨酸通过Waters Symmetry ShieldRP18色谱柱分离。用不同比例混合的甲醇-七氟丁酸溶液作流动相进行梯度洗脱,用电感耦合等离子体质谱法在线测定洗脱液中各种形态硒的含量。5种形态硒的质量浓度均在100μg·L-1以内呈线性。应用此法分析了富硒酵母样品,结果表明:无机硒占总硒量的1.63%,65%的硒以易被人体吸收的硒代蛋氨酸形式存在。

【Abstract】 Sample of selenium-enriched yeast was extracted with Tris-HCl buffer solution of pH 7.5 by ultrasonic smashing for 30 min.Protein hydrolytic enzyme Pronase E was added to the extract,which was stirred at 37 ℃ for 18 h and separated by high speed centrifugation.The supernatant was taken and filtrated with Millipore Corp super-filtering membrane.Waters Symmetry Shield RP18 chromatographic column was used for separation of inorganic selenium(Ⅵ and Ⅳ) and organic selenium compounds of Selenocysteine,Selenourea and Seleno-methionine.Mixtures of methanol and heptafluoro-butanoic acid mixed in different ratios were used as mobile phase in gradient elution for the 5 species of selenium.The eluates were used for ICP-MS determination.Linear relationships of the 5 species of selenium were kept in the same range within 100 μg·L-1.The proposed method was applied to the analysis of sample of selenium-enriched yeast.It was found that: the amount of inorganic selenium was only amounting to 1.63% of the total selenium.The major species of selenium existed in yeast was selenomethionine,amounting to 65%,which was easily absorbed and utilized by the human body.

【基金】 国家自然科学基金(20777075;20931160430)
  • 【文献出处】 理化检验(化学分册) ,Physical Testing and Chemical Analysis(Part B:Chemical Analysis) , 编辑部邮箱 ,2013年06期
  • 【分类号】TS207.4;O657.63
  • 【被引频次】11
  • 【下载频次】315
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