富硒大米中硒的种态分析研究.docVIP

富硒酵母中含硒的提取和硒种态分析 , 蒲云霞1,2, 彭晓敏1, 吴刚2 (1. 中国科学院高能物理研究所核分析技术重点实验室，北京 1000492. 包头医学院基础医学部，包头 014040） 摘 要 富硒酵母品质与所含硒的种态关。为了评价富硒酵母品质，用超声法破碎细胞壁，用考马斯亮兰法测定提取液中的蛋白浓度，用酶解提取法提取酵母含硒组分，用电感耦合等离子体质谱（ICP-MS）测定硒含量，通过提取液中的硒含量评估提取，用反相离子对高效液相色谱与电感耦合等离子体质谱联用技术（HPLC-ICP-MS）酶解液中的小分子硒。酶解提取法硒回收率为80.97%，其中，SeO32-和SeO42-占总硒的1.63 %，硒以易被人体吸收的硒代蛋氨酸形式存在于蛋白分子中。结果表明：酶解提取法适合于富硒酵母，所研究富硒酵母中硒主要以易被人体吸收的含硒蛋白形式存在。 关键词: 富硒酵母，种态分析，HPLC-ICP-MS 中图分类号：R194.5，O652 Extraction and Speciation of Selenium-Containing Species in Selenium-enriched Yeast GAO Yu-xi1, PU Yun-xia1,2, PENG Xiao-min1, WU Gang2 (1. Key Laboratory of Nuclear Analytical Techniques, Institute of High Energy Physics, Chinese Academy of Sciences, Beijing 100049 China; 2. School of Basic medicine, Baotou Medical College, Baotou 014040, China) Abstract: Se-enriched yeast is a common nurture for selenium complement. Its quality depends on the Se-containing species. To evaluate the quality of selenium-enriched yeast, the Se-containing species were analyzed as following: the walls of Se-enriched yeast were disrupted by an ultrasonic assisted procedure, and then the Se-containing species were extracted with an enzymolysis procedure. The validity of disruption was estimated based on the protein concentration in the disrupting buffer measured with the Bradford method. The extraction efficiency was evaluated based on the selenium content in the extract measured with ICP-MS. The selenium-containing species in enzymolysis supernatant was analyzed with reversed-phase ion-pairing HPLC-ICP-MS to assess the quality of selenium-enriched yeast. The Se recovery for the enzymolysis procedure was 80.97%, among them, the inorganic forms of Se (SeO32- and SeO42-) was only 1.63% of total Se. A majority of selenium existed in the yeast was in the form of selenomethionine which can be easily absorbed and utilized by humans. The results indicate that the enzymolysis extraction after ultrasonic disruption is a suitable technique of sample preparation for assessing th