高效液相色谱-电感耦合等离子体质谱法测定富硒蛋白多糖中纳米单质硒含量  

Measurement of Nano Elemental Selenium in Selenium-Enriched Polysaccharides Using High Performance Liquid Chromatography Coupled With Inductivity Coupled Plasma Mass Spectrometry Method

作　　者：王凤芹[1] 程远之 肖肖 李笑笑[1] 宋德广 路则庆[1] 汪以真[1] WANG Fengqin　;CHENG Yuanzhi　;XIAO Xiao　;LI Xiaoxiao　;SONG Deguang;LU Zeqing　;WANG Yizhen(Feed Science Institute of Zhejiang University,Hangzhou 310058,China;National Engineering Laboratory of Bio-Feed Safety and Pollution Prevention,Hangzhou 310058,China;Key Laboratory of Animal Nutrition and Feed Science(Huadong),Ministry of Agriculture,Hangzhou 310058,China)

机构地区：[1]浙江大学饲料科学研究所生物饲料安全与污染防控国家工程实验室农业部华东动物营养与饲料重点实验室,杭州310058

出　　处：《动物营养学报》2018年第3期1155-1161,共7页CHINESE JOURNAL OF ANIMAL NUTRITION

基　　金：现代农业生猪产业体系(CARS-36)；浙江大学实验技术研究项目(SJS201711)

摘　　要：本试验旨在建立高效、准确、快速测定富硒蛋白多糖中纳米单质硒含量的方法。将25 mg阴沟肠杆菌(Enterobacter cloacae)Z0206产富硒蛋白多糖样品悬浮在3 mL蒸馏水中,在3 mgⅩⅣ型蛋白酶作用下超声处理30 min(37℃),将酶解液离心得到的沉淀物(纳米单质硒)在常温下经双氧水-盐酸(H_2O_2-HCl)体系消解,所得溶液直接经高效液相色谱-电感耦合等离子体质谱(HPLC-ICP-MS)分析[以PRP X100阴离子色谱柱对样品中的亚硒根离子(SeO_2-3)进行分离,流动相组成为5 mmol/L柠檬酸水溶液(pH 4.5)],测得样品中纳米单质硒含量为2 439μg/g。以国家标准(GB/T 13883—2008)方法[样品以硝酸-高氯酸(HNO_3-HClO_4)体系消解处理,经氢化物发生-原子荧光(HG-AFS)分析]为对照,测得样品中纳米单质硒含量为2 450μg/g。2种方法的测定结果高度一致。与GB/T 13883—2008中描述的方法相比,采用本试验建立的氧化体系条件更温和、快速并且易于控制,适用于纳米单质硒的快速检测。This experiment was conducted to establish a novel method which could efficiently,accurately and quickly determine the nano elemental selenium(Nano-Se)content extracted from selenium-enriched polysaccharides.Approximately 3 mg of proteaseⅩⅣwas added into 3 mL of H 2O contained 25 mg selenium-enriched polysaccharides which produced by Enterobacter cloacae Z0206,and the resulting mixture was maintained at 37℃in an ultrasonic bath for 30 min.After centrifugation,the precipitate(Nano-Se)was oxidized by HCl-H2O2 system at room temperature.Then the consequent solution was directly analyzed by high performance liquid chromatography coupled with inductivity coupled plasma mass spectrometry(HPLC-ICP-MS)(equipped with PRP-X100 anion column and the mobile phase was 5 mmol/L citric acid with a pH of 4.5)and the content of Nano-Se was 2 439μg/g.As a contrast,the same samples were oxidized by HNO 3-HClO 4 system and determined by the method described in GB/T 13883-2008 using hydride generation atomic fluorescence spectrometry(HG-AFS)and the content of Nano-Se was 2450μg/g which was similar to the result above.Compare with the method described in GB/T13883-2008,the oxidation system in the current method is much milder and faster and easy to control which is suitable for Nano-Se detection.

关 键 词：高效液相色谱 电感耦合等离子体质谱 富硒蛋白多糖 纳米单质硒 

分 类 号：Q657.32[生物学—生物物理学] Q936