利用分散液-液微萃取和反相高效液相色谱-荧光检测器测定硒蛋白多糖中硒含量

本试验介绍了一种利用分散液-液微萃取前处理并将一种新型反相色谱柱应用于高效液相色谱-荧光检测器(HPLC-FLD)测定硒蛋白多糖中硒含量的方法。该方法以2,3-二氨基萘(DAN)为螯合剂,400μL乙腈为分散剂,120μL氯苯为萃取剂,硒(Ⅳ)与2,3-二氨基萘的螯合物(Se-DAN螯合物)经乙腈稀释后进行高效液相色谱分析。采用PFP色谱柱分离,乙腈为流动相,用荧光检测器在激发波长为376 nm、发射波长为520 nm条件下测定荧光强度,外标法定量。结果显示:该方法成功地用于测定硒蛋白多糖中硒含量,硒的检出限为2.5μg/L,线性范围为5.0~250.0μg/L。采用该方法测定的硒蛋白多糖中硒含量为1337.0μg/g,与荧光分光光度法(国标法)测定结果高度一致;并且,该方法也适用于酵母硒中硒含量的测定。本试验成功建立了检测硒蛋白多糖中硒含量的方法,该方法具有操作简便、易普及、试剂消耗少、分离度好、回收率高和适用性强等特点。

Determination of Selenium Content in Selenium-Enriched Polysaccharides by Dispersive Liquid-Liquid Microextraction Coupled to Reversed Phase High-Performance Liquid Chromatography with Fluorescence Detector

This study described a new method using dispersive liquid-liquid microextraction prior to reverse phase high-performance liquid chromatography with fluorescence detector(HPLC-FLD)on a new reversedphase column for the determination of selenium content in selenium-enriched polysaccharides.In the procedure,2,3-diaminonaphthalene(DAN)was used as the chelating reagent,400μL acetonitrile was used as the dispersive solvent and 120μL chlorobenzene was used as the extraction solvent.The chelate of selenium(Ⅳ)and DAN(Se-DAN chelate)was diluted by acetonitrile and then was analyzed by high-performance liquid chromatography.Using acetonitrile as mobile phase,the Se-DAN chelate was separated by PFP column,and the fluorescence intensity was determined by fluorescence detector(excitation wavelength at 376 nm and emission wavelength at 520 nm)and quantitative analysis by external standard method.The sample was quantified by external standard method.The results showed that this method was successfully applied to determine the selenium content in selenium-enriched polysaccharides with the detection limit of selenium was 2.5μg/L,and the linear range was 0.5 to 250.0μg/L.In this experiment,a method for determination of selenium content in selenium-enriched polysaccharides is successfully set,and this method has the advantages of simple and fast operation,low reagent consumption and high recovery rate and good applicability.