高效液相色谱-荧光检测法测定食品中维生素K_2的含量

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英文篇名：Determination of Vitamin K_2 in Food by High Performance Liquid Chromatography with Fluorescence Detection 作者：邓梦雅 ; 彭祖茂 ; 朱丽 ; 杨俊 ; 杨世添 ; 张协光 英文作者：DENG Meng-ya;PENG Zu-mao;ZHU Li;YANG Jun;YANG Shi-tian;ZHANG Xie-guang;Shenzhen Academy of Metrology & Quality Inspection,National Nutritional Food Testing Center(Guangdong); 关键词：高效液相色谱-荧光检测法(HPLC-FLD) ; 食品 ; 维生素K_2 ; 四烯甲萘醌(MK-4) ; 七烯甲萘醌(MK-7) 英文关键词：high performance liquid chromatography-fluorescence detection(HPLC-FLD);;food;;vitamin K_2;;menatetrenone 4(MK-4);;menatetrenone 7(MK-7) 中文刊名：食品工业科技 英文刊名：Science and Technology of Food Industry 机构：深圳市计量质量检测研究院国家营养食品质量监督检验中心(广东); 出版日期：2019-06-03 09:41 出版单位：食品工业科技 年：2019 期：19 基金：深圳市计量质量检测研究院科研项目(2018-YA19) 语种：中文; 页：246-250+256 页数：6 CN：11-1759/TS ISSN：1002-0306 分类号：O657.3;TS207.3

摘要

建立了高效液相色谱-荧光检测法(HPLC-FLD)同时快速分离检测猪肉、猪肝、纯牛奶、酸奶、黄油、鸡蛋、豆豉、纳豆8种食品中2种维生素K_2的方法。样品经酶解、皂化、萃取、氮吹复溶处理后,用Agilent ZORBAX SB-C_(18)色谱柱(150 mm×4.6 mm,5μm)分离,柱后锌粉还原柱(35 mm×4.6 mm)衍生,以甲醇(含有二氯甲烷10%、冰醋酸0.03%、氯化锌1.5 g/L、无水乙酸钠0.5 g/L)为流动相,进入荧光检测器进行检测。结果表明,四烯甲萘醌(Menatetrenone4,MK-4)、七烯甲萘醌(Menatetrenone7,MK-7)在15 min内分离出峰。2种维生素K_2在各自的线性范围内关系良好,相关系数r=1.0000。当取样量为2 g,定容2 mL时,MK-4的检出限为0.01μg/100 g,定量限为0.04μg/100 g;MK-7的检出限为0.06μg/100 g,定量限为0.2μg/100 g。当取样量为10 g,定容2 mL时,MK-4的检出限为0.002μg/100 g,定量限为0.008μg/100 g;MK-7的检出限为0.012μg/100 g,定量限为0.04μg/100 g。精密度(RSD)均不超过3.56%,回收率为80.6%～95.5%。8种食品中,猪肉、纯牛奶、酸奶、黄油、鸡蛋中仅含有MK-4,其中纯牛奶中含量最低,仅为(0.34±0.02)μg/100 g,鸡蛋中含量最高,达(25.38±0.85)μg/100 g。猪肝、豆豉、纳豆中仅含MK-7,其中猪肝含量较低,为(25.45±0.83)μg/100 g,纳豆中含量最为丰富,高达(923.20±11.45)μg/100 g。该法准确、可靠、高效,可用于食品中维生素K_2的高通量检测。
        A method for simultaneous determination of two kinds of vitamin K_2 in 8 kinds of food,including pork,pork liver,pure milk,yogurt,butter,eggs,lobster sauce and natto,by high performance liquid chromatography-fluorescence detector(HPLC-FLD)was developed. After enzymatic hydrolysis,saponification,extraction,nitrogen blowing and resolving treatment,the sample was separated by an Agilent ZORBAX SB-C_(18)column(150 mm×4.6 mm,5 μm)and derivatized with a post-column zinc powder reduction column(35 mm×4.6 mm). Methanol(containing 10% dichloromethane,0.03% glacial acetic acid,1.5 g/L zinc chloride and 0.5 g/L anhydrous sodium acetate)was used as mobile phase for detection by fluorescence detector. Results showed that the chromatographic peaks of menatetrenone 4(MK-4)and menatetrenone7(MK-7)were found within 15 min. A good respective linearity of two kinds of vitamin K_2was obtained with the correlation coefficient(r=1.0000).When the sample size was 2 g and the volume was 2 mL,the detection limit of MK-4 was 0.01 μg/100 g and the quantitative limit was 0.04 μg/100 g.The detection limit of MK-7 was 0.06 μg/100 g and the quantitative limit was 0.2 μg/100 g. When the sample size was 10 g and the volume was 2 mL,the detection limit of MK-4 was 0.002 μg/100 g and the quantitative limit was 0.008 μg/100 g.The detection limit of MK-7 was 0.012 μg/100 g and the quantitative limit was 0.04 μg/100 g.The relative standard deviation(RSD)was no more than 3.56%,and the recoveries ranged from 80.6% to 95.5%.Among the 8 kinds of food,pork,pure milk,yogurt,butter and eggs only contained MK-4,among which the content of pure milk was the lowest(0.34±0.02) μg/100 g and the content of egg was the highest(25.38±0.85) μg/100 g.There was only MK-7 in pork liver,lobster sauce and natto,among which the content of pork liver was low(25.45±0.83) μg/100 g and the content of natto was the most abundant,up to(923.20±11.45) μg/100 g.The method is accurate,reliable,efficient,and can be used for high-throughput determination of vitamin K_2 in food.

引文

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