柱前衍生结合高效液相色谱荧光检测法快速测定奶粉中1-脱氧野尻霉素  

Rapid Determination of 1-Deoxynojirimycin in Milk Powder by Precolumn Derivatization Combined with High Performance Liquid Chromatography with Fluorescence Detection

作　　者：邓思思 覃浩 熊梅瑾 赵红阳 刘玲 黄玉英 鲍蕾 李绍波 DENG Sisi;QIN Hao;XIONG Meijin;ZHAO Hongyang;LIU Ling;HUANG Yuying;BAO Lei;LI Shaobo(Chengdu Institute of Food Inspection,Chengdu 611130,China;NestléR&D(China)Ltd.,Beijing 100016,China;Key Laboratory of Chemical Metrology and Applications on Nutrition and Health for State Market Regulation,Beijing 100029,China)

机构地区：[1]成都市食品检验研究院,四川成都611130 [2]雀巢研发(中国)有限公司,北京100016 [3]国家市场监管重点实验室(营养与健康化学计量及应用),北京100029

出　　处：《食品科学》2025年第4期278-284,共7页Food Science

基　　金：国家市场监督管理总局科技计划项目(2023MK156);“十四五”国家重点研发计划重点专项(2022YFF0710405);四川省科技计划重点研发项目(2023YFS0402;2022YFS0511)。

摘　　要：建立功能性奶粉中1-脱氧野尻霉素含量柱前衍生结合高效液相色谱荧光快速检测方法。奶粉经60%甲醇水溶液提取,在pH 8.5的硼酸盐缓冲液条件下,加入邻苯二甲醛衍生剂,去除伯胺类物质的干扰后,样品中的1-脱氧野尻霉素与9-芴甲氧羰酰氯反应生成具有荧光吸收的络合物。采用C18液相色谱柱(150 mm×4.6 mm,2.7μm),流动相A为10 mmol/L乙酸钠缓冲溶液,流动相B为乙腈,梯度洗脱,流速1.5 mL/min,柱温45℃,激发波长275 nm,发射波长320 nm。结果表明:1-脱氧野尻霉素与其他组分分离效果良好,方法检出限为4 ng/mL,在0.01~50μg/mL范围内线性关系良好,决定系数R2>0.999,回收率为101.8%~104.1%。方法验证结果表明该方法稳定可靠,可作为测定奶粉中DNJ含量的有效方法。A rapid precolumn derivatization combined with high performance liquid chromatography(HPLC)with fluorescence detection method for the determination of the 1-deoxynojirimycin(DNJ)content in functional milk powder.Samples were extracted with 60%methanol in water.In pH 8.5 borate buffer solution,o-phthalaldehyde was added as a derivatizing agent to remove the interference from primary amines.DNJ in samples reacted with 9-fluorenylmethoxycarbonyl chloride to form a complex with fluorescence absorption.The separation was performed on a C18column(150 mm×4.6 mm,2.7μm).Mobile phase A was 10 mmol/L sodium acetate buffer solution,and mobile phase B was acetonitrile.Gradient elution was performed at a flow rate of 1.5 mL/min.The column temperature was 45℃.The excitation wavelength was 275 nm.The emission wavelength was 320 nm.Good separation of DNJ from other components was achieved.The detection limit was 4 ng/mL,and the calibration curve was linear in the range of 0.01 to 50μg/mL,with a determination coefficient(R2)>0.999.The recovery rates ranged from 101.8%to 104.1%.The method was validated to be stable and reliable,and could be used as an effective method for determining the content of DNJ in milk powder.

关 键 词：奶粉 1-脱氧野尻霉素 高效液相色谱 荧光检测 

分 类 号：TS207[轻工技术与工程—食品科学]