液相色谱-荧光法测定乳粉中4种母乳低聚糖  

Determination of 4 kinds of breast milk oligosaccharides in milk powder by liquid chromatography-fluorescence method

作　　者：邓思怡 黄伟乾 吴俊发 覃天福 陈祖健 余梓恒 李镇杰 DENG Si-Yi;HUANG Wei-Qian;WU Jun-Fa;QIN Tian-Fu;CHEN Zu-Jian;YU Zi-Heng;LI Zhen-Jie(Guangzhou Inspection Testing and Certification Group Co.,Ltd.,Guangzhou 511447,China)

机构地区：[1]广州检验检测认证集团有限公司,广州511447

出　　处：《食品安全质量检测学报》2025年第5期32-37,共6页Journal of Food Safety and Quality

摘　　要：目的 建立液相色谱-荧光法同时测定乳粉中4种母乳低聚糖[2'-岩藻糖基乳糖(2'-fucosyllactose,2'-FL)、3'-唾液酸乳糖(3'-sialyllactose,3'-SL)、6'-唾液酸乳糖(6'-sialyllactose,6'-SL)和乳糖-N-新四糖(lacto-N-neo-tetraose,LNnT)]的含量分析方法。方法 样品加水溶解后,使用淀粉葡萄糖苷酶或和β-半乳糖苷酶酶解,用2-氨基苯甲酰胺(2-aminobenzamide,2-AB)和2-甲基吡啶硼烷(2-methylpyridine borane,2-PB)衍生后,采用酰胺键合色谱柱分离,荧光检测器检测和内标法定量。结果 4种母乳低聚糖在10~600μg/mL质量浓度范围均有良好的线性关系,相关系数均为r2>0.999;4种母乳低聚糖的检出限和定量限分别为0.94~2.31 mg/100 g、3.12~7.69mg/100g;4种母乳低聚糖加标回收率为97.7%~101.5%,相对标准偏差(relative standard deviation,RSD)(n=7)为0.53%~3.09%。结论 该方法对测定乳粉中的4种母乳低聚糖含量的检测人员、设备和环境要求不高、前处理操作简单,且具有良好的准确度和精密度,可为乳粉中母乳低聚糖质量控制提供方法参考。Objective To establish a method for simultaneous determination the content of 4 kinds of human milk oligosaccharides[2’-fucosyllactose(2’-FL),3’-sialic acid(3’-SL),6’-sialic acid(6’-SL)and lactose-N-neotetraose(LNnT)]in milk powder by liquid chromatography-fluorescence method.Methods The samples were dissolved in water,enzymolized with amyloglucosidase orβ-galactosidase,derived with 2-aminobenzamide(2-AB)and 2-methylpyridine borane(2-PB),and separated by amide bonding column,detected by fluorescence detector,and quantitated by internal standard methods.Results The 4 kinds of milk oligosaccharides had a good linear relationship in the concentration range of 10–600μg/mL,and the correlation coefficients(r2)were more than 0.999.The limits of detection and quantification of 4 kinds of milk oligosaccharides were 0.94–2.31 mg/100 g and 3.12–7.69 mg/100 g,respectively;the recovery rates of 4 kinds of milk oligosaccharides were 97.7%–101.5%,and the relative standard deviations(RSD)(n=7)were 0.53%–3.09%.Conclusion This method does not have high requirements for people,equipments and environment for determining the content of 4 kinds of milk oligosaccharides in milk powder.The pre-treatment operation is simple,and it has good accuracy and precision.It can provide a reference for the quality control method of human milk oligosaccharides in milk powder.

关 键 词：2'-岩藻糖基乳糖 3'-唾液酸乳糖 6'-唾液酸乳糖 乳糖-N-新四糖 液相色谱-荧光法 乳粉 

分 类 号：O657[理学—分析化学] TS252.7[理学—化学]