采用脉冲安培检测的高效阴离子交换色谱法(HPAEC-PAD)测定2'-岩藻糖基乳糖(2'-FL)、3-岩藻糖基乳糖(3-FL)、6'-唾液乳糖(6'-SL)、3'-唾液乳糖(3'-SL)、乳糖-N-四糖(LNT)和乳糖-N-新四糖(LNnT)的方法:第一步2022.04。

IF 1.7 4区 农林科学 Q3 CHEMISTRY, ANALYTICAL Journal of AOAC International Pub Date : 2023-09-01 DOI:10.1093/jaoacint/qsad072
Philip Haselberger, Fang Tian, Renée Erney, Shuang Liu, Shuo Wang, Qi Lin, Yi Ding
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引用次数: 0

摘要

背景:介绍了一种同时测定六种母乳低聚糖(HMO)的方法。HMO包括2'-岩藻糖基乳糖(2'-FL,CAS编号41263-94-9)、3-岩藻糖基乳糖(3-FL,CAS编号41312-47-4)、6'-唾液乳糖(6'-SL,CAS编号35890-39-2)、3'-唾液乳糖(3'-SL,CAS编号35890-38-1)、乳糖-N-四糖(LNT,CAS编号14116-68-8)和乳糖-N-新四糖(LNnT,CAS编号13007-32-4)。该方法的设计分别符合各自的标准方法性能要求(SMPR®;见表1)。目的:该方法对婴儿配方奶粉和成人营养基质中的六种HMO有效,包括含有完整蛋白质的样品、蛋白质水解物、不含完整蛋白质的元素配方和SMPR中定义范围内的米粉(见表2)。该方法不适用于二葡糖醛酸酶(DFL/DiFL)的测定。方法:对于大多数样品,用水重建,然后过滤。对于含有干扰物质(果聚糖和麦芽糊精)的产品,使用酶水解。制备后,使用脉冲安培检测的高效阴离子交换色谱法(HPAEC-PAD)分析样品。该方法可分离婴儿配方奶粉和成人营养产品中常见的六种HMO和其他碳水化合物(如乳糖、蔗糖和GOS)。结果:该研究包括来自全球多个实验室评估的多个基质的数据。RSDr在0.0068至4.8%之间,刺突回收率在89.4%至109%之间。用二次曲线进行最佳校准拟合;交替线性拟合对数据没有显示出统计学上的显著影响(当相关性通过时)。结论:AOAC SPIFAN专家审查小组(ERP)对该方法进行了审查,并确定该方法符合六个著名HMO的SMPR。亮点:该方法被授予第一行动官方方法SM状态。
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Method for the Determination of 2'-Fucosyllactose (2'-FL), 3-Fucosyllactose (3-FL), 6'-Sialyllactose (6'-SL), 3'-Sialyllactose (3'-SL), Lacto-N-Tetraose (LNT), and Lacto-N-neoTetraose (LNnT) by High-Performance Anion-Exchange Chromatography With Pulsed Amperometric Detection (HPAEC-PAD): First Action 2022.04.

Background: A method for simultaneous determination of six human milk oligosaccharides (HMOs) is described. The HMOs include 2'-fucosyllactose (2'-FL, CAS number 41263-94-9), 3-fucosyllactose (3-FL, CAS number 41312-47-4), 6'-sialyllactose (6'-SL, CAS number 35890-39-2), 3'-sialyllactose (3'-SL, CAS number 35890-38-1), lacto-N-tetraose (LNT, CAS number 14116-68-8), and lacto-N-neotetraose (LNnT, CAS number 13007-32-4). The method was designed to comply with the respective Standard Method Performance Requirements (SMPR®; seeTable 1), respectively.

Objective: The method is valid for six HMOs in infant formula and adult nutritional matrixes, including samples with intact protein, protein hydrolysates, elemental formulations free of intact protein, and rice flour over the ranges defined in the SMPR (seeTable 2). The method is not valid for determination of difucosyllactose (DFL/DiFL).

Method: For most samples, reconstitution with water followed by filtration. For products containing interferences (fructans and maltodextrins), hydrolysis with enzymes is used. After preparation, samples are analyzed using high-performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD). The method provides for separation of six HMOs and other carbohydrates commonly found in infant formula and adult nutritional products (e.g., lactose, sucrose, and GOS).

Results: This study includes data from multiple matrixes evaluated by multiple laboratories globally. RSDr ranged from 0.0068 to 4.8% RSDr, and spike recovery results ranged from 89.4 to 109%. Calibration fit optimal with a quadratic curve; alternately linear fit showed no statistically significant impact to data (when correlation passes).

Conclusions: This method was reviewed by the AOAC SPIFAN Expert Review Panel (ERP) and determined to meet the SMPRs for the six noted HMOs.

Highlights: The method was granted First Action Official MethodsSM status.

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来源期刊
Journal of AOAC International
Journal of AOAC International 医学-分析化学
CiteScore
3.10
自引率
12.50%
发文量
144
审稿时长
2.7 months
期刊介绍: The Journal of AOAC INTERNATIONAL publishes the latest in basic and applied research in analytical sciences related to foods, drugs, agriculture, the environment, and more. The Journal is the method researchers'' forum for exchanging information and keeping informed of new technology and techniques pertinent to regulatory agencies and regulated industries.
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