离子色谱法测定母乳中的寡聚糖与游离唾液酸
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Determination of Oligosaccharides and Free Sialic Acid in Human Milk by Ion Chromatography
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    摘要:

    母乳寡聚糖(HMOs)是母乳中第三大营养素,其含量仅次于乳糖与脂肪,是婴儿健康成长的重要营养来源。本研究采用离子色谱法(IC)建立并验证了测定母乳中6种主要母乳寡聚糖含量的方法,包括3-岩藻糖基乳糖(3-FL)、2’-岩藻糖基乳糖(2’-FL)、乳糖-N-新四糖(LNnT)、乳糖-N-四糖(LNT)、6’-唾液酸乳糖(6’-SL)及3'-唾液酸乳糖(3’-SL)。另外,母乳中另一种活性成分唾液酸(Neu5Ac)也可以同时测定。将母乳样用超纯水稀释,微孔滤膜过滤后进样分析。以水-氢氧化钠溶液-乙酸钠溶液为流动相,选用CarboPacTMPA-1分析柱进行色谱分离,用脉冲安培检测器检测。所测7种分析组分在该试验条件下能很好分离,且在较宽的质量浓度范围内与其峰面积呈良好的线性关系(r>0.999)。方法的检出限(3 s/n)分别为5.2,0.4,2.8,2.5,1.0,1.8,0.5 mg/L母乳,定量限分别为19.8,9.8,19.9,19.7,9.9,9.9,5.0 mg/L母乳。通过测定分泌型与非分泌型的3段乳得出该方法的线性范围与定量限能够满足各类母乳样品的需求。以母乳作为基质,测得各分析组分的加标回收率(n=6)在91.8%~106.1%之间,母乳基质与加标样测定值的相对标准偏差(n=6)小于3.7%。本方法样品处理简单快速,具有较低的检出限与定量限,较宽的线性范围,良好的重现性与回收率,适用于母乳样品中6种主要寡聚糖与游离唾液酸的准确测定。

    Abstract:

    After lactose and fat, human milk oligosaccharides (HMOs) are the third most abundant nutrient in human milk. They are very important for the healthy growth of infants. A quantitative method has been developed and validated for the determination of 6 main HMOs in human milk, including 3-fucosyllactose (3-FL), 2’-fucosyllactose (2’-FL), lacto-N-neotetraose (LNnT), lacto-N-tetraose (LNT), 6’-sialyllactose (6’-SL) and 3’-sialyllactose (3’-SL), by ion chromatography(IC). In addition, another active compound sialic acid (Neu5Ac) in human milk could be determined at the same time. A portion of human milk sample was diluted by water and subjected to IC analysis after filtration. Water, sodium hydroxide solution, and sodium acetate solution were used as mobile phase. CarboPacTM PA-1 analytical column and pulsed amperometric detector (PAD) were adopted for IC separation and detection, respectively. The method validation results showed a good linearity(r>0.999) in the analytical range for all the 7 analytes. The limit of detection (LOD, 3 s/n) and limit of quantitation(LOQ) of the method for each analyte were 5.2, 0.4, 2.8, 2.5, 1.0, 1.8, 0.5 mg/L human milk and 19.8, 9.8, 19.9, 19.7, 9.9, 9.9, 5.0 mg/L human milk, respectively. The analytical range and LOQ meet detection requirements of all types of human milk samples including the secretory and non-secretory three-stage human milk. The recovery of each analyte in human milk were all within 90.3%-107.3% (n=6) with a good precision (RSD%<3.7%, n=6). This method provides a simple and fast approach for the accurate determination of HMOs and free sialic acid in human milk.

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陈磊;PhilipHaselberger;田芳;蔡小堃;毛颖异;刘爽;赵艳荣;王硕;关岩;.离子色谱法测定母乳中的寡聚糖与游离唾液酸[J].中国食品学报,2019,19(10):227-234

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  • 在线发布日期: 2019-11-21
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