食品科学 ›› 2019, Vol. 40 ›› Issue (16): 152-159.doi: 10.7506/spkx1002-6630-20181008-039

• 成分分析 • 上一篇    下一篇

羊乳低聚糖超高效液相色谱-质谱鉴定中样品前处理方法及液相色谱条件优化

张 艳,芦 晶,张书文,逄晓阳,王军辉,吕加平   

  1. 1.合肥工业大学食品与生物工程学院,安徽 合肥 230009;2.中国农业科学院农产品加工研究所,北京 100193
  • 出版日期:2019-08-25 发布日期:2019-08-26
  • 基金资助:
    国家自然科学基金面上项目(31671878);国家自然科学基金青年科学基金项目(31401514);教育部留学回国人员科研启动经费项目;现代农业产业技术体系北京市奶牛产业创新团队项目;“十三五”国家重点研发计划重点专项(2018YFD0400900)

Optimization of Sample Pretreatment and Chromatographic Separation Conditions for Identification of Goat Milk Oligosaccharides by Ultra Performance Liquid Chromatography-Q-Exactive Focus Tandem Mass Spectrometry

ZHANG Yan, LU Jing, ZHANG Shuwen, PANG Xiaoyang, WANG Junhui, Lü Jiaping   

  1. 1. School of Food and Biological Engineering, Hefei University of Technology, HeFei 230009, China; 2. Institute of Agro-Products Processing Science and Technology, Chinese Academy of Agricultural Sciences, Beijing 100193, China
  • Online:2019-08-25 Published:2019-08-26

摘要: 通过Sep-Pak C18与石墨化碳柱法、超滤法、透析法和葡萄糖凝胶层析法4 种不同的前处理方法,以离子交换色谱法检测乳糖的去除率和低聚糖的损失率为评价指标,最终确定葡聚糖凝胶层析法为最优前处理方法。然后,采用超高效液相色谱-串联Q-Exactive Focus质谱对羊乳中的低聚糖进行分析及结构鉴定,以低聚糖的总离子流图为评价指标,对液相洗脱条件进行优化,最终优化条件为:0~20 min,95%~78% A(乙腈);20~35 min,78%~73% A;35~38 min,73~62% A;38~45 min,62%~50% A。结果显示,采用凝胶层析法前处理样品测得低聚糖种类显著高于未去除乳糖的样品,在最优液相色谱分离条件下,羊乳低聚糖中同分异构体得到较好分离,对羊乳中低聚糖的结构解析具有重要参考价值。

关键词: 羊乳, 低聚糖, 超高效液相色谱-串联Q-Exactive Focus质谱

Abstract: Four different sample pretreatment procedures, sequential chromatography on Sep-Pak C18 and graphitized carbon columns, ultrafiltration, dialysis and Sephadex-G10 gel column chromatography, were used to purify oligosaccharides to remove lactose from crude extracts. Ion exchange chromatography was used to determine the removal of lactose and the loss of oligosaccharides. The Sephadex-G10 method was shown to be the best of the four methods. Oligosaccharides in goat milk were identified and structurally analyzed by ultra performance liquid chromatography-Q-Exactive Focus tandem mass spectrometry. The total ion current (TIC) chromatogram was recorded, and based on it, the gradient elution program was determined as follows: 0–20 min, 95%–78% A (acetonitrile); 20–35 min, 78%–73% A; 35–38 min, 73%–62% A; 38–45 min, 62%–50% A. A significantly larger number of oligosaccharides were identified in the samples purified by gel filtration chromatography than the crude extract samples. The oligosaccharide isomers in goat milk were separated effectively under the optimal chromatographic conditions. This method could be considered as an important tool for structural analysis of goat milk oligosaccharides.

Key words: goat milk, oligosaccharides, ultra performance liquid chromatography-Q-Exactive Focus tandem mass spectrometry

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