食品科学 ›› 2021, Vol. 42 ›› Issue (4): 181-189.doi: 10.7506/spkx1002-6630-20190623-273

• 成分分析 • 上一篇    下一篇

基于UPLC-Triple-TOF MS/MS对巴氏杀菌乳中磷脂成分的分析

曹雪,任皓威,王筱迪,许铵颀,商佳琦,刘宁   

  1. (乳品科学教育部重点实验室,东北农业大学食品学院,黑龙江?哈尔滨 150030)
  • 出版日期:2021-02-25 发布日期:2021-02-25
  • 基金资助:
    “十三五”国家重点研发计划重点专项(2018YFC1604304-01)

Identifying and Quantifying Phospholipids in Pasteurized Milk Using UPLC-Triple-TOF MS/MS

CAO Xue, REN Haowei, WANG Xiaodi, XU Anqi, SHANG Jiaqi, LIU Ning   

  1. (Key Laboratory of Dairy Science, Ministry of Education, College of Food Science, Northeast Agricultural University, Harbin 150030, China)
  • Online:2021-02-25 Published:2021-02-25

摘要: 基于超高效液相色谱-串联四极杆飞行时间高分辨质谱(ultra-high performance liquid chromatography-triple quadrupole-time of flight tandem mass spectrometry,UPLC-Triple-TOF MS/MS)法对巴氏杀菌乳中的磷脂成分进行分析鉴定。采用色谱柱ACQUITY UPLC? BEH C18(2.1 mm×100 mm,1.7 μm),以流动相A水-甲醇-乙腈(1∶1∶1,V/V)和流动相B异丙醇-乙腈(1∶1,V/V)进行梯度洗脱,在电喷雾电离正模式下采集数据。依据高分辨质谱提供的保留时间、准分子离子、二级碎片等脂质分析数据,并结合标准品碎裂方式等信息,共鉴定出磷脂分子68 种。其中,磷脂酰胆碱22 种,占比7.98%;磷脂酰乙醇胺17 种,占比56.60%;磷脂酰丝氨酸10 种,占比1.70%;磷脂酰肌醇7 种,占比1.33%;鞘磷脂12 种,占比32.39%。本研究采用灵敏度高、精密度及稳定性良好的UPLC-Triple-TOF MS/MS方法实现了对牛乳中磷脂的全面定性与定量分析。此方法检测出的牛乳磷脂信息完整、磷脂种类多于其他检测技术,可为牛乳的磷脂研究提供数据基础。

关键词: 巴氏杀菌乳;磷脂;磷脂组学;超高效液相色谱-串联四极杆飞行时间质谱

Abstract: The phospholipids in pasteurized milk samples were analyzed and identified by ultra-high performance liquid chromatography-triple quadrupole-time of flight tandem mass spectrometry (UPLC-Triple-TOF MS/MS). The chromatographic separation was performed on an ACQUITY UPLC? BEH C18 (2.1 mm × 100 mm, 1.7 μm) column using gradient elution with water/methanol/acetonitrile (1:1:1, V/V) as mobile phase A and isopropanol/acetonitrile (1:1, V/V) as mobile phase B. Data acquisition was done in the positive ion mode using electrospray ionization (ESI). According to the high-resolution mass spectral data of retention time, quasi-molecular ions and secondary fragments combined with information on the mass spectral fragmentation pathways of reference standards, a total of 68 phospholipid molecules were identified including 22 phosphatidylcholine (PC, 7.98%), 17 phosphatidylethanolamine (PE, 56.60%), 10 phosphatidylserine (PS, 1.70%), 7 phosphatidylinositol (PI, 1.33%), and 12 sphingomyelin (SM, 32.39%). This study achieved comprehensive qualitative and quantitative determination of phospholipids in milk by using the UPLC-Triple-TOF MS/MS method with high sensitivity, precision and stability. This method could detect more phospholipids in milk than other detection techniques and thus provide more basic data for research on milk phospholipids.

Key words: pasteurized milk; phospholipid; phospholipidomics; ultra-high perfomance liquid chromatography-triple quadrupole-time of flight tandem mass spectrometry

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