基于SPE-HILIC-QTOF-MS技术鉴定 巴氏杀菌乳中的磷脂成分Identification of phospholipids in pasteurized milk using SPE-HILIC-QTOF-MS

• Published in: Zhongguo you zhi Vol. 50; no. 5; pp. 141 - 149
• Main Author: 顾竞一，李夏冰，廖光琴，王天彩，王子双，贾琪，张星联，钱永忠，邱静 GU Jingyi, LI Xiabing, LIAO Guangqin, WANG Tiancai, WANG Zishuang, JIA Qi, ZHANG Xinglian, QIAN Yongzhong, QIU Jing
• Format: Journal Article
• Language: English
• Published: 中粮工科（西安）国际工程有限公司 01.05.2025
• Subjects: phospholipid; solid-phase extraction; hydrophilic interaction liquid chromatography-quadrupole-time-of-flight-mass spectrometry; 磷脂；固相萃取；亲水作用液相色谱-四极杆飞行时间-质谱
• ISSN: 1003-7969
• DOI: 10.19902/j.cnki.zgyz.1003-7969.240058

为了全面认识巴氏杀菌乳的磷脂组成，并为乳磷脂研究提供数据基础，建立了一种固相萃取(SPE)结合亲水作用液相色谱(HILIC)-四极杆飞行时间（QTOF）-质谱（MS）技术鉴定巴氏杀菌乳中磷脂的方法，对脂质提取、SPE以及MS条件进行优化，并对4种哺乳动物（骆驼、山羊、水牛、奶牛）巴氏杀菌乳磷脂组成进行了分析。结果表明：选取4 mL二氯甲烷-甲醇(体积比2∶ 1)溶剂提取1 mL样品中的脂质，通过10 mL正己烷和10 mL异丙醇-二氯甲烷(体积比3∶ 7)的两步淋洗与2 mL甲醇一步洗脱分离磷脂组分；选择3次迭代采集样品MS/MS信息；在4种哺乳动物巴氏杀菌乳中共鉴定出涵盖7个磷脂亚类的580个磷脂分子，包括116个鞘磷脂、18个心磷脂、181个磷脂酰乙醇胺、262个磷脂酰胆碱、1个磷脂酰甘油、1个磷脂酰肌醇和1个磷脂酰丝氨酸。综上，SPE-HILIC-QTOF-MS分析方法可有效鉴定乳品中的磷脂组成。 In order to comprehensively understand of pasteurized milk phospholipids composition and provide a data base for milk phospholipid research, a solid-phase extraction (SPE) combined with hydrophilic interaction liquid chromatography (HILIC)-quadrupole- time-of-flight (QTOF)-mass spectrometry (MS) technique was developed for the identification of phospholipids in pasteurized milk. Lipid extraction, SPE, and MS conditions were optimized and the phospholipid composition of pasteurized milk from four mammalian species (camel, goat, buffalo and cow) was analyzed. The results showed that 4 mL of dichloromethane-methanol （volume ratio 2∶ 1） was selected to extract the lipid from 1 mL of the sample, and the phospholipid fractions were separated by a two-step washing of 10 mL of n-hexane and 10 mL of isopropanol-dichloromethane （volume ratio 3∶ 7） with a one-step elution of 2 mL of methanol; three iterations were selected to collect the MS/MS information of the sample. A total of 580 phospholipid molecules covering 7 subclasses were identified, including 116 sphingomyelins, 18 cardiolipins, 181 phosphatidylethanolamines, 262 phosphatidylcholines, 1 phosphatidylglycerol, 1 phosphatidylinositol and 1 phosphatidylserine. In conclusion, the SPE-HILIC-QTOF-MS analytical method can effectively identify the composition of phospholipids in dairy products.