Hydrophilic interaction chromatography based solid-phase extraction and MALDI TOF mass spectrometry for revealing the influence of Pseudomonas fluorescens on phospholipids in salmon fillet
Hydrophilic interaction chromatography based solid-phase extraction and MALDI TOF mass spectrometry for revealing the influence of Pseudomonas fluorescens on phospholipids in salmon fillet
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DOI：
10.1007/s00216-014-8365-8
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发表时间：
2015-02-01
影响因子：
4.3
通讯作者：
Cheung, Hon-Yeung
中科院分区：
文献类型：
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作者：
Shen, Qing;Yang, Qi;Cheung, Hon-Yeung
鲑鱼是一种受欢迎的食物，但它很容易受到微生物污染而变质。本研究建立了一种基于亲水相互作用色谱（HILIC）的固相萃取（SPE）和基质辅助激光解吸电离飞行时间/飞行时间质谱技术，通过测定鲑鱼鱼片中磷脂酰胆碱和磷脂酰乙醇胺的含量变化，揭示了荧光假单胞菌对鲑鱼鱼片保质期的影响。新鲜样品用荧光假单胞菌（106 cfu g(-1)）接种30 s，在0、24、48和72 h提取脂质。在质谱分析之前，使用自制的固相萃取盒，包装HILIC吸附剂（1,2-二羟基丙烷衍生的二氧化硅）进行基质清理。共检测和鉴定了30种磷脂和16种溶血磷脂。结果表明，随着冷藏时间的延长，磷脂含量显著降低，溶血磷脂含量先升高，后逐渐降低。荧光假单胞菌污染对新鲜鲑鱼的品质没有明显的生理变化，但对人体健康有潜在的威胁。这项研究表明，这种行之有效的方法可以用于检测鲑鱼鱼片中的磷脂，也许也可以用于其他食物。
Salmon is a popular food but it is easily susceptible to spoilage by contamination with microorganisms. In this study, a method using hydrophilic interaction chromatography (HILIC)-based solid-phase extraction (SPE) and matrix-assisted laser desorption and ionization time-of-flight/time-of-flight mass spectrometry was developed and applied to reveal the effect of Pseudomonas fluorescens on salmon fillet during the shelf-life period by measuring the changes in the levels of phosphatidylcholine and phosphatidylethanolamine. Fresh samples were inoculated with P. fluorescens (106 cfu g(-1)) for 30 s, and lipids were extracted at 0, 24, 48, and 72 h. A homemade SPE cartridge packed with HILIC sorbent (silica derivatized with 1,2-dihydroxypropane) was used for matrix cleanup prior to analysis by mass spectrometry. In total, 30 phospholipids and 16 lysophospholipids were detected and elucidated. The results revealed that the content of phospholipids decreased significantly, whereas that of lysophospholipids increased initially, followed by a gradual reduction as the cold storage time increased. The contamination by P. fluorescens negatively affected the quality of fresh salmon without obvious physical changes, but it posed a potential threat to human health. This study suggests that the well-established method could be used for detecting phospholipids in salmon fillet and perhaps other foods as well.