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基于亲水相互作用色谱的固相萃取和基质辅助激光解吸电离飞行时间质谱法用于揭示荧光假单胞菌对鲑鱼片磷脂的影响。

Hydrophilic interaction chromatography based solid-phase extraction and MALDI TOF mass spectrometry for revealing the influence of Pseudomonas fluorescens on phospholipids in salmon fillet.

作者信息

Shen Qing, Yang Qi, Cheung Hon-Yeung

机构信息

Department of Biomedical Sciences, City University of Hong Kong, Tat Chee Avenue, Kowloon Tong, Hong Kong Special Administrative Region, China.

出版信息

Anal Bioanal Chem. 2015 Feb;407(5):1475-84. doi: 10.1007/s00216-014-8365-8. Epub 2014 Dec 10.

Abstract

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 (10(6) 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.

摘要

三文鱼是一种受欢迎的食物,但它很容易因微生物污染而变质。在本研究中,开发了一种基于亲水相互作用色谱(HILIC)的固相萃取(SPE)以及基质辅助激光解吸电离飞行时间/飞行时间质谱联用的方法,并通过测量磷脂酰胆碱和磷脂酰乙醇胺水平的变化,来揭示荧光假单胞菌在保质期内对三文鱼片的影响。将新鲜样品接种荧光假单胞菌(10(6) cfu g(-1))30秒,分别在0、24、48和72小时提取脂质。在通过质谱分析之前,使用装有HILIC吸附剂(用1,2 - 二羟基丙烷衍生化的硅胶)的自制SPE柱进行基质净化。总共检测并鉴定了30种磷脂和16种溶血磷脂。结果表明,磷脂含量显著下降,而溶血磷脂含量最初增加,随后随着冷藏时间的增加逐渐减少。荧光假单胞菌的污染对新鲜三文鱼的品质产生了负面影响,且无明显物理变化,但对人类健康构成了潜在威胁。本研究表明,该成熟方法可用于检测三文鱼片中的磷脂,或许也可用于其他食品。

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