Profiling and Qualitative Assessment of Enzymatically and Thermally Oxidized Egg Yolk Phospholipids using a Two‐Step High‐Performance Liquid Chromatography Protocol
Profiling and Qualitative Assessment of Enzymatically and Thermally Oxidized Egg Yolk Phospholipids using a Two‐Step High‐Performance Liquid Chromatography Protocol
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DOI：
10.1002/aocs.12218
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发表时间：
2019-06
期刊：
影响因子：
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通讯作者：
K. Parchem;B. Kusznierewicz;T. Chmiel;Paulina Maciołek;A. Bartoszek
中科院分区：
文献类型：
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作者：
K. Parchem;B. Kusznierewicz;T. Chmiel;Paulina Maciołek;A. Bartoszek
建立了食品中氧化磷脂（oxPL）的两步高效液相色谱（HPLC）分析和定性评价方法。验证了两步高效液相色谱法分离酶和热氧化蛋黄磷脂作为相关食品模型的适用性。第一步，使用亲水性相互作用液相色谱（HILIC）分离7个单独的PL类。将收集到的两种主要的蛋黄PL类——磷脂酰乙醇胺和磷脂酰胆碱的组分进一步进行第二步分离，目的是分析和定性评价它们的氧化形态。为此，采用了反相色谱法（RP）耦合带电气溶胶，紫外检测（UV）和质谱检测。建立了一个潜在的氧化产物数据库，包括初级氧化产物（氢过氧化物）和长链次级氧化产物（环氧化物、醇类和酮类）以及它们的一些可能组合。此外，还将结果与N，N -二甲基-对苯二胺可视化薄层色谱（TLC）得到的PL氢过氧化物谱进行了比较。
A two‐step high‐performance liquid chromatography (HPLC) method for the profiling and qualitative assessment of oxidized phospholipids (oxPL) present in foods was developed. The applicability of the investigated two‐step HPLC protocol was verified for separation of enzymatically and thermally oxidized hen egg yolk phospholipids (PL) as a relevant food model. In the first step, seven individual PL classes were separated using hydrophilic interaction liquid chromatography (HILIC). The collected fractions of two main egg yolk PL classes—phosphatidylethanolamines and phosphatidylcholines—were further directed to the second step of separation aimed at profiling and qualitative assessment of their oxidized species. For this purpose, the reverse phase (RP) chromatography coupled to charged aerosol, ultraviolet detection (UV) and mass spectrometry detection were employed. A database of potential oxPL including primary (hydroperoxides) and long‐chain secondary PL oxidation products (epoxides, alcohols, and ketones) as well as some of their possible combinations was created. Additionally, the results were compared with the profiles of PL hydroperoxides obtained using thin layer chromatography (TLC) with N,N‐dimethyl‐p‐phenylenediamine visualization.