Quantification of free fatty acids in human stratum corneum using tandem mass spectrometry and surrogate analyte approach
Quantification of free fatty acids in human stratum corneum using tandem mass spectrometry and surrogate analyte approach
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DOI：
10.1002/bmc.4056
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发表时间：
2018-02-01
影响因子：
1.8
通讯作者：
Jakasa, Ivone
中科院分区：
文献类型：
--
作者：
Dapic, Irena;Kobetic, Renata;Jakasa, Ivone
游离脂肪酸（FFA）是皮肤最上层角质层（SC）中脂质的主要成分之一。FFAs的相对组成已被提议作为特应性皮炎（AD）患者皮肤屏障状态的生物标志物。在这里，我们开发了一种LC-ESI-MS/MS方法，用于同时定量的范围内的FFAs与长链和非常长的链长的SC收集胶带（D-Squame）。该方法，基于衍生化与2-溴-1-甲基吡啶碘化物和3-甲醇-1-甲基吡啶碘化物，允许高灵敏度的检测和定量的FFA使用多反应监测。对于定量，我们应用了替代分析物方法和使用同位素标记的FFA衍生物的内部标准化。胶带显示存在几种FFA，这些FFA也存在于SC中，这是以前研究中遇到的问题。因此，使用C12：0校正SC中的FFA水平，C12：0存在于胶带上，但在SC中未检测到。该方法应用于特应性皮炎患者和健康受试者的SC样品。使用多反应监测的定量允许足够的灵敏度来分析仅在一个胶带条上收集的SC中的链长C16-C28的FFA。
The free fatty acids (FFAs) are one of the major components of the lipids in the stratum corneum (SC), the uppermost layer of the skin. Relative composition of FFAs has been proposed as a biomarker of the skin barrier status in patients with atopic dermatitis (AD). Here, we developed an LC-ESI-MS/MS method for simultaneous quantification of a range of FFAs with long and very long chain length in the SC collected by adhesive tape (D-Squame). The method, based on derivatization with 2-bromo-1-methylpyridinium iodide and 3-carbinol-1-methylpyridinium iodide, allowed highly sensitive detection and quantification of FFAs using multiple reaction monitoring. For the quantification, we applied a surrogate analyte approach and internal standardization using isotope labeled derivatives of FFAs. Adhesive tapes showed the presence of several FFAs, which are also present in the SC, a problem encountered in previous studies. Therefore, the levels of FFAs in the SC were corrected using C12:0, which was present on the adhesive tape, but not detected in the SC. The method was applied to SC samples from patients with atopic dermatitis and healthy subjects. Quantification using multiple reaction monitoring allowed sufficient sensitivity to analyze FFAs of chain lengths C16-C28 in the SC collected on only one tape strip.