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Updated: Jun 10, 2025
Cellular Lipid Extraction for Targeted Stable Isotope Dilution Liquid Chromatography-Mass Spectrometry Analysis
Published on: November 17, 2011
A lipidomic approach towards identifying the effects of fragrance hydroperoxides on keratinocytes
Aaron I Moore1, Ana S P Moreira2,3, Inês M S Guerra2,3
1School of Pharmacy and Pharmaceutical Sciences, Panoz Institute and Trinity Biomedical Sciences Institute, Trinity College Dublin, The University of Dublin, Dublin, Ireland.
Background:
Limonene and linalool are used in cosmetic products for their floral scents, but their oxidation products are strong contact allergens whose mechanisms of action are still not fully understood.
Objectives:
The effects of limonene hydroperoxide (Lim-2-OOH) and linalool hydroperoxides (Lin-6/7-OOH) on the lipid profile of a human keratinocyte cell line (HaCaT) were evaluated. 2,4-Dinitrofluorobenzene (DNFB) was also included.
Methods:
Lim-2-OOH and Lin-6/7-OOH were synthesised according to previous methods. HaCaT cells were treated with allergens (10 μM) for 24 h and the cellular lipid extracts were analysed by C18 liquid chromatography with tandem mass spectrometry (LC-MS/MS). Data analysis was performed using Lipostar software. Statistical analysis was carried out using Metaboanalyst and R software.
Results:
All three sensitisers used caused significant changes in the lipidome of HaCaT cells in a similar trend. There was an upregulation in several plasmanyl/plasmenyl phospholipids (O-/P-phosphatidylcholines [PC] and O-/P-phosphatidylethanolamines [PE]), sphingolipids (HexCer) and triacylglycerol lipid species, and a decrease in some polyunsaturated fatty acids-containing phospholipid (PE and PC) species suggesting oxidative stress and inflammation.
Conclusions:
This study is the first to evaluate the plasticity of the HaCaT cell lipidome in response to allylic hydroperoxide allergens Lim-2-OOH and Lin-6/7-OOH, together with the experimental contact allergen DNFB. These allergens are able to upregulate and downregulate certain lipid classes to a varying degree.
