Development of a Quantitative Method for Epoxyeicosanoids in Mouse Brain Microorgan

User Benefits

- Enables highly sensitive analysis of epoxy-eicosanoids - Enables 15-minute analysis cycle time - Use to search for new biomarkers or explore new physiological functions

Introduction

Epoxy-eicosanoids (epoxy-eicosatrienoic acids or EETs) are known as a group of bioactive lipids derived from arachidonic acid, in addition to prostaglandins and leukotrienes. EETs are formed from arachidonic acid by cytochrome-P450 (CYP). 5(6)-EET has been reported as a lipid mediator with physiological functions such as vasorelaxation. The LC/MS/MS Method Package for Lipid Mediators Ver. 3 enables comprehensive analysis of 196 metabolites of polyunsaturated fatty acids. EETs elute after 18 minutes of the 20-minute linear gradient time specified in the comprehensive method. This article describes the development of a method for quantitative analysis of EETs in tiny mouse brain tissue (organum vasculosum laminae terminalis, or OVLT). A heptacosyl group-based column was used to develop the method, which elutes EETs in 8 to 9 minutes. The peak resolution of EETs was significantly improved by using a curved gradient. Using this method, we were able to quantitatively analyze 5(6)-EET and 8(9)-EET in less than 1 mg of the OVLT of mice brain tissue samples.

April 30, 2021 GMT