Shimadzu Europa:
Unique LC/MS/MS Method Package for Lipid Mediators

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  • Published: Mar 17, 2014
  • Categories: Base Peak
thumbnail image: <font size=3>Shimadzu Europa</font>:<br />Unique LC/MS/MS Method Package for Lipid Mediators

In the last decade, comprehensive monitoring of lipid mediators using LC/MS has been developed as key technology for disease studies or biomarker exploring. Designed especially for lipid mediator profiling analysis in various biological samples, Shimadzu’s LC/MS/MS method package provides a valuable tool for establishing this type of analysis. The method package offers a complete solution, from sample pretreatment to data acquisition including separation conditions, MRM optimization, MS parameter settings and quantitative analysis.

Lipidomics is a relatively new research field and is engaged in identification, characterization and quantitation of cellular lipids and their metabolic pathways in biological systems. The word lipidome is used to describe the highly complex complete lipid profile within a cell, tissue or organism. The analysis of lipids and lipid mediators, meaning biological active lipids e.g. as messengers, is of growing interest due to their role in lifestyle related diseases such as hypertension, diabetes, obesity, arteriosclerosis and allergy diseases.

The major lipid mediators are eicosanoids such as prostaglandins, leukotrienes and recently discovered resolvin D1, which result from arachidonic acid and are associated with various physiological and pathological roles (Figure: Lipid mediator and physiology). Due to the same origin, lipid mediators and their related substances have a similar chemical structure, resulting in same MRM transitions and often the same retention times as well.

Optimized HPLC conditions for 130 components

Method optimization for these substances requires plenty of time. The LC/MS/MS method package contains optimized HPLC conditions for 130 components, so even prostaglandins (PGA2 and PGJ2), isoprostanes and stereoisomers of leukotrienes, all of which show exactly the same MRM transitions can be separated chromatographically.

For analysis of fatty acids, ESI negative ionization is commonly used for measurement, but in the case of special leukotrienes, ethanolamides and lactones ESI positive ionization is preferred because of better sensitivity. Traditional LCMS is limited to one polarity but thanks to Shimadzu’s UFMS technology and the high speed polarity switching (5 msec) of the Shimadzu LCMS-8050, it is possible to analyze both polarities in a single run, achieving the best sensitivity for every compound.

New injection method

The two major metabolites of prostaglandine in urine, tetranor PGEM and tetranor PGDM, are highly polar and show worse peak shape when injected by a conventional injection autosampler. A new injection method named Co-Injection was developed especially for this method package, reducing peak-shape related problems and allowing batch analysis also for highly polar compounds.

For quantitative analysis the 130 components of the method package are divided into 13 groups according to their physical properties and an appropriate internal standard is selected for each group. In this way it is possible to correct quantitative value and retention time so as to support accurate peak identification.

Shimadzu’s unique LC/MS/MS Method Package provides all necessary knowledge for performing the complicated analysis of lipid mediators, including new injection method for polar compounds (Co-Injection), separation conditions, MRM optimization and measurement parameter settings.

RUO: Research Use Only

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