Ben Figard (Presenter)
Shimadzu Scientific Instruments
Authorship: Masaki Yamada(1), Tairo Ogura(2), Chris Gilles (2), Scott Kuzdzal(2), Jorge Smith(3) and Ben Figard (3)
(1) Global Applications Development Group, Shimadzu Corporation, Kyoto, Japan. (2) Shimadzu Scientific Instruments, Columbia, Maryland. (3) Shimadzu Scientific Instruments, Houston, Texas
| Short Abstract Lipid mediators are bioactive lipids which play a role in many biological functions, including immune dysfunction after severe inflammation. Recent development of a high sensitivity ultra-fast mass spectrometer enables lower detection limits for lipid mediator species. A comprehensive and highly sensitive application for the analysis of lipid mediators and their metabolites using a triple quadrupole mass spectrometer is presented here. Method conditions were developed to simultaneously analyze 158 lipid mediator-related compounds. A single chromatographic analysis is capable of separating a wide range of positive and negative species such as hydrophilic metabolites, tetranor-PGs and hydrophobic arachidonic acid. |
Long Abstract
Lipid mediators are bioactive lipids which play a role in many biological functions, including immune dysfunction after severe inflammation. Recent development of a high sensitivity ultra-fast mass spectrometer enables lower detection limits for lipid mediator species. A comprehensive and highly sensitive application for the analysis of lipid mediators and their metabolites using a triple quadrupole mass spectrometer is presented here. Method conditions were developed to simultaneously analyze 158 lipid mediator-related compounds. A single chromatographic analysis is capable of separating a wide range of positive and negative species such as hydrophilic metabolites, tetranor-PGs and hydrophobic arachidonic acid (Fig. 1).
The method was applied to analysis of lipid extracts from human sera. One mL of methanol and 10 ìL of internal standard mixture were added to 200 ìL of human serum and mixed for three minutes. After centrifugation, solid phase extraction (SPE) was .conducted following elution with 300 ìL of methanol An analysis of a 10-fold concentrated SPE eluent resulted in the detection of 85 peaks with signal-to noise ratio (S/N) of greater than 3 and a retention time difference of less than 1 second from primary standards. Internal standard quantitation showed that the concentrations of 5-HETE, the most abundant of all metabolites, and 12-HHT, the least abundant, were 1 ìM and 0.5 nM, respectively. 8-iso-PGF2á, an oxidative stress marker, was 0.1 nM. Replicate analyses in human sera were performed for three example analytes. Results indicate that triple quadrupole MS is capable of quantitative profiling in a wide dynamic range from sub-nM to ìM. Because of the potential for interfering peaks, MRM analysis often requires confirmation ions. This technique enables ultra-fast MRM analysis, allowing the acquisition of multiple MRM transitions while maintaining high data quality.
References & Acknowledgements:
| Description | Y/N | Source |
| Grants | no | |
| Salary | yes | Shimadzu |
| Board Member | no | |
| Stock | no | |
| Expenses | no |
IP Royalty: no
| Planning to mention or discuss specific products or technology of the company(ies) listed above: | yes |