In the last 2 decades, lipidomics has become one of the fastest expanding scientific disciplines in biomedical research. With an increasing number of new research groups to the field, it is even more important to design guidelines for assuring high standards of data quality. The Lipidomics Standards Initiative is a community-based endeavor for the coordination of development of these best practice guidelines in lipidomics and is embedded within the International Lipidomics Society. It is the intention of this review to highlight the most quality-relevant aspects of the lipidomics workflow, including preanalytics, sample preparation, MS, and lipid species identification and quantitation. Furthermore, this review just does not only highlights examples of best practice but also sheds light on strengths, drawbacks, and pitfalls in the lipidomic analysis workflow. While this review is neither designed to be a step-by-step protocol by itself nor dedicated to a specific application of lipidomics, it should nevertheless provide the interested reader with links and original publications to obtain a comprehensive overview concerning the state-of-the-art practices in the field.

• Keywords
• LC-MS, MS, chromatography, ion mobility spectrometry, lipid identification, lipidomics, metabolomics, phospholipids, sphingolipids,
• MeSH
• Mass Spectrometry MeSH
• Humans MeSH
• Lipidomics * standards   MeSH
• Lipids analysis   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Review MeSH
• Research Support, N.I.H., Extramural MeSH
• Names of Substances
• Lipids MeSH

Lipids form a significant part of animal organs and they are responsible for important biological functions, such as semi-permeability and fluidity of membranes, signaling activity, anti-inflammatory processes, etc. We have performed a comprehensive nontargeted lipidomic characterization of porcine brain, heart, kidney, liver, lung, spinal cord, spleen, and stomach using hydrophilic interaction liquid chromatography (HILIC) coupled to electrospray ionization mass spectrometry (ESI/MS) to describe the representation of individual lipid classes in these organs. Detailed information on identified lipid species inside classes are obtained based on relative abundances of deprotonated molecules [M-H](-) in the negative-ion ESI mass spectra, which provides important knowledge on phosphatidylethanolamines and their different forms of fatty acyl linkage (ethers and plasmalogens), phosphatidylinositols, and hexosylceramides containing nonhydroxy- and hydroxy-fatty acyls. The detailed analysis of identified lipid classes using reversed-phase liquid chromatography in the second dimension was performed for porcine brain to determine more than 160 individual lipid species containing attached fatty acyls of different acyl chain length, double-bond number, and positions on the glycerol skeleton. The fatty acid composition of porcine organs is determined by gas chromatography with flame ionization detection after the transesterification with sodium methoxide.

• MeSH
• Chromatography, Liquid methods   MeSH
• Chromatography, Gas methods   MeSH
• Phosphatidylethanolamines analysis   MeSH
• Phospholipids analysis   MeSH
• Spectrometry, Mass, Electrospray Ionization methods   MeSH
• Hydrophobic and Hydrophilic Interactions MeSH
• Liver chemistry   MeSH
• Kidney chemistry   MeSH
• Lipids analysis   chemistry   MeSH
• Fatty Acids analysis   MeSH
• Spinal Cord chemistry   MeSH
• Brain Chemistry MeSH
• Myocardium chemistry   MeSH
• Plasmalogens analysis   MeSH
• Lung chemistry   MeSH
• Swine MeSH
• Spleen chemistry   MeSH
• Stomach chemistry   MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Phosphatidylethanolamines MeSH
• Phospholipids MeSH
• Lipids MeSH
• Fatty Acids MeSH
• Plasmalogens MeSH