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Lipid Analysis Using Supercritical Fluid Technology





              Data                                                                                                                                                                                                                             Subcutaneous Tissue  Mass Imaging of
                                                                                                                                      Supercritical fluids are fluids that are above their critical point. They have
                                                                                                                                      properties of both gases and liquids, such as low viscosity, high diffusivity, and
                  Bioactive lipids with a lysophospholipid skeleton are referred to as lysophospholipid mediators and are attracting attention as   high solubility. Generally, carbon dioxide is used for supercritical fluids.
                  second-generation bioactive lipids. One typical lysophospholipid mediator is sphingosine-1-phosphate. Fingolimod, a drug released   Supercritical fluid extraction (SFE) uses a supercritical fluid to extract               Analysis of
                  in 2011 for treating multiple sclerosis and used as an active ingredient of pharmaceuticals, is presumed to function as a   components, which offers the advantage of easier solvent removal than                              Comprehensive
                  sphingosine-1-phosphate receptor agonist. Lysophosphatidic acid is a lysophospholipid with an extremely simple      conventional solvent extraction methods. Supercritical fluid chromatography                            Glycerophospholipids
                  phosphate-glycerol-fatty acid structure, but an extremely diverse range of its effects have been reported. These include cell   uses supercritical fluid as a mobile phase. Due to properties of supercritical
                  proliferation, cell locomotion, platelet coagulation, and smooth muscle contraction.                                fluids, it achieves higher speed and higher separation for simultaneous analysis.





                                                Peaks                                                                                                                                                                                        Technology  Supercritical Fluid
                                              1. Phosphatidyl Choline                                                                                                                                                                            Lipid Analysis Using
                                              2. Lyso Phosphatidyl Choline
                                              3. Phosphatidyl Ethanolamine
                                              4. Lyso Phosphatidyl Ethanolamine    Dried Blood Spots
                                              5. Phosphatidyl Glycerol
                                              6. Phosphatidyl Inositol
                                                                                                                                            Supercritical Fluid Technology                                                                       Analysis of

                                                                                                                                            Nexera UC improves the analytical workflow by utilizing a completely new separation   Unified Chromatography  Glycerophospholipids
                                                                                                                                            technology, Unified Chromatography, which unites sample separation, analysis with
                                                                                                                                            various separation modes, and high-sensitivity detection.
                                                                                                                                                                                                                         SFC
                                                                                                                                            The Nexera UC on-line SFE-SFC system is a revolutionary system that combines
                                                                                                                                            supercritical fluid extraction (SFE) with supercritical fluid chromatography (SFC). By   LC
                                                                                                                                            automating the process of target compound extraction from solid samples and analysis,   GC       Blood Serum
                      0      2.5     5.0    7.5     10.0    12.5  min                                                                       the system offers a variety of new solutions.                                                      Mediators in Human   Analysis of Lipid
                        Fig. 1   Results from DBS Analysis of Blood Plasma Spiked with Phospholipids  Photo 1   DBS Extraction Vessel
                                                                                                                                            • Automates process from target component extraction to analysis
                                                                                                                                             Automatically extracts components from solid samples using a supercritical fluid and then analyzes them on-line, without any
                                                                                                                                             intervention.
                  Dried Blood Spotting (DBS) is attracting attention even from clinical research or biomarker search fields due to the ease of shipping
                                                                                                                                            • Analyzes unstable compounds without pretreatment
                  specimens. DBS requires using a solvent to extract target molecules from filter paper and then analyzing the extract. However,
                                                                                                                                             Extraction is performed automatically in an environment shielded from light and that inhibits oxidation, which makes it possible
                  manual processing is required for add the solvent and extract the targets, which limits productivity.                                                                                                                        of Glycolipids
                                                                                                                                             to analyze even unstable compounds that were previously decomposed in the pretreatment stage.       Structural Analysis
                  In contrast, analysis using supercritical fluid only requires placing the dried filter into the extraction vessel (Photo 1), with the   • Achieves the ultimate in high speed, high separation, and high sensitivity
                  remaining steps from extraction to analysis performed automatically. Furthermore, extraction is performed automatically in an   Using supercritical fluid provides high-separation, high-sensitivity analysis that allows shorter analysis times for simultaneous
                  environment shielded from light and filled with carbon dioxide to inhibit oxidation, which makes it well suited to analyzing even   analysis of multiple components.
                  unstable compounds.

                  Various phospholipids, including lysophospholipids, were added to blood on DBS, supercritical fluid (SFE) was used to extract them,                                                                                            Analysis of Fatty Acid
                  and then the extract was analyzed by supercritical fluid chromatography-mass spectrometry (SFC-MS). Fig. 1 shows the results and                                                                                           Content of Human ES Cells  Composition in Overall Lipid
                  indicates that six types of phospholipids were separated, including lysophosphatidylcholine. The reverse-phase LC-MS using a C18
                  column enables separation mainly based on differences in the chain length of fatty acids, but not based on differences in polar
                  groups, whereas separation by supercritical fluid chromatography is based on differences in polar groups.




                                                                                                                                                                       Supercritical Fluid Extraction/Chromatograph System                   Esters by GC  Acid Methyl   Analysis of Fatty
                                                                                                                                                                                  Nexera UC

                                                                                                                                            Reference: Nexera UC brochure (C190-E185)




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