CD BioGlyco provides reliable and effective Lipidomics Analysis Services by mass spectrometry (MS). We expect to become customers' research assistants in Sphingolipid Analysis Services.
GD3 is a minor ganglioside present in most normal tissues but plays a critical role in brain development. The expression of GD3 is significantly reduced in adults. However, GD3 expression is increased under pathological conditions, such as cancer and neurodegenerative diseases.
GD3 mediates many functions in cancer through different mechanisms. GD3 expression is upregulated in tumor cells, where it mediates cancer invasion and survival. Interestingly, GD3 also regulates other biological functions such as cell proliferation and differentiation by stimulating the expression of vascular endothelial growth factor (VEGF). GD3 is also found to be closely related to immunity. GD3 is identified as a suppressor of the innate immune response in ovarian cancer, and VEGF enhances GD3-mediated immune suppression. Therefore, GD3 is a drug target for cancer therapy.
Fig.1 GD3 regulates phagocytosis of a stressed neuron following global cerebral ischemia. (Wang, et al., 2021)
CD BioGlyco has developed a complete solution for Disialoganglioside (GD) Analysis Service. We offer a method for the quantitative determination of GD3 in milk and other dairy products and side-stream using multiple reaction monitoring (MRM) ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). This is the quantification of gangliosides by standard addition using advanced UHPLC coupled to triple quadrupole MS. This measurement helps identify processing streams rich in bioactive gangliosides, allowing the dairy industry to strategically capture previously unavailable milk components that could be rapidly adapted into new functional foods.
GD3 is separated from neutral lipids in all samples by two consecutive chloroform and methanol extractions. A liquid milk sample is mixed with methanol:chloroform:water in a certain ratio. A sample of cream and butter and a sample of buttermilk are also dissolved in the solvent solutions. The solutions are sonicated and centrifuged. Subsequently, 2 mL of water is added for phase separation and collect the upper aqueous phase. A mixture of methanol, chloroform, and water is added to the bottom layer for secondary extraction, shaking, and centrifuge. The supernatant is collected and combined with the supernatant from the first extraction. Then the combined solution is dried by a concentrator and the lyophilized sample is resuspended in a methanol-water solution. Gangliosides are enriched using a C8 solid-phase extraction cartridge. The eluant is divided into five aliquots. For quantification, five concentration levels of GD3 standards are added. Samples are dried by the concentrator, and the residue is dissolved in liquid chromatography (LC) starting solvent. One microliter of the sample is injected for MRM analysis.
Reverse phase analysis is performed on LC. A C18 analytical column is used for UHPLC separation. Analytes are detected and quantified using a triple quadrupole mass spectrometer operating in negative electrospray ionization mode.
GD3 Is quantified by the standard addition method. The peak areas of each GD3 in the samples are summed and the sum of the areas is used to draw a regression line to estimate the content of gangliosides in the samples. Calculating the concentration in the original sample by dividing by the dilution factor. Percent recovery is determined by subtracting the amount of ganglioside measured in the spiked sample from the amount of ganglioside measured in the spiked sample and dividing by the amount spiked.
CD BioGlyco has been working on Ganglioside Analysis Service for many years. We provide comprehensive GD3 analysis service through UHPLC-MS/MS method. If you are interested in our services, please contact us for more details without any hesitation.
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