여름정기학술대회
2022여름초록
발표자 및 발표 내용
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접수자
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Glycolipids enriched on cell membrane microdomains
play significant roles in cell signaling and cell communication. Experimental
evidence for quantitative changes of specific glycolipids in various
neurological diseases such as Alzheimer's disease, Parkinson's disease, and
epilepsy strongly supports the importance of glycolipids in disease
pathogenesis and disease. Therefore, analytical methods for absolute
quantitation of disease-related target glycolipids are essential for biomarker
discovery and disease monitoring. However, glycolipids are amphiphilic
substances consisting of a hydrophilic carbohydrate chain and a hydrophobic
lipid moiety, which makes their separation and analysis extremely challenging.
Here, we developed a highly sensitive and highly selective absolute
quantitation platform based on reversed-phase LC/MRM-MS. First, the organic
modifier content of the diluent was increased to improve the solubility of
glycolipids, which directly affects data repeatability. We also optimized the
ratio of isopropanol in the mobile phase solvent to enhance the instrument
repeatability and minimize the increase of column pressure due to isopropanol.
As a result, we could determine high sensitivity analysis at the femtomole
level and achieve high repeatability within 8% of the CV. The calibration curve
was quantitatively linear with a correlation coefficient (R2) greater
than 0.99. This analytical method enables the detection of ng levels of glycolipids
per μg of protein in brain tissue. Our glycolipid quantitation platform will be
utilized with a high potential for diagnosis and monitoring of glycolipid-related
diseases
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