Kate Zhang
Biopharmaceutical Development,USA
Title: A Simple, High-Throughput Method for Analysis of Ceramide, Glucosylceramide, and Ceramide Trihexoside in Dried Blood Spots by LC/MS/MS
Biography
Biography: Kate Zhang
Abstract
A unique monophasic extraction system coupled with LC/MS/MS to reduce matrix effects for sphingolipid analysis was developed. A solvent mixture of methanol, acetonitrile, and water was identified to simultaneously extract multiple sphingolipids with broad polarity range. To reduce matrix effects, the targeted sphingolipids were analyzed by liquid chromatography tandem mass spectrometry (LC/MS/MS). The extraction solvent was used as an isocratic mobile phase in chromatographic separation to eliminate solvent exchange steps and enable high-throughput multiple lipid assay. The assay is linear for ceramide from 0.6 to 9 µg/mL with bias <15%. The intra assay coefficient of variation is less than 10% for concentrations from 1.2 to 9 µg/mL, and less than 25% for concentrations below 1.2 µg/mL. For glucosylceramide and ceramide trihexoside the linear range is 0.05 to 3 µg/mL with biases <10% and <20%, respectively. The intra assay coefficient of variation for these analytes is less than 10% at concentrations from 0.4 to 3 µg/mL, and less than 25% for concentrations below 0.4 µg/mL.