A Novel Approach for Quantitation of Glucosylceramide in Human Dried Blood Spot Using LC–MS/MS

• DOI: 10.4155/bio.15.77
• Publication Date: 2015-07-13T15:01:48Z
• Authors: Allena Ji Ji; Haixing Wang; Enida Ziso-Qejvanaj; Kefei Zheng; Lee Lee Chung; Timothy Foley; Wei-Lien Chuang; Susan Richards; Crystal Sung
• Keywords: Quality Control; Gaucher Disease; Tandem Mass Spectrometry; Humans; Dried Blood Spot Testing; Glucosylceramides; 01 natural sciences; Biomarkers; Chromatography, High Pressure Liquid; 0104 chemical sciences

Glucosylceramide, an efficacy biomarker for Gaucher Type 1 disease, exhibits poor solubility in polar solvents and whole blood which makes it difficult to prepare a homogenous blood standard.We developed a novel method using standard addition approach by spiking a small volume of analyte solution on the surface of prespotted dried blood spot. The whole spots were punched out for subsequent extraction and LC-MS/MS analysis. The assay performance met all validation acceptance criteria. Glucosylceramide concentrations in 50 paired plasma and dry blood spot samples obtained from Gaucher Type 1 patients were tested and the results demonstrated the feasibility of using the DBS method for clinical biomarker monitoring.The new approach greatly improves assay precision and accuracy.