A novel approach for quantitation of glucosylceramide in human dried blood spot using LC-MS/MS.
Allena Ji Ji, Haixing Wang, Enida Ziso-Qejvanaj, Kefei Zheng, Lee Lee Chung, Timothy Foley, Wei-Lien Chuang, Susan Richards, Crystal Sung
Index: Bioanalysis 7 , 1483-96, (2015)
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Abstract
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.
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