A HILIC-MS/MS method for simultaneous quantification of the lysosomal disease markers galactosylsphingosine and glucosylsphingosine in mouse serum

Rohini Sidhu, Christina R. Mikulka, Hideji Fujiwara, Mark S. Sands, Jean E. Schaffer, Daniel S. Ory, Xuntian Jiang

Research output: Contribution to journalArticlepeer-review

11 Scopus citations

Abstract

Deficiencies of galactosylceramidase and glucocerebrosidase result in the accumulation of galactosylsphingosine (GalSph) and glucosylsphingosine (GluSph) in Krabbe and Gaucher diseases, respectively. GalSph and GluSph are useful biomarkers for both diagnosis and monitoring of treatment effects. We have developed and validated a sensitive, accurate, high-throughput assay for simultaneous determination of the concentration of GalSph and GluSph in mouse serum. GalSph and GluSph and their deuterated internal standards were extracted by protein precipitation in quantitative recoveries, baseline separated by hydrophilic interaction chromatography and detected by positive-ion electrospray mass spectrometry in multiple reaction monitoring mode. Total run time was 7 min. The lower limit of quantification was 0.2 ng/mL for both GalSph and GluSph. Sample stability, assay precision and accuracy, and method robustness were demonstrated. This method has been successfully applied to measurement of these lipid biomarkers in a natural history study in twitcher (Krabbe) mice.

Original languageEnglish
Article numbere4235
JournalBiomedical Chromatography
Volume32
Issue number7
DOIs
StatePublished - Jul 2018

Keywords

  • Gaucher disease
  • Krabbe disease
  • galactosylsphingosine
  • glucosylsphingosine
  • hydrophilic interaction chromatography–tandem mass spectrometry

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