title: Quantification of psychosine in the serum of twitcher mouse by LC-ESI-tandem-MS analysis.
abstract: Globoid cell leukodystrophy or Krabbe disease is an inherited autosomal recessive disorder caused by mutations in the galactosylceramidase (GALC) gene. Deficiency of GALC results in the accumulation of a highly cytotoxic metabolite galactosylsphingosine (psychosine). In the present study, we describe the development and validation of a sensitive and specific LC-ESI-tandem-MS method for the determination of psychosine in the serum of twitcher mice, the naturally occurring animal model of this disease. The method was validated in terms of accuracy, precision, specificity, linearity and sensitivity. Calibration plots were linear over the concentration range of 2.5-50ng/mL. Recovery of psychosine from serum was in the range 94.20-98.02%. The results of this study show that in the affected mice the concentration of psychosine (ranging from 2.53 to 33.27ng/mL) increased significantly with the progression of the disease. The maximum level (33.27ng/mL) was detected in the serum of one of the twitcher mice sacrificed at 40PND. Psychosine was not detected at significant levels in wild type mice. These results clearly demonstrate that this noninvasive, rapid, and highly sensitive LC-ESI-tandem-MS method is a very useful approach for the detection of psychosine in serum.
authors.affiliation: Dipartimento di Biotecnologie, Chimica e Farmacia, Università degli Studi di Siena, Via A. Moro, 2, 53100 Siena, Italy.
authors.forename: Assunta
authors.initials: A
authors.lastname: Zanfini
authors.forename: Elena
authors.initials: E
authors.lastname: Dreassi
authors.forename: Anna
authors.initials: A
authors.lastname: Berardi
authors.forename: Laura
authors.initials: L
authors.lastname: Governini
authors.forename: Gianfranco
authors.initials: G
authors.lastname: Corbini
authors.forename: Elvira
authors.initials: E
authors.lastname: Costantino-Ceccarini
authors.forename: Paolo
authors.initials: P
authors.lastname: Balestri
authors.forename: Alice
authors.initials: A
authors.lastname: Luddi
mesh_terms: D000818:Animals; D002138:Calibration; D002853:Chromatography, Liquid / Q000379:methods*; D004195:Disease Models, Animal; D018450:Disease Progression; D007965:Leukodystrophy, Globoid Cell / Q000503:physiopathology*; D051379:Mice; D008810:Mice, Inbred C57BL; D011609:Psychosine / Q000097:blood*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D011609:Psychosine