Novel procedure for measuring psychosine derivatives by an HPLC method.
We developed a sensitive and simple method to determine galactosylsphingosine and glucosylsphingosine as a 4-fluoro-7-nitrobenzofurazan autofluorescent compound, using HPLC equipped with a Showdex sugar column. Amounts of galactosylsphingosine were successfully measured in the picomole range. This novel procedure is more stable and simpler than the previous method using o-phthalaldehyde. It was applied to tissues from the twitcher mouse, an animal model of human globoid cell leukodystrophy. The amount of galactosylsphingosine was 34-102 micrograms/kg of wet tissues in control cerebrum and cerebellum, whereas in twitcher mice the range was 2,251-4,228 micrograms/kg of wet tissues. The psychosine concentration was also increased in the liver and kidney of twitcher mice, respectively, 1,513 micrograms and 1,106 micrograms/kg of wet tissue (normal liver, 125 micrograms; normal kidney, 74 micrograms/kg of wet tissue). This novel procedure is useful for the pathochemical evaluation of lysosphingolipids in various sphingolipidoses as well as in other neuropathological and cellular conditions.
Nozawa
M
M
Department of Pediatrics, Tokyo Jikei University School of Medicine, Japan.
Iwamoto
T
T
Tokoro
T
T
Eto
Y
Y
D009327:4-Chloro-7-nitrobenzofurazan / Q000031:analogs & derivatives; D000818:Animals; D002531:Cerebellum / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D007668:Kidney / Q000737:chemistry; D008099:Liver / Q000737:chemistry; D051379:Mice; D011609:Psychosine / Q000031:analogs & derivatives* / Q000032:analysis / Q000378:metabolism
D011609:Psychosine; C032930:7-fluoro-4-nitrobenzo-2-oxa-1,3-diazole; C035742:sphingosyl beta-glucoside; D009327:4-Chloro-7-nitrobenzofurazan