Synthetic sulfogalactosylceramide (sulfatide) and its use for the mass spectrometric quantitative urinary determination in metachromatic leukodystrophies

Abstract

3-O-Sulfogalactosylceramides (sulfatides) accumulate in the genetic disease metachromatic leukodystrophy which is due to a defect in the catabolic enzyme, arylsulfatase A. Clinical diagnosis is usually confirmed by in vitro enzymatic deficiency of arylsulfatase A activity. The diagnosis may be complicated because of arylsulfatase A pseudo-deficiencies and another cause of MLD, sphingolipid activator B deficiency. As large quantities of sulfatides can be found in the urine in this disease, sulfatiduria appears as an extremely useful test. As recently enzyme replacement is underway, the quantitative determination, using an internal standard, appears particularly useful as a follow-up. Thus a non-physiological sulfatide was synthesized for this purpose, i.e. 3-O-sulfo-beta-D-C17 galactosylceramide (3-O-Sulfo-D: -Galactosyl-beta1'-->1-N-Heptadecanoyl-D-erythro-Sphingosine). It has been prepared through condensation of an azidosphingosine derivative with a protected D-galactopyranosyltrichloroacetimidate. Reduction of the azide was followed by acylation of a C-17 fatty acid. The key step was achieved by selective sulfation of the desired hydroxyl group on the sugar residue of the galactosylceramide using the stannylene methodology to give a 3'-sulfated beta-galactosyl C-17 ceramide.

Quantification of sulfatides and lysosulfatides in tissues and body fluids by liquid chromatography-tandem mass spectrometry

“…The accumulation of sulfatides in kidney leads to an increased excretion of sulfatides in urine. Determination of sulfatides in urine is a convenient diagnostic tool to confi rm MLD in patients with arylsulfatase A defi ciency or saposin B defi ciency (13)(14)(15). In addition to mutation analysis of the ASA gene, it is also used to confi rm a pseudodefi ciency of arylsulfatase A.…”

“…Several methods have been used to quantify sulfatides, including TLC ( 19 ), HPLC ( 14,20 ), and, more recently, MS ( 4,13,15,(21)(22)(23)(24)(25). Here we describe a LC/MS/MS method to quantify sulfatides and their molecular species over a wide range of concentrations in normal and diseased tissues, plasma, and urine.…”

Contribution of tandem mass spectrometry to the diagnosis of lysosomal storage disorders

“…The result of Gb 3 or sulfatide measurement is the sum of all isoforms. Despite the commercial availability of C18-D 3 -sulfatide (Matreya, State College, PA, USA), many reports refer to endogenous [C16 sulfatide (Whitfield et al 2001, Tan et al 2010); C18-sulfatide (Kuchař et al 2013)] or synthetic [C12 sulfatide (Saville et al 2017); C17-sulfatide (Cui et al 2008,Barcenasetal.2014a] IS species.…”

Role of sulfatide in normal and pathological cells and tissues

“…This leads to progressive demyelination ( 45 ). Large quantities of sulfatides can be found in the urine of MLD patients ( 46 ). There was no difference in quantitative levels of ceramide compositions (d18:1-C22:1, sphingadienine d18:2-C22:0, d18:1-C22:0 h, d18:1-C24:0, d18:1-C24:0 h, d18:2-C23:0 h, and d18:2-C24:0 h) of sulfatide in the urine samples from patients with two forms of clinical manifestation: motor form (homozygous for the ASA mutation P426L) and psychotic form (heterozygous for the ASA mutation I179S) ( 47 ).…”