title: Butanolysis derivatization: improved sensitivity in LC-MS/MS quantitation of heparan sulfate in urine from mucopolysaccharidosis patients.
abstract: Heparan sulfate (HS) is a complex oligosaccharide that is a marker of a number of diseases, most notably several of the mucopolysaccharidoses (MPS). It is a very heterogeneous compound and its quantification at physiological concentrations in patient samples is challenging. Here, we demonstrate novel derivatization chemistry for depolymerization/desulfation and alkylation of HS based on butanolysis. The resultant alkylated disaccharides are quantifiable by LC-MS/MS. This new method is at least 70-fold more sensitive than a previously published methanolysis method. Disaccharide yield over time is compared for methanolysis, ethanolysis, and butanolysis. Maximum disaccharide concentration was observed after 2 h with butanolysis and 18 h with ethanolysis whereas a maximum was not reached over the 24 h of the experiment with methanolysis. The sensitivity of the new technique is illustrated by the quantification of HS in 5 μL urine samples from MPS patients and healthy controls. HS was quantifiable in all samples including controls. Disaccharide reaction products were further characterized using exact mass MS/MS.
authors: Lysosomal Diseases Research Unit, South Australian Health and Medical Research Institute (SAHMRI) , North Terrace, Adelaide, South Australia 5000, Australia.
authors: Paul J
authors: PJ
authors: Trim
authors: Lysosomal Diseases Research Unit, South Australian Health and Medical Research Institute (SAHMRI) , North Terrace, Adelaide, South Australia 5000, Australia.
authors: John J
authors: JJ
authors: Hopwood
authors: Lysosomal Diseases Research Unit, South Australian Health and Medical Research Institute (SAHMRI) , North Terrace, Adelaide, South Australia 5000, Australia.
authors: Marten F
authors: MF
authors: Snel
mesh_terms: D000293:Adolescent; D000440:Butanols / Q000737:chemistry*; D002648:Child; D002675:Child, Preschool; D002853:Chromatography, Liquid; D005260:Female; D006497:Heparan Sulfate / Q000737:chemistry* / Q000652:urine*; D006801:Humans; D007223:Infant; D057230:Limit of Detection; D008297:Male; D000432:Methanol / Q000737:chemistry; D009083:Mucopolysaccharidoses / Q000652:urine*; D053719:Tandem Mass Spectrometry; D016482:Urinalysis / Q000379:methods*; D055815:Young Adult
chemical_list: D000440:Butanols; D006497:Heparan Sulfate; D000432:Methanol