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|Title:||Butanolysis derivatization: improved sensitivity in LC-MS/MS quantitation of heparan sulfate in urine from mucopolysaccharidosis patients|
|Citation:||Analytical Chemistry, 2015; 87(18):9243-9250|
|Publisher:||American Chemical Society|
|Paul J. Trim, John J. Hopwood, and Marten F. Snel|
|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.|
|Keywords:||Humans; Mucopolysaccharidoses; Butanols; Methanol; Heparitin Sulfate; Urinalysis; Chromatography, Liquid; Adolescent; Child; Child, Preschool; Infant; Female; Male; Tandem Mass Spectrometry; Young Adult; Limit of Detection|
|Rights:||© 2015 American Chemical Society|
|Appears in Collections:||Medicine publications|
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