Profiling oligosaccharidurias by electrospray tandem mass spectrometry: Quantifying reducing oligosaccharides (2025)

Analysis of urinary oligosaccharides in lysosomal storage disorders by capillary high-performance anion-exchange chromatography–mass spectrometry

Analytical and Bioanalytical Chemistry, 2012

Many lysosomal storage diseases are characterized by an increased urinary excretion of glycoconjugates and oligosaccharides that are characteristic for the underlying enzymatic defect. Here, we have used capillary highperformance anion-exchange chromatography (HPAEC) hyphenated to mass spectrometry to analyze free oligosaccharides from urine samples of patients suffering from the lysosomal storage disorders fucosidosis, α-mannosidosis, G M1 -gangliosidosis, G M2 -gangliosidosis, and sialidosis. Glycan fingerprints were registered, and the patterns of accumulated oligosaccharides were found to reflect the specific blockages of the catabolic pathway. Our analytical approach allowed structural analysis of the excreted oligosaccharides and revealed several previously unpublished oligosaccharides. In conclusion, using online coupling of HPAEC with mass spectrometric detection, our study provides characteristic urinary oligosaccharide fingerprints with diagnostic potential for lysosomal storage disorders.

Glycan profiling of urine, amniotic fluid and ascitic fluid from galactosialidosis patients reveals novel oligosaccharides with reducing end hexose and aldohexonic acid residues

FEBS Journal, 2010

Screening of Inherited Oligosaccharidurias Among Mentally Retarded Patients in Northern Finland

Journal of Intellectual Disability Research, 2008

A new UHPLC-MS/MS method for the screening of urinary oligosaccharides expands the detection of storage disorders

Orphanet Journal of Rare Diseases

Background Oligosaccharidoses are storage disorders due to enzymatic defects involved in the breakdown of the oligosaccharidic component of glycosylated proteins. The defect cause the accumulation of oligosaccharides (OS) and, depending on the lacking enzyme, results in characteristic profiles which are helpful for the diagnosis. We developed a new tandem mass spectrometry method for the screening of urinary OS which was applied to identify a large panel of storage disorders. Methods The method was set-up in urine and dried urine spots (DUS). Samples were analysed, without derivatization and using maltoheptaose as internal standard, by UHPLC-MS/MS with MRM acquisition of target OS transitions, including Glc4, the biomarker of Pompe disease. The chromatographic run was

A quantitative and comprehensive method to analyze human milk oligosaccharide structures in the urine and feces of infants

Analytical and Bioanalytical Chemistry, 2013

Human milk oligosaccharides (HMOs), though non-nutritive to the infant, shape the intestinal microbiota and protect against pathogens during early growth and development. Infant formulas with added galacto-oligosaccharides have been developed to mimic the beneficial effects of HMOs. Premature infants have an immature immune system and a leaky gut and are thus highly susceptible to opportunistic infections. A method employing nanoflow liquid chromatography time-of-flight mass spectrometry (MS) is presented to simultaneously identify and quantify HMOs in the feces and urine of infants, of which 75 HMOs have previously been fully structurally elucidated. Matrix-assisted laser desorption/ionization Fourier transform ion cyclotron resonance MS was employed for high-resolution and rapid compositional profiling. To demonstrate this novel method, samples from mother-infant dyads as well as samples from infants receiving infant formula fortified with dietary galacto-oligosaccharides or probiotic bifidobacteria were analyzed. Ingested oligosaccharides are demonstrated in high abundance in the infant feces and urine. While the method was developed to examine specimens from preterm infants, it is of general utility and can be used to monitor oligosaccharide consumption and utilization in term infants, children and adults. This method may therefore provide diagnostic and therapeutic opportunities.

Differential diagnosis of mucopolysaccharidosis and oligosaccharidosis of a sample of Egyptian children

Bulletin of Faculty of Pharmacy, Cairo University, 2018

Mucopolysaccharidosis (MPS) and oligosaccharidosis are lysosomal storage disorders (LSDs) that share many clinical features. The present study aimed to establish a protocol for the biochemical diagnosis of these disorders and their subtypes in affected Egyptian children as well as in pregnant females, in order to prepare children or fetus for enzyme replacement therapy. Urine, plasma and leukocyte samples were collected from 280 children with symptoms suggestive of LSDs. Fourteen amniotic fluid samples were collected from pregnant females having positive family history or having one affected sibling. Assessment of urinary glycosaminoglycans (GAGs) followed by two dimensional electrophoresis (2-DEP), thin layer chromatographic (TLC) for separation of oligosaccharides and plasma or leukocyte enzyme activity were performed. Six of pregnancies were diagnosed to have affected fetuses. 84 children had abnormal 2-DEP and classified as 26 MPS I, 10 MPS II, 24 MPS III and 24 MPS VI. Two were diagnosed as α-mannosidosis and 2 as GM 1 gangliosidosis. In conclusion; MPS should be excluded before suspecting oligosaccharidosis. 2-DEP and TLC alone cannot rule out the diagnosis of either MPS or oligosaccharidosis and confirmation must be done by specific lysosomal enzymatic assay. Analysis of GAGs by 2-DEP in amniotic fluid can be promising method for prenatal diagnosis of MPS.

Electron ionization mass spectra of reduced and permethylated urinary oligosaccharides from patients with mannosidosis

Biological Mass Spectrometry, 1984

The electron ionization mass spectra of reduced and permethylated isomeric mixtures of the major urinary tri-to deca-oligosaccharides of patients with mannosidosis are reported. Many of the oligosaccharide isomers can be differentiated in the mixtures on the basis of their distinct fragmentation patterns.

Detection of milk oligosaccharides in plasma of infants

Analytical and Bioanalytical Chemistry, 2014

Human milk oligosaccharides (HMO) are one of the major components of human milk. HMO are nondigestible by the human gut, where they are known to play important functions as prebiotics and decoys for binding pathogens. Moreover, it has been proposed that HMO may provide sialic acids to the infant that are important in brain development, however this would require absorption of HMO into the bloodstream. HMO have consistently been found in the urine of humans and other mammals, suggesting systemic absorption. Here, we present a procedure for the profiling of milk oligosaccharides (MO) in plasma samples obtained from 13 term infants hospitalized for surgery for congenital heart disease. The method comprises protein denaturation, oligosaccharide reduction, and porous graphitized carbon solid phase extraction for purification followed by analysis using nHPLC-PGC-chip-TOF-MS. Approximately 15 free MO were typically observed in the plasma of human infants, including LNT, LDFP, LNFT, 3′SL, 6′SL, 3′SLN, and 6′SLN, of which the presence was confirmed using fragmentation studies. A novel third isomer of SLN, not found in human or bovine milk was also consistently detected. Differences in the free MO profiles were observed between infants that were totally formula-fed and infants that received at least some part breast milk. Our results indicate that free MO similar in structure to those found in human milk and urine are present in the blood of infants. The method and results presented here will facilitate further research toward the possible roles of free MO in the development of the infant.

Oligosaccharide excretion in adult Gaucher disease

Journal of Inherited Metabolic Disease, 1998

Normalization of glycosaminoglycan-derived disaccharides detected by tandem mass spectrometry assay for the diagnosis of mucopolysaccharidosis

Scientific Reports

Mucopolysaccharidosis (MPS) is caused by the deficiency of a specific hydrolytic enzyme that catalyzes the step-wise degradation of glycosaminoglycans (GAGs). In this study, we propose an empirical method to calculate levels of GAG-derived disaccharides based on the quantity (peak areas) of chondroitin sulfate (Cs) with the aim of making a diagnosis of Mps more accurate and reducing the occurrence of false positive and false negative results. In this study, levels of urinary GAG-derived disaccharides were measured in 67 patients with different types of MPS and 165 controls without MPS using a tandem mass spectrometry assay. Two different methods of reporting GAG-derived disaccharides were assessed; normalization to urinary Cs (in μg/mL), and normalization to μg/ mg creatinine. Cs-normalization yielded more consistent values than creatinine-normalization. In particular, levels of urinary dermatan sulfate (DS), heparan sulfate (HS), and keratan sulfate (KS) significantly varied because of changes in urine creatinine levels, which were proportional to age but inversely proportional to DS, HS, and KS measurements. Using CS-normalization revealed the actual status of DS, HS, and KS without the influence of factors such as age, urine creatinine, and other physiological conditions. It could discriminate between the patients with Mps and controls without MPS, and also to evaluate changes in GAG levels pre-and post-enzyme replacement therapy. Mucopolysaccharidoses (MPSs) are a group of lysosomal storage disorders (LSDs) caused by deficiency in one specific enzyme that catalyzes the stepwise degradation of glycosaminoglycans (GAGs). Depending on the type of MPS, this deficiency leads to excessive lysosomal storage of either chondroitin sulfate (CS), dermatan sulfate (DS), heparin sulfate (HS), or keratan sulfate (KS) and results in devastating manifestations such as coarse facial features, developmental delay and decline, gibbus, hepatosplenomegaly, cardiac valve disease, umbilical and inguinal hernias, joint deformity with a restricted range of motion, airway dysfunction with complications, sleep apnea, recurrent otitis media, and premature death 1-5. The clinical manifestations of MPS are chronic and progressive, and the initial onset of clinical signs and symptoms of MPS emerge between the ages of 18 months and 4 years, depending on disease severity 6-8. Enzyme replacement therapy (ERT) is widely used and available for MPS I, MPS II, MPS IVA and MPS VI 5,9-14 , and trials are currently ongoing for MPS IIIB. Achieving optimal benefits from ERT requires commencing treatment before the onset of irreversible clinical presentations 5,15,16 .