A gas chromatographic method for the quantitative detemination of hexuronic acids in alginic acid

Vadas, L.; Prihar, Harry S.; Pugashetti, Balappa K.; Feingold, David S.

doi:10.1016/0003-2697(81)90484-x

Cited by 30 publications

(11 citation statements)

References 12 publications

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“…In these three cases in which this was done, 13C-NMR gave consistantly higher guluronic acid values than our gas chromatography analytical procedure. We have recent evidence that the reduction method via the carbodiimide adduct, which is a step in the procedure for guluronic-and mannuronic-acid determination in alginic acid (29), may be responsible for this discrepancy. Molecular weight parameters were determined by gel permeation-high-pressure liquid chromatography.…”

Section: Discussionmentioning

confidence: 99%

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Exopolysaccharides of the phytopathogen Pseudomonas syringae pv. glycinea

Osman¹,

Fett²,

Fishman³

1986

J Bacteriol

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Exopolysaccharides (EPS) of the soybean pathogen Pseudomonas syringae pv. glycinea were isolated from culture ifitrates and infected soybean leaves. Levan (a polyfructan with a C-2 --C-6 backbone and C-2 -+ C-1 branching) or acetylated alginate (a linear polyuronide of C-1-+ C-4-linked mannuronic and guluronic acids) was isolated from culture filtrates when bacterial strains were grown in a semisynthetic medium containing sucrose or glucose, respectively, as the primary carbon source. Acetylated alginate was the only EPS isolated from soybean [Glycine max (L.) Merr.] leaves inoculated with compatible (disease-inducing) strains of P. syringae pv. glycinea. The acetyl content of the P. syringae pv. glycinea alginates varied from 3 to 14%, and the amount of guluronic acid varied from less than 1 to 20%. The P. syringae pv. glycinea alginates from in vitro batch cultures were of lower molecular weight and polydispersity thakm those from in planta cultures, and both were of lower molecular weights than alginates produced by Pseudomonas aeruginosa.

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Section: Discussionmentioning

confidence: 99%

“…b Determined by a modification of the method described by Vadas et al (29). % mannuronic acid = 100 -% guluronic acid.…”

mentioning

confidence: 99%

Exopolysaccharides of the phytopathogen Pseudomonas syringae pv. glycinea

Osman¹,

Fett²,

Fishman³

1986

J Bacteriol

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“…Purified alginic acid was dialyzed against water at 4°C for 2 days and then dried in a 65°C oven. The dry weight was obtained, and triplicate samples (10 to 20 mg) of the alginic acid were converted to a mixture of D-mannitol and n-glucitol by the procedure of Vadas et al (40). Xylitol (50 ,ug) was added to the samples as an internal standard.…”

Section: Methodsmentioning

confidence: 99%

Phosphorylcholine stimulates capsule formation of phosphate-limited mucoid Pseudomonas aeruginosa

1988

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Production of both alginic acid and lipopolysaccharide by a mucoid strain of Pseudomonas aeruginosa, SRM-3, was studied in a chemostat system during growth under nutrient-limiting conditions chosen to reflect the chronic growth conditions in the lungs of cystic fibrosis patients. Since mucoid strains have been shown to elaborate extracellular proteases and phospholipase C, nitrogen and phosphate limitation were selected for analysis. A modified alginate-promoting medium containing either 1 mM glutamate or 0.05 mM K2HPO4 as limiting nutrient and doubling times of 1.6 to 15.7 h were used. Under nitrogen limitation, strain SRM-3 produced 1.4 mg of uronic acid per mg (dry weight) of cells at all doubling times studied. However, phosphate limitation resulted in the synthesis of only 0.4 mg of uronic acid per mg (dry weight) of cells. The role of phosphate in alginic acid polysaccharide production was further investigated by using phosphorylcholine, a product of phospholipase C activity on phosphatidylcholine, the major lung surfactant. Not only were mucoid cells capable of utilizing phosphorylcholine for growth, but a highly specific interaction occurred among phosphorylcholine, alginate, and whole cells, resulting in greatly enhanced culture viscosity. Electron micrographs showed the gradual formation of a capsule during growth on phosphorylcholine, indicating that the mucoid strain has the ability to utilize surfactant not only as a nutrient source but also for constructing a capsule with greatly enhanced adhesive properties.

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“…Although each of these GAGs has a predominant disaccharide component, heterogeneity does exist in the sugars presented in the make-up of any given class of GAG. In this study, the identification of acid MPS by the modified LC-MS/MS method of the predominant disaccharide component of GAGs after methanolysis [14–18] is proposed and validated for MPS type determination [19,20]. …”

Section: Introductionmentioning

confidence: 99%

A modified liquid chromatography/tandem mass spectrometry method for predominant disaccharide units of urinary glycosaminoglycans in patients with mucopolysaccharidoses

Chen

Lin

Wang

et al. 2014

Orphanet J Rare Dis

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BackgroundThe identification of acid mucopolysaccharide by the liquid chromatography/tandem mass spectrometry method (LC-MS/MS) of the predominant disaccharide units of glycosaminoglycans (GAGs) (chondroitin sulfate, CS; dermatan sulfate, DS; heparan sulfate, HS) after methanolysis is validated and applicable for mucopolysaccharidosis (MPS) type determination.MethodsA total of 76 urine samples were collected and analyzed, from nine MPS I patients, 13 MPS II patients, seven MPS III patients, eight MPS VI patients, and 39 normal controls. Urinary GAG was first precipitated by the Alcian blue method followed by a treatment of 3 N HCl methanol. The protonated species of the methylated disaccharide products were detected by using a multiple reaction monitoring experiment. Internal standards, [2H6] CS, [2H6] DS and [2H6] HS, were prepared in-house by deuteriomethanolysis of CS, DS and HS.ResultsOne particular disaccharide for each GAG was selected, in which the parent ion and its daughter ion after collision were m/z 426.1 → 236.2 for DS (m/z 432 → 239 for dimers derived from [2H6] CS and [2H6] DS) and m/z 384.2 → 161.9 for HS (m/z 390.4 → 162.5 for the [2H6] HS dimer). The quantities of DS and HS were determined, which varied from one MPS type to the other. The results can be used to evaluate the severity of MPS subgroups, as well as urinary GAG amelioration at follow-up after enzyme replacement therapy (ERT).ConclusionsThe modified LC–MS/MS method for MPS type determination is specific, sensitive, validated, accurate, and applicable for simultaneous quantifications of urinary DS and HS. This method can help to make correct diagnosis of MPS patients and evaluate the effectiveness of ERT.

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A gas chromatographic method for the quantitative detemination of hexuronic acids in alginic acid

Cited by 30 publications

References 12 publications

Exopolysaccharides of the phytopathogen Pseudomonas syringae pv. glycinea

Exopolysaccharides of the phytopathogen Pseudomonas syringae pv. glycinea

Phosphorylcholine stimulates capsule formation of phosphate-limited mucoid Pseudomonas aeruginosa

A modified liquid chromatography/tandem mass spectrometry method for predominant disaccharide units of urinary glycosaminoglycans in patients with mucopolysaccharidoses

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