Determination Of Amino Sugars Research Articles

Determination of Amino Sugars in Blood Serum Samples by RP-HPLC Method

Determination of Amino Sugars in Blood Serum Samples by RP-HPLC Method

Specificity of Lectins Labeled with Colloidal Gold to the Exopolymeric Matrix Carbohydrates of the Sulfate-Reducing Bacteria Biofilm Formed on Steel

The studies of the carbohydrate composition of the sulfate-reducing bacteria (SRB) biofilms formed on the steel surface, which are a factor of microbial corrosion, are significant. Since exopolymers synthesized by bacteria could activate corrosive processes. The aim of the study was to investigate the specificity of commercial lectins, labeled with colloidal gold to carbohydrates in the biofilm exopolymeric matrix produced by the corrosive-relevant SRB strains from man-caused ecotopes. Methods. Microbiological methods (obtaining of the SRB biofilms during cultivation in liquid Postgate B media under microaerophilic conditions), biochemical methods (lectin-binding analysis of 10 commercial lectins, labeled with colloidal gold), transmission electron microscopy using JEM-1400 JEOL. Results. It was shown using transmission electron microscopy that the binding of lectins with carbohydrates in the biofilm of the studied SRB strains occurred directly in the exopolymerіс matrix, as well as on the surfaces of bacterial cells, as seen by the presence of colloidal gold particles. For detection of the neutral carbohydrates (D-glucose and D-mannose) in the biofilm of almost all studied bacterial strains PSA lectin was the most specific. This lectin binding in biofilms of Desulfotomaculum sp. К1/3 and Desulfovibrio sp. 10 strains was higher in 90.8% and 94.4%, respectively, then for ConA lectin. The presence of fucose in the SRB biofilms was detected using LABA lectin, that showed specificity to the biofilm EPS of all the studied strains. LBA lectin was the most specific to N-аcetyl-D-galactosamine for determination of amino sugars in the biofilm. The amount of this lectin binding in D. vulgaris DSM644 biofilm was 30.3, 10.1 and 9.3 times higher than SBA, SNA and PNA lectins, respectively. STA, LVA and WGA lectins were used to detect the N-acetyl-Dglucosamine and sialic acid in the biofilm. WGA lectin showed specificity to N-acetyl-D-glucosamine in the biofilm of all the studied SRB; maximum number of bounded colloidal gold particles (175 particles/μm2) was found in the Desulfotomaculum sp. TC3 biofilm. STA lectin was interacted most actively with N-acetyl-D-glucosamine in Desulfotomaculum sp. TC3 and Desulfomicrobium sp. TC4 biofilms. The number of bounded colloidal gold particles was in 9.2 and 7.4 times higher, respectively, than using LVA lectin. The lowest binding of colloidal gold particles was observed for LVA lectin. Conclusions. It was identified the individual specificity of the 10 commercial lectins to the carbohydrates of biofilm matrix on the steel surface, produced by SRB. It was estimated that lectins with identical carbohydrates specificity had variation in binding to the biofilm carbohydrates of different SRB strains. Establishing of the lectin range selected for each culture lead to the reduction of the scope of studies and labor time in the researching of the peculiarities of exopolymeric matrix composition of biofilms formed by corrosiverelevant SRB.

Development of a precolumn derivatization HPLC method with diode-array detection for the determination of amino sugars in peat and soil humic acids.

The work is focused on the development of a high-performance liquid chromatography method with diode-array detection for the separation and quantitation of the three most abundant amino sugars; d-glucosamine, d-galactosamine, and d-mannosamine. The high-performance liquid chromatography separation was carried out by reversed-phase chromatography on Chromolith Performance RP-18e monolithic column after acid hydrolysis (5M HCl) and precolumn derivatization of samples using diethyl ethoxymethylenemalonate. Gradient elution and a mobile phase composed of ammonium formate buffer solution (10mmol/L, pH 3.60) and methanol with flow rate of 1.0mL/min were used. The monitoring wavelength was set at 280nm. The limits of detection and quantitation for analytes ranged from 0.017 to 0.122mg/L and from 0.057 to 0.407mg/L, respectively. The proposed method was successfully applied for the determination of amino sugars in samples of humic acids isolated from different soils and peat.

Comparison of HPLC Methods for the Determination of Amino Sugars in Soil Hydrolysates

A study on the suitability of chromatographic techniques such as high performance anion exchange chromatography (HPAEC) with fluorescence detection (FL) and pulsed amperometric detection (PAD) and reversed phase (RP) chromatography for the determination of galactosamine, glucosamine, mannosamine, and muramic acid in soil hydrolysates was carried out. The reversed phase fluorescence method was rapid, provided good validation parameters, and employed relatively inexpensive instrumentation. The HPAEC methods had slightly higher limits of quantification, 0.6–5.0 µmol L−1 (HPAEC-FL) and 1.0–10.0 µmol L−1 (HPAEC-PAD), compared to the reversed phase fluorescence method (0.5–5.0 µmol L−1). Various sample pretreatment methods and chromatographic methods were investigated and the advantages and disadvantages of the HPLC methods are discussed.

Determination of amino sugars in environmental samples with high salt content by high-performance anion-exchange chromatography and pulsed amperometric detection.

Amino sugars were determined in natural samples, including seawater, using high-performance anion-exchange chromatography with pulsed amperometric detection and a new-off-line sample cleanup procedure. Samples were hydrolyzed with 3 M HCl for 5 h (100 degrees C) and neutralized with anion retardation resin. Before injection, salts and organic contaminants were removed with a strong cation exchanger in the Na+ form. Detection limits for amino sugars were between 1 and 4 nM (signal-to-noise ratio 3), allowing for the first time quantification of amino sugars in seawater without preconcentration. Precision was 2-11% at the 20 nM level. The relatively simple and rapid sample preparation makes it suitable for routine analyses.

Determination of amino sugars in synthetic glycopeptides during the conditions of amino acid analysis utilizing precolumn derivatization and high-performance liquid chromatographic analysis

The growing number of synthetic glycopeptides required an in-house method for the analysis of the final products. A reversed-phase high-performance liquid chromatographic protocol was developed to verify the presence of N-acetylglucosamine and N-acetylgalactosamine in synthetic glycopeptides after hydrolysis and derivatization with 4-dimethylaminoazobenzene-4′- sulphonyl chloride. Different sugar and glycoamino acid standards were used to separate the two carbohydrate moieties, and the location of the derivatized 2-aminoglucose was established by the use of radiolabeled sugar. The utility of the approach is demonstrated by the amino acid analysis of N- and O-glycosylated synthetic peptides and the method could provide an alternative for sugar analysis of glycoproteins.

Determination of amino sugars and amino acids in glycoconjugates using precolumn derivatization with o-phthalaldehyde

This report examines the RP-HPLC separation of o-phthalaldehyde derivatives of amino acids, amino sugars, and amino sugar alcohols using either 2-mercaptoethanol or 3-mercaptopropionic acid. A method with pmol sensitivity for the analysis of N-acetylamino sugars of glycoconjugates was elaborated. Upon hydrolysis, amino sugars are reduced with borohyride. Automated precolumn derivatization and chromatographic conditions for the resulting hexosaminitols are the same as those used for the analysis of amino acids. The method has been tested with as little as 2 μg of bovine fetuin, with a glycopeptide from bromelain and with an oligosaccharide after periodate oxidation.

A simplified procedure for the photometric determination of amino sugars in soil

A simplified procedure for the photometric determination of amino sugars in soil

Improvements in the high-pressure liquid chromatographic determination of amino sugars and .alpha.-galactosides in faba bean, lupine, and pea

Presentation d'une methode simplifiee utilisable pour les analyses de routine des glucosides et α-D-galactosides (raffinose, stachyose et verbascose), composes responsables de la flatulence

Use of an extraction-photometric method in the quantitative determination of amino sugars for investigating natural polymers

Use of an extraction-photometric method in the quantitative determination of amino sugars for investigating natural polymers

Structures of the major carbohydrates of natural human interleukin-2.

Purified human interleukin-2 secreted by peripheral blood lymphocytes from healthy donors was found to exist in several forms. These forms were (partially) resolved by reversed-phase high-performance liquid chromatography and sodium dodecyl sulfate/polyacrylamide gel electrophoresis. Two major polypeptide species (interleukin-2 N1 and N2, 16.5 kDa) were shown to be glycosylated on the basis of [3H]galactose/[3H]glucosamine incorporation and determination of amino sugars after acid hydrolysis. A third component (interleukin-2 M, 14.5 kDa) represents a nonglycosylated form. The amino acid composition and the NH2-terminal sequence of both forms are consistent with the data deduced from the cDNA coding for interleukin-2 after removal of a leader peptide of 20 amino acids. Carbohydrates are O-linked to the IL-2 protein via threonine-3 of the polypeptide chain. The oligosaccharides were released by reductive beta-elimination and were purified by gel filtration and high-performance liquid chromatography. Applying methylation analysis, exoglycosidase digestion and fast atom bombardment mass spectrometry the following major carbohydrate structures were identified: N1, NeuAc(alpha 2-3)Gal(beta 1-3)GalNAc-ol; and N2, NeuAc(alpha 2-3)Gal(beta 1-3)[NeuAc(alpha 2-6)]GalNAc-ol.

Quantittive determination of amino sugars in bacterial lipopolysaccharides

A modification of the method of quantitatively determining amino sugars in polysaccharide preparations of bacterial origin is described which excludes the influence of the mixture of amino acids and monosaccharides on the result of the analysis. The modification is based on differences in the extraction capacity of organic solvents in relation to the various chromophores obtained in the Elson-Morgan reaction from amino sugars and from the mixture of amino acids and monosaccharides. It has been found that the modification described is distinguished by a high sensitivity, specificity, and reproducibility of the results. The standard deviation calculation for 10 determinations does not exceed 4%. The method is simple to perform and it requires uncomplicated equipment and small amounts of reagents.

A fluorometric assay for the quantitation of aminosugars

A method is presented for the quantitative determination of aminosugars in glycoproteins. Glycoproteins are acid hydrolyzed and reacted with the fluorogenic reagent 1-dimethylaminonaphthalene-5-sulfonyl chloride. Following cellulose thin layer electrophoresis in pyridine/acetic acid (pH 4.4), 5 h at 500 V, appropriate areas of the plate are scraped and the dansyl hexosamines are eluted with 95% ethanol. Aliquots are then applied to a silica gel tlc plate and subjected to ascending thin layer chromatography in cyclohexane/ethylacetate/ethanol (6/4/3). After spraying the plate with triethanolamine/isopropanol ( 1 4 ), fluorescent intensities are measured by in situ scanning. The aminosugar content of the glycoprotein is determined from a curve generated from a series of standards run concurrently on each plate. The method clearly resolves glucosamine from galactosamine, and is sensitive for the detection of aminosugars in the subnanomole range.

An automatic analyzer for the specific determination of amino sugars

The techniques previously employed for the extraction and determination of amino acids from different matrices are not necessarily optimal for the determination of the amino sugars. An analytical system is described which is a hybrid between the conventional amino acid analyzer and the liquid chromatographic system for the detection of reducing sugars. The major, naturally occurring amino sugars are separated in about 40 min, with sensitivites lying under the nanomole range, without interference from other co-extracted compounds such as amino acids and sugars. The reagent employed is noncorrosive and stable over long periods of time. The amino sugar analyzer can be readily constructed by simple modification of a conventional phenylketonuria or amino acid analyzer.

A simplified method for determination of amino sugars in glycoproteins

A simplified method for determination of amino sugars in glycoproteins

The pattern of urinary glycosaminoglycans excretion in relation to age in man: II. Isolation, proportions, and properties of the fraction resistant to testicular hyaluronidase

Crude urinary glycosaminoglycan preparations were fractionated by a procedure consisting of a digestion with testicular hyaluronidase, a gel filtration through a column of Sephadex G-75, a papain digestion of the peak emerging first and a gel filtration of the papain digest. This procedure permitted the separation of the glycosaminaglycans resistant to testicular hyaluronidase from those susceptible to the enzyme and from breakdown products of small molecular weight and was applied in the study of age-related differences in the composition of preparations isolated from urines of seven healthy males aged from 3 to 83 years. In terms of uronic acid the glycosaminoglycans resistant to hyaluronidase represented for the various individuals 6–45% of the total and increased with advancing age, thus confirming our previous findings obtained by a series of different techniques. Determination of aminosugars showed that glucosamine-, uronic acid-containing glycosaminoglycan (s), likely heparan sulfate, appears to be mainly responsible for the increase with age of this fraction of urinary glycosaminoglycans.

A novel fluorophotometric method for determination of hexosamines using pyridoxal and zinc(II) ion

A novel fluorophotometric method for the detection and determination of amino sugars was established utilizing pyridoxal-HCl and Zn(II) ion. This method was highly sensitive; its limit of determination was 1 × 10 −11 mole of amino sugars. The fluorogen of this fluorescence reaction was Zn(II) chelate of N-pyridoxylidene amino sugar containing a 1:1 ratio of amino sugar to Zn(II) ion. The hexosamine content of chondroitin sulfate A and C, hyaluronic acid, and heparin estimated by this method was higher than that obtained by the Elson-Morgan method. Only 2–3 hr of hydrolysis with 2 N HCl was sufficient for the assay of chondroitin sulfate and heparin.

Determination of amino sugars in mixtures containing glucosamine, galactosamine and muramic acid.

A colorimetric method is described whereby the direct quantitative determination of glucosamine, galactosamine and muramic acid can be achieved without previous treatment of the cell-wall hydrolysate, for example by column chromatography. Molar ratios of hexosamines in cell-wall preparations, from a number of bacterial species, determined by this method were found to be in general agreement with previously published results.

Immunochemical studies on polysaccharide A of Staphylococcus aureus. 5. Identification of amino sugars and amino acids.

Acta Pathologica Microbiologica ScandinavicaVolume 55, Issue 3 p. 308-318 IMMUNOCHEMICAL STUDIES ON POLYSACCHARIDE A OF STAPHYLOCOCCUS AUREUS 5.Identification of Amino Sugars. Quantitative Determination of Amino Sugars and Amino Acids. Gunnar Haukenes, Gunnar Haukenes THE UNIVERSITY OF BERGEN, SCHOOL OF MEDICINE THE GADE INSTITUTE, DEPARTMENT OF MICROBIOLOGY, BERGEN, NORWAYSearch for more papers by this author Gunnar Haukenes, Gunnar Haukenes THE UNIVERSITY OF BERGEN, SCHOOL OF MEDICINE THE GADE INSTITUTE, DEPARTMENT OF MICROBIOLOGY, BERGEN, NORWAYSearch for more papers by this author First published: July 1962 https://doi.org/10.1111/j.1699-0463.1962.tb04130.xCitations: 7AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat Citing Literature Volume55, Issue3July 1962Pages 308-318 RelatedInformation

The determination of amino sugars in the presence of amino acids and glucose.

Research Article| December 01 1960 The determination of amino sugars in the presence of amino acids and glucose C. Cessi; C. Cessi 1Istituto di Patologia Generale, Università, Bologna, Italy Search for other works by this author on: This Site PubMed Google Scholar Franca Piliego Franca Piliego 1Istituto di Patologia Generale, Università, Bologna, Italy Search for other works by this author on: This Site PubMed Google Scholar Biochem J (1960) 77 (3): 508–510. https://doi.org/10.1042/bj0770508 Views Icon Views Article contents Figures & tables Video Audio Supplementary Data Peer Review Share Icon Share MailTo Twitter LinkedIn Cite Icon Cite Get Permissions Citation C. Cessi, Franca Piliego; The determination of amino sugars in the presence of amino acids and glucose. Biochem J 1 December 1960; 77 (3): 508–510. doi: https://doi.org/10.1042/bj0770508 Download citation file: Ris (Zotero) Reference Manager EasyBib Bookends Mendeley Papers EndNote RefWorks BibTex toolbar search Search Dropdown Menu toolbar search search input Search input auto suggest filter your search All ContentAll JournalsBiochemical Journal Search Advanced Search This content is only available as a PDF. © 1960 CAMBRIDGE UNIVERSITY PRESS1960 Article PDF first page preview Close Modal You do not currently have access to this content.