Amino acid distribution in soil humic acids fractionated by tandem size exclusion chromatography polyacrylamide gel electrophoresis

Trubetskaya, O. E.; Reznikova, O. I.; Afanas'eva, G. V.; Markova, L.F.; Muranova, T.A.; Trubetskoj, Oleg

doi:10.1016/s0160-4120(98)00036-1

Cited by 30 publications

(13 citation statements)

References 11 publications

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“…fractions. In addition, Trubetskaya et al (1998) analyzed the amino acid content of the HA fractions obtained by the same method after acid hydrolysis and found that the amino acid content increased with increasing molecular size. This indicated that the proportion of proteinous moieties increased with the increase in the molecular size of HA.…”

Section: Infrared Spectroscopy Of the Fractionsmentioning

confidence: 99%

“…Trubetskoj et al (1997) developed a method for the fractionation of soil HAs based on the combined use of size-exclusion chromatography and PAGE in the presence of concentrated urea. They have been applying this method to the investigation of soil HAs (Trubetskaya et al 1998(Trubetskaya et al , 2002(Trubetskaya et al , 2011Trubetskoj et al 1999;Aguer et al 2001;Saiz-Jimenez et al 2006;Sanchez-Cortes et al 2006;Guyot et al 2007;Richard et al 2007;Francioso et al 2009). The advantage of this method is that it allows one to collect a sufficient amount of the fractions with different electrophoretic behavior for characterization.…”

Section: Introductionmentioning

confidence: 99%

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Fractionation of soil humic acids by preparative polyacrylamide gel electrophoresis in the presence of concentrated urea

Karim

Aoyama

2013

Soil Science and Plant Nutrition

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To separate soil humic acids (HAs) into their constituents and characterize them, polyacrylamide gel electrophoresis (PAGE) was carried out in the presence of 7 M urea using a preparative electrophoresis system. Two types of soil HAs were fractionated into nine fractions by PAGE. The dark-colored constituents were recovered from the electrophoretic fractions by precipitation on acidification, and the brown-colored constituents dissolved in the acidic solution of fast-moving fractions were recovered by adsorption onto DAX-8 resin. High-performance size-exclusion chromatography (HPSEC) confirmed that the constituents of the HAs were separated based on their molecular size by PAGE. The dark-colored constituents exhibited higher degrees of humification than did the corresponding unfractionated HAs, except for the constituents remaining in the electrophoretic gels at the end of electrophoresis. Diffuse reflectance infrared spectroscopy revealed that the chemical properties of the dark-colored constituents changed regularly: the content of carboxyl groups decreased and the proportions of proteinous, aliphatic and polysaccharide moieties increased with increasing molecular size. The humification degrees of the constituents adsorbed onto DAX-8 resin were considerably lower than those of the corresponding unfractionated HAs. The chemical properties of the DAX-8-adsorbed constituents were different from those of the dark-colored constituents. Observation of electrophoretic fractions under blue light (470 nm) and HPSEC with fluorescence detection at an excitation wavelength of 460 nm and an emission wavelength of 520 nm showed that green fluorescent substances were largely concentrated in the smallest molecular size fractions and were partitioned into both the dark-colored precipitates and DAX-8-adsorbed fractions. The proportion of organic carbon recovered by precipitation and adsorption onto DAX-8 resin was 45-63%, indicating that substantial parts of the HA constituents were missing. The unrecovered constituents were considered to be acid-soluble, nearly colorless substances. The dissociation of the acid-soluble constituents from the acid-insoluble dark-colored constituents during the preparative PAGE of soil HAs was ascribed to disruption of hydrogen bonding and hydrophobic interactions caused by concentrated urea.

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Section: Infrared Spectroscopy Of the Fractionsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Fractionation of soil humic acids by preparative polyacrylamide gel electrophoresis in the presence of concentrated urea

Karim

Aoyama

2013

Soil Science and Plant Nutrition

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“…By contrast, little attention has been given to the analysis of hydrophilic constituents of humic acids. Only limited studies have been devoted to carbohydrate composition of soil humic acids (Ogner, 1980;Coelho et al, 1988;Tsutsuki and Kuwatsuka, 1989) and to amino acid composition of soil (Saharinen and Schnitzer, 1989;Scheller, 1996;Szajdak and Ö sterberg, 1996;Trubetskaya et al, 1998;Szajdak et al, 2003) and lignite (Damian et al, 1985) humic acids.…”

Section: Introductionmentioning

confidence: 99%

A comparative study on the chemical composition of humic acids from forest soil, agricultural soil and lignite deposit

Allard

2006

Geoderma

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“…In our preliminary investigation, SEC-PAGE allowed the fractionation of several soil HAs of different genesis and we obtained HA fractions with clear differences in terms of ultraviolet (UV)-visible spectra, fluorescence properties, photochemical activity, and amino acid content. 7,8 Spectroscopic and structural analyses of HS have experienced a great deal of growth over recent years due to the development of high field magnets and an array of multidimensional techniques. The use of heteronuclear and multidimensional nuclear magnetic resonance (NMR) spectroscopy for mapping the chemical structures and organization of HS provided considerable progress on these complex environmental samples.…”

Section: Introductionmentioning

confidence: 99%

Surface-Enhanced Raman Spectroscopy of Chernozem Humic Acid and Their Fractions Obtained by Coupled Size Exclusion Chromatography—Polyacrylamide Gel Electrophoresis (SEC-PAGE)

et al. 2006

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A humic acid extracted from a chernozem soil was fractionated combining size exclusion chromatography and polyacrylamide electrophoresis (SEC-PAGE). Three fractions named A, B, and CþD, with different electrophoretic mobilities and molecular sizes (MS), were obtained and subsequently characterized by thermochemolysis and surface-enhanced Raman spectroscopy (SERS). The data confirmed that fraction A, with the higher MS, was more aliphatic than fractions B and CþD and, in turn, fractions with lower MS (B and CþD) denoted an enrichment in lignin residues. These structural features explain conformational changes when varying the pH in the humic fraction A and indicated that combination of the two techniques is a good approach for characterizing humic substances.

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Amino acid distribution in soil humic acids fractionated by tandem size exclusion chromatography polyacrylamide gel electrophoresis

Cited by 30 publications

References 11 publications

Fractionation of soil humic acids by preparative polyacrylamide gel electrophoresis in the presence of concentrated urea

Fractionation of soil humic acids by preparative polyacrylamide gel electrophoresis in the presence of concentrated urea

A comparative study on the chemical composition of humic acids from forest soil, agricultural soil and lignite deposit

Surface-Enhanced Raman Spectroscopy of Chernozem Humic Acid and Their Fractions Obtained by Coupled Size Exclusion Chromatography—Polyacrylamide Gel Electrophoresis (SEC-PAGE)

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