The composition of non-cellulosic polysaccharides was determined according to Foster et al. (2010) with modifications. Due to shortage of plant material, analysis was performed only for samples harvested at the heading stage and maturity. De-starched AIR material (5 mg) was hydrolysed in 2 M TFA for 90 min at 121 °C. TFA was removed by drying under vaccum. TFA breaks down the cell wall network to release primarily non-cellulosic polysaccharides, and some fractions of cellulose that contain kinks and chain dislocations (amorphous form). The composition of monosaccharides in the filtrate was determined and quantified by high-performance anion-exchange chromatography coupled with integrated pulsed amperometric detection (HPAEC-iPAD) using Thermo Scientific Dionex ICS5000 as described previously by Głazowska et al. (2018) (link). The system was calibrated with standards (l -Ara, d -Xyl, d -Gal, d -Glc, d -GlcA, and d -GalA) (Sigma). All calculations were done in Chromeleon CDS software.
D glca
Manufactured by Merck Group
Sourced in United States, United Kingdom
D-GlcA is a chemical compound used as a laboratory reagent. It is the glucuronic acid form of glucose. D-GlcA is commonly utilized in various analytical and research applications, but a detailed description of its core function is not available while maintaining an unbiased and factual approach.
Automatically generated - may contain errors
Lab products found in correlation
D gal, by Merck Group (3 mentions)
D xyl, by Merck Group (3 mentions)
D glc, by Merck Group (3 mentions)
L ara, by Merck Group (3 mentions)
D gala, by Merck Group (2 mentions)
L fuc, by Merck Group (2 mentions)
L rha, by Merck Group (2 mentions)
Dionex ics 5000, by Thermo Fisher Scientific (1 mentions)
Carbopac pa20 column, by Thermo Fisher Scientific (1 mentions)
Pa20 column, by Thermo Fisher Scientific (1 mentions)
3 protocols using d glca
1
Quantifying Non-Cellulosic Polysaccharides
2
Monosaccharide Composition Analysis of Plant Cell Walls
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To determine monosaccharide composition, 3 replicates of cell walls from 10 plants were used. 1 mg of dry de-starched cell wall was hydrolyzed with 2 N trifluoroacetic acid at 120 °C for 2 h. The hydrolysates were dried at 50 °C, re-dissolved in water, and analyzed by high-performance anion-exchange chromatography with pulsed-amperometric detection using a CarboPac PA-20 column (3 mm × 150 mm; Dionex, Sunnyvale, CA, USA) as described earlier [99 (link)]. Monosaccharides were separated using a gradient of 100 mM NaOH in water at 0.5 mL min−1 under the following conditions: 0–0.05 min—12 mM NaOH; 0.05–26 min—0.65 mM NaOH; 26–46 min—300 mM NaOH; 46–55 min—12 mM NaOH. Monosaccharide standards included L-Fuc, L-Rha, L-Ara, D-Gal, D-Glc, D-Xyl, D Man, D-GalA, and D-GlcA (all from Sigma–Aldrich, St. Louis, MO, USA). To determine response factors, standard curves were created using mixtures of all standard monosaccharides at different concentrations.
3
Monosaccharide Profiling by HPAEC-PAD
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Cell wall monosaccharide was determined by HPAEC-PAD of hydrolysed material. A PA20 column (Dionex, USA) was used at a flow rate of 0.5 ml/min. The column was washed with 200 mM NaOH for 10 min before the injection of each sample and then equilibrated with 10 mM NaOH for 10 min.
The elution programme consisted of an isocratic elution with 10 mM NaOH from 0 to 37 min, followed by a linear gradient up to 800 mM NaOH from 37 to 43 min, and finally down to 10 mM NaOH from 43 to 45 min. Monosaccharide standards included L-Fuc, L-Rha, L-Ara, D-Gal, D-Glc, D-Xyl, D-Man, D-GalUA and D-GlcA (Sigma, UK); the standard mixture concentration ranges from 0.001 μg/10 μl to 0.1 μg/10 μl. A standard mixture run was performed before analysis of a batch of samples for verification of the response factors.
The elution programme consisted of an isocratic elution with 10 mM NaOH from 0 to 37 min, followed by a linear gradient up to 800 mM NaOH from 37 to 43 min, and finally down to 10 mM NaOH from 43 to 45 min. Monosaccharide standards included L-Fuc, L-Rha, L-Ara, D-Gal, D-Glc, D-Xyl, D-Man, D-GalUA and D-GlcA (Sigma, UK); the standard mixture concentration ranges from 0.001 μg/10 μl to 0.1 μg/10 μl. A standard mixture run was performed before analysis of a batch of samples for verification of the response factors.
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