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Analysis of ascorbate in plant tissues by high-performance capillary zone electrophoresis.
Anal Biochem. 1996 Jul 15; 239(1):8-19.AB

Abstract

We describe here a simple and rapid capillary electrophoresis method for the determination of ascorbic acid (L-AA) and isoascorbic acid (D-AA) in vegetative tissues. For optimal yields and stabilization, samples are extracted with cold 3% metaphosphoric acid. Hydrophobic contaminants are then removed by passage through a C18 solid-phase extraction cartridge. The analysis itself is performed on a fused silica capillary with 200 mM borate, pH 9, as the carrier electrolyte, using on-line diode array detection over the range 190-350 nm. Quantitation was performed at 260 nm, the uv-absorption maximum for ascorbate at this pH. This method has a minimum detection limit of 84 fmol/injection and linearity of detector response was observed up to at least 12 pmol/injection. We also describe the influence of electrolyte concentration, pH, and the presence of detergent on separations of L-AA, D-AA, and L-galacturonic acid-1,4-lactone. The protocol has been demonstrated to be suitable for the analysis of L-AA in Arabidopsis, parsley, and mushroom. The method has superior resolution to comparable HPLC separations, a comparable analysis time, but lower sensitivity because of the concentration limitations of the detection system.

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Authors+Show Affiliations

Davey MW
Department of Genetics, Flanders Interuniversity Institute for Biotechnology, Universiteit Gent, Gent, Belgium.
Bauw G
No affiliation info available
Montagu MV
No affiliation info available

MeSH

ArabidopsisAscorbate OxidaseAscorbic AcidBasidiomycotaChromatography, High Pressure LiquidElectrophoresis, CapillaryHydrogen-Ion ConcentrationPlantsReproducibility of ResultsSodium Dodecyl Sulfate

Pub Type(s)

Comparative Study
Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

8660619

Citation

Davey, M W., et al. "Analysis of Ascorbate in Plant Tissues By High-performance Capillary Zone Electrophoresis." Analytical Biochemistry, vol. 239, no. 1, 1996, pp. 8-19.
Davey MW, Bauw G, Montagu MV. Analysis of ascorbate in plant tissues by high-performance capillary zone electrophoresis. Anal Biochem. 1996;239(1):8-19.
Davey, M. W., Bauw, G., & Montagu, M. V. (1996). Analysis of ascorbate in plant tissues by high-performance capillary zone electrophoresis. Analytical Biochemistry, 239(1), 8-19.
Davey MW, Bauw G, Montagu MV. Analysis of Ascorbate in Plant Tissues By High-performance Capillary Zone Electrophoresis. Anal Biochem. 1996 Jul 15;239(1):8-19. PubMed PMID: 8660619.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Analysis of ascorbate in plant tissues by high-performance capillary zone electrophoresis. AU - Davey,M W, AU - Bauw,G, AU - Montagu,M V, PY - 1996/7/15/pubmed PY - 1996/7/15/medline PY - 1996/7/15/entrez SP - 8 EP - 19 JF - Analytical biochemistry JO - Anal Biochem VL - 239 IS - 1 N2 - We describe here a simple and rapid capillary electrophoresis method for the determination of ascorbic acid (L-AA) and isoascorbic acid (D-AA) in vegetative tissues. For optimal yields and stabilization, samples are extracted with cold 3% metaphosphoric acid. Hydrophobic contaminants are then removed by passage through a C18 solid-phase extraction cartridge. The analysis itself is performed on a fused silica capillary with 200 mM borate, pH 9, as the carrier electrolyte, using on-line diode array detection over the range 190-350 nm. Quantitation was performed at 260 nm, the uv-absorption maximum for ascorbate at this pH. This method has a minimum detection limit of 84 fmol/injection and linearity of detector response was observed up to at least 12 pmol/injection. We also describe the influence of electrolyte concentration, pH, and the presence of detergent on separations of L-AA, D-AA, and L-galacturonic acid-1,4-lactone. The protocol has been demonstrated to be suitable for the analysis of L-AA in Arabidopsis, parsley, and mushroom. The method has superior resolution to comparable HPLC separations, a comparable analysis time, but lower sensitivity because of the concentration limitations of the detection system. SN - 0003-2697 UR - https://www.unboundmedicine.com/medline/citation/8660619/Analysis_of_ascorbate_in_plant_tissues_by_high_performance_capillary_zone_electrophoresis_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0003-2697(96)90284-5 DB - PRIME DP - Unbound Medicine ER -