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  • 1 Hungarian Academy of Sciences Department of Plant Physiology, Agricultural Institute, Centre for Agricultural Research Brunszvik u. 2 H-2462 Martonvásár Hungary
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Plants have developed various mechanisms to protect themselves against oxidative stress. One of the most important non-enzymatic antioxidants is ascorbic acid. There is thus a need for a rapid, sensitive method for the analysis of the reduced and oxidised forms of ascorbic acid in crop plants. In this paper a simple, economic, selective, precise and stable HPLC method is presented for the detection of ascorbate in plant tissue. The sensitivity, the short retention time and the simple isocratic elution mean that the method is suitable for the routine quantification of ascorbate in a high daily sample number. The method has been found to be better than previously reported methods, because of the use of an economical, readily available mobile phase, UV detection and the lack of complicated extraction procedures. The method has been tested on Arabidopsis plants with different ascorbate levels and on wheat plants during Cd stress.

  • Agarwal, S. (2007) Increased antioxidant activity in Cassia seedlings under UV-B radiation. Biol. Plant. 51, 157–160.

    Agarwal S. , 'Increased antioxidant activity in Cassia seedlings under UV-B radiation ' (2007 ) 51 Biol. Plant. : 157 -160.

    • Search Google Scholar
  • Arya, S. P., Mahajan, M., Jain, P. (2000) Non-spectrophotometric methods for determination of Vitamin C. Anal. Chim. Acta 417, 1–14.

    Jain P. , 'Non-spectrophotometric methods for determination of Vitamin C ' (2000 ) 417 Anal. Chim. Acta : 1 -14.

    • Search Google Scholar
  • Attolico, A. D., De Tullio, M. C. (2006) Increased ascorbate content delays flowering in long-day grown Arabidopsis thaliana (L.) Heynh. Plant Physiol. Biochem. 44, 462–466.

    Tullio M. C. , 'Increased ascorbate content delays flowering in long-day grown Arabidopsis thaliana (L.) Heynh. ' (2006 ) 44 Plant Physiol. Biochem. : 462 -466.

    • Search Google Scholar
  • Bartoli, C. G., Pastori, G. M., Foyer, C. H. (2000) Ascorbate biosynthesis in mitochondria is linked to the electron transport chain between complexes III and IV. Plant Physiol. 123, 335–343.

    Foyer C. H. , 'Ascorbate biosynthesis in mitochondria is linked to the electron transport chain between complexes III and IV ' (2000 ) 123 Plant Physiol. : 335 -343.

    • Search Google Scholar
  • Chen, T. H. H., Murata, N. (2002) Enhancement of tolerance of abiotic stress by metabolic engineering of betaines and other compatible solutes. Curr. Opin. Plant Biol. 5, 250–257.

    Murata N. , 'Enhancement of tolerance of abiotic stress by metabolic engineering of betaines and other compatible solutes ' (2002 ) 5 Curr. Opin. Plant Biol. : 250 -257.

    • Search Google Scholar
  • Conklin, P. L., Williams, E. H., Last, R. L. (1996) Environmental stress sensitivity of an ascorbic acid-deficient Arabidopsis mutant (ozone/ultraviolet B/sulfur dioxide/reactive oxygen detoxification/vitamin C). Proc. Natl. Acad. Sci USA 93, 9970–9974.

    Last R. L. , 'Environmental stress sensitivity of an ascorbic acid-deficient Arabidopsis mutant (ozone/ultraviolet B/sulfur dioxide/reactive oxygen detoxification/vitamin C) ' (1996 ) 93 Proc. Natl. Acad. Sci USA : 9970 -9974.

    • Search Google Scholar
  • Davey, M. W., Van Montagu, M., Inze, D., Sanmartin, M., Kanellis, A., Smirnoff, N., Benzie, I. J. J., Strain, J. J., Favell, D., Fletcher, J. (2000) Plant L-ascorbic acid: chemistry, function, metabolism, bioavailability and effects of processing. J. Sci. Food Agric. 80, 825–860.

    Fletcher J. , 'Plant L-ascorbic acid: chemistry, function, metabolism, bioavailability and effects of processing ' (2000 ) 80 J. Sci. Food Agric. : 825 -860.

    • Search Google Scholar
  • Demirevska-Kepova, K., Simova-Stoilova, L., Stoyanova Z. P., Feller, U. (2006) Cadmium stress in barley: growth, leaf pigment, and protein composition and detoxification of reactive oxygen species. J. Plant Nutr. 29, 451–468.

    Feller U. , 'Cadmium stress in barley: growth, leaf pigment, and protein composition and detoxification of reactive oxygen species ' (2006 ) 29 J. Plant Nutr. : 451 -468.

    • Search Google Scholar
  • Fercha, A., Hocine, G., Mebarek, B. (2011) Improvement of salt tolerance in durum wheat by ascorbic acid application. J. Stress Physiol. Biochem. 7, 27–37.

    Mebarek B. , 'Improvement of salt tolerance in durum wheat by ascorbic acid application ' (2011 ) 7 J. Stress Physiol. Biochem. : 27 -37.

    • Search Google Scholar
  • Foyer, C. H., Noctor, G. (2005) Redox homeostis and antioxidant signaling: a metabolic interface between stress perception and physiological responses. Plant Cell 17, 1866–1875.

    Noctor G. , 'Redox homeostis and antioxidant signaling: a metabolic interface between stress perception and physiological responses ' (2005 ) 17 Plant Cell : 1866 -1875.

    • Search Google Scholar
  • Furusawa, N. (2001) Rapid high-performance liquid chromatographic identification/quantification of total vitamin C in fruit drinks. Food Control 12, 27–29.

    Furusawa N. , 'Rapid high-performance liquid chromatographic identification/quantification of total vitamin C in fruit drinks ' (2001 ) 12 Food Control : 27 -29.

    • Search Google Scholar
  • Gallie, D. R. (2013) l-Ascorbic acid: A multifunctional molecule supporting plant growth and development. Scientifica, 2013, Article ID 795964.

    Gallie D. R. , '', in Scientifica, 2013 , (2013 ) -.

  • Gazdik, Z., Zitka, O., Petrlova, J., Adam, V., Zehnalek, J., Horna, A., Reznicek, V., Beklova, M., Kizek, R. (2008) Determination of vitamin C (ascorbic acid) using high performance liquid chromatography coupled with electrochemical detection. Sensors 8, 7097–7112.

    Kizek R. , 'Determination of vitamin C (ascorbic acid) using high performance liquid chromatography coupled with electrochemical detection ' (2008 ) 8 Sensors : 7097 -7112.

    • Search Google Scholar
  • Gest, N., Gautier, H., Stevens, R. (2013) Ascorbate as seen through plant evolution: the rise of a successful molecule? J. Exp. Bot. 64, 33–53.

    Stevens R. , 'Ascorbate as seen through plant evolution: the rise of a successful molecule? ' (2013 ) 64 J. Exp. Bot. : 33 -53.

    • Search Google Scholar
  • Karau, G. M., Njagi, E. N. M., Machocho, A. K., Wangai, L. N. (2012) Phytonutrient, mineral composition and in vitro antioxidant activity of leaf and stem bark powders of Pappea capensis (L.). Pak. J. Nutrition 11, 123–132.

    Wangai L. N. , 'Phytonutrient, mineral composition and in vitro antioxidant activity of leaf and stem bark powders of Pappea capensis (L.). Pak ' (2012 ) 11 J. Nutrition : 123 -132.

    • Search Google Scholar
  • Khan, T. A., Mazid, M., Mohammad, F. (2012) Potential of ascorbic acid against oxidative burst in plants under biotic stress: A review. J. Ind. Res. & Technology 2, 72–80.

    Mohammad F. , 'Potential of ascorbic acid against oxidative burst in plants under biotic stress: A review ' (2012 ) 2 J. Ind. Res. & Technology : 72 -80.

    • Search Google Scholar
  • Kotchoni, S. O., Larrimore, K. E., Mukherjee, M., Kempinski, C. F., Barth, C. (2009) Alterations in the endogenous ascorbic acid content affect flowering time in Arabidopsis. Plant Physiol. 149, 803–815.

    Barth C. , 'Alterations in the endogenous ascorbic acid content affect flowering time in Arabidopsis ' (2009 ) 149 Plant Physiol. : 803 -815.

    • Search Google Scholar
  • Li, Q., Krauss, M. R. Hempfling, W. P. (2006) Wounding of root or basal stalk prior to harvest affects pre-harvest antioxidant accumulation and tobacco-specific nitrosamine formation during air curing of burley tobacco (Nicotiana tabacum L.). J. Agr. Crop Sci. 192, 267–277.

    Hempfling W. P. , 'Wounding of root or basal stalk prior to harvest affects pre-harvest antioxidant accumulation and tobacco-specific nitrosamine formation during air curing of burley tobacco (Nicotiana tabacum L.) ' (2006 ) 192 J. Agr. Crop Sci. : 267 -277.

    • Search Google Scholar
  • Li, Q., Li, Y., Li, C., Yu, X. (2012) Enhanced ascorbic acid accumulation through overexpression of dehydroascorbate reductase confers tolerance to methyl viologen and salt stresses in tomato. Czech J. Genet. Plant Breed. 48, 74–86.

    Yu X. , 'Enhanced ascorbic acid accumulation through overexpression of dehydroascorbate reductase confers tolerance to methyl viologen and salt stresses in tomato ' (2012 ) 48 Czech J. Genet. Plant Breed : 74 -86.

    • Search Google Scholar
  • López-Carbonell, M., Munné-Bosch, S., Alegre, L. (2006) The ascorbate-deficient vtc-1 Arabidopsis mutant shows altered ABA accumulation in leaves and chloroplasts. J. Plant Growth Regul. 25, 137–144.

    Alegre L. , 'The ascorbate-deficient vtc-1 Arabidopsis mutant shows altered ABA accumulation in leaves and chloroplasts ' (2006 ) 25 J. Plant Growth Regul. : 137 -144.

    • Search Google Scholar
  • Nayyar, H., Chander, S. (2004) Protective effects of polyamines against oxidative stress induced by water and cold stress in chickpea. J. Agr. Crop Sci. 190, 355–365.

    Chander S. , 'Protective effects of polyamines against oxidative stress induced by water and cold stress in chickpea ' (2004 ) 190 J. Agr. Crop Sci. : 355 -365.

    • Search Google Scholar
  • Niu, Y., Wang, Y., Li, P., Zhang, F., Liu, H., Zheng, G. (2012) Drought stress induces oxidative stress and the antioxidant defense system in ascorbate-deficient vtc1 mutants of Arabidopsis thaliana. Acta Physiol. Plant. 35, 1189–1200.

    Zheng G. , 'Drought stress induces oxidative stress and the antioxidant defense system in ascorbate-deficient vtc1 mutants of Arabidopsis thaliana ' (2012 ) 35 Acta Physiol. Plant. : 1189 -1200.

    • Search Google Scholar
  • Novákova, L., Solich, P., Solichova, D. (2008) HPLC methods for simultaneous determination of ascorbic and dehydroascorbic acids. Trends Anal. Chem. 27, 942–958.

    Solichova D. , 'HPLC methods for simultaneous determination of ascorbic and dehydroascorbic acids ' (2008 ) 27 Trends Anal. Chem. : 942 -958.

    • Search Google Scholar
  • Panda, S. K., Khan, M. H. (2009) Growth, oxidative damage and antioxidant responses in Greengram (Vigna radiata L.) under short-term salinity stress and its recovery. J. Agr. Crop Sci. 195, 442–454.

    Khan M. H. , 'Growth, oxidative damage and antioxidant responses in Greengram (Vigna radiata L.) under short-term salinity stress and its recovery ' (2009 ) 195 J. Agr. Crop Sci. : 442 -454.

    • Search Google Scholar
  • Pisoschi, A. M., Danet, A. F., Kalinowski, S. (2008) Ascorbic acid determination in commercial fruit juice samples by cyclic voltammetry. J. Autom. Methods Manag. Chem. 2008, Article ID 937651, doi: 10.1155/2008/937651

    Kalinowski S. , '', in J. Autom. Methods Manag. Chem , (2008 ) -.

  • Prasad, B. B., Tiwari, K., Singh, M., Sharma, P. S., Patel, A. K., Srivastava, S. (2008) Molecularly imprinted polymer-based solid-phase microextraction fiber coupled with molecularly imprinted polymer-based sensor for ultratrace analysis of ascorbic acid. J. Chromatogr. A1198, 59–66.

    Srivastava S. , 'Molecularly imprinted polymer-based solid-phase microextraction fiber coupled with molecularly imprinted polymer-based sensor for ultratrace analysis of ascorbic acid ' (2008 ) A1198 J. Chromatogr. : 59 -66.

    • Search Google Scholar
  • Ribeiro, C., Cambraia, J., Peixoto, P. H. P., da Fonseca, J. É. M. (2012) Antioxidant system response induced by aluminum in two rice cultivars. Braz. J. Plant Physiol. 24, 107–116.

    Fonseca J. É. M. M. , 'Antioxidant system response induced by aluminum in two rice cultivars ' (2012 ) 24 Braz. J. Plant Physiol. : 107 -116.

    • Search Google Scholar
  • Sawant, L., Prabhakar, B., Mahajan, A., Pai, N., Pandita, N. (2011) Development and validation of HPLC method for quantification of phytoconstituents in Phyllanthus emblica. J. Chem. Pharm. Res. 3, 937–944.

    Pandita N. , 'Development and validation of HPLC method for quantification of phytoconstituents in Phyllanthus emblica ' (2011 ) 3 J. Chem. Pharm. Res. : 937 -944.

    • Search Google Scholar
  • Shekhovtsova, T. N., Muginova, S. V., Luchinina, J. A., Galimova, A. Z. (2006) Enzymatic methods in food analysis: determination of ascorbic acid. Anal. Chim. Acta 573–574, 125–132.

    Galimova A. Z. , 'Enzymatic methods in food analysis: determination of ascorbic acid ' (2006 ) 573–574 Anal. Chim. Acta : 125 -132.

    • Search Google Scholar
  • Smirnoff, N. (1996) The function and metabolism of ascorbic acid in plants. Annals Bot. 78, 661–669.

    Smirnoff N. , 'The function and metabolism of ascorbic acid in plants ' (1996 ) 78 Annals Bot. : 661 -669.

    • Search Google Scholar
  • Smirnoff, N. (2000) Ascorbic acid: metabolism and functions of a multi-facetted molecule. Curr. Opin. Plant Biol. 3, 229–235.

    Smirnoff N. , 'Ascorbic acid: metabolism and functions of a multi-facetted molecule ' (2000 ) 3 Curr. Opin. Plant Biol. : 229 -235.

    • Search Google Scholar
  • Sommano, S., Caffin, N., McDonald, J., Kerven, G. (2011) Measurement of ascorbic acid in Australian native plants. IFRJ 18, 1017–1020.

    Kerven G. , 'Measurement of ascorbic acid in Australian native plants ' (2011 ) 18 IFRJ : 1017 -1020.

    • Search Google Scholar
  • Soni, H., Singhai, A. K., Sharma, S. (2012) Quantification of ascorbic acid in leaves of Annona squamosa. Int. J. Pharm. Pharm. Sci. 4, 144–147.

    Sharma S. , 'Quantification of ascorbic acid in leaves of Annona squamosa ' (2012 ) 4 Int. J. Pharm. Pharm. Sci. : 144 -147.

    • Search Google Scholar
  • Soni, H., Singhai, A. K., Sharma, S., Nayak, G., Swarnkar, P. (2012) Quantification of ascorbic acid in salad components. Int. J. Curr. Pharm. Res. 4, 43–47.

    Swarnkar P. , 'Quantification of ascorbic acid in salad components ' (2012 ) 4 Int. J. Curr. Pharm. Res. : 43 -47.

    • Search Google Scholar
  • Streb, P., Aubert, S., Gout, E., Bligny R. (2003) Reversibility of cold- and light-stress tolerance and accompanying changes of metabolite and antioxidant levels in the two high mountain plant species Soldanella alpina and Ranunculus glacialis. J. Exp. Bot. 54, 405–418.

    Bligny R. , 'Reversibility of cold- and light-stress tolerance and accompanying changes of metabolite and antioxidant levels in the two high mountain plant species Soldanella alpina and Ranunculus glacialis ' (2003 ) 54 J. Exp. Bot. : 405 -418.

    • Search Google Scholar
  • Streb, P., Feierabend, J., Bligny, R. (1997) Resistance of photoinhibition of photosystem II and catalase and antioxidative protection in high mountain plants. Plant Cell Environ. 20, 1030–1040.

    Bligny R. , 'Resistance of photoinhibition of photosystem II and catalase and antioxidative protection in high mountain plants ' (1997 ) 20 Plant Cell Environ. : 1030 -1040.

    • Search Google Scholar
  • Szarka, A., Tomasskovics, B., Bánhegyi, G. (2012) The ascorbate-glutathione-α-tocopherol triad in abiotic stress response. Int. J. Mol. Sci. 13, 4458–4483.

    Bánhegyi G. , 'The ascorbate-glutathione-α-tocopherol triad in abiotic stress response ' (2012 ) 13 Int. J. Mol. Sci. : 4458 -4483.

    • Search Google Scholar
  • Vermeir, S., Hertog, M. L., Schenk, A., Beullens, K., Nicolai, B. M., Lammertyn, J. (2008) Evaluation and optimization of high-throughput enzymatic assays for fast L-ascorbic acid quantification in fruit and vegetables. Anal. Chim. Acta 618, 94–101.

    Lammertyn J. , 'Evaluation and optimization of high-throughput enzymatic assays for fast L-ascorbic acid quantification in fruit and vegetables ' (2008 ) 618 Anal. Chim. Acta : 94 -101.

    • Search Google Scholar
  • Wingsle, G., Moritz, T. (1997) Analysis of ascorbate and dehydroascorbate in plant extracts by high-resolution selected ion monitoring gas chromatography-mass spectrometry. J. Chromatogr. A782, 95–103.

    Moritz T. , 'Analysis of ascorbate and dehydroascorbate in plant extracts by high-resolution selected ion monitoring gas chromatography-mass spectrometry ' (1997 ) A782 J. Chromatogr. : 95 -103.

    • Search Google Scholar
  • Yang, Y., Han, C., Liu, Q., Lin, B., Wang, J. (2008) Effect of drought and low light on growth and enzymatic antioxidant system of Picea asperata seedlings. Acta Physiol. Plant. 30, 433–440.

    Wang J. , 'Effect of drought and low light on growth and enzymatic antioxidant system of Picea asperata seedlings ' (2008 ) 30 Acta Physiol. Plant. : 433 -440.

    • Search Google Scholar
  • Zhang, J., Kirkham, M. B. (1996) Antioxidant responses to drought in sunflower and sorghum seedlings. New Phytol. 132, 361–373.

    Kirkham M. B. , 'Antioxidant responses to drought in sunflower and sorghum seedlings ' (1996 ) 132 New Phytol. : 361 -373.

    • Search Google Scholar
  • Zhou, Y., Tao, Q. C., Wang, Z. N., Fan, R., Li, Y., Sun, X. F., Tang, K. X. (2012) Engineering ascorbic acid biosynthetic pathway in Arabidopsis leaves by single and double gene transformation. Biol. Plant. 56, 451–457.

    Tang K. X. , 'Engineering ascorbic acid biosynthetic pathway in Arabidopsis leaves by single and double gene transformation ' (2012 ) 56 Biol. Plant. : 451 -457.

    • Search Google Scholar

Editorial Board

      Csányi, Vilmos (Göd)
      Dudits, Dénes (Szeged)
      Falus, András (Budapest)
      Fischer, Ernő (Pécs)
      Gábriel, Róbert (Pécs)
      Gulya, Károly (Szeged)
      Gulyás, Balázs (Stockholm)
      Hajós, Ferenc (Budapest)
      Hámori, József (Budapest)
      Heszky, László (Gödöllő)
      Hideg, Éva (Szeged)
      E. Ito (Sanuki)
      Janda, Tibor (Martonvásár)
      Kavanaugh, Michael P. (Missoula)
      Kása, Péter (Szeged)
      Klein, Éva (Stockholm)
      Kovács, János (Budapest)
      Brigitte Mauch-Mani (Neuchâtel)
      Nässel, Dick R. (Stockholm)
      Nemcsók, János (Szeged)
      Péczely, Péter (Gödöllő)
      Roberts, D. F. (Newcastle-upon-Tyne)
      Sakharov, Dimitri A. (Moscow)
      Singh, Meharvan (Fort Worth)
      Sipiczky, Mátyás (Debrecen)
      Szeberényi, József (Pécs)
      Székely, György (Debrecen)
      Tari, Irma (Szeged)
      Vágvölgyi, Csaba (Szeged),
      L. Zaborszky (Newark)

 

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Acta Biologica Hungarica
Language English
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1950
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Founder Magyar Tudományos Akadémia
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ISSN 0236-5383 (Print)
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