Development of a Rapid and Efficient Liquid Chromatography Method for Determination of Gibberellin A4 in Plant Tissue, with Solid Phase Extraction for Purification and Quantification

Abstract

A new, rapid and efficient reverse phase Liquid Chromatography (RP-LC) method was developed for determination of Gibberellin A4 (GA4) in samples of flower stalk of Dasylirion cedrosanum and vegetative tissue of Epithelantha micromeris. Purification of GA4 was carried out by solid phase extraction (SPE), in Epithelantha micromeris. In the chromatography method was obtaining a retention time of 2.1 min, using Hypersil GOLD C-18 column (100 × 4.6 mm dim and size particle 5 μ), mobile phase 50/50 acetonitrile/water and a flow 1.0 ml/min. Detection was carried out by a UV detector set at 205 nm, and a quantization limit of 0.4 mg/L. The obtained correlation coefficient was 0.995.

Share and Cite:

Macías, J. , Pournavab, R. , Reyes-Valdés, M. and Benavides-Mendoza, A. (2014) Development of a Rapid and Efficient Liquid Chromatography Method for Determination of Gibberellin A4 in Plant Tissue, with Solid Phase Extraction for Purification and Quantification. American Journal of Plant Sciences, 5, 573-583. doi: 10.4236/ajps.2014.55072.

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1] Lale, G. and Gadre, R. (2010) Enhanced Production of Gibberellin A4 (GA4) by a Mutant of Gibberella fujikuroi in Wheat Gluten Medium. Journal of Industrial Microbiology & Biotechnology, 37, 297-306.
http://dx.doi.org/10.1007/s10295-009-0673-1
[2] Macmillan, J. (2002) Occurrence of Gibberellins in Vascular Plants, Fungi, and Bacteria. Journal of Plant Growth Regulation, 20, 387-442. http://dx.doi.org/10.1007/s003440010038
[3] Hedden, P. and Phillip, A.L. (2000) Gibberellin Metabolism: New Insights Revealed by the Genes. Trends in Plant Science, 5, 523-530. http://dx.doi.org/10.1016/S1360-1385(00)01790-8
[4] Olszewski, N., Sun, T. P. and Gubler, F. (2002) Gibberellin Signaling: Biosynthesis, Catabolism, and Response Pathways. The Plant Cell, 14, S61-S80.
[5] Mander, L. (1992) The Chemistry of Gibberellins: An Overview. Chemical Reviews, 92, 573-612.
http://dx.doi.org/10.1021/cr00012a005
[6] Dunberg, A. and Odén, P.C. (1983) Gibberellins and Conifers. In: Crozier, A., Ed., The Biochemistry and Physiology of Gibberellins, 2nd Edition, Praeger, New York, 221-295.
[7] Fernández, H., Fraga, M.F., Bernard, P. and Revilla, M.A. (2003) Quantification of GA1, GA3, GA4, GA7, GA9, and GA20 in Vegetative and Male Cone Buds from Juvenile and Mature Trees of Pinus radiate. Plant Growth Regulation, 40, 185-188. http://dx.doi.org/10.1023/A:1025070707899
[8] Dobrev, P.I. and Kaminek, M. (2002) Fast and Efficient Separation of Cytokinins from Auxin and Abscisic Acid and Their Purification Using Mixed-Mode Solid-Phase Extraction. Journal of Chromatography A, 950, 21-29.
[9] Mortiz, T., Philipson, J. and Odén, C. (1990) Quantization of Gibberellins A1 , A3, A4, A9 and an A9-Conjugate in Good- and Poor-Flowering Clones of Sitka spruce (Picea sitchensis) during the Period of Flower-Bud Differentiation. Planta, 181, 538-542.
[10] Gallazzo, J.L. and Lee, M.D. (2001) Production of High Titers of Gibberellins, GA4 and GA7 by Strain. LTB-1027. US Patent 6287800.
[11] Bruckne, B. and Blechschmid, D. (1990) The Gibberellin Fermentation. Critical Reviews in Biotechnology, 11, 163-192.
[12] Kim, H.-J. and Millar, B.W. (2009) GA4+7 plus BA Enhances Postproduction Quality in Pot Tulips. Postharvest Biology and Technology, 51, 272-277. http://dx.doi.org/10.1016/j.postharvbio.2008.07.002
[13] Batlang, U.L. (2009) Benzyladenine plus Gibberellins GA4+7 Increase Fruit Size and Yield in Greenhouse-Grown Hot Pepper (Capsicum annuum L.). Journal of Biological Sciences, 8, 659-662. http://dx.doi.org/10.3923/jbs.2008.659.662
[14] Dawes, P., Dawes, E., Difeo, O.D. and Wayyne, P. (2007) On-Line and Off-Line Application of Micro-SPE (MEPS) SGE. Analytical Science. http://www.sge.com/uploads/Kq/J3/KqJ3-rqH9_4kF680zX7uKQ/TP-0158-S_MEPS-A2.pdf
[15] Bhalla, K., Bala, S. and Agarwal, S.R. (2010) Quantitative Determination of Gibberellins by High Performance Liquid Chromatography from Various Gibberellins Producing Fusarium strains. Environmental Monitoring and Assessment, 167, 515-520. http://dx.doi.org/10.1007/s10661-009-1068-5
[16] du Preez, J.C., Qian, X.-M. and Kilian, S.G. (1993) Stability and Bioactivity of Gibberellic Acid in Different Solvents. Biotechnology Techniques, 7, 391-396. http://dx.doi.org/10.1007/BF00152547
[17] Nefed’eva, E.E and Mazey, N.G. (2009) Gibberellin A3 Detection in Plants with High-Performance Liquid Chromatography. Applied Biochemistry and Microbiology, 45, 454-458. http://dx.doi.org/10.1134/S000368380904019X
[18] Kelen, M. (2004) Separation of Abscisic Acid, Indole-3-Acetic Acid, Gibberellic Acid in 99 R (Vitis berlandieri x Vitis rupestris) and Rose Oil (Rosa damascena Mill.) by Reversed Phase Liquid Chromatography. Turkish Journal of Chemistry, 28, 603 -610.
[19] Ayele, B.T., Magnus, V., Mihaljevic, S.A., Prebeg, T., CoZ-Rakovac, R., Ozga, J.A., Reinecke, M.D., Mander, L.N., Kamiya, Y., Yamaguchi, S. and Salopek-Sondi, B. (2009) Endogenous Gibberellin Profile during Christmas Rose (Helleborus niger L.) Flower and Fruit Development. Journal of Plant Growth Regulation, 29, 194-209.
http://dx.doi.org/10.1007/s00344-009-9124-5
[20] Stephan, M., Bangerth, F. and Schneider, G. (1999) Quantification of Endogenous Gibberellins in Exudates from Fruits of Malus domestica. Plant Growth Regulation, 28, 55-58. http://dx.doi.org/10.1023/A:1006211309707
[21] King, R.W., Pharis, R.P and Mander, L.N. (1987) Gibberellins in Relation to Growth and Flowering in Pharbitis nil Chois. Plant Physiology, 84, 1126-1131. http://dx.doi.org/10.1104/pp.84.4.1126
[22] Tudzynski, B. (2005) Gibberellin Biosynthesis in Fungi: Genes, Enzymes, Evolution and Impact on Biotechnology. Applied Microbiology and Biotechnology, 66, 597-611.
[23] Menéndez, V., Revilla, M.A., Bernard, P., Gotor, V. and Fernández, H. (2006) Gibberellins and Antheridiogen on Sex in Blechnum spicant L.. Plant Cell Reports, 25, 1104-1110. http://dx.doi.org/10.1007/s00299-006-0149-y
[24] Chen, J.G., Zhou, X. and Zhang, Y.Z. (1998) Gibberellin-Responding and Non-Responding Dwarf Mutants in Foxtail Millet. Plant Growth Regulation, 26, 19-24. http://dx.doi.org/10.1007/s00299-006-0149-y
[25] Kim, S.K., Lee, S.C.H., Shin, D.H., Jang, S.W., Nam, W.J., Park, T.S. and Lee, L.G. (2003) Quantification of Endogenous Gibberellins in Leaves and Tubers of Chinese yam, Dioscorea opposita Thunb cv. Tsukune during Tuber Enlargement. Plant Growth Regulation, 39, 125-130. http://dx.doi.org/10.1023/A:1022569506883
[26] Oden, P.C., Schwenen, L. and Graebe, J.E. (1987) Identification of Gibberellins in Norway Spruce (Picea abies [L.J Karst.) by Combined Gas Chromatography-Mass Spectrometry. Plant Physiology, 84, 516-519. http://dx.doi.org/10.1104/pp.84.2.516
[27] Meijón, M., Cañal, M.J., Fernández, H., Rodríguez, A., Fernández, B., Rodríguez, R. and Feito, I. (2011) Hormonal Profile in Vegetative and Floral Buds of Azalea: Levels of Polyamines, Gibberellins, and Cytokinins. Journal of Plant Growth Regulation, 30, 74-82. http://dx.doi.org/10.1007/s00344-010-9169-5
[28] Sigma Aldrich Co. (1999) Guide to Solid Phase Extraction, Bulletin, 919.
[29] Kim, S.K., Sohn, E.Y., Kang, S.M. and Lee, I.J. (2009) Quantification of Endogenous Gibberellins in Two Flax (Linum usitatissimum L.) Cultivars during Seed Development. Journal of Crop Science and Biotechnology, 12, 43-46.

Copyright © 2024 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.