Validating a High-performance Liquid Chromatography method for the quantification of gibberellic acid in germinating maize seeds
https://orcid.org/0000-0003-1335-5620
Javier Torres
https://orcid.org/0000-0002-9962-5769
Yaned M Correa-Navarro
https://orcid.org/0000-0001-9236-209X
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Gibberellic acid is a phytohormone that triggers the germination of seeds in a state of dormancy. Through the quantification of this hormone, the physiological condition of seeds of economic importance can be studded. In this work we validated a High-Performance Liquid Chromatography method to quantify gibberellic acid in germinated maize (Zea mays L.) seeds. Chromatographic conditions included the use of a C-18 reversed-phase column, acetonitrile-formic acid (1 : 9 %) as the mobile phase, flow of 0.5 mL·min-1, and detection at 195 nm. We evaluated our method for seven analytical parameters. The method was linear for gibberellic acid concentrations from1.0 mg·kg-1 to 50.0 mg·kg-1. The method’s limits were 0.3 mg·kg-1 and
1.0 mg·kg-1 for detection and quantification, respectively. The method was highly precise; we obtained variable but low relative standard deviations (2.62 % - 12.66 %) for the studied gibberellic acid concentrations. We assessed accuracy through recovery percentages, ranging from 52.85 % - 63.68 %, for three gibberellic acid concentrations. We conclude that our analytical method can be used to measure gibberellic acid during the early stages of maize germination. In addition, the method could be used for the analysis of other types of plant matrices.
High-Performance Liquid Chromatography, Dynamic sonication-assisted solvent extraction (DSASE), phytohormones, gibberellic acid, validation, concentration, quantification
doi: 10.1242/dev.087650
[2] Miransari M, Smith DL. Plant hormones and seed germination. Environmental and Experimental Botany, 99: 110-121, 2014.
doi: 10.1016/j.envexpbot.2013.11.005
[3] UNODC, Directrices para la validación de métodos analíticos y la calibración del equipo utilizado para el análisis de drogas ilícitas en materiales incautados y especímenes biológicos. Naciones Unidas, New York, USA 2010.
[4] Urbanová T, Tarkowská D, Novák O, Hedden P, Strnad M. Analysis of gibberellins as free acids by ultra performance liquid chromatography-tandem mass spectrometry. Talanta, 112: 85-94, 2013.
doi: 10.1016/j.talanta.2013.03.068
[5] Hamayun M, Khan SA, Khan AL, Ahmad N, Nawaz Y, Sher H, Lee I. Gibberellin producing Neosartorya sp. CC8 reprograms Chinese cabbage to higher growth. Scientia Horticulturae, 129: 347-352, 2011.
doi: 10.1016/j.scienta.2011.03.046
[6]Wu Q, Wu D, Duan C, Shen Z, Guan Y. Hollow fiber-based liquidliquid- liquid micro-extraction with osmosis: II. Application to quantification of endogenous gibberellins in rice plant. Journal of Chromatography A, 1265: 17-23, 2012.
doi: 10.1016/j.chroma.2012.09.066
[7] Xie W, Han C, Zheng Z, Chen X, Qian X, Ding H, Shi L, Lv C. Determination of Gibberellin A3 residue in fruit samples by liquid chromatography-tandem mass spectrometry. Food Chemistry, 127: 890-892, 2011.
doi: 10.1016/j.foodchem.2011.01.018
[8] Correa YM, Buriticá LM, Rivera JD, Penagos JP, Torres JI. Optimización del protocolo para la extracción y la cuantificación de proteínas totales en semillas germinadas de maíz (Zea mays L.). Revista Facultad de. Ciencias Basicas, 13(1): 60-64, 2017.
[9] Rivera JD, Correa YM, Penagos JP. Evaluación de métodos de extracción para la obtención del ácido giberélico en semillas germinadas de maíz (Zea mays L.). Revista Colombiana de Química, 46(2): 45-50, 2017.
doi: 10.15446/rev.colomb.quim.v46n2.63015
[10] Bhalla K, Singh SB, Agarwal R. Quantitative determination of gibberellins by high performance liquid chromatography from various gibberellins producing Fusarium strains. Environmental Monitoring and Assessment, 167: 515-520, 2010.
doi: 10.1007/s10661-009-1068-5
[11] Instituto de Salud Pública. Validación de métodos y determinación de la incertidumbre de la medición. Santiago, Chile 2010.
[12] Miller JN, Miller JC. Estadística Y Quimiometría Para Química Analítica. Prentice Hall, Madrid, Spain 2002.
[13] Danzer K, Currie LA. Guideline for calibration in analytical chemistry. Part 1. Fundamentals and single component calibration. Pure and Applied Chemistry, 70(4): 993-1014, 1998.
doi: 324 10.1016/j.cell.2014.03.046
[14] Olivieri AC. Practical guidelines for reporting results in singleand multi-component analytical calibration: A tutorial. Analytica Chimica Acta, 868: 10-22, 2015.
doi: 10.1016/j.aca.2015.01.017
[15] Ge L, Peh CYC, Yong JWH, Tan SN, Hua L, Ong ES. Analyses of gibberellins by capillary electrophoresis-mass spectrometry combined with solid-phase extraction. Journal of Chromatography A, 1159: 242-249, 2007.
doi: 10.1016/j.chroma.2007.05.041
[16] Manzi M, Gómez A, Arbona V. Rapid and reproducible determination of active gibberellins in citrus tissues by UPLC/ESI-MS/MS. Plant Physiology and Biochemistry, 94: 1-9, 2015.
doi: 10.1016/j.plaphy.2015.04.015