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.

ESTANDARIZACIÓN DE UN MÉTODO CROMATOGRÁFICO PARA LA IDENTIFICACIÓN DEL ÁCIDO GIBERÉLICO EN SEMILLAS DE MAÍZ (Zea mays L.)

“…Bajo estas condiciones se encontró al patrón de ácido giberélico (GA 3 ) a una longitud de onda máxima de absorción de 195 nm. ; por debajo de lo reportado por Macías et al, [14], Bhalla et al, [13] La curva de Van Deemter (Figura 2) muestra la relación entre la velocidad lineal (u) de la fase móvil y la altura equivalente de plato teórico (HETP). En esta se observa que la menor altura de equivalente de plato teórico (0.0025 mm.)…”

“…Sin embargo, el uso de estos protocolos se ha dejado de lado a medida que se han desarrollado técnicas físicas de medida que permiten la identificación y la cuantificación precisa de giberelinas específicas a partir de pequeñas cantidades de tejido vegetal (del orden de mg a g) [11]. En la actualidad se emplea con más frecuencia la cromatografía líquida de alta eficiencia con detector de arreglo de diodos (HPLC-DAD) [12]- [14] y la cromatografía líquida de alta eficiencia acoplada a espectrometría de masas (HPLC-MS) [5], [7], [15], Estas técnicas tienen como ventajas mayor eficiencia en la separación y alta especificidad en la detección del analito de estudio [16]. Otro método menos utilizado es la electroforesis capilar acoplada a espectrometría de masas (CE-MS) [17] y la cromatografía de gases acoplada a espectrometría de masas (GC-MS) [18]- [20], sin embargo, en esta última técnica cromatográfica la muestra a analizar debe ser derivatizada previamente lo cual alarga el proceso de análisis y no asegura la modificación de la muestra [21].…”

Multifamily Determination of Phytohormones and Acidic Herbicides in Fruits and Vegetables by Liquid Chromatography–Tandem Mass Spectrometry under Accredited Conditions

“…LC coupled to mass spectrometry (MS) provides a powerful tool to analyze both phytohormones [ 4 , 22 ] and acidic herbicides [ 33 ] in food matrices. Furthermore, LC can be coupled to other detection systems due to the presence of chromophores of the analytes under UV–vis conditions, such as diode arrays (DADs) ultraviolet (UV) [ 34 , 35 ] and fluorescence detectors (FLDs), [ 36 ], but these have less applicability in this field due to their technical limitations.…”

Phytohormone Profiling Method for Rice: Effects of GA20ox Mutation on the Gibberellin Content of Japonica Rice Varieties

“…Gibberellins, a large hormone category, are a large group of tetracyclic diterpenoid carboxylic acids, which were first identified as secondary metabolites of the fungus Gibberella fujikuroi (Hedden and Thomas, 2012). Nowadays, more than 136 different gibberellin structures have been found, but four of them are highly bioactive: GA 1 , GA 3 , GA 4 , and GA 7 (Hedden and Phillips, 2000; Yamaguchi, 2008; Macías et al, 2014). Moreover, these four bioactive GAs have been detected in different rice developmental stages (Ma et al, 2011; Wu et al, 2012).…”

“…Crucial points during gibberellin analysis are extraction and cleaning steps, which should ensure high presence of GAs and low presence of other molecules (Urbanová et al, 2013). The majority of current GAs extraction methods use the classic liquid-liquid extraction and solid phase extraction (SPE) with reverse phase C-18 cartridge for sample concentration and clean up (Macías et al, 2014). Nowadays, HPLC-MS/MS is the standard and routine technique for GAs separation and detection (Urbanová et al, 2013; Macías et al, 2014) mainly using triple quadrupole instruments for their quantification at trace levels.…”