Optimization of an HPLC [high performance liquid chromatography] method for gibberellic acid level determination in mango
1997
Protacio, C.M. | Molinyawe, G.G. | Bugante, R.D. Jr. (Philippines Univ. Los Banos, College, Laguna (Philippines). Dept. of Horticulture)
In order to understand the physiological changes accompanying observed phenological markers of flowering in the mango tree levels of gibberellins and carbohydrates were measured. This preliminary study details the optimization procedure for assaying gibberellic acid by high-performance liquid chromatography (HPLC) as reported by Barendse et al. (1980. Plant Physiol 198:449-455). Essentially, different available GA standards in methanol were passed through C18 cartridges to simulate sample clean up. The mixture of standards was then chromotographed through a Waters TM HPLC system using reversed phase columns of different pore sizes (4 to 10 micro mole) to determine if they can be resolved individually using an isocratic water-methanol mobile phase. Once the individual peaks for each standard were satisfactorily resolved and their retention times determined, then sample extracts were run. Fractions corresponding to various gibberellins were collected and their biological activities checked by dwarf rice leaf sheath elongation bioassay using the var. Tan-ginboru (Murakami, 1970. Bot. Mag. 83:312-324)
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