Normal-phase liquid chromatography of plant hormones using reversed cholic acid micelles as the mobile phase.

When several plant hormones with similar structures are present in a sample, conventional analytical methods are not sufficiently selective to effectively differentiate them. However, the excellent selectivity of micellar liquid chromatography allows similarly structured compounds to be easily differentiated. We used reversed cholic acid micelles dissolved in tetrahydrofuran as the mobile phase in normal-phase chromatography of seven plant hormones. Our results conformed closely to those predicted by the pseudophase micellar liquid chromatography theory. The chromatographic efficiencies were calculated from the peak width and the retention times, and we compared these values with those obtained by reversed-phase chromatography using sodium dodecyl sulfate mobile phase in water. To test the efficiency of this selective method for the analysis of plant tissues that may also contain contaminants, we applied it to homogeneous suspensions of spiked maize roots. Recoveries ranged from 91 to 110%, the relative standard deviations were between 0.46 and 10.03%, and the detection limits were between 0.08 and 0.80 micrograms g-1.