High-resolution method for regulatory control of Echinacea species in Nutraceuticals by CD-MEKC

One problem in the international regulatory control of Echinacea, a therapeutic Nutraceutical, is recognition of caffeoyl solutes and alkamides in different products. Cyclodextrin-modified micellar electrokinetic chromatography (CD-MEKC) has been applied to Echinacea spp. in combination with pattern recognition of some caffeoyl solutes. A novel metric based on relative migration time (RMT) data has been developed in CE to address the problem of variable reported migration time. The CD-MEKC method of Gotti's group using hydroxypropyl-beta-cyclodexrin (HP-beta-CD; 100 mM) with sodium dodecyl sulphate (SDS; 110 mM), in a triacid background electrolyte (10 mM, pH 8) under 19 kV was adapted to identify two key hydrophilic solutes: chlorogenic acid and cichoric acid present in all commercial products. Two internal markers were taken as reference points to calculate the RMT of any target peak: RMT=t(m (target))/t(m (marker)). The RMT method was robust to temperature change from 20 to 40 degrees C, but sensitive to pH. The lateral shift and reproducibility of the target peak t(m (target)) were significantly reduced by this novel transformation. In the worst cases migration time variability ranged up to 12% (n=6); the RMT algorithm reduced this to less than 1%. In general, the RMT transformation reduced the variability of migration time data by a factor of 2-5. For systematic comparison of electrophoretic profiles for test sample and standard, a new pattern recognition algorithm permits sequential peak-by-peak comparison using specified segments of the electropherograms for comparison of test and Echinacea purpurea (root product) as a standard. This algorithm was capable of rapidly characterising the similarity of target peaks in a test sample relative to those in the reference standard. Combination of the RMT algorithm and pattern recognition in CE is expected to offer a robust approach to international regulatory characterisation and control of Nutraceuticals.