A description of the three main solvent systems which are suitable for gradient CCC separation of compounds.

Final choice of which solvent is to be used is based on their elution position on a 100% water ( buffered to pH 6.8 with 10mM ammonium formate ) to 100% acetonitrile; reverse phase hplc gradient. The L. trifolia extract corresponded to region B, which suggests mid-polarity compounds, and a preferred choice of a hexane: ethyl acetate: methanol: water biphasic solvent system. Next research was undertaken to find the best solvent ratios in order to develop a gradient system."

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