<div>Cyclic loading begins with the crude methanolic extract and a reverse-phase HP20 column. The methanolic extract is passed through the HP20 column, where the relatively non-polar metabolites will adhere to the column. The eluent is then diluted by 50% to make it more polar and is then passed through the HP20 column again. Again, non-polar metabolites will adhere to the column, eliminating things such as salts. The eluent is diluted again by 50% and passed through one more time. The column is then stripped with varying concentrations of acetone to create three main fractions; 30% acetone, 75 and 100%. Since a mix of acetone and water is notorious to evaporate, the 30% and 75% fractions are back loaded onto the HP20 column (cyclic loaded again) and then eluted with 100% acetone.&nbsp; This in-house method was developed to combat the problems with traditional isolation methods, using liquid-liquid partitioning, and other reactions which further decreased the yield of the compounds obtained. 
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