Technical Abstract: A preparative method for decigram-scale polishing of limonin glucoside with regard to removing traces of limonin, the main bitter principle in Citrus, is reported. During the method development and up-scaling stages, in which sample purity, sample size, solvent amounts, and drying conditions were varied, a rapid isocratic HPLC method with UV absorbance detection was used to monitor the elution behavior of limonin and subsequently, fractions obtained using the finalized method were analyzed for their limonoid glucoside and aglycone content in a single HPLC run using a binary gradient HPLC method and an evaporative light scattering detector. The resulting polishing method consists of a simple step gradient that utilizes food-grade solvents, ethanol and water at 15% EtOH and 50% EtOH steps. Using a 75 x 300 mm C-18 column, this method is capable of processing twenty-grams of material per run in less than 3 h. Recovery of the purified limonin glucoside following evaporation of the solvent was 93.5% (+/-2.8, n=6) and the limonin concentrations in the resulting materials were found to be reduced 10- to 15-fold.