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Abstract
The isolation of milligram amounts of the enantiomers of a benzodiazepinone derivative was performed on an analytical cellulose tribenzoate-based column by multiple repetitive injections. An enantiomeric purity greater than 98% was required. First, an analytical method was developed to maximize the resolution by adjusting the mobile phase composition, flow-rate and most importantly the column temperature. Then the preparative separation was optimized by adjusting the sample size and detecting the sample where its UV absorbance was low. The locations of the cut points were determined by use of detector response levels. The method development, preparative separations and analytical assays of the fractions obtained were all performed on analytical columns.
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