Hyoscyamine is a natural plant belladonna alkaloid derivative and the levorotatory form of racemic atropine. It functions as a non-selective, competitive antagonist of muscarinic receptors and is used to treat a variety of stomach pains and intestinal problems as well as to decrease side effects of certain medications and insecticides. The drug does that by decreasing the acid production in the stomach and slowing down the natural movements of the gut. Primesep 200, a reverse phase column, contains embedded acidic ionizable groups and can retain Hyoscyamine. The method is UV compatible and can be used as a general approach for analyzing similar compounds.