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Montelukast is a leukotriene receptor antagonist (LTRA) used for the maintenance treatment of asthma and to relieve symptoms of seasonal allergies. It is effectively used to treat asthma attacks. Montelukast is hydrophobic and zwitterionic in nature. It contains carboxylic acid and quinoline fragments. Montelukast and related impurities were analyzed and separated by mixed-mode chromatography using the Heritage MA mixed-mode column. Montelukast is retained and separated from other impurities by strong hydrophobic and strong cation-exclusion interactions. Retention time, selectivity, and resolution between peaks can be adjusted by modification of amount of ACN, buffer and buffer pH. Various mobile phases can be used with Heritage MA column to accommodate different detection techniques (UV, MS, CAD, ELSD detection).