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CIT/MIT biocides are widely used as preservatives in variety of applications, which include household products, personal care products and polymer materials. Apart of their antimicrobial activity, CIT/MIT preservatives may cause allergy reactions. Due to this reason, the preservatives additives should be used at the lowest possible concentration and strictly controlled. This application shows the determination of CIT/MIT preservative in rhamnolipids production. ARION® C8 column shows good resolution and peaks symmetry for CIT/MIT components at low concentrations.