HPLC Separation of Tylosin and Albendazole on Zodiac HST TH Column

Tylosin is an antibiotic widely used in veterinary medicine and occurs naturally as a fermentation product. Due to the presence of multiple sugar moieties and amino groups, tylosin and other aminoglycosides cannot be effectively analyzed using conventional reversed-phase HPLC under LC/MS-compatible conditions. Mixed-mode chromatography overcomes this limitation by enabling retention and separation of such polar, basic compounds. The Zodiac HST TH column was used for this purpose, offering efficient analysis of aminosugar-based antibiotics including streptomycin, neomycin, amikacin, arbekacin, dibekacin, framycetin, kanamycin, gentamicin, verdamicin, astromicin, hygromycin, and others.

For molecules with multiple amino groups, enhanced elution is achieved using higher buffer concentrations and a lower pH (3.0–3.5). This method avoids the use of ion-pairing reagents and serves as a modern alternative to traditional reversed-phase and ion-pairing chromatography approaches. Albendazole was employed as an internal standard to quantify aminoglycosides.