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This service provides a novel capillary electrophoresis – capacitively coupled contactless conductivity detection (CE-C4D) method for the separation of 12 antibiotics, including four types of aminoglycosides, three types of fluoroquinolones, two types of tetracyclines, and three types of macrolides. Half of these antibiotics were not determined by ultraviolet (UV) because of their lack of UV-absorbing groups. Formic acid (FA) (pH 2.50) with low conductivity was employed as the background electrolyte (BGE) in comparison with three BGE systems (i.e., HAc, HCl and H3PO4), which not only allowed complete protonation and electrophoresis separation but provided more cost-effectiveness and shorter analysis time. Under these conditions, a UV detector was employed as an additional detection mode to evaluate the qualitative analysis of 6 antibiotics possessing UV absorbing groups.