Isocratic high-performance liquid chromatography on a poly(styrene-diviny1benzene)copolymer column allows the complete separation and resolution of tetracycline, 4-epitetracycline, anhydrotetracycline and 4 -e p i a n h y d r o t e t r a c y c l i n e .A fermentation impurity, 2-acetyl-2-decarboxamidotetracycline is also resolved from tetracycline. The mobile phase combines tert.-butanol, water and phosphate buffer, tetrabutylammonium sulphate and sodium ethylenediaminetetraacetate at a pH of 9.0 for elution at a temperature of 60 OC. The method was used to analyse official standards and commercial samples.

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