Identification and Determination of Oxytetracycline, Tiamulin, Lincomycin, and Spectinomycin in Veterinary Preparations by Thin-Layer Chromatography/Densitometry

A thin-layer chromatographic/densitometric method was developed for the identification and quantitation of oxytetracycline, tiamulin, lincomycin, and spectinomycin in veterinary preparations. Silica gel-coated thin layer chromatography plates and 2 mobile phases were used to separate these constituents. The appropriate compositions of the suitable mobile phases were established: 10% citric acid solution–n-hexane–ethanol (80 + 1 + 1, v/v) and n–butanol–ethanol–chloroform– 25% ammonia (4 + 5 + 2 + 5, v/v). Along with Rf values and spot colors, direct UV and visual densitometric measurements were used for identification. Similar measuring ranges were used for quantitative analysis to obtain repeatable and reliable results for the preparations examined. The results of the quantitative analysis are characterized by a small confidence interval and are close to the declared contents of active constituents: oxytetracycline 30.01 ± 0.38 g at λ = 350 nm and 30.24 ± 0.86 g at λ = 430 nm; tiamulin, 10.19 ± 0.86 g at λ = 450 nm; lincomycin, 2.27 ± 0.08 g at λ = 278 nm; and spectinomycin, 2.18 ± 0.07 g at λ = 421 nm. The recoveries for all antibiotics ranged from 100.01 to 102.54%.