Effects of dietary n-3 polyunsaturated fatty acids and ... - FINS

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1 st WorkshopXIII International Feed Technology Symposiumexplains the smaller slower elution larger concentration of salinomycin applied to silicagel column, and is more efficient elution samples with high content of salinomycinapplication required greater volume of methanol.Table 1. Results of the determination of contents of salinomycin in premixes and broilerfeed by spectrophotometric methodAdded[µg/g feed]Found ± SD[µg/g feed]Efficacy of extraction(% ) ± SD10 10,4±0,7 103,5±6,820 18,6±0,5 92,8±5,430 26,9±0,9 89,5±8,640 34,3±0,8 85,7±7,560 50,3±0,7 83,8±5,4Based on the results we can conclude that our original extraction method is suitable toextract salinomycine from premixes and poultry feed samples., and that thespectrophotometric method can be successfully applied in monitoring the content ofsalinomycin in eluates, with detection limit of 0,5 mg/kg poultry feed. This means thatthe described method allows the determination of 120 times lower concentrations ofpreventive doses of salinomycine in broiler feed, which is 60 mg/kg feed.For the quantitative determination of salinomycin content in the poultry feed samplescomparative method was elaborated liquid chromatography with post-columnderivatisation with vanillin reagent. Salinomycin from samples extracted by columnchromatographyand eluated with methanol. Eluents was centrifuged and filtered.Separation was carried out with TSK ODS-120T, 10µm column with a mobile phaseconsisting of methanol- water – acetic acid (94:6:0,1, v/v/v) and than using post-columnderivatisation with vanillin reagent. Detection was performed with a spectrophotometricdetector with tungsten lamp at 520 nm. Retention time for salinomycine, under theseconditions of the determination, is 12,0 min. Results of the determination of contents ofsalinomycin in premixes and broiler feed by high performance liquid chromatographyare shown in table 2. Based on the results shown in table 2, we notice that the applicationof the described HPLC procedure is possible to determine the salinomyin content in thepoultry feed samples with high reproducibility rates, ranging from 96,3±2,6 % to112,7±2,7% and detection limit of 15 µg/kg of examined poultry feed.279
1 st WorkshopXIII International Feed Technology SymposiumTable 2. Results of the determination of contents of salinomycin in premixes and broilerfeed by liquid chromatographyAdded[µg/g feed]Found ± SD[µg/g feed]Efficacy of extraction(% ) ± SD10 11,3±0,3 112,7±2,720 20,8±0,2 103,8±1,830 29,6±0,5 98,5±4,940 33,2±0,6 98,0±5,460 57,8±0,3 96,3±2,6CONCLUSIONDeveloped extraction method by column-chromatography and method ofspectrophotometric determination in the visible area and the method of liquidchromatography can be applied successfully to determination of salinomycin content inpremixes and final fodder mixtures for fattening broilers. The obtained detection limitsare far below the doses used for coccidiostatic salinomycin in broilers feed (60 mg/kg offeed). Recovery of both procedures studied using the standard addition method was over80%. Applied spectrophotometric method is faster, simpler and cheaper method of liquidchromatographic method, in term of recovery, accuracy and detection limit are notbehind precise, but more expensive and durable liquid chromatographic method.Therefore, spectrophotometry should be a routine method of choice in determining thesalinomycin content in the poultry feed samples.REFERENCES1. Asuhabe H., Sasaki T., Harada K.I., Suzuki M.: Fluorodensitometricdetermination of polyether antibiotics, J. of Chromatography, 295 (1984), 453-461.2. Blanchlower W.J., Desmond A.R., Hamilton J.T.G.: Simultaneous highperformance liquid chromatographic determination of monensin, narasin andsalinomycin in feeds using post-column derivatisation, Analyst, 110 (1985),1283-1287.3. Blood D.C., Henderson J.A., Radostits O.M.: Veterinary Medicine. 5th Ed.Bailliere Tindall, London 1979, p 39.4. Brander G.C., Pugh D.M., Bywater R.J.: Veterinary Applied Pharmacologyand Therapeutics. Bailliere Tindall, London, ( 1982 ), p.785-795.5. Braunius W.W.: Coccidiosis in Broilers: the Effective use of Anticoccidialdrugs, Worlds Poultry Science Journal, 38 (1982), 176-185 CambridgeUniversity Press.280
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Effects of dietary n-3 polyunsaturated fatty acids and ... - FINS

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