Optimization of the experimental conditions for macrolide antibiotics in high performance liquid chromatography by using response surface methodology and determination of tylosin in milk samples |
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Authors: | Senem ?anli ?smail M Palabiyik Nurullah ?anli Zeynep B Guzel-Seydim Güleren Alsancak |
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Institution: | 1.Department of Chemistry, Faculty of Science and Arts,Hitit University,?orum,Turkey;2.Department of Analytical Chemistry, Faculty of Pharmacy,Ankara University,Ankara,Turkey;3.Department of Food Engineering, Faculty of Engineering and Architecture,Süleyman Demirel University,Isparta,Turkey;4.Department of Chemistry, Faculty of Science and Arts,Süleyman Demirel University,Isparta,Turkey |
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Abstract: | The simple, rapid and sensitive liquid chromatographic separation of five macrolides (tilmicosin, erythromycin, tylosin, roxithromycin
and josamycin) widely used in food producing animals was developed. Response surface methodology was used as an optimization
method of mobile phase, column temperature and pH to provide the best resolution of these analytes. The separation was performed
by using an end-capped X-Terra RP-18 column (250 × 4.6 mm I.D × 5 m) with an isocratic system of 15 mM hydrochloric acid (pH
2.5)-acetonitrile as the mobile phase at a temperature of 30°C and flow-rate of 1.0 mL/min. The suitability of the method
for multi-residue determination of the macrolides is demonstrated by the analysis of milk samples spiked with tylosin. Roxithromycin
was used as internal standard. The recovery of tylosin was quite good as 90.8%. The limit of quantification and detection
limit were 0.024 and 0.007 μg/mL, respectively. The method was successfully applied to determination of macrolides at levels
below the maximum concentration legally allowed in milk samples. |
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