A liquid chromatography method with UV detection for determination of oxytetracycline (OTC) in honey has been developed. The samples were extracted with the solution of oxalic acid. The clean-up procedure was performed by solid phase extraction (SPE) using polymeric Strata X and carboxylic acid cartridges. Chromatographic separation was carried out on the Luna C8 analytical column with mobile phase consisting of acetonitrile-0.02 M oxalic acid. The method has been successfully validated according to the requirements of the European Decision 2002/657/EC and this method is used in routine control of oxytetracycline in honey samples. The limit of detection (LOD) and limit of quantification (LOQ) of the presented method were 10 and 12.5 μg/kg, respectively. The developed method has also been verified in quantitative determination of oxytetracycline residues in honey after experimental treatment with this product in bee colonies.
Argauer R. J., Moats W. A. (1991) - Degradation of oxytetracycline in honey as measured by fluorescence and liquid chromatographic assays. Apidologie, 22: 109-115.
Commission Decision 2002/657/EC of 12 August 2002 implementing Council Directive 96/23/EC concerning the performance of analytical methods and the interpretation of results. Off. J. Eur. Commun. 2002, L 221, pp. 8-36.
Commission Regulation (EU) No 37/2010 of 22 December 2009 on pharmacologically active substances and their classification regarding maximum residue limits in foodstuffs of animal origin. Off. J. Eur. Commun. 2010, L 15, pp. 1-72.
Dinkov D., Kanelov I., Zhelyazkova I., Vashin I. (2005) - Persistence of tetracycline and oxytetracycline in bee honey after improper application on bee families. Bulg. J. Vet. Med., 8(3): 205-509.
European Medicines Agency Veterinary Medicines and Inspections: Summary opinion of the committee for medicinal products for veterinary use on the establishment of Maximum Residue Limits - Oxytetracycline (Extension to honey bees), EMEA/CVMP/581586/2008.
Fujita K., Ito H., Ishihara M., Inukai S., Tanaka H., Taniguchi M. (2008) - Analysis of trace residues of tetracyclines in dark-colored honeys by highperformance liquid chromatography using polymeric cartridge and metal chelate affinity chromatography. J. Food Hyg. Soc. Japan, 49(3): 196-203.
Gojmerac W. L. (1980) - Enemies of honey bees, in: Gojmerac W. L. (Ed.) Bees, beekeeping, honey and pollination, Avi Pub. Co., Westport, pp. 127-137.
Huq S., Garriques M., Kallury K. M. R. (2006) - Role of zwitterionic structures in the solid-phase extraction based method development for clean up of tetracycline and oxytetracycline from honey. J. Chromatogr. A., 1135: 12-18.
Khong S. P., Hammel Y. A, Guy P. A. (2005) - Analysis of tetracyclines in honey by high-performance liquid chromatography/ tandem mass spectrometry. Rapid Commun. Mass Spectrom., 19: 493-502.
Martel A. C., Zeggane S., Drajnudel P., Faucon J. P., Aubert M. (2006) - Tetracyclines residues in honey after hive treatment. Food Addit. Contam., 23(3): 265-273.
Neils H. J., De Leenheer A. P. (1980) - Retention mechanisms of tetracyclines on a C8 reversed-phase material. J. Chromatogr., 195: 35-42.
Oka H., Ikai Y., Kawamura N., Uno K., Yamada M. (1987) - A simple method for residual tetracyclines analysis in honey using a tandem cartridge clean-up system. J. Chromatogr., 389: 417-426.
Pena A., Pelantova N., Lino C. M., Silveira M. I. N., Solich P. (2005) - Validation of an analytical methodology for determination of oxytetracycline and tetracycline residues in honey by HPLC with fluorescence detection. J. Agric. Food Chem., 53: 3784-3788.
Peres G. T., Rath S., Reyes Reyes F. G. (2010) - A HPLC with fluorescence detection method for the determination of tetracyclines residues and evaluation of their stability in honey. Food Control, 21: 620-325.
Regulation (EC) No 470/2009 of the European Parliament and of the Council of 6 May 2009 laying down Community procedures for the establishment of residue limits of pharmacologically active substances in foodstuffs of animal origin, repealing Council Regulation (EEC) No 2377/90 and amending Directive 2001/82/EC of the European Parliament and of the Council and Regulation and of the Council and Regulation (EC) No 726/2004 of the European Parliament and of the Council. Off. J. Eur. Commun. 2009, L 152, pp. 11-22.
Sporns P., Kwan S., Roth L. A. (1986) - HPLC analysis of oxytetracycline residues in honey. J. Food Prot., 49(5): 383-388.
Thompson H. M., Waite R. J., Wilkins S., Brown M. A., Bigwood T., Shaw M., Ridgway Ch., Sharman M. (2005) - Effects of European foulbrood treatment regime on oxytetracycline levels in honey extracted from treated honeybee (Apismelifera) colonies and toxicity to brood. FoodAddit. Contam., 22(6): 573-578.
USFDA (US Food and Drug Administration). Center for veterinary medicine database of approved animal drugs. Animal drugs @ FDA. October 1, 2008. [online] http://www.accessdata.fda.gov/scripts/animaldrugsatfda/ (accessed on September 2009).
Victorita B., Marghitas L. A., Dezmirean D., Moise A., Bobis O., Maghear O. (2007) - Optimization of HPLC method for quantifying tetracycline residue in honey. Bull. USAMV-CN, 64(1-2).
Vinas P., Balsalobre N., Lopez- Erroz C., Hernandez-Cordoba M. (2004) - Liquid chromatography with ultraviolet detection for the analysis of tetracycline residues in honey. J. Chromatogr. A, 1022: 125-129.
Zhou J., Xue X., Zhang Y. L. J., Chen F., Wu L., Chen L., Zhao J. (2009) - Multiresidue determination of tetracycline antibiotics in propolis by using HPLC-UV detection with ultrasonic- assisted extraction and two-step solid phase extraction. Food Chem., 115: 1074-1080.