Investigation on amitraz, coumaphos and thymol concentrations in honey produced by Slovenian beekeepers in 2020

Helena BAŠA ČESNIK, Veronika KMECL


A survey on concentrations of veterinary drug residues amitraz, coumaphos and thymol in honey, produced in year 2020 by Slovenian beekeepers, was conducted. 100 samples were analysed: 22 from organic and 78 from conventional production, with two analytical methods. In method for determination of coumaphos and thymol samples were extracted with acetone, petroleumether and dichlorometane. In method for determination of amitraz and its degradation products, samples were hydrolisated with HCl and NaOH, extractied with n-hexane and derivatisated with heptafluorobutyric anhydride. Determination in both methods was performed with gas chromatograph coupled with mass spectrometer. Measured concentrations of amitraz, coumaphos and thymol were in the range of 0.01-0.12 mg kg-1, 0.02-0.06 mg kg-1 and 0.08-0.17 mg kg-1, respectively. In 61 % of samples analysed no residues of amitraz, thymol and coumaphos were found. Data obtained were compared with the data from literature. Chronic and acute exposure were calculated for consumers. Maximum chronic exposure for amitraz and thymol was 0.1 % and 0.05 % of acceptable daily intake, respectively. Maximum acute exposure for amitraz and thymol was 4 % and 0.8 % of acute reference dose, respectively


acaricide residues; GC-MS; amitraz; coumaphos; thymol; honey; consumer exposure

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Anderson, D.L., Trueman, J.W.H. (2000). Varroa jacobsoni (Acari: Varroidae) is more than one species. Experimental and Applied Acarology, 24, 165-189.

Anklam, E. (1998). A review of the analytical methods to determine the geographical and botanical origin of honey. Food Chemistry, 63, 549-562.

Bargańska, Ż., Ślebioda, M., Namieśnik, J. (2013). Pesticide residues levels in honey from apiaries located of Northern Poland. Food Control, 31, 196-201.

Baša Česnik, H., Kmecl, V. (2017). Vsebnost nekaterih akaricidov v medu slovenskih čebelarjev v letu 2017. Kemija v šoli in družbi, year 29, no. 1, pp 1-5,

Baša Česnik, H., Kmecl, V. (2018). Vsebnost nekaterih akaricidov v medu slovenskih čebelarjev v letu 2018. Kemija v šoli in družbi, year 2018, no. 1, pp. 1-4.

Baša Česnik, H., Kmecl, V. (2019). Vsebnost nekaterih akaricidov v medu slovenskih čebelarjev v letu 2019. Kemija v šoli in družbi, year 2019, no. 1, pp. 1-5.

Baša Česnik, H., Kmecl, V., Velikonja Bolta, Š. (2019). Pesticide and veterinary drug residues in honey - validation of methods and a survey of organic and conventional honeys from Slovenia. Food Additives and Contaminants Part A, 36, 1358-1375.

Bogdanov, S., Imdorf, A., Kilchenmann, V. (1998). Residues in wax and honey after Apilife VAR treatment. Apidologie, 29, 513-524.

Chiesa, L.M., Labella, G.F., Giorgi, A., Panseri, S., Pavlovic, R., Bonacci, S., Arioli, F. (2016). The occurrence of pesticides and persistent organic pollutants in Italian organic honeys from different productive areas in relation to potential environmental pollution. Chemosphere, 154, 482-490.

Del Carlo, M., Pepe, A., Sergi, M., Mascini, M., Tarentini, A., Compagnone, D. (2010). Detection of coumaphos in honey using a screening method based on an electrochemical acetylcholinestearase bioassay. Talanta, 81: 76-81.

Fernandez-Muiño, M.A., Sancho, M.T., Muniategui, S., Huidobro, J.F., Simal-Lozano, J. (1995). Acaricide residues in honey: analytical methods and levels found, Journal of Food Protection, 58, 449-454.

Gaweƚ, M., Kiljanek, T., Niewiadowska, A., Semeniuk, S., Goliszek, M., Burek, O., Posyniak, A. (2019). Determination of neonicotinoids and 199 other pesticide residues in honey by liquid and gas chromatography coupled with tandem mass spectrometry. Food Chemistry, 282, 36-47.

Juan-Borrás, M., Domench, E., Escriche, I. (2016). Mixture-risk-assessment of pesticide residues in retail polyfloral honey. Food Control, 67, 127-134.

Kmecl, V., Baša Česnik, H. (2011). A survey of total amitraz residues in honey produced in Slovenia. In: J. Pulkrabová (Ed.), M. Tomaniová (Ed.). Book of abstracts, 5th International Symposium on Recent Advances in Food Analysis (pp. 407). Prague: Institute of Chemical Technology.

Korta, E., Bakkali, A., Berrueta, L.A., Gallo, B., Vicente, F., Kilchenmann, V., Bogdanov, S. (2001). Study on acaricide stability in honey. Characterization of amitraz degradation products in honey and beeswax. Journal of Agricultural and food Chemistry, 49, 5835-5842.

Lozano, A., Hernando, M.D., Uclés, S., Hakme, E., Fernádez-Alba, A.R. (2019). Identification and measurement of veterinary drug residues in beehive products. Food Chemistry, 274, 61-70.

Martel, A.C. & Zeggane S. (2002). Determination of acaricides in honey by high-performance liquid chromatography with photodiode array detection. Journal of Chromatography A, 954, 173-180.

Nozal, M.J., Bernal, J.L., Jiménez, J.J., González, M.J., Higes, M. (2002). Extraction of thymol, eucalyptol, menthol, and camphor residues from honey and beeswax. Determination by gas chromatography with flame ionization detection. Journal of Chromatography A, 954, 207-215.

Rial-Otero, R., Gaspar, E.M., Moura, I., Capelo, J.L. (2007). Gas chromatography mass spectrometry determination of acaricides from honey after a new fast ultrasonic-based soild phase micro-extraction sample treatment. Talanta, 71, 1906-1914.

SANTE/11813/2017. Guidance document on analytical quality control and method validation procedures for pesticide residues analysis in food and feed. DG SANTE, European Comission, 2017.

Viñas, P., Soler-Romera, M.J., Hernández-Córdoba, M. (2006). Liquid chromatographic determination of phenol, thymol and carvacrol in honey using fluorimetric detection. Talanta, 69, 1063-1067.

Wiest, L., Buleté, A., Giroud, B., Fratta, C., Amic, S., Lambert, O., Pouliquen, H., Arnaudguilhem, C. (2011). Multi-residue analysis of 80 environmental contaminants in honeys, honeybees and pollens by one extraction procedure followed by liquid and gas chromatography coupled with mass spectrometric detection. Journal of Chromatography A, 1218, 5743-5756.



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