Plant-derived compounds have been recognized by the feed industry as important supplements for livestock welfare and health. In this context, Citrus aurantium L. extract and Origanum vulgare L. essential oil have been demonstrated to have strong anti-inflammatory and antioxidant effects on animals. Being the composition of plant-derived extracts extremely influenced by the environmental and growing conditions of the plants, quality control is necessary in terms of the concentration of the active compounds to assure the reproducibility of natural feed additives. The present work aimed at the validation of the extraction procedure from feed additives of Hesperidin (HES) and Carvacrol (CAR), the main active compounds of Citrus aurantium and Origanum vulgare extracts. Then, the quantification method of both the analytes was developed and validated by reversed high-performance liquid chromatography coupled with a UV detector. The validated method was tested on premixtures and final feed additives supplied by a local feed factory to supervise the production chain. The extraction method with methanol resulted to be efficient and highly reproducible, with recovery higher than 90% for both the analytes. The chromatographic method has been demonstrated to be accurate, precise (relative standard deviation percent lower than 2.06%), and linear in the tested range concentrations, with regression coefficients equal to 0.995 and 0.999 for HES and CAR respectively. The method demonstrated that the feed additives prepared by the factory by diluting the premixtures were less concentrated than what was declared on the label.
Silbergeld, E.K. (2019) One Health and the Agricultural Transition in Food Animal Production. Global Transitions, 1, 83-92. https://doi.org/10.1016/j.glt.2019.01.003
Elsasser, T.H., Klasing, K.C., Filipov, N. and Thompson, F. (2009) The Metabolic Consequences of Stress: Targets for Stress and Priorities of Nutrient Use. In: Moberg, G.P. and Mench, J.A., The Biology of Animal Stress: Basic Principles and Implications for Animal Welfare, Centre for Agriculture and Bioscience International, Wallingford, 77-110. https://doi.org/10.1079/9780851993591.0077
Blokhuis, H.J., Hopster, H., Geverink, N.A., Korte, S.M. and Van Reenen, C.G. (1998) Studies of Stress in Farm Animals. Comparative Haematology International, 8, 94-101. https://doi.org/10.1007/BF02642498
Falowo, A.B. and Akimoladun, O.F. (2020) Veterinary Drug Residues in Meat and Meat Products: Occurrence, Detection and Implications. In: Bekoe, S.O. and Saravanan, M., Eds., Veterinary Medicine and Pharmaceuticals, IntechOpen, London, 1-18. https://doi.org/10.5772/intechopen.83616
European Food Safety Authority (2020) Report for 2018 on the Results from the Monitoring of Veterinary Medicinal Product Residues and Other Substances in Live Animals and Animal Products. EFSA Supporting Publications, 17, Article No. 1775E. https://doi.org/10.2903/sp.efsa.2020.EN-1775
Sanz, D., Razquin, P., Condón, S., Juan, T., Juan, T., Herraiz, B., et al. (2015) Incidence of Antimicrobial Residues in Meat Using a Broad Spectrum Screening Strategy. European Journal of Nutrition & Food Safety, 5, 156-165. https://doi.org/10.9734/EJNFS/2015/13795
Rana, M.S., Lee, S.Y., Kang, H.J. and Hur, S.J. (2019) Reducing Veterinary Drug Residues in Animal Products: A Review. Food Science of Animal Resources, 39, 687-703. https://doi.org/10.5851/kosfa.2019.e65
Kang, J.W., Park, H.C., Gedi, V., Park, S.J., Kim, M.A., Kim, M.K., et al. (2015) Veterinary Drug Residues in Domestic and Imported Foods of Animal Origin in the Republic of Korea. Food Additives and Contaminants: Part B Surveillance, 8, 106-112. https://doi.org/10.1080/19393210.2014.1001795
Gallo, P., Fabbrocino, S., Dowling, G., Salini, M., Fiori, M., Perretta, G., et al. (2010) Confirmatory Analysis of Non-Steroidal Anti-Inflammatory Drugs in Bovine Milk by High-Performance Liquid Chromatography with Fluorescence Detection. Journal of Chromatography A, 1217, 2832-2839. https://doi.org/10.1016/j.chroma.2010.02.047
Mehtabuddin, A.A., Ahmad, T., Nadeem, S., Tanveer, Z.I. and Arshad, J. (2012) Sulfonamide Residues Determination in Commercial Poultry Meat and Eggs. Undefined, 22, 473-478.
Martin, G.B. and Ferasyi, T.R. (2016) Clean, Green, Ethical (CGE) Management: What Research Do We Really Need? The International Journal of Tropical Veterinary and Biomedical Research, 1, 1-8. https://doi.org/10.21157/ijtvbr.v1i1.5066
European Parliament (2003) Regulation (EC) No 1831/2003 on the European Parliament and of the Council of 22 September 2003. On Additives for Use in Animal Nutrition. Official Journal of the European Communities, 268, 29-43.
Surai, P.F. (2014) Polyphenol Compounds in the Chicken/Animal Diet: From the Past to the Future. Journal of Animal Physiology and Animal Nutrition, 98, 19-31. https://doi.org/10.1111/jpn.12070
Tsiplakou, E., Pitino, R., Manuelian, C.L., Simoni, M., Mitsiopoulou, C., De Marchi, M., et al. (2021) Plant Feed Additives as Natural Alternatives to the Use of Synthetic Antioxidant Vitamins in Livestock Animal Products Yield, Quality, and Oxidative Status: A Review. Antioxidants, 10, Article No. 780. https://doi.org/10.3390/antiox10050780
Stevanovic, Z.D., Bosnjak-Neumüller, J., Pajic-Lijakovic, I., Raj, J. and Vasiljevic, M. (2018) Essential Oils as Feed Additives—Future Perspectives. Molecules, 23, Article No. 1717. https://doi.org/10.3390/molecules23071717
Gadde, U., Kim, W.H., Oh, S.T. and Lillehoj, H.S. (2017) Alternatives to Antibiotics for Maximizing Growth Performance and Feed Efficiency in Poultry: A Review. Animal Health Research Reviews, 18, 26-45. https://doi.org/10.1017/S1466252316000207
Hashemi, S.R. and Davoodi, H. (2011) Herbal Plants and Their Derivatives as Growth and Health Promoters in Animal Nutrition. Veterinary Research Communications, 35, 169-180. https://doi.org/10.1007/s11259-010-9458-2
Elferink, E.V., Nonhebel, S. and Moll, H.C. (2008) Feeding Livestock Food Residue and the Consequences for the Environmental Impact of Meat. Journal of Cleaner Production, 16, 1227-1233. https://doi.org/10.1016/j.jclepro.2007.06.008
European Parliament (2018) Regulation (EU) 2018/848 of the European Parl of 30 May 2018 on Organic Production and Labelling of Organic Products and Repealing Council Regulation (EC) No 834/2007. Official Journal of the European Communities, 150, 1-92.
de la Rosa, J.D.P., Ruiz-Palomino, P., Arriola-Guevara, E., García-Fajardo, J., Sandoval, G. and Guatemala-Morales, G.M. (2018) A Green Process for the Extraction and Purification of Hesperidin from Mexican Lime Peel (Citrus aurantifolia Swingle) That Is Extendible to the Citrus Genus. Processes, 6, Article No. 266. https://doi.org/10.3390/pr6120266
Di Mauro, A., Fallico, B., Passerini, A. and Maccarone, E. (2000) Waste Water from Citrus Processing as a Source of Hesperidin by Concentration on Styrene—Divinylbenzene Resin. Journal of Agricultural and Food Chemistry, 48, 2291-2295. https://doi.org/10.1021/jf990992w
Russo, M., Bonaccorsi, I., Inferrera, V., Dugo, P. and Mondello, L. (2015) Underestimated Sources of Flavonoids, Limonoids and Dietary Fiber: Availability in Orange’s By-Products. Journal of Functional Foods, 12, 150-157. https://doi.org/10.1016/j.jff.2014.11.008
Tadros, F.J. and Andrade, J.M. (2021) Impact of Hesperidin in 100% Orange Juice on Chronic Disease Biomarkers: A Narrative Systematic Review and Gap Analysis. Critical Reviews in Food Science and Nutrition. https://doi.org/10.1080/10408398.2021.1927976
Kamboh, A.A., Memon, A.M., Mughal, M.J., Memon, J. and Bakhetgul, M. (2018) Dietary Effects of Soy and Citrus Flavonoid on Antioxidation and Microbial Quality of Meat in Broilers. Journal of Animal Physiology and Animal Nutrition, 102, 235-240. https://doi.org/10.1111/jpn.12683
Hager-Theodorides, A.L., Massouras, T., Simitzis, P.E., Moschou, K., Zoidis, E., Sfakianaki, E., et al. (2021) Hesperidin and Naringin Improve Broiler Meat Fatty Acid Profile and Modulate the Expression of Genes Involved in Fatty Acid β-Oxidation and Anti-oxidant Defense in a Dose Dependent Manner. Foods, 10, Article No. 739. https://doi.org/10.3390/foods10040739
Goliomytis, M., Kartsonas, N., Charismiadou, M., Symeon, G., Simitzis, P. and Deligeorgis, S. (2015) The Influence of Naringin or Hesperidin Dietary Supplementation on Broiler Meat Quality and Oxidative Stability. PLoS ONE, 10, Article ID: e0141652. https://doi.org/10.1371/journal.pone.0141652
Goliomytis, M., Simitzis, P., Papalexi, A., Veneti, N., Hager-Theodorides, A., Charismiadou, M., et al. (2019) Influence of Citrus Flavonoids on Laying Hen Performance, Inflammatory Immune Response, Egg Quality and Yolk Oxidative Stability. British Poultry Science, 60, 272-278. https://doi.org/10.1080/00071668.2019.1587150
Fotakis, C., Lantzouraki, D., Goliomytis, M., Simitzis, P., Charismiadou, M., Deligeorgis, S., et al. (2017) NMR Metabolomics Investigates the Influence of Flavonoid-Enriched Rations on Chicken Plasma. Journal of AOAC International, 100, 315-322. https://doi.org/10.5740/jaoacint.16-0405
Jugreet, B.S., Suroowan, S., Rengasamy, R.R.K. and Mahomoodally, M.F. (2020) Chemistry, Bioactivities, Mode of Action and Industrial Applications of Essential Oils. Trends in Food Science & Technology, 101, 89-105. https://doi.org/10.1016/j.tifs.2020.04.025
Institut de Recerca i Tecnologia Agroalimentàries (2015) Review of Immune Stimulator Substances/Agents That Are Susceptible of Being Used as Feed Additives: Mode of Action and Identification of End-Points for Efficacy Assessment. EFSA Supporting Publications, 12, Article No. 905E. https://doi.org/10.2903/sp.efsa.2015.EN-905
Castillo, M., Martín-Orúe, S.M., Roca, M., Manzanilla, E.G., Badiola, I., Perez, J.F., et al. (2006) The Response of Gastrointestinal Microbiota to Avilamycin, Butyrate, and Plant Extracts in Early-Weaned Pigs. Journal of Animal Science, 84, 2725-2734. https://doi.org/10.2527/jas.2004-556
Veterinary Medicines Evaluation Unit (1997) VICH Topic GL2 (Validation: Methodology) Guideline on Validation of Analytical Procedures: Methodology Date for Coming into Operation by Vich gl2 (Validation Methodology).
Mueller-Harvey, I. (2004) Modern Techniques for Feed Analysis. In: Assesing Quality and Safety of Animal Feeds, Food and Agriculture Organization of the United Nations, Rome, 1-38.
Victor, M.M., David, J.M., Cortez, M.V.M., Leite, J.L. and da Silva, G.S.B. (2021) A High-Yield Process for Extraction of Hesperidin from Orange (Citrus sinensis L. osbeck) Peels Waste, and Its Transformation to Diosmetin, A Valuable and Bioactive Flavonoid. Waste and Biomass Valorization, 12, 313-320. https://doi.org/10.1007/s12649-020-00982-x
Ma, Y., Ye, X., Hao, Y., Xu, G., Xu, G. and Liu, D. (2008) Ultrasound-Assisted Extraction of Hesperidin from Penggan (Citrus reticulata) Peel. Ultrasonics Sonochemistry, 15, 227-232. https://doi.org/10.1016/j.ultsonch.2007.03.006
Pensel, P.E., Maggiore, M.A., Gende, L.B., Eguaras, M.J., Denegri, M.G. and Elissondo, M.C. (2014) Efficacy of Essential Oils of Thymus vulgaris and Origanum vulgare on Echinococcus granulosus. Interdisciplinary Perspectives on Infectious Diseases, 2014, Article ID: 693289. https://doi.org/10.1155/2014/693289
Optimization and Validation of a High-Performance Liquid Chromatography Method for the Analysis of Hesperidin and Carvacrol for Veterinary Use — Oak Academic Publishing