Food Processing: Techniques and Technology Food Processing: Techniques and Technology Техника и технология пищевых производств 2074-9414 2313-1748 89004 10.21603/2074-9414-2024-3-2523 ОРИГИНАЛЬНАЯ СТАТЬЯ ORIGINAL ARTICLE ОРИГИНАЛЬНАЯ СТАТЬЯ Light Stable Isotopes and Their Ratios in Honey Components Анализ отношений стабильных изотопов легких элементов в отдельных компонентах мёда https://orcid.org/0000-0001-8195-4292 Оганесянц Лев Арсенович Oganesyants Lev A. https://orcid.org/0000-0002-5502-7951 Панасюк Александр Львович Panasyuk Alexander L. https://orcid.org/0000-0002-5595-0455 Свиридов Дмитрий Александрович Sviridov Dmitriy A. labvin@yandex.ru https://orcid.org/0000-0003-0518-1181 Ганин Михаил Юрьевич Ganin Mikhail Yu. https://orcid.org/0000-0003-1223-0703 Шилкин Алексей Александрович Schilkin Alexei A. https://orcid.org/0000-0002-8337-2322 Серебрякова Оксана Владимировна Serebryakova Oksana V. Всероссийский научно-исследовательский институт пивоваренной, безалкогольной и винодельческой промышленности Москва Россия All-Russian Scientific Research Institute of Brewing, Beverage and Wine Industry Moscow Russian Federation Всероссийский научно-исследовательский институт пивоваренной, безалкогольной и винодельческой промышленности Москва Россия All-Russian Scientific Research Institute of Brewing, Beverage and Wine Industry Moscow Russian Federation Всероссийский научно-исследовательский институт пивоваренной, безалкогольной и винодельческой промышленности Москва Россия All-Russian Scientific Research Institute of Brewing, Beverage and Wine Industry Moscow Russian Federation Всероссийский научно-исследовательский институт пивоваренной, безалкогольной и винодельческой промышленности Москва Россия All-Russian Scientific Research Institute of Brewing, Beverage and Wine Industry Moscow Russian Federation Всероссийский научно-исследовательский институт пивоваренной, безалкогольной и винодельческой промышленности Москва Россия All-Russian Research Institute of Brewing, Non-Alcoholic and Wine Industry Moscow Russian Federation Научный центр пчеловодства Рыбное Россия Scientific Center of Beekeeping Rybnoye Russian Federation 02 10 2024 02 10 2024 54 3 522 531 10 04 2024 04 06 2024 https://fptt.ru/en/issues/22856/22829/

Мёд, благодаря своим органолептическим качествам и питательным свойствам, является ценным ингредиентом в пищевой промышленности. Однако мёд стал одним из самых фальсифицируемых продуктов. Нарушение технологии при производстве мёда является одной из главных проблем, угрожающих стабильному развитию и деятельности пчеловодческой отрасли. Разработка способов определения подлинности мёда и продуктов на его основе является востребованным направлением исследований. С целью идентификации мёда авторами проведен анализ отношений стабильных изотопов легких элементов в его отдельных компонентах. В ходе исследования изучили 36 образцов мёда различного географического и ботанического происхождения, а также 5 образцов сахарных сиропов из различного сырья. Для установления числовых значений показателей изотопных характеристик легких элементов применяли изотопный масс-спектрометр Delta Advantage V (США – Германия), с дополнительными модулями Flash IRMS и Conflo IV. В исследуемых образцах измерили значения показателей отношений стабильных изотопов углерода: в мёде (брутто) δ13С и азота δ15N в его белковой фракции, а также показатели δ13C, δ18О и δ2Н этанола, выделенного из ферментированного мёда. Анализ значений показателей δ13С брутто и δ13С белковой части меда позволяет рассчитать количество внесенных сахаров, полученных из кукурузы и тростника. В двух образцах были обнаружены экзогенные сахара в количестве 6,5 и 18 % в пересчёте на тростниковый сахар. Проведенные исследования показали, что использование метода изотопной масс-спектрометрии позволяет выявить образцы мёда, содержащие экзогенные сахара. На основании методики AOAC 998.12., в 2 образцах мёда из 36 было выявлено присутствие экзогенных сахаров из С4-типа растений. Однако данный метод не позволяет выявить присутствие внесенных сахаросодержащих веществ из С3-типа растений. Были проведены исследования значений показателей δ13C, δ18О и δ2Н этанола, выделенного из ферментированного мёда. Установлено, что показатель δ18O может выступать в качестве идентификационного критерия при установлении присутствия сахаров из С3 типа растений в меде. Изучение отношений изотопов азота в белковой части мёда может быть полезным инструментом для определения его подлинности и дополнительным критерием идентификации пчеловодческой продукции. В ходе работы был получен патент (RU2809285C1) по определению экзогенных сахаросодержащих веществ в пчелином мёде.

Honey possesses excellent sensory and nutritional properties, which makes it a valuable food ingredient. However, the same qualities make it one of the most often adulterated products in the world. Constant violations of processing technology threaten the beekeeping industry. In this regard, new authenticity criteria are a popular area of honey studies. The article introduces a method for establishing ratios of light stable isotopes in honey. The study featured 36 samples of honey of various geographical origins and botanical profiles, as well as five samples of sugar syrups from various raw materials. The quantitative profiles were obtained using a Delta Advantage V isotope mass spectrometer (USA – Germany) with additional Flash IRMS and Conflo IV modules. The experiment involved the ratios of stable carbon isotopes in honey δ13C (gross) and nitrogen δ15N in its protein fraction, as well as the values of δ13C, δ18O, and δ2H of ethanol isolated from fermented honey. The values of δ13C (gross) and δ13C of honey proteins made it possible to calculate the amount of added sugar of corn and cane origin. Exogenous sugars in the amounts of 6.5% and 18% cane sugar were detected in two samples. Isotope mass spectrometry was able to identify honey samples with exogenous sugars of C4-plant origin. However, the method failed to detect sugar-containing substances from C3-type plants. The δ18O index demonstrated some prospects as an identification criterion for sugars from C3-plants in honey. Nitrogen isotope ratios in honey proteins proved to be an efficient tool for determining honey authenticity and an additional criterion for identifying bee products. The research resulted in a patent (RU2809285C1) for a new method of determining exogenous sugar-containing substances in honey

 Мёд продукты пчеловодства идентификация фальсификация изотопная масс-спектрометрия легких элементов Honey bee products identification falsification isotope mass spectrometry

Khatun MA, Yoshimura J, Yoshida M, Suzuki Y, Huque R, Kelly SD, et al. Isotopic characteristics (δ13C, δ15N, and δ18O) of honey from Bangladesh retail markets: Investigating sugar manipulation, botanical and geographical authentication. Food Chemistry. 2024;435:137612. https://doi.org/10.1016/j.foodchem.2023.137612 Khatun MA, Yoshimura J, Yoshida M, Suzuki Y, Huque R, Kelly SD, et al. Isotopic characteristics (δ13C, δ15N, and δ18O) of honey from Bangladesh retail markets: Investigating sugar manipulation, botanical and geographical authentication. Food Chemistry. 2024;435:137612. https://doi.org/10.1016/j.foodchem.2023.137612 Ždiniaková T, Lörchner C, De Rudder O, Dimitrova T, Kaklamanos G, Breidbach A, et al. EU Coordinated action to deter certain fraudulent practices in the honey sector. JRC Technical Report. 2023. https://data.europa.eu/doi/10.2760/184511 Ždiniaková T, Lörchner C, De Rudder O, Dimitrova T, Kaklamanos G, Breidbach A, et al. EU Coordinated action to deter certain fraudulent practices in the honey sector. JRC Technical Report. 2023. https://data.europa.eu/doi/10.2760/184511 Ковалев А. М. Учебник пчеловода: Учебник для сельского профессионального технического училища. Москва: Колос, 1973. 432 с. Kovalev AM. Beekeeper's Textbook: Textbook for rural vocational technical school. Moscow: Kolos; 1973. 432 p. (In Russ.). 70 percent of honey in Russia may be adulterated: how to recognize deception [Internet]. [cited 2024 Jan 15]. Available from: https://www.gastronom.ru/text/nepravilnyj-med-do-70-meda-v-rossii-mozhet-okazatsya-falsificirovannym-1017587. 70 percent of honey in Russia may be adulterated: how to recognize deception [Internet]. [cited 2024 Jan 15]. Available from: https://www.gastronom.ru/text/nepravilnyj-med-do-70-meda-v-rossii-mozhet-okazatsya-falsificirovannym-1017587. Tosun M. Detection of adulteration in honey samples added various sugar syrups with 13C/12C isotope ratio analysis method. Food Chemistry. 2013;138(2–3):1629–1632. https://doi.org/10.1016/j.foodchem.2012.11.068. Tosun M. Detection of adulteration in honey samples added various sugar syrups with 13C/12C isotope ratio analysis method. Food Chemistry. 2013;138(2–3):1629–1632. https://doi.org/10.1016/j.foodchem.2012.11.068. Ajlouni S, Sujirapinyokul P. Hydroxymethylfurfuraldehyde and amylase contents in Australian honey. Food Chemistry. 2010;119(3):1000–1005. https://doi.org/10.1016/j.foodchem.2009.07.057 Ajlouni S, Sujirapinyokul P. Hydroxymethylfurfuraldehyde and amylase contents in Australian honey. Food Chemistry. 2010;119(3):1000–1005. https://doi.org/10.1016/j.foodchem.2009.07.057 Chen C-T, Chen B-Y, Nai Y-S, Chang Y-M, Chen K-H, Chen Y-W. Novel inspection of sugar residue and origin in honey based on the 13C/12C isotopic ratio and protein content. Journal of Food and Drug Analysis. 2019;27(1): 175–183. https://doi.org/10.1016/j.jfda.2018.08.004 Chen C-T, Chen B-Y, Nai Y-S, Chang Y-M, Chen K-H, Chen Y-W. Novel inspection of sugar residue and origin in honey based on the 13C/12C isotopic ratio and protein content. Journal of Food and Drug Analysis. 2019;27(1): 175–183. https://doi.org/10.1016/j.jfda.2018.08.004 Kawashima H, Suto M, Suto N. Stable carbon isotope ratios for organic acids in commercial honey samples. Food Chemistry. 2019;289:49–55. https://doi.org/10.1016/j.foodchem.2019.03.053 Kawashima H, Suto M, Suto N. Stable carbon isotope ratios for organic acids in commercial honey samples. Food Chemistry. 2019;289:49–55. https://doi.org/10.1016/j.foodchem.2019.03.053 Cabañero AI, Recio JL, Rupérez M. Liquid Chromatography Coupled to Isotope Ratio Mass Spectrometry: A New Perspective on Honey Adulteration Detection. Journal of Agricultural and Food Chemistry. 2006;54(26):9719–9727. https://doi.org/10.1021/jf062067x Cabañero AI, Recio JL, Rupérez M. Liquid Chromatography Coupled to Isotope Ratio Mass Spectrometry: A New Perspective on Honey Adulteration Detection. Journal of Agricultural and Food Chemistry. 2006;54(26):9719–9727. https://doi.org/10.1021/jf062067x Xu JZ, Liu X, Wu B, Cao YZ. A comprehensive analysis of 13C isotope ratios data of authentic honey types produced in China using the EA-IRMS and LC-IRMS. Journal Food Science Technology. 2020;57(4):1216–1232. https://doi.org/10.1007/s13197-019-04153-2 Xu JZ, Liu X, Wu B, Cao YZ. A comprehensive analysis of 13C isotope ratios data of authentic honey types produced in China using the EA-IRMS and LC-IRMS. Journal Food Science Technology. 2020;57(4):1216–1232. https://doi.org/10.1007/s13197-019-04153-2 Vetrova OV, Melkov VN, Simonova GV, Kalashnikova DA. Detection of honey adulterations with sugar syrups by stable isotope mass spectrometry. Journal of Analytical Chemistry. 2017;72(7):756–760. https://doi.org/10.1134/S1061934817070152; https://www.elibrary.ru/XNGJKQ Vetrova OV, Melkov VN, Simonova GV, Kalashnikova DA. Detection of honey adulterations with sugar syrups by stable isotope mass spectrometry. Journal of Analytical Chemistry. 2017;72(7):756–760. https://doi.org/10.1134/S1061934817070152; https://www.elibrary.ru/XNGJKQ Tsagkaris AS, Koulis GA, Danezis GP, Martakos I, Dasenaki M, Georgiouc CA, et al. Honey authenticity: analytical techniques, state of the art and challenges. RSC Advances. 2021;11:11273–11294. https://doi.org/10.1039/d1ra00069a Tsagkaris AS, Koulis GA, Danezis GP, Martakos I, Dasenaki M, Georgiouc CA, et al. Honey authenticity: analytical techniques, state of the art and challenges. RSC Advances. 2021;11:11273–11294. https://doi.org/10.1039/d1ra00069a Oganesyants LA, Panasyuk AL, Kuzmina EI, Kharlamova LN. Determination of the carbon isotope 13C/12C in ethanol of fruit wines in order to define identification characteristics. Food and raw materials. 2016;4(1):141–147. https://doi.org/10.21179/2308-4057-2016-1-141-147 Oganesyants LA, Panasyuk AL, Kuzmina EI, Kharlamova LN. Determination of the carbon isotope 13C/12C in ethanol of fruit wines in order to define identification characteristics. Food and raw materials. 2016;4(1):141–147. https://doi.org/10.21179/2308-4057-2016-1-141-147 Oganesyants LA, Panasyuk AL, Kuzmina EI, Sviridov DA, Ganin MYu, Shilkin AA. Traditional siders and perry identification by isotope mass spectrometry. Food Industry. 2021;4:55–57. (In Russ.). https://doi.org/10.24412/0235-2486-2021-4-0036 ; https://www.elibrary.ru/XLXEAU Oganesyants LA, Panasyuk AL, Kuzmina EI, Sviridov DA, Ganin MYu, Shilkin AA. Traditional siders and perry identification by isotope mass spectrometry. Food Industry. 2021;4:55–57. (In Russ.). https://doi.org/10.24412/0235-2486-2021-4-0036 ; https://www.elibrary.ru/XLXEAU Oganesyants LA, Panasyuk AL, Kuzmina EI, Ganin MYu. Isotopes of Carbon, Oxygen, and Hydrogen Ethanol in Fruit Wines. Food Processing: Techniques and Technology. 2020;50(4):717–725. (In Russ.). https://doi.org/10.21603/2074-9414-2020-4-717-725; https://www.elibrary.ru/TRNZXN Oganesyants LA, Panasyuk AL, Kuzmina EI, Ganin MYu. Isotopes of Carbon, Oxygen, and Hydrogen Ethanol in Fruit Wines. Food Processing: Techniques and Technology. 2020;50(4):717–725. (In Russ.). https://doi.org/10.21603/2074-9414-2020-4-717-725; https://www.elibrary.ru/TRNZXN Oganesyants LA, Panasyuk AL, Kuzmina EI, Sviridov DA. Geographical place of origin influence on isotope characteristics of ethanol elements and wine aquatic component. Food industry. 2020;(12):78–80. (In Russ.). https://doi.org/10.24411/0235-2486-2020-10152; https://www.elibrary.ru/MVIQNZ Oganesyants LA, Panasyuk AL, Kuzmina EI, Sviridov DA. Geographical place of origin influence on isotope characteristics of ethanol elements and wine aquatic component. Food industry. 2020;(12):78–80. (In Russ.). https://doi.org/10.24411/0235-2486-2020-10152; https://www.elibrary.ru/MVIQNZ Oganesyants LA, Panasyuk AL, Kuzmina EI, Peschanskaya VA. Аpplication features of isotopic mass spectrometry when analyzing carbon ethanol in cognac and cognac distillates. Winemaking and viticulture. 2016;3:4–7. (In Russ.). https://doi.org/10.24412/0235-2486-2021-4-0036; https://www.elibrary.ru/WFSBFD Oganesyants LA, Panasyuk AL, Kuzmina EI, Peschanskaya VA. Application features of isotopic mass spectrometry when analyzing carbon ethanol in cognac and cognac distillates. Winemaking and viticulture. 2016;3:4–7. (In Russ.). https://doi.org/10.24412/0235-2486-2021-4-0036; https://www.elibrary.ru/WFSBFD Талибова А., Колеснов А. Оценка качества и безопасности пищевой продукции методом изотопной масс-спектрометрии // Аналитика. 2011. № 1. С. 44–48. https://www.elibrary.ru/PIEIFJ Talibova A, Kolesnov A. Quality and safety assessment of food products by isotope mass spectrometry. Analytics. 2011;(1):44–48. (In Russ.). https://www.elibrary.ru/PIEIFJ Oganesyants LA, Panasyuk AL, Kuzmina EI, Sviridov DA, Ganin MYu, Shilkin AA. Using the isotope mass spectrometry method to detect the presence of exogenous sugars in honey. Food Industry. 2023;(12):105–113. (In Russ.). https://doi.org/10.52653/PPI.2023.12.12.021; https://www.elibrary.ru/BHHQJQ Oganesyants LA, Panasyuk AL, Kuzmina EI, Sviridov DA, Ganin MYu, Shilkin AA. Using the isotope mass spectrometry method to detect the presence of exogenous sugars in honey. Food Industry. 2023;(12):105–113. (In Russ.). https://doi.org/10.52653/PPI.2023.12.12.021; https://www.elibrary.ru/BHHQJQ AOAC official methods of analysis. Method 998.12: C-4 plant sugars in honey, internal standard stable carbon isotope ratio method, AOAC Int. Gaithersburg MD USA. 1999;44:27–30. AOAC official methods of analysis. Method 998.12: C-4 plant sugars in honey, internal standard stable carbon isotope ratio method, AOAC Int. Gaithersburg MD USA. 1999;44:27–30. Rogers KM, Cook JM, Krueger D, Beckmann K. Modification of AOAC Official Method SM 998.12 to Add Filtration and/or Centrifugation: Interlaboratory Comparison Exercis. Journal of AOAC International. 2013; 96(3):607–614. https://doi.org/10.5740/jaoacint.12-386 Rogers KM, Cook JM, Krueger D, Beckmann K. Modification of AOAC Official Method SM 998.12 to Add Filtration and/or Centrifugation: Interlaboratory Comparison Exercis. Journal of AOAC International. 2013; 96(3):607–614. https://doi.org/10.5740/jaoacint.12-386 Bocian A, Buczkowicz J, Jaromin M, Hus KK, Legáth J. An Effective Method of Isolating Honey Proteins. Molecules. 2019;24(13):2399. https://doi.org/10.3390/molecules24132399 Bocian A, Buczkowicz J, Jaromin M, Hus KK, Legáth J. An Effective Method of Isolating Honey Proteins. Molecules. 2019;24(13):2399. https://doi.org/10.3390/molecules24132399 White JW, Doner LW. The 13C/12C ratio in honey. Journal of Apicultural Research.1978;17:94–99. https://doi.org/10.1080/00218839.1978.11099910 White JW, Doner LW. The 13C/12C ratio in honey. Journal of Apicultural Research.1978;17:94–99. https://doi.org/10.1080/00218839.1978.11099910 Trifkovic J, Andric F, Ristivojevic P, Guzelmeric E, Yestilada E. Analytical Methods in Tracing Honey Authenticity. Journal of AOAC in International. 2017;100(4):827–839. https://doi.org/10.5740/jaoacint.17-0142 Trifkovic J, Andric F, Ristivojevic P, Guzelmeric E, Yestilada E. Analytical Methods in Tracing Honey Authenticity. Journal of AOAC in International. 2017;100(4):827–839. https://doi.org/10.5740/jaoacint.17-0142 Kalashnikova DA, Simonova GV. Ratios of stable isotopes 13C/12C and 15N/14N in samples of dead honey bees and beekeeping products. Journal of Analytical Chemistry. 2021;76(4):359–368. (In Russ.). https://doi.org/10.31857/S004445022104006X; https://www.elibrary.ru/JHBSUQ Kalashnikova DA, Simonova GV. Ratios of stable isotopes 13C/12C and 15N/14N in samples of dead honey bees and beekeeping products. Journal of Analytical Chemistry. 2021;76(4):359–368. (In Russ.). https://doi.org/10.31857/S004445022104006X; https://www.elibrary.ru/JHBSUQ Taki H, Ikeda H, Nagamitsu T, Yasuda M, Sugiura S, Maeto K, et al. Stable nitrogen and carbon isotope ratios in wild native honeybees: the influence of land use and climate. Biodivers Conserv. 2017;26:3157–3166 https://doi.org/10.1007/s10531-016-1114-x Taki H, Ikeda H, Nagamitsu T, Yasuda M, Sugiura S, Maeto K, et al. Stable nitrogen and carbon isotope ratios in wild native honeybees: the influence of land use and climate. Biodivers Conserv. 2017;26:3157–3166 https://doi.org/10.1007/s10531-016-1114-x Stocker A, Rossmann A, Kettrup A, Bengsch E. Detection of royal jelly adulteration using carbon and nitrogen stable isotope ratio analysis. Rapid Communications in Mass Spectrometry. 2005;20(2):181–184. https://doi.org/10.1002/rcm.2287 Stocker A, Rossmann A, Kettrup A, Bengsch E. Detection of royal jelly adulteration using carbon and nitrogen stable isotope ratio analysis. Rapid Communications in Mass Spectrometry. 2005;20(2):181–184. https://doi.org/10.1002/rcm.2287 Dinca O-R, Ionete RE, Popescu R, Costinel D, Radu G-L. Geographical and Botanical Origin Discrimination of Romanian Honey Using Complex Stable Isotope Data and Chemometrics. Food Analitical Methods. 2015;8:401–412.https://doi.org/10.1007/s12161-014-9903-x Dinca O-R, Ionete RE, Popescu R, Costinel D, Radu G-L. Geographical and Botanical Origin Discrimination of Romanian Honey Using Complex Stable Isotope Data and Chemometrics. Food Analitical Methods. 2015;8:401–412.https://doi.org/10.1007/s12161-014-9903-x Izol E, Kaya E, Karahan D. Investigation of Some Metals in Honey Samples Produced in Different Regions of Bingöl Province by ICP-MS. Mellifera. 2021;21:1–17. Izol E, Kaya E, Karahan D. Investigation of Some Metals in Honey Samples Produced in Different Regions of Bingöl Province by ICP-MS. Mellifera. 2021;21:1–17.