Volume 2, Issue 4, 2019 ПИЩЕВЫЕ СИСТЕМЫ ПИЩЕВЫЕ СИСТЕМЫ ПИЩЕВЫЕ СИСТЕМЫ FOOD SYSTEMS An international science journal FOOD SYSTEMS FOOD SYSTEMS FOOD SYSTEMS FOOD SYSTEMS FOOD SYSTEMS FOOD SYSTEMS FOOD SYSTEMSISSN 2618-9771 (Print) ISSN 2618-7272 (On line) FOOD SYSTEMShttp://www.fsjour.com

ПИЩЕВЫЕ СИСТЕМЫ | Том 2 № 4 | 2019 FOOD SYSTEMS | Volume 2 № 4 | 2019

Федеральное агентство научных организаций РЕДАКЦИОННАЯ КОЛЛЕГИЯ Абрамова Любовь Сергеевна — ​доктор технических наук, профессор, Федеральное государственное бюджетное научное помощник директора ФГБНУ «Всероссийский научно-исследователь- учреждение «Федеральный научный центр ский институт рыбного хозяйства и океанографии», Москва, Россия пищевых систем им. В.М. Горбатова» РАН Баженова Баяна Анатольевна — ​доктор технических наук, доцент, профессор кафедры «Технология мясных и консервированных продук- тов» ФГБОУ ВО «Восточно-Сибирский университет технологии и управ- ПИЩЕВЫЕ СИСТЕМЫ ления», Улан-Удэ, Россия Галстян Арам Генрихович — ​доктор технических наук, профессор РАН, FOOD SYSTEMS член-корреспондент РАН, заведующий Межотраслевым научно-техни- ческим центром мониторинга качества пищевых продуктов, Всерос- Учредитель и издатель сийский научно-исследовательский институт пивоваренной, безалко- Федеральное государственное бюджетное гольной и винодельческой промышленности — ​филиал Федерального научное учреждение «Федеральный научный государственного бюджетного научного учреждения «Федеральный на- центр пищевых систем им. В.М. Горбатова» РАН учный центр пищевых систем им. В.М. Горбатова» РАН, Москва, Россия Горлов Иван Федорович — ​доктор сельскохозяйственных наук, профес- Главный редактор сор, академик РАН, Научный руководитель ФГБНУ «Поволжский научно- Кузнецова Оксана Александровна — ​ исследовательский институт производства и переработки мясомолоч- ной продукции», Волгоград, Россия Доктор технических наук, Врио директора ФГБНУ Донник Ирина Михайловна — доктор​ биологических наук, профессор, «Федеральный научный центр пищевых систем Академик РАН, Вице-президент РАН, Москва, Россия им. В.М. Горбатова» РАН, Москва, Россия Евдокимов Иван Алексеевич — ​доктор технических наук, профессор, Заведующий кафедрой «Технология молока и молочных продуктов» Заместитель Главного редактора ФГАОУ ВО «Северо-Кавказский федеральный университет» Ставрополь, Лисицын Андрей Борисович — ​Доктор Россия технических наук, профессор, Академик РАН, Ежкова Галина Олеговна — ​доктор биологических наук, профессор, заведующий кафедрой «Технология мясных и молочных продуктов» научный руководитель ФГБНУ «Федеральный ­ФГБОУ ВО «Казанский национальный исследовательский технологиче- научный центр пищевых систем им. В.М. Горбатова» ский университет», Казань, Россия РАН, Москва, Россия Замаратская Галия — ​кандидат технических наук, доцент, Научный ра- ботник Шведского университета аграрных наук, Упсала, Швеция Научный редактор Иванкин Андрей Николаевич — ​доктор химических наук, профессор, Горбунова Наталия Анатольевна — ​Кандидат Заведующий кафедрой «Химия», Мытищинский филиал Московского государственного технического университета им. Н.Э. Баумана, Мыти- технических наук, ученый секретарь, ФГБНУ щи, Московская область, Россия «Федеральный научный центр пищевых систем Кочеткова Алла Алексеевна — ​доктор технических наук, профессор, им. В.М. Горбатова» РАН, Москва, Россия Руководитель лаборатории пищевых биотехнологий и специализиро- ванных продуктов, ФГБУН «Федеральный исследовательский центр пи- Научный редактор тания, биотехнологии и безопасности пищи», Москва, Россия Банникова Анна Владимировна — ​ Машенцева Наталья Геннадиевна- доктор технических наук, доцент, Доктор технических наук, Заведующий профессор РАН, заведующий кафедрой «Биотехнология и технология продуктов биоорганического синтеза» ФБГОУ ВО «Московский государ- учебно-научно-испытательной лабораторией, ственный университет пищевых производств», Москва, Россия ФГБОУ ВО «Саратовский государственный аграрный Мирошников Сергей Александрович — ​доктор биологических наук, университет имени Н.И. Вавилова», профессор, Директор ФГБНУ «Всероссийский научно-исследовательский Саратов, Россия институт мясного скотоводства», Оренбург, Россия Римарева Любовь Вячеславовна — ​доктор технических наук, профес- Выпускающий редактор сор, Академик РАН, Заместитель директора Всероссийского научно-ис- Захаров Александр Николаевич — ​Кандидат следовательского института пищевой биотехнологии — ​филиал ФГБУН «Федеральный исследовательский центр питания, биотехнологии и без- технических наук, старший научный сотрудник, опасности пищи», Москва, Россия заведующий редакционно-издательским отделом, Настасиевич Иван — ​доктор, адъюнкт-директор, Институт гигиены ФГБНУ «Федеральный научный центр пищевых и технологии мяса, Белград, Сербия систем им. В.М. Горбатова» РАН, Москва, Россия Петров Андрей Николаевич — ​доктор технических наук, профессор, Академик РАН, Директор Всероссийский научно-исследовательский ин- ститут технологии консервирования — ​филиал Федерального государ- АДРЕС РЕДАКЦИИ И ТИПОГРАФИИ: ственного бюджетного научного учреждения «Федеральный научный 109316, Россия, Москва, Талалихина, 26, центр пищевых систем им. В.М. Горбатова» РАН, Видное, Московская Федеральный научный центр пищевых систем область, Россия им. В.М. Горбатова РАН. Просеков Александр Юрьевич — ​доктор технических наук, профессор, профессор РАН, Ректор ФГБОУ ВО «Кемеровский государственный уни- www.fsjour.com верситет», Кемерово, Россия Ребезов Максим Борисович — ​доктор сельскохозяйственных наук, Журнал зарегистрирован в Роскомнадзоре профессор, Заведующий кафедрой прикладной биотехнологии ФГАОУ Регистрационные данные: ВО «Южно-Уральский государственный университет (национальный ис- следовательский университет), Челябинск, Россия ПИ № ФС77–71610 от 13.11.2017 Такеда Широ, — ​адьюнкт- профессор, Профессор лаборатории науки ЭЛ № ФС 77–72022 от 26.12.2017 о пище, Институт ветеринарной медицины, Университет Азабу, Сагами- Периодичность — ​4 номера в год. хара, Япония Издается с 2018 года. Тимошенко Николай Васильевич- доктор технических наук, профес- сор, Заведующий кафедрой технология хранения и переработки живот- Подписано в печать 18.12.2019. новодческой продукции, ФГБОУ ВО «Кубанский государственный аграр- Тираж 300 экз. Заказ № 249. ный университет», Краснодар, Россия Типография ФНЦПС. Чернуха Ирина Михайловна — ​доктор технических наук, профессор, член-корреспондент РАН, Руководитель научного направления ФГБНУ «Федеральный научный центр пищевых систем им. В.М. Горбатова» РАН, © ФНЦПС, 2019 Москва, Россия ISSN 2618-9771 (Print) Швегеле Фреди — ​доктор, Директор Института Макса Рубнера, Куль- ISSN 2618-7272 (Online) мбах, Федеративная Республика Германия

1 ПИЩЕВЫЕ СИСТЕМЫ | Том 2 № 4 | 2019 FOOD SYSTEMS | Volume 2 № 4 | 2019

Federal Agency of Scientific Organizations EDITORIAL BOARD (FANO of Russia) Liubov S. Abramova, doctor of technical sciences, professor, assistant direc- V.M. Gorbatov Federal Research Center for Food tor of Russian Federation Research Institute of Fishery and Oceanography, Systems of Russian Academy of Sciences Moscow, Russia (Gorbatov Research Center for Food Systems) Baiana A. Bazhenova, doctor of technical sciences, docent, professor of the chair «Meat and canned product technology» of East Siberia State University of Technology and Management, Ulan-Ude, Russia ПИЩЕВЫЕ СИСТЕМЫ Aram G. Galstyan, Doctor of Technical Science, Professor of RAS, Corre- FOOD SYSTEMS sponding Member of RAS, Head of the Interbranch Scientific and Technical Center for Food Quality Monitoring of All-Russian Scientific Research Insti- Founder and publisher: tute of Brewing, Beverage and Wine Industry — ​Branch of V.M. Gorbatov Fed- V.M. Gorbatov Federal Research Center for Food eral Research Center for Food Systems of RAS, Moscow, Russia Systems of Russian Academy of Sciences Ivan F. Gorlov, doctor of agricultural sciences, professor, academician of Editor-in-Chief: RAS, Scientific supervisor of Povolzhskiy Research Institute of Production Oxana A. Kuznetsova, doctor of technical sciences, and Processing of Meat and Dairy Products, Volgograd, Russia Director of V.M. Gorbatov Federal Research Center Irina M. Donnik, doctor of biological sciences, professor, academician of for Food Systems of Russian Academy of Sciences, RAS, Vice president of RAS, Moscow, Russia Moscow, Russia Ivan A. Evdokimov, doctor of technical sciences, professor, Head of the chair «Technology of milk and dairy products» of North-Caucasus Federal Univer- Deputy Editor-in-Chief: sity, Stavropol, Russia Andrey B. Lisitsyn, doctor of technical sciences, Galina O.Ezhkova, doctor of biological sciences, professor, the head of the professor, Academician of RAS, Scientific supervisor chair «Technology of meat and dairy products» of Kazan National Research Of V.M. Gorbatov Federal Research Center Technological University, Kazan, Russia for Food Systems of Russian Academy of Sciences, Galia Zamaratskaya, candidate of technical sciences, docent, research work- Moscow, Russia er, the Swedish University of Agricultural Sciences, Uppsala, Sweden Science editor: Andrey N. Ivankin, doctor of Chemical Sciences, professor, the head of the chair of Chemistry of Mytishchi branch of Bauman Moscow State Technical Natalia A. Gorbunova, candidate University, Mytischi, Moscow region, Russia of technical sciences, Academic Secretary of Alla A. Kochetkova, doctor of technical sciences, professor, the head of the V.M. Gorbatov Federal Research Center for Food «Laboratory of food biotechologies and specialized products», Federal Re- Systems of Russian Academy of Sciences, Moscow, search Centre of nutrition, biotechnology and food safety, Moscow, Russia Russia Natal’ya G. Mashentseva, doctor of technical sciences, professor RAS, the Science editor: head of the chair of Biotechnology and Technology of Products of Bioorganic Anna V. Bannikova, doctor of technical sciences, Synthesis of Moscow State University of Food Production, Moscow, Russia Head of the educational-scientific-testing laboratory, Sergey A. Miroshhikov, doctor of biological sciences, professor, director of Saratov State Vavilov Agrarian University, Saratov, The All-Russian Research Institute of Beef Cattle, Orenburg, Russia Russia Liubov V. Rimareva, doctor of technical sciences, professor, academician of RAS, deputy director of The All-Russian Scientific Research Institute of Food Production editor: Biotechnology — ​branch Federal Research Centre of nutrition, biotechnology Aleksandr N. Zakharov, candidate of technical and food safety, Moscow, Russia sciences, senior ressearch worker, Head of the Editorial Nastasijevic Ivan, doctor, Associate Director of the Institute of Meat Hygiene and Publishing Department of V.M. Gorbatov Federal and Technology, Belgrad, Serbia Research Center for Food Systems of Russian Academy Andrey N. Petrov, doctor of technical sciences, professor, academician of of Sciences», Moscow, Russia RAS, Director of All-Russian Research Institute of Canning Technology —​ Branch of V.M. Gorbatov Federal Research Center for Food Systems of RAS, PRINTING OFFICE: Vidnoe, Moscow region, Russia 109316, Talalikhina str. 26, Moscow, Russia, Aleksandr Yu. Prosekov, doctor of technical sciences, professor, professor of Gorbatov Research Center for Food Systems. RAS, Rector of Kemerovo State University, Kemerovo, Russia www.fsjour.com Maxim B. Rebezov, doctor of agricultural sciences, professor, the head of the chair of Applied Biotechnology of South Ural State University (national The journal is registered in ROSKOMNADZOR. research university), Chelyabinsk, Russia Registration data: ShiroTakeda, associate Professor, Laboratory of Food Science School of ПИ № ФС77–71610 from 13.11.2017 Veterinary Medicine, Azabu University, Sagamihara, Japan ЭЛ № ФС 77–72022 from 26.12.2017 Nikolai V. Timoshenko, doctor of technical sciences, professor, the head of Frequency — ​4 issues a year. the chair «Technology of storage and processing of animal products» of the Published in 2018. Kuban State Agrarian University, Krasnodar, Russia Signed print 18.12.2019. Irina M. Chernukha, doctor of technical sciences, professor, corresponding Circulation — ​300 copies. Order № 249. members of RAS, head of the scientific direction of V.M. Gorbatov Federal Printing house — ​FNCPS. Research Center for Food Systems of Russian Academy of Sciences, Moscow, © FNCPS, 2019 Russia ISSN 2618-9771 (Print) Fredi Schwagele, doctor, Director of Max Rubner-Institut, Kulmbach, ISSN 2618-7272 (Online) Germany.

2 ПИЩЕВЫЕ СИСТЕМЫ | Том 2 № 4 | 2019 FOOD SYSTEMS | Volume 2 № 4 | 2019

CONTENTS

Liubov S. Abramova, Andrey V. Kozin, Alexander S. Shashkov NMR — ​SPECTROSCOPY TECHNIQUE FOR SALMON FISH SPECIES QUALITY ASSESSMENT ...... 4

Marina V. Gernet, Irina N. Gribkova, Olga A. Borisenko THE INCREASING THE DURATION OF FUNCTIONAL BEVERAGES METHODS ...... 10

Vera V. Eveleva, Tatyana M. Cherpalova INNOVATIVE DECISIONS TO IMPROVE FOOD QUALITY AND SAFETY ...... 14

Irina S. Vitol, Natalia A. Igoryanova, Elena P. Meleshkina BIOCONVERSION OF SECONDARY PRODUCTS OF PROCESSING OF GRAIN CEREALS CROPS ...... 18

Yuri F. Markov, Alexandra N. Buriak, Larisa G. Eresko EQUIPMENT AND SCIENTIFIC STUDIES OF EXPERIMENTAL DATA ON STORAGE OF WHEAT GRAIN ...... 25

Artem V. Samoilov, Natalia M. Suraeva, Sergey V. Koptsev, Sergey V. Glazkov DEVELOPING THE METHODS OF FOOD PRESERVATIVES EXTRACTION FROM COMPLEX MATRICES FOR BIOASSAY PURPOSE ...... 31

Elena V. Topnikova, Valentina A. Mordvinova, Galina M. Sviridenko, Ekaterina S. Danilova STUDY OF FATTY ACID COMPOSITION OF MILK FOR CHEESE PRODUCTION ...... 34

Tatyana V. Savenkova, Alexander R. Karimov, Mikhail A. Taleysnik, Timofei V. Gerasimov, Nikolai B. Kondratev MECHANISMS OF DESTRUCTION AND SYNTHESIS OF LIQUID MEDIA, USED IN THE FOOD INDUSTRY UNDER NON-EQUILIBRIUM CONDITIONS ...... 38

Svetlana V. Avilova, Vladimir N. Kornienko, Aleksey A. Gryzunov, Anna A. Vankova AN EFFECT OF STORAGE AND TRANSPORTATION TEMPERATURE ON QUANTITATIVE AND QUALITATIVE COMPOSITION OF MICROFLORA OF PLANT PRODUCTS ...... 42

Galina P. Pokudina, Marina V. Trishkaneva, Raisa A. Volkova DEVELOPMENT OF PASTERIZATION MODES FOR HIGH-SUGAR CANS IN CONTINUOUS ACTING PASTEURIZERS ...... 48

3 ПИЩЕВЫЕ СИСТЕМЫ | Том 2 № 4 | 2019 FOOD SYSTEMS | Volume 2 № 4 | 2019

UDC [664.951.014:543.42]:639.211 DOI: 10/21323/2618–9771–2019–2–4–4–9

The Original Scientific Article NMR — ​SPECTROSCOPY TECHNIQUE FOR SALMON FISH SPECIES QUALITY ASSESSMENT Liubov S. Abramova*1, Andrey V. Kozin1, Alexander S. Shashkov2 1 Research Institute of Fisheries and Oceanography, Moscow, Russia 2 N.D. Zelinsky Institute of Organic Chemistry Russian Academy of Sciences, Moscow, Russia

KEY WORDS: ABSTRACT salmon fish species, NMR‑spectroscopy was used for development of the criteria which characterize the chilled and frozen fish quality. NMR — ​spectroscopy, quality It has been shown that 1H-NMR experiments can be used as quality factor to measure the concentration of inosine, factor K1, quality factor H hypoxanthine and inosine‑5’-monophosphate generated during the fish storage. The quality factor is expressed

by the K1 correlates well with the sensory quality of chilled Atlantic salmon (Salmo Salar), whereas, quality fac- tor H is more sensitive for measuring the quality characteristics of frozen pink salmon (Oncorhynchus gorbuscha), chum salmon (Oncorhynchus keta), sockeye salmon (Oncorhynchus nerka).

1. Introduction of fish freshness. The quality factor K is defined by the following One of the fundamental tasks of the Fisheries Industries is to equation [6,7,8]: ensure the preservation of the high-quality fishery products, the + manufacturing of products with the predetermined customer = 100, % (1) (1) properties and the guaranteed shelf life. + + + +++ + == 𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼 𝐻𝐻𝐻𝐻𝐻𝐻𝐻𝐻 100100,,%% ((11)) In order to comply with these fundamental task prerequi- It𝐾𝐾𝐾𝐾 has been suggested++ that the++ determination++ ++ of adenosine∗ ++ ‑5’- sites, it is obligatory to be familiar with the processes occurring triphosphateIt has 𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴been degradation suggested𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴 𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴 productsthat the𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴 contentdetermination𝐻𝐻𝐻𝐻𝐻𝐻𝐻𝐻𝐻𝐻𝐻𝐻𝐻𝐻𝐻𝐻𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼 in the𝐻𝐻𝐻𝐻𝐻𝐻𝐻𝐻 fish of canadenosine be used- 5'-triphosphate degradation products content𝐾𝐾𝐾𝐾 in the fish can be used as the basis of the quality∗ factor K calculation, when during the fish storage and processing, to know how to manage as the basisIt 𝐾𝐾𝐾𝐾ofhas the 𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴been quality suggested𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴 factor 𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴 Kthat calculation, the𝐼𝐼𝐼𝐼𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴 determination when𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼 evaluating𝐻𝐻𝐻𝐻𝐻𝐻𝐻𝐻 of∗ adenosine -5'-triphosphate degradation evaluating postIt- mortemhas 𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴been fish suggested quality𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴 changes𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴 that the during𝐼𝐼𝐼𝐼𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴 determination fish𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼 storage𝐻𝐻𝐻𝐻𝐻𝐻𝐻𝐻 of [8 ]adenosine. -5'-triphosphate degradation them and to have an opportunity to control the basic indexes post-mortemproductsproducts contentfishcontent quality inin thethe changes fishfish cancan during bebe usedusedfish asstorageas thethe basisbasis[8]. ofof thethe qualityquality factorfactor KK calculation,calculation, whenwhen characterizing its quality. AsAsevaluating a a result, result, post it became-becamemortem considerably considerablyfish quality changes interesting interesting during not not fish only only storage to to [analyze8]. the content of metabolitesevaluating in the chilledpost-mortem fish during fish quality their storage changes but during also fishto characterize storage [8]. the raw fish quality, During fishing auctions in the European Community coun- analyze theAsAs contentaa result,result, of it metabolitesit becamebecame considerably considerably in the chilled interestinginteresting fish during notnot onlyonly toto analyzeanalyze thethe contentcontent ofof tries, the sensory quality of fish (fish quality index) is used forfrom their whichmetabolites storage the frozen but in also productthe tochilled characterize was fish manufactured. during the rawtheir fish storage quality, but fromalso to characterize the raw fish quality, Accordingmetabolites to Karube in the chilled [9] for fishsome during fish speciestheir storage ATP decomposes but also to characterizevery quickly theto IMPraw fishand quality, the evaluation of the freshness grade of the fishes. This quality whichfromfrom the whichwhich frozen thethe product frozenfrozen was productproduct manufactured. waswas manufactured.manufactured. index can also be used at fish unloading points, for calculationthe definition of quality factor K can be modified by excluding the adenosine phosphates and AccordingAccordingAccording to Karube toto KarubeKarube [9] for [ some[99]] fforor fish ssoomeme fishspeciesfish speciesspecies ATP ATPATP decom - decomposesdecomposes veryvery quicklyquickly toto IMPIMP andand of the shelf life and for the storage conditions which are used suggestedposes a very new quickly calculation to IMP quality and factor the definition К1 defined ofas: quality factor thethe definitiondefinition ofof qualityquality factorfactor KK cancan bebe modifiedmodified byby excludingexcluding thethe adenosineadenosine phosphatesphosphates andand for sorting before further processing [1]. Therefore, the sensory K can be modified by excluding the adenosine phosphates and suggestedsuggested aa newnew calculationcalculation qualityquality factorfactor КК1 defidefinedned as:as: properties such as: taste, sight, smell, and touch are evaluated suggested a new calculation+ quality factor К defined1 as: = 100, % 1 (2) 1 + + for the appearance, flavor, mucus condition, eyes, gills and ab- ++ dominal cavity of the whole or gutted fish. On the basis of these 1𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼== 𝐻𝐻𝐻𝐻𝐻𝐻𝐻𝐻 100100,,%% (2) ((22)) 𝐾𝐾𝐾𝐾 1 ++ ∗ ++ Luong [10]𝐼𝐼𝐼𝐼 𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴proved𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼 that, 𝐻𝐻𝐻𝐻𝐻𝐻𝐻𝐻for some species, quality factor K and K1 do not adequately indexes, the fish is referred to E (Extra) grade —​that is the high- 𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼 𝐻𝐻𝐻𝐻𝐻𝐻𝐻𝐻𝐻𝐻𝐻𝐻𝐻𝐻𝐻𝐻 est freshness grade, А, В or С grades (runoff product). reflect theLuong alterations [10] proved𝐾𝐾𝐾𝐾𝐾𝐾𝐾𝐾 and that, proposed for some the species, ∗quality∗ quality factor factor H based K and on Нх concentration to be LuongLuong [[1010]]𝐼𝐼𝐼𝐼 𝐼𝐼𝐼𝐼 𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴provedproved𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼 that,that, 𝐻𝐻𝐻𝐻𝐻𝐻𝐻𝐻 𝐻𝐻𝐻𝐻𝐻𝐻𝐻𝐻forfor somesome species,species, qualityquality factorfactor KK andand KK11 dodo notnot adequatelyadequately The rapid test quality index method (QIM) is used for the consideredK do notas a adequately good indicator reflect of fish the freshness alterations under physiologicaland proposed and the sensory points of view: 1 reflectreflect thethe alterationsalterations andand proposedproposed thethe qualityquality factorfactor HH basedbased onon НхНх concentrationconcentration toto bebe chilled fish [2]. The present method is based on consideration qualityconsideredconsidered factor H as asbased aa goodgood on indicatorНхindicator concentration ofof fishfish freshnessfreshness to be considered underunder physiologicalphysiological as a andand sensorysensory pointspoints ofof viewview:: of the specific changes in the fish, namely appearance, flavor good indicator of fish freshness under physiological and sensory and consistence changes. The penalty points from 0 to 3 are points of view: = 100, % (3) calculated for each index, then the points are summarized by + + all the indexes and the total sensory estimate is obtained, that =𝐻𝐻𝐻𝐻𝐻𝐻𝐻𝐻 100, % (3) (3) 𝐻𝐻𝐻𝐻 = ∗ 100, % (3) is known as the “quality index”, linearly increasing with the ++ ++ In recent years,𝐼𝐼𝐼𝐼𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴 it has𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼 been𝐻𝐻𝐻𝐻𝐻𝐻𝐻𝐻𝐻𝐻𝐻𝐻𝐻𝐻𝐻𝐻𝐻𝐻𝐻𝐻𝐻𝐻𝐻𝐻 shown that NMR spectroscopy is a good tool for assessing the fish storage time. Under modern conditions the method men-quality Inof recentfish raw years, 𝐻𝐻𝐻𝐻materials it has been [7,8]. shown The thatfish∗ NMRquality spectroscopy factor can isbe a calculated by analyzing the In recent𝐻𝐻𝐻𝐻 years,𝐼𝐼𝐼𝐼𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴 it has𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼 been𝐻𝐻𝐻𝐻𝐻𝐻𝐻𝐻 shown∗ that NMR spectroscopy is a good tool for assessing the tioned above allows for quick and objective evaluation of the NMRgood spectra tool andforIn recentassessing determining years,𝐼𝐼𝐼𝐼𝐴𝐴𝐴𝐴𝐴𝐴𝐴𝐴 the itthequality has𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼𝐼 concentration been of𝐻𝐻𝐻𝐻𝐻𝐻𝐻𝐻 fish shown raw of that materialsATP NMR decay [7,8]. spectroscopy products. The is a good tool for assessing the fish quality by means of the special program, adapted for dif- fishquality qualityquality of offactor fishfish canrawraw be materialsmaterials calculated [7,8].[7,8]. by analyzing TheThe fishfish thequalityquality NMR factor factorspec- cancan bebe calculatedcalculated bbyy analyzinganalyzing thethe In NMRthe work, spectra NMR and spectroscopydetermining thewas concentration used to assess of theATP quality decay ofproducts. salmon species of fish. ferent computer-based systems. Evaluation can be performed The traresearch andNMR determining w spectraas conducted and the determining onconcentration chilled the Atlantic concentration of ATP salmo decayn (ofSalmo products. ATP Salardecay) products.as the most massive fish, at any stage: during the fishery products delivery at production In the InInwork, thethe NMRwork,work, spectroscopy NMRNMR spectroscopyspectroscopy was used waswas to assessusedused toto the assass qualessess -thethe qualityquality ofof salmonsalmon speciesspecies ofof fish.fish. sold throughThe research retail chains was conductedas chilled onand chilled frozen A pinctlantic salmon salmo (Oncorhynchusn (Salmo Salar )gorbuscha) as the most, chummassive fish, site, fish storage or sale. The disadvantage of QIM is that it issalmon ity of(TheOncorhynchus salmon research species w ketaas conducted of), fish.sockeye onThe salmon chilled research (Oncorhynchus Atlantic was s almo conducted nerkan (Salmo) on which Salar are) usedas the in most trade massive and fish, sold through retail chains as chilled and frozen pinc salmon (Oncorhynchus gorbuscha), chum applied only to the proven testing groups and it can be used processingchilledsold asAtlantic throughfrozen. salmon retail chains(Salmo asSalar chilled) as theand most frozen massive pinc salmon fish, (Oncorhynchus gorbuscha), chum salmonsalmon ((OncorhynchusOncorhynchus ketaketa),), sockeyesockeye salmonsalmon ((OncorhynchusOncorhynchus nerkanerka)) whichwhich areare usedused inin tradetrade andand only for whole fish. sold through retail chains as chilled and frozen pinc salmon (On‑ It is well known that autolysis and bacterial deterioration corhynchusprocessingprocessing gorbuscha) asas frozen.frozen., chum salmon (Oncorhynchus keta), sock- lead to changes in concentrations of adenosine‑5’-triphosphate eye2. salmonMaterials (Oncorhynchus and methods nerka ) which are used in trade and [АТP], adenosine‑5’-diphosphate [АDP], adenosine‑5’-mono- processing as frozen. 2.1. Samples preparation phosphate [АМP] and inosine‑5’-monophosphate [ІМР], which 2.2. MaterialsMaterials andand methodsmethods The chilled Atlantic Salmon (Salmo Salar) with storage duration of 9 days produced on the convert quantitatively in inosine [Ino] and hypoxanthine [Нх]. 2. Materials2.1.2.1. SamplesSamplesand methods preparationpreparation Faroe Islands (starting from the preparation date) and on the 18 day of storage (from the It has been reported that there was a good correlation relating 2.1. SampleThe preparation chilled Atlantic Salmon (Salmo Salar) with storage duration of 9 days produced on the preparation date)The at chilled the temperature Atlantic Salmon of approximately (Salmo Salar 0) оwithС, frozen storage pinc duration salmon of (Oncorhynchus9 days produced on the the decrease of fish freshness with the increase in nucleotides TheFaroeFaroe chilled IslandsIslands Atlantic (starting(starting Salmon fromfrom (Salmo thethe preparationSalarpreparation) with storage date)date) and anddura on-on thethe 1818 dayday ofof storagestorage (from(from thethe formation for a large number of fish species [3,4,5]. In this con-gorbuscha)tion of ,9 chumdays producedsalmon ( onOncorhynchus the Faroe Islands keta), (starting sockeye fromsalmon theоо (Oncorhynchus nerka) with preparationpreparation date)date) atat thethe temperaturetemperature ofof approximatelyapproximately 00 СС,, frozenfrozen pincpinc salmonsalmonо (Oncorhynchus(Oncorhynchus text the quality factor K was used for the quantitative evaluation storagepreparation duration date)about and three on monththe 18 sday at ofthe storage temperature (from theof approximately prepara- minus 18 С were used gorbuscha)gorbuscha),, chumchum salmonsalmon ((OncorhynchusOncorhynchus ketaketa),), sockeyesockeye salmonsalmon ((OncorhynchusOncorhynchus nerkanerka)) withwith as objects for research. For the analysis were used average muscular samples of different parts,о FOR CITATION: Abramova L. S., Kozin A. V., Shashkov A. S. NMR — ​spectroscopy techniquestorage for salmon dur ationfish species about quality three assessment. monthsFood at the systems. temperature 2019; 2(4): 4–??.of approximately minus 18 оС were used of severalstorage chilled dur andation frozen about fish. three months at the temperature of approximately minus 18 С were used DOI: 10.21323/2618–9771–2019–2–4–4–9 asas objectsobjects forfor research.research. ForFor thethe analysisanalysis werewere usedused averageaverage muscularmuscular samplessamples ofof differentdifferent parts,parts, Theof proteinseveral chilledcontent and was frozen determined fish. on the autoanalyzer «Kjeltec» Foss-2300 (Sweden) accordingof to several the Kjeldahl chilled andmethod frozen [11] fish.. Moisture content was determined in according to GOST 4 TheThe proteinprotein contentcontent waswas determineddetermined onon thethe autoanalyzerautoanalyzer «Kjeltec»«Kjeltec» FossFoss--23002300 ((SwedenSweden)) 7636 [11according]. Lipids tocontent the Kjeldahl was defined method on [11] the .automated Moisture contentextractor was SER determined 148/6 manufactured in according by to GOST VELP companyaccording according to the Kjeldahl to the Soxhletmethod method[11]. Moisture. The content content of wasTotal determined Volatile Basic in according Nitrogen to GOST 76367636 [1[111]].. LLipidsipids contentcontent waswas defineddefined onon thethe automatedautomated extractorextractor SERSER 148/6148/6 manufacturedmanufactured byby (TVB-N)VELP was companydetermined according on the to semi the -Soxhletautomatic method distillation. The content unit K of-355 Total (Buchi Volatile company Basic Nitrogen, SwitzerlandVELP). company according to the Soxhlet method. The content of Total Volatile Basic Nitrogen (TVB-N) was determined on the semi-automatic distillation unit K-355 (Buchi company, 2.2.(TVB Samples-N) preparationwas determined for NMR on -thespectroscopy semi-automatic distillation unit K-355 (Buchi company, SSwitzerlandwitzerland).). 2.2.2.2. SamplesSamples preparationpreparation forfor NMRNMR--spectroscopyspectroscopy

2

22

ПИЩЕВЫЕ СИСТЕМЫ | Том 2 № 4 | 2019 FOOD SYSTEMS | Volume 2 № 4 | 2019 tion date) at the temperature of approximately 0 °С, frozen pinc The TVB-N index was 31 g/100 g. According to Commission salmon (Oncorhynchus gorbuscha), chum salmon (Oncorhynchus Regulation EC No 1022/2008/ЕС of 17 Dec. 2008, which makes keta), sockeye salmon (Oncorhynchus nerka) with storage dura- amendments in Regulation ЕС № 2074/2005 in respect of the tion about three months at the temperature of approximately critical concentrations of Total Volatile Basic Nitrogen (­TVB-N) minus 18 оС were used as objects for research. For the analysis [13] the critical content of TVB-N should be 35 mg/100 g for were used average muscular samples of different parts, of several salmon fishes. When the present value exceeds 35 mg/100 g, chilled and frozen fish. the product is considered inedible and unsuitable for indus- The protein content was determined on the autoanalyzer trial processing. «Kjeltec» Foss‑2300 (Sweden) according to the Kjeldahl method Table 2 [11]. Moisture content was determined in according to GOST Sensory quality of fish muscle and TVB-N content 7636 [11]. Lipids content was defined on the automated extrac- tor SER148/6 manufactured by VELP company according to the Soxhlet method. The content of Total Volatile Basic Nitrogen Organoleptic muscular Title (TVB-N) was determined on the semi-automatic distillation unit indicators K‑355 (Buchi company, Switzerland). Storage days duration, Storage °С temperature, TVB-N content, mg/100 g

2.2. Sample preparation for NMR‑spectroscopy Atlantic Salmon Bright color, no evidence 9 0 22.4 The water soluble polar metabolites of fish samples were ex- (Salmo Salar) of oxidation, no off-odor tracted by 7.5% solution of trichloroacetic acid (TCA), as it was Color with shade of described in the paper [7]. For this purpose 25 g of fish muscle Atlantic Salmon orange, showing signs of 18 0 31.1 was added to 50 mL of 7.5% TCA and homogenized using a verti- (Salmo Salar) oxidation, the off-odor of cal homogenizer. The homogenate was filtered through the pa- rank fish per filter. The filtrate was neutralized by 9 М solution of КОН up After thawing the color Atlantic Salmon is bright, no evidence of 9 minus 20 24.1 to рН value of 7.8. The solution was filtered through the paper (Salmo Salar) oxidation, no off-odor filter (№ 1) and it was stored at the temperature minus 40 оС un- til the measurements conduction. After thawing the color Pinc salmon is characteristic of the (Oncorhynchus species, there is no 90 minus 18 14.0 2.3. NMR‑spectroscopy performance gorbuscha) evidence of oxidation and For NMR‑spectral data collection the samples were thawed no off-odor and dissolved in D2O. The NMR‑spectral data was collected After thawing the color with Bruker AV‑600 spectrometer (Germany) at 30 °С with TSP is characteristic of the Chum salmon species, there is no 95 minus 18 9.8 (3-(trimethilsilyl)-propionic‑2,2,3,3-d acid sodium salt) taken (Oncorhynchus keta) 4 evidence of oxidation and as the internal standard (δН 0.0, δС –1.6). All metabolites marked no off-odor at spectra were identified with the use of1 1D( Н, 13C) and 2D 1 1 1 13 After thawing the color ( H, H COSY, TOCSY, ROESY и H, C HSQC, HMBC) NMR‑spec- Sockeye salmon is bright red, there is no 98 minus 18 12.6 troscopy, and also on the basis of literature values [7,8,12]. Spec- (Oncorhynchus nerka) evidence of oxidation and tra processing was performed with the use of standard software no off-odor package of the Bruker company (TopSpin 3.6.1) According to the TVB-N index chilled Atlantic Salmon with 3. Results and discussion storage duration of 18 days at 0 °С met the requirements appli- 3.1. Proximate analysis cable to food fishery products suitable for industrial process- Proximate analysis chemical composition of the fish muscle ing and direct consumption. But at the same time on the basis of Atlantic salmon (Salmo Salar), pink salmon (Oncorhynchus of the sensory quality of fish it could not be recommended for gorbuscha), chum salmon (Oncorhynchus keta), sockeye salmon direct consumption. Consequently, the TVB-N index which is (Oncorhynchus nerka) are presented in Table 1. recommended to be determined in case of discrepancies dur- Table 1 ing evaluation of the sensory quality of the chilled or frozen Chemical composition of the fish muscle fishery products, sometimes characterizes the product quality incorrectly. Content,% Title moisture lipids ashes protein 3.2. NMR‑spectra 1 Atlantic Salmon The Н-NMR spectra of chilled Atlantic salmon (Salmon Sa- 60.20 ± 0.07 17.70 ± 0.10 1.90 ± 0,12 20.20 ± 0.10 (Salmo Salar) lar) with 9 and 18 days storage duration at 0 °С muscle TCA ex- 1 Pinc salmon tract is presented in Figure 1. The similar Н-NMR spectra were (Oncorhynchus 71.46 ± 0.05 5.32 ± 0.02 1.71 ± 0.10 21.51 ± 0.11 recorded for all fish samples. gorbuscha) The low-field region 1Н-NMR spectra of Pinc salmon (On‑ Chum salmon corhynchus gorbuscha), Chum salmon (Oncorhynchus keta), 73.70 ± 0.07 4.08 ± 0.02 1.55 ± 0.08 20.67 ± 0.08 (Oncorhynchus keta) Sockeye salmon (Oncorhynchus nerka) muscle TCA extract are Sockeye salmon presented in Figure 2. The signals of the following metabo- 69.40 ± 0.05 7.32 ± 0.02 1.67 ± 0.11 21.61 ± 0.10 (Oncorhynchus nerka) lites were assigned: lactic acid (LA), anserin (Ans), creatine (Crt), trimethylamine (TMA), choline (Cho), trimethylamine The quality estimation of the fish samples studied were ob- N‑oxide (TMAO), adenosine‑5’-triphosphate (АТP), ade- tained from different conditions of storage (duration) and the nosine‑5’-diphosphate (АDP), adenosine‑5’-monophosphate sensory quality of fish were estimated and TVB-N values deter- (АМP), inosine (Ino), hypoxanthine (Hx) and inosine‑5’-mo- mined (see the results in Table 2). nophosphate (IMP). The structural formulas and chemical As it is seen from the presented data on 18 day of stor- shifts of 1H and 13C of the metabolites mentioned above are age the chilled fish had the off-odor and changed the color. presented in ­Table 3.

5 ПИЩЕВЫЕ СИСТЕМЫ | Том 2 № 4 | 2019 FOOD SYSTEMS | Volume 2 № 4 | 2019

Figure 1. 1Н-NMR spectrum of chilled Atlantic Salmon (Salmo Salar) muscle TCA extract with 9 days (black color) and 18 days (red color) storage duration at 0 °С

Figure 2. The low-field region1 Н-NMR spectra of fish muscle TCA extract: A) Pinc salmon (Oncorhynchus gorbuscha), B) Chum salmon (Oncorhynchus keta), C) Sockeye salmon (Oncorhynchus nerka)

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Table 3 Chemical shifts of 1H and 13C of the metabolites identified in1 Н — ​1 3 С HSQC NMR spectra of Salmon fish species muscle TCA extract

Carbon Chemical shifts Metabolite Chemical Structures atom proton δН, ppm Multiplicity* Carbon atom δС, ppm 1 183.7 2 4.12 d 69.7 Lactic acid (LA) 3 1.33 d 21.5

2 7.84 s 138.2 4 131.3 5 6.91 s 123.1 6 3.77 s 33.6 Ancerin 7 3.22, 3.06 dd 27.3 (Ans) 8 4.47 dd 54.8 9 178.1 11 172.8 12 2.71 m 33.3 13 3.26 dd 37.0 1 175.6 Creatine 2 3.94 s 55.2 (Crt) 4 158.2 5 3.06 s 38.4

Trimethylamine 1,2,3 2.90 s 47.3 (ТMA)

Choline 3,4,5 3.24 s 54.8 (Cho)

Trimethylamine N‑oxide 1,2,3 3.27 s 62.2 (ТМАО)

2 8.34 147.5 4 149.5 5 125.7 6 160.0 Inosin‑5’-phosphate 8 8.58 s 141.2 (IMP) 1’ 6.14 s 89.1 2’ 4.70 m 76.2 3’ 4.49 dd 71.5 4’ 4.36 m 85.5 5’ 4.06, 4.01 m 64.8 2 8.34 s 147.5 4 149.5 5 125.7 6 160.0 Inosine 8 8.24 s 141.5 (Ino) 1’ 6.09 d 89.9 2’ 4.77 s 75.3 3’ 4.45 dd 71.5 4’ 4.29 dd 86.7 5’ 3.95, 3.86 dd 62.6 2 8.21 s 146.7 4 153.9 5 120.7 Hypoxantin (Hx) 6 158.6

8 8.19 s 143.1

* s — ​singlet; d — ​doublet; dd — ​doublet of doublet; m — ​multiplet.

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3.3. Determination of adenosine‑5’-triphosphate degradation high, which did not correlate with satisfactory sensory quality of products’ content in fishery products fish and the TVB-N indexes for these samples. For these reasons The analysis of 1Н-NMR weak field spectra has allowed us and as proposed by others [10,12,14] the quality factor H was used to establish that all the samples exhibited the absence of ade- for characteristics of frozen fish, estimated under the formula 3. nosine‑5’-triphosphate, adenosine‑5’-diphosphate and ade- Obtained results in table 4 show that quality factor H more nosine‑5’-monophosphate signals, whereas they were charac- objectively reflects quality of frozen samples. It can be conclud- terized by the diagnostic signals of inosine, hypoxanthine and ed that Pinc salmon (Oncorhynchus gorbuscha), chum salmon inosine‑5’-monophosphate. (Oncorhynchus keta), sockeye salmon (Oncorhynchus nerka) were This obtained data was in good agreement with the results frozen immediately after catching, as they have high sensory published in the literature [14,15]. Indeed, these authors estab- quality of fish and consequently low of quality factor H. lished that after post-mortem changes, the total decomposition It allows to conclude that the following methodical approach of АТP occurred and the analyzed fish contained metabolites can be used for characteristics of not only frozen fish but also such as inosine‑5’-monophosphate, inosine and hypoxanthine. for raw fish from which frozen pinc salmon (Oncorhynchus gor‑ Therefore, in our manuscript the quality estimation of fish, re- buscha), chum salmon (Oncorhynchus keta), sockeye salmon lied on the identification of three metabolites: inosine- 5’-mo- (Oncorhynchus nerka) were produced. Thus the NMR‑spectros- nophosphate, inosine and hypoxanthine. copy allows for objective evaluation of the fish chemical qual- On the basis of 1Н-NMR spectra analysis, it is concluded that ity. Consequently, reliable information concerning the product the value of the quality factor K for stored chilled and frozen can be obtained, the shelf life can be predicted and the tailor- raw materials can be calculated by the formula 2. The results are made products manufacturing processes management can be presented in Table 4. performed on the basis of the comprehensive research and cor- It follows from the presented data that the chilled Atlantic relation with the organoleptic analysis as well as the nutritional salmon which were kept in storage for a duration of 18 days at 0оC quality and the processing properties evaluation. gave unsatisfactory sensory quality (signs of oxidation and off- odor of rank fish) that had quality factor1 К equal to 90%. This 4. Conclusion quality factor K1, which defines the fish quality, was indicative It has been established that the fish quality index can be ex- for the biochemical changes occurring in fish due to their storing pressed by the quality factor K1 which is calculated by means of conditions. Consequently, the higher the biochemical changes, analysis of 1Н-NMR spectra and concentration measurement of and consequently the higher value of quality factor K1, indicate inosine, hypoxanthine and inosine‑5’-monophosphate gener- about loss of fish quality. This correlates well with degradation of ated during the fish storage. The quality factor 1K correlates well the sensory quality of fish and the test of spoiled fish. The quality with the sensory quality of chilled fish. For quality estimation factor K1 of more than 80% is the threshold quality value, when of frozen pinc salmon (Oncorhynchus gorbuscha), chum salmon the product is considered to be unfit for food. (Oncorhynchus keta), sockeye salmon (Oncorhynchus nerka) qual-

It is important to point out that quality factor K1 for frozen ity factor H is more informative, which correlate with low indica- samples of fish estimated under given above formula was too tors of TVB-N and high sensory quality of fish. Table 4

Calculation quality factors К1 and Н for Salmon fish species samples of different storage duration Inosine- Storage duration, Hypoxanthine Hx Inosine Ino Title 5’-monophosphate К , % Н, % days (8.21 ppm), a.u.* (8.34 ppm), a.u. 1 IMP (8.58 ppm) a.u. Atlantic Salmon 9 0.334 1.453 1.326 57.4 10.7 (Salmo Salar) 0 °С Atlantic Salmon 18 0.773 1.753 0.270 90.3 27.7 (Salmo Salar) 0 °С Atlantic Salmon 9 0.389 1.526 1.312 59.3 12.0 (Salmo Salar) –20 °С Pinc salmon 90 0.436 1.208 8.277 16.6 4.4 (Oncorhynchus gorbuscha) –18 °С Chum salmon 95 0.420 3.313 1.041 78.2 8.8 (Oncorhynchus keta) –18 °С Sockeye salmon 98 0.234 1.732 2.667 42.4 5.0 (Oncorhynchus nerka) –18 °С * a. u. — ​unit of area

REFERENCES 1. Council Regulation (EC) No 2406/96 of 26 November 1996 laying down relation with quality parameters. Journal of Food Science, 72(7), 356–362. common marketing standards for certain fishery products. Official Jour- DOI: 10.1111/j.1750–3841.2007.00468.x nal L 334, 23/12/1996 P. 0001–0015. 5. Mohan, C.O., Ravishankar, C.N., Srinivasa Gopal, T.K., Ashok Kumar, K. 2. Luten, J. B., Oehlenschläger, J., Olafsdottir, G. (2003). Quality of Fish from (2009). Nucleotide breakdown products of seer fish (Scomberomorus Catch to Consumer: Labelling, Monitoring and Traceability. The Nether- commerson) steaks stored in O2 scavenger packs during chilled storage. lands: Wageningen Academic. —265 p. ISBN: 978–90–76998–14–5, DOI: Innovative Food Science and Emerging Technologies, 10(2), 272–278. DOI: 10.3920/ 978–90–8686–510–9 10.1016/j.ifset.2008.11.012 3. Mendes, R., Quinta, R., Nunes, M.L. (2001). Changes in baseline levels of 6. Saito, T., Arai, K.-I., Matsuyoshi, M. (1959). A new method for estimating nucleotides during ice storage of fish and crustaceans from the Portu- the freshness of fish. Nippon Suisan Gakkaishi (Japanese Edition), 24(9), guese coast. European Food Research Technology, 212(2), 141–146. DOI: 749–750. DOI: 10.2331/suisan.24.749 10.1007/s002170000222 7. Ciampa, A., Picone, G., Laghi, L., Nikzad, H., Capozzi, F. (2012). Changes 4. Márquez-Ríos, E., Morán-Palacio, E.F., M E Lugo-Sánchez, M.E., Ocano- in the amino acid composition of Bogue (Boops boops) fish during stor- Higuera, V.M., Pacheco-Aguilar, R. (2007). Postmortem biochemical be- age at different temperatures by 1H-NMR spectroscopy. Nutrients, 4(6), havior of giant squid (Dosidicus gigas) mantle muscle stored in ice and its 542–553. DOI: 10.3390/nu4060542

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8. Shumilina, E., Ciampa, A., Capozzi, F., Rustad, T., Dikiy, A. (2015). NMR 12. Shumilina, E., Slizyte, R., Mozuraityte, R., Dykyy, A., Stein, T.A., Dikiy, A. approach for monitoring post-mortem changes in Atlantic salmon fillets (2016) Quality changes of salmon by-products during storage: Assess- stored at 0 and 4 oC. Food Chemistry, 184, 12–22. DOI: 10.1016/j.food- ment and quantification by NMR. Food Chemistry, 211, 803–811. DOI: chem.2015.03.037 10.1016/j.foodchem.2016.05.088 9. Karube, I., Matsuoka, H., Suzuki, S., Watanabe, E., Toyama, K. (1984). De- 13. Commission Regulation (EC) No 1022/2008 of 17 October 2008 amending termination of fish freshness with an enzyme sensor system.Journal of Ag‑ Regulation (EC) No 2074/2005 as regards the total volatile basic nitrogen ricultural and Food Chemistry, 32(2), 314–319. DOI: 10.1021/jf00122a034 (TVB-N) limits. Official Journal of the European. L.227, 18–20. 10. Luong, J.L.T., Male, K.B., Masson, C., Nguyen, A. L. Hypoxanthine Ratio 14. Hattula, T. (1997). Adenosine triphosphate breakdown products as a Determination in Fish Extract Using Capillary Electrophoresis and Im- freshness indicator of some fish species and fish products. Technical mobilized Enzymes. (1992) Journal of Food Science, 57(1), 77–81. DOI: Research Center of Finland: VTT Publications. —48 p. ISBN: 951–38– 10.1111/j.1365–2621.1992.tb05429.x 4955–4 11. GOST 7636–85 «Fish, marine mammals, marine invertebrates and prod- 15. Hamada-Sato, N., Usui, K., Kobayashi, T., Imada, C., Watanabe, E. (2005) ucts of their processing. Methods of analysis». Moscow: Standartinform. Quality assurance of raw fish based on HACCP concept. Food Control, 1986. — 124 p. (In Russian) 16(4), 301–307. DOI: 10.1016/j.foodcont.2004.02.001

AUTHOR INFORMATION Liubov S. Abramova — ​Doctor of Technical Sciences, Professor, Advisor on the quality of fish products of the Department of Habitat Monitoring,- Aquat ic Biological Resources and Products of Their Processing, Research Institute of Fisheries and Oceanography, 107140, Moscow, V. Krasnoselskaya street, 17, Tel.: +7–499–264–35–91, e-mail: [email protected] *corresponding author Andrey V. Kozin — ​Candidate of Chemical Sciences, Senior Researcher of the Department of Habitat Monitoring, Aquatic Biological Resources and Prod- ucts of Their Processing, Research Institute of Fisheries and Oceanography, 107140, Moscow, V. Krasnoselskaya street, 17, Tel.: +7–499–264–18–33, e-mail: [email protected]. Alexander S. Shashkov — ​Doctor of Chemical Sciences, Professor, Leading Researcher, Laboratory of Metal-Complex and Nanoscale Catalysts, N. D. Zelinsky Institute of Organic Chemistry Russian Academy of Sciences, 119334 Moscow, Leninsky prospect, 47, Tel.: +7–499–135–90–94, e-mail: [email protected]. All authors bear responsibility for the work and presented data. All authors made an equal contribution to the work. The authors were equally involved in writing the manuscript and bear the equal responsibility for plagiarism. The authors declare no conflict of interest. Received 17.10.2019 Accepted in revised 10.11.2019 Accepted for publication 30.11.2019

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UDK 663.053 DOI: 10/21323/2618–9771–2019–2–4–10–13 Review paper THE INCREASING THE DURATION OF FUNCTIONAL BEVERAGES METHODS Marina V. Gernet1,2, Irina N. Gribkova1*, Olga A. Borisenko1 1 All-Russian Scientific Research Institute of the Brewing, Non-Alcoholic and Wine Industry — Branch of the V. M. Gorbatov Federal Research Center for Food Systems of RAS, Moscow, Russia 2 Moscow State University of Food Production, Moscow, Russia

KEY WORDS: ABSTRACT functional drinks, shelf life, In the Russia beverage market, functional beverages are increasingly popular with the population due to their a group of functional compounds, intensive lifestyle and worsening environmental situation. Of great importance is the shelf life of drinks, since processing method, flavanoids they determine the presence in the composition of useful nutrients that affect the various systems of functioning of the human body. The main groups of functional compounds are given. Ways to increase the shelf life of bever- ages, all aspects and their impact on the safety of the functional components of beverages are considered. The role of flavonoids as one of the groups of functional compounds has been evaluated. Recent studies on the effect of certain functional compounds on each other are given.

1. Introduction There are various functional beverage divisions [4,5]. This is Drinks are the most accessible food form relative to nutrients due to the direction of particular product useful nutrients action entering the human body, providing the body’s needs for various for different groups of the population. compounds. This is explained by the possibility of having nutrients However, in our opinion, the functional drinks division can be in a dissolved state for their better absorption. supplemented with fermentation and blending beverages produced There are many types of beverages and this is related to the by mixing the ingredients in the prescribed manner. production technology (carbonated / non-carbonated, fermented Plant material is a rich source of useful nutrients, it is enriched beverages / non-alcoholic beverages obtained by blending ingre- with such biologically active substances as anthocyanins, poly- dients, etc.), raw materials (vegetable / animal origin), functional phenols, carotenoids, vitamins, oligosaccharides, amino acids [6] orientation (content in its composition ingredients responsible The presence of the fermentation as technological stage makes for the biological value of the product), etc. it possible to enrich the drink with the vital activity products of Recently, in connection with the deteriorating environment microorganisms that are also related to functional compounds — it has become more significant to lead a healthy lifestyle. This is these are alcohols, ethers, carbonyl compounds, organic acids, due, inter alia, to the consumption of beverages enriched with vitamins, amino acids, etc., which can also enhance the beneficial compounds, which allow maintaining body functions and level- effect of each other. ing the negative effect of negative environmental factors on the Table 1 presents functional food compounds main categories, whole on health. obtained either directly from vegetable raw materials, or during An important issue regarding the quality of such beverages fermentation [7]. is the shelf life. The trading network is more willing to work with Table 1 manufacturers who guarantee sufficiently long storage periods Dietary Fiber Vitamins for their products. Therefore, there is a reasonable problem of preserving functional compounds quantity and quality in bever- Оligosaccharides Choline ages during long periods of beverages storage. Sugar alcohols Bifidobacterium The purpose of the work is to analyze the market of beverages, including functional ones, with regard to ways to increase shelf Amino acids, peptides, proteids Mineral components life without affecting the quantitative and qualitative composition Polyunsaturated Fatty Acids, Glycosides of functional compounds. Antioxidants Alcohols Cytamines 2. Main part It is known that functional drinks are enriched with useful Organic Acids Vegetable Enzymes nutrients groups of general or directed action product for various groups of the population, which make it possible to improve the Among listed above functional nutrients, almost all are ther- human body immunoresistance to diseases arising from either mally unstable and undergo chemical changes associated with unfavorable ecological conditions or related to the failure of physi- oxidation under the influence of various factors (elevated tem- ological processes [1]. perature, changes in the acidity of the environment, the presence This group of drinks owes to vitamins, microelements, dietary of oxygen, ultraviolet, etc.) [8,9,10,11,12,13,14]. fibers, probiotics, amino acids, volatile compounds, organic ac- Therefore, it is very important, especially in the case of enriched ids, antioxidants, etc. presence in its composition for functional with useful nutrients beverages production, to apply technological properties [2]. methods that allow to keep functional compounds in an active The beer and soft drinks industry regulatory documents spell form for the longest possible time. out clear requirements as to whether a drink can be attributed to a There are several ways that solved the biological stability functional group: this is possible if one or more functional compo- problem of the beverage composition in the classical technology nents are present in an amount from 10 to 50% of the daily intake [3]. of beverages production, including functional ones:

FOR CITATION: Gernet М. V., Gribkova I. N., Borisenko O. A. The increasing the duration of functional beverages methods. Food systems. 2019; 2(4): 10–13. DOI: 10.21323/2618–9771–2019–2–4–10–13

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‰‰ pasteurization (processing temperature not higher than in all oxidative processes with their participation. That is, flavo- 70 °C); noids are capable of acting both antioxidants and prooxidants, ‰‰ hot bottling; and the mechanisms of the course of these reactions have not ‰‰ the safety compounds use (benzoic, sorbic acids and their yet been clarified. salts — sodium benzoate and potassium sorbate, as well But the flavonoid presence in the drink is not enough, since as hydroxy derivatives 1,4 naphthoquinone — juglone and there are various factors affecting their bioavailability for the plumbagin) [15]. All of them are aimed to suppressing the human body (ability to be absorbed by the human intestine and extraneous microflora development, due to which stability of absorbed in the body), because fermentation drinks are character- the physicochemical beverage composition is achieved. ized by a richer composition of functional compounds extracted In addition to the classical methods of enhancing the biologi- from raw materials. Such factors include daily intake, absorption, cal beverages stability, there are other, that including innovative biotransformation and excretion [48]. methods associated with vacuum evaporation, the use of pulsation, Scientists provide different information about the quantitative ultrafiltration, cryopreservation, etc. [16,17,18,19]. native polyphenol absorption in the intestine — from 1% [49] to The concentration method is acceptable because it comfort- 50–70% [50,51] of the eaten polyphenols. In their opinion, it is the able to beverage producers under conditions of mini-production, small intestine that serves as a place of absorption, in particular, when large production areas are not required. However, from the of flavonoids. The degree of flavonoids absorption is different and point of view of biological stability, this technique is very doubtful, depends on their form — aglycones of flavonoids and isoflavones because it connected with impact on useful nutrients, leading to are easily absorbed [52,53], and glycosides of flavonols, flavones their inactivation or loss of functional properties [10]. and isoflavones should be pre-hydrolyzed to aglycones. Another way to increase beverages shelf life is cold filtration In the colon, the polyphenols are further processed by bacterial [20], ultrafiltration [21], the ultrasound use with the anthocy- enzymes with cleavage of a heterocyclic ring containing oxygen, anogen raw material confirmed preservation [22], the radiation dehydroxylation and decarboxylation [54,55,56]. technology use [23,24]. It is also necessary to note the fact that the presence of con- However, these methods, as already mentioned, will lead to comitant other compounds in the drink can either enhance the additional costs for the manufacturer in terms of energy consump- effect of polyphenols on absorption, or suppress. tion, specialized personnel for equipment maintenance, as well Thus, proteins reduce the polyphenol adsorption capacity, as will take away some of the production space. since they bind to them, and fats and alcohols intensify them [57]. Increasing shelf life research is also aimed at the use of nano- Green tea catechins, grape seed oligomeric proanthocyanidins structured particles based preservatives new types. Preservatives, as and milk thistle silibilins have been shown to be better absorbed is well known, suppress the development of extraneous microflora in the form of phospholipid complexes [57]. In confirmation of in food products, in particular beverages. Thus, there are studies the better absorbability of polyphenols against the background on the use of lactic acid associates with silver nanoparticles [25], of the presence of alcohols, one can cite the fact that red wine as well as with lichen nanoparticles [26]. polyphenols are absorbed more intensively compared to soft However, there is evidence of the microflora development sup- drinks [58,59,60]. pression by nanochayats duration, in particular silver, only for 8 Thus, the flavonoids, as substances with anti-radical activity, months in the literature [27]. The researchers also noted that it is are also labile and for their protection it is also necessary to choose important to take into account the dispersion medium (beverage) the modes of processing products. nature and the nanoparticle suspensions storage time in order to effectively use them as preservatives [28]. 3. Conclusion Researchers are trying to apply raw materials new types for The functional drinks market is developing in terms of raw the functional compounds enrichment of products and the exten- materials variety that enrich the drinks with useful nutrients; sion shelf life periods [29,30,31]. There is a technique when high technological practices that contribute to the extension of shelf content of antioxidants plant materials are used as raw materials life, and related to the physico-chemical processes of processing of in technology [2,32,33,39]. finished products. Accumulated and conducted scientific research Among the plant antioxidants, phenolic compounds, in par- aimed at clarifying the mechanisms of action and interaction of ticular flavonoids, are widely represented. These compounds, in compounds responsible for the functional profile of products; addition to direct antiradical action, bind metals with a transi- on the development of technological methods that contribute to tion valence, due to which inhibition of free radical processes in the preservation of nutritional properties of compounds during cells occurs [40]. the storage period. It was also found that flavonoids also affect signaling processes Thus, to solve the issue of creating a drink with the useful in living systems, due to specific interaction with proteins that properties of a long shelf life, it is necessary to solve a whole perform regulatory functions [41]. By binding to protein and non- range of tasks: protein structures, flavonoids change the functional state of the ‰‰ pick up vegetable raw materials that are rich in beneficial cell and the whole organism. compounds; The mechanism of flavonoids and ascorbic acid interaction ‰‰ create a technology for processing raw materials that pro- has been discovered, which allows maintaining the flavonoid in a mote the extraction of useful compounds in easily digestible reduced state, associated with a reciprocal reaction, and direction bioavailable form; of which reaction depends entirely on the ORP of the flavonoid ‰‰ have the concept of the presence of substances that can react interacting with the ascorbate radical [42]. with antioxidants or other nutrients during the production There is evidence that flavonoids are capable to oxidation in and storage of the drink; the literature, that is, the manifestation of prooxidant properties ‰‰ to process finished products (using various methods) for the [43,44,45,46,47]. The intermediate products of the flavonoid oxida- prolonged action of useful nutrients in the composition of tion, which are formed during such reactions, appear practically beverages.

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52. Hollman, P.C., Trijpa, J. M.P., Mengelers, M., Vriesb, J.H.M., Katanb, M. 56. Vitaglione, P., Donnarumma, G., Napolitano, A., Galvano, F., Gallo, A., B. (1997) Bioavailability of the dietary antioxidant flavonol querce- Scalfi, L., Fogliano, V. Protocatechuic acid is the major human metabolite tin in man. Cancer Letters, 114(1–2), 139–140. DOI: 10.1016/s0304– of cyanidin-glucosides. The Journal of Nutrition, 137(9), 2043–2048. DOI: 3835(97)04644–2 10.1093/jn/137.9.2043 53. Vitaglione, P., Donnarumma, G., Napolitano, A., Galvano, F., Gallo, A., 57. Hollman, P.C.H. (2004). Absorption, bioavailability, and metabo- Scalfi, L., Fogliano, V. (2007). Protocatechuic acid is the major human lism of flavonoids. Pharmaceutical Biology, 42(1), 74–83. DOI: metabolite of cyanidin-glucosides. Journal of Nutrition, 137(9), 2043– 10.3109/13880200490893492 2048. 58. Pietta, P.G. (2000). Flavonoids as antioxidants. Journal of Natural Products 54. Izumi, T., Piskula, M.K., Osawa, S., Obata, A., Tobe, K., Saito, M., Kataoka, 59. 63(7), 1035–1042. DOI: 10.1021/np9904509 S., Kubota, Y., Kikuchi, M. (2000). Soy isoflavone aglycones are absorbed 60. Serafini, M., Maiani, G., Ferro-Luzzi, A. (1997). Effect of ethanol on red faster and in higher amounts than their glucosides in humans. Journal of wine tannin-protein (BSA) interactions. Journal of Agricultural and Food Nutrition, 130(7), 1695–1699. Chemistry, 459(8), 3148–3151. DOI: 10.1021/jf960864x 55. Spencer, J.P.E., Chowrimootoo, G., Choudhury, R., Debnam, E.S., Srai, 61. Booyse, F.M., Pan, W., Grenett, H.E., Parks, D.A., Darley-Usmar, V.M., S.K., Rice-Evans, C. (1999). The small intestine can both absorb and Bradley, K.M., Tabengwa, E. M. (2007). Mechanism by which alcohol and glucuronidate luminal flavonoids. FEBS Letters, 458(2), 224–230. DOI: wine polyphenols affect coronary heart disease risk. Annals of Epidemiol‑ 10.1016/s0014–5793(99)01160–6 ogy, 7(5), S24–S31. DOI: 10.1016/j.annepidem.2007.01.006

AUTНOR INFORMATION Marina V. Gernet — professor, doctor of technical sciences, head of brewing technology department All-Russian Scientific Research Institute of Brewing, Beverage and Wine Industry — Branch of V. M. Gorbatov Federal Research Center for Food Systems of RAS, 119021, Moscow, Rossolimo str., 7. Tel.: +7–499–245– 10–79, e-mail: [email protected] Irina N. Gribkova — candidate of technical sciences, senior researcher of brewing technology department, All-Russian Scientific Research Institute of Brewing, Beverage and Wine Industry — Branch of V. M. Gorbatov Federal Research Center for Food Systems of RAS, 119021, Moscow, Rossolimo str., 7. Tel.: +7–499–246– 04–47, e-mail: [email protected] *corresponding author Olga A. Borisenko — senior researcher of brewing technology department, All-Russian Scientific Research Institute of Brewing, Beverage and Wine Industry — Branch of V. M. Gorbatov Federal Research Center for Food Systems of RAS, 119021, Moscow, Rossolimo str., 7. Теl.: +7–499–246–01–96, e-mail: institut-beer@ mail.ru All authors bear responsibility for the work and presented data. All authors made an equal contribution to the work. The authors were equally involved in writing the manuscript and bear the equal responsibility for plagiarism. The authors declare no conflict of interest. Received 30.04.2019 Accepted in revised 14.11.2019 Accepted for publication 26.11.2019

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UDC 663.05 DOI: 10.21323/2618–9771–2019–2–4–14–17 Original scientific paper INNOVATIVE DECISIONS TO IMPROVE FOOD QUALITY AND SAFETY Vera V. Eveleva*, Tatyana M. Cherpalova All-Russia Research Institute for Food Additives — ​Branch of V. M. Gorbatov Federal Research Center for Food Systems of RAS, St. Petersburg, Russia

KEY WORDS: ABSTRACT complex lactate-containing food The scope of the project consisted in research-based development of new complex food additives from lactic additives, antimicrobial effect, acid and its derivatives for enhancing microbiological safety and shelf life extension of healthy food products. food shelf life The object of research included: trial samples of complex food additives from lactic acids and its derivatives. The samples were obtained chemically using the following basic components: food grade lactic acid with 79.6% base substance mass fraction; acetic acid with 99.8% base substance mass fraction; propionic acid with 99.6% base substance mass fraction; neutralizing agents for synthesis of salts of the acids used, and propylene glycol with 99.8% base substance mass fraction. The optimal balance of the ingredients in the formula of the additive delivers the optimal level of true acidity combined with antimicrobial and antioxidant effect, and enhances organoleptical performance and process parameters of food products. This complex food additive containing lactate and acetate features high counter-regulatory effect on testing cultures of pathogenic organisms of rope spoilage of wheat bread and can be used for production of non-perishable products. The use of complex food additive in the produc- tion of dressed fish preserves activates biochemical processes related to fish maturation and delivers improved product quality and extended shelf life.

1. Introduction ing Pseudomonas spp., Enterobacteriaceae, Staphylococcus and Transition to accomplishment of new priority goals in the Salmonella [4]. The results of microbiological studies quoted field of research and hi-tech development of the Russian Federa- herein are supported by process trials demonstrating that ad- tion, which consist in innovative development of the domestic dition of 2.5% of sodium lactate to raw material for half-smoke food market, efficient processing of agricultural products, devel- charcuterie production extended their shelf life to 30 days. [5]. opment of safe and quality food products, including functional Shelf life extension of mechanically deboned poultry meat food, requires creation of new research and hi-tech solutions was achieved through poultry breast soaking for 10 minutes in and improvement of the existing processes. 0.2% and 0.3% lactic acid solution or in 1% and 2% sodium lac- Consumer focus shift to healthy nutrition puts in the fore- tate solution [6]. Along with shelf life extension, broiler fillet front deployment of food additives from agricultural compo- treatment with 1% lactic acid solution inhibits biogenic amine nents or using microbiological synthesis processes. Such ad- synthesis and rancidity [7]. The performance of antimicrobial ditives include lactate-containing additives that are generally formulas based on lactate containing additives was found to recognized as safe and widely used in global and national pro- be related to the formation in the process of their synthesis of duction processes for food quality and safety enhancement. compounds consisting of cation-active surfactants and anions Despite the progress made, food safety remains an unre- of lactic, acetic and propionic acids that are beneficial to prod- solved issue, since the risks related to potential in-process con- uct storage stability and microbiological safety [8]. tamination with dangerous and inducing spoilage pathogens. To The recorded tendencies for preservative agent and edible this effect, various research efforts in the field of use of anti- salt content reduction in fish products along with reduction of microbial ingredients for food toxic infections prevention have curing agent content in unsterilized canned fish products are re- been undertaken. lated to the fact that glucono-delta-lactone and enzyme formu- Maximization of the benefits of the use of food additives las and glucono-delta-lactone, citric acid and lactose formulas in food production can be achieved through the synergistic ef- used in unsterilized canned fish production facilitate matura- fect of their ingredients. The survey of antimicrobial effect of tion processes and reduce finished product shelf life. However, food additives on Listeria monocytogenes bacteria inoculated traditional pickling processes require extended maturation on hotdog surface has demonstrated significant reduction of when lactic acid bacteria present in fish belly produce lactic acid their propagation in the samples containing lactate and ac- ensuring microbiological purity of the product and enhancing etate combination [1]. The analysis of the effect of antimicro- enzyme activation. Besides, lactic acid features high penetration bial additive injections in steaks superficially contaminated capability compared to edible salt and promotes its faster and with Escherichia coli K12 has shown that the formula contain- more even distribution in meat fiber, thus accelerating the pick- ing sodium lactate and sodium diacetate is the most active in- ling processes. Food grade lactic acid is identical to natural lactic hibitor of the growth of these bacteria [2]. Similar results were acid synthesized in fish meat tissues during maturation and has obtained based on the analysis of the capacity of potassium a similar effect when added with brine. Combined with phytonu- lactate (PURASAL Hi Pure P) and combination of potassium trients, lactic acid produces functional unsterilized canned fish lactate and sodium diacetate (PURASAL Opti. Form PD4) as products from clupeidae for people suffering from metabolic and inhibitors of Listeria monocytogenes growth in sliced ham in gastrointestinal disorders [9]. modified atmosphere packaging [3]. The analysis of sodium Consumer demand for bakery products with zero or low lactate and lactic acid effect on microbiological parameters content of chemical preservatives generates interest in natural of marinated chicken meat has shown the efficiency of these antimicrobial agents for bread. Publication [10] demonstrates additives against various spoilage microorganisms, includ- that the formulas containing acetic acid, lactic acid and calcium

FOR CITATION: Eveleva V. V., Cherpalova T. M. Innovative decisions to improve food quality and safety. Food systems. 2019; 2(4): 14–17. DOI: 10.21323/2618–9771– 2019–2–4–14–17

14 ПИЩЕВЫЕ СИСТЕМЫ | Том 2 № 4 | 2019 FOOD SYSTEMS | Volume 2 № 4 | 2019 lactate inhibited propagation of Bacillus subtilis and Bacillus li‑ 3. Results and discussion cheniformis bacteria, and bread spoilage mold. At the same time, A developmental research of new complex food additives they inhibited yeast activity considerably less than calcium pro- containing lactate, acetate, propionate and propylene glycol pionate that is traditionally used in baking. Ripened sourdough ingredients for improvement of microbiological safety of food and lactic acid starters introduction in the dough at the knead- products and their shelf life extension was conducted. ing phase has been found to successfully prevent rope spoilage A complex processing aid for non-perishable wheat bread of bread [11]. Surveys of antifungal effect of lactic acid bacteria production under AL‑1 reference name was developed. The sur- obtained from grain varieties have demonstrated its relation to vey demonstrated high counter-regulatory capacity of the ad- antifungal peptide synthesis [12]. ditive to testing cultures of rope spoilage agents compared to The quoted analysis of reference data shows the efficiency of the effect of individual additives, i. e. lactic and acetic acid, in the use of lactic acid, sodium and potassium lactates and related dosages delivering equal titrated dough acidity and regulatory formulas in poultry meat and meat product manufacturing. The bread acidity values (Table 1). Relation between acidity of dough benefits of use of lactate-containing additives in fish products and wheat bread and ratio of AL‑1 ingredients was established. and bread have also been identified. Although the all-purpose With the increase of the ratio of mass fractions of the aggregate antimicrobial preservative agents that have been used so far amount of lactic and acetic acid to mass fraction of sodium lac- inhibit propagation of microbial food spoilage organisms, they tate in the additive, the acidity of dough and bread increases in- also inhibit biosynthetic activity of microorganisms used in food significantly even in the case of the maximum dosage of the ad- processes, specifically, in fermented milk and bread production. ditive in the amount of 0.5% flour weight and the maximum total This leads to lower product quality and impedes healthy food acid content. Based on the analysis of dough and bread quality production. indicator variation, the following specifications to the complex The scope of the project consisted in research-based devel- food additive were defined: active acidity — ​from 4.0 to 4.4 pH, opment of new complex food additives from lactic acid and its acidity — ​from 230 to 310 deg., dosage — ​0.5% flour weight. derivatives for enhancement of microbiological safety and shelf Table 1 life extension of food products. Counter-regulatory activity of complex food additive and its components 2. Materials and methods Area of testing culture Additive dose, % The object of research included: test samples of complex Additive inhibition, mm flour weight food additives from lactic acid and its derivatives consisting of B. pumilis B. subtilis liquids with different components and physical and chemical Lactic acid 0.01 Not detected Not detected parameters. Acetic acid 0.01 Not detected Not detected The samples were obtained chemically using the following AL‑1 0.50 25.0 19.0 basic components: food grade lactic acid with 79.6% base sub- stance mass fraction; acetic acid with 99.8% base substance The results of testing of AL‑1 additive counter-regulatory mass fraction; propionic acid with 99.6% base substance mass effect on B. pumilis and B. subtilis by baking test method have fraction; neutralizing agents for synthesis of salts of the acids shown that additive application increases the shelf life of wheat used, and propylene glycol with 99.8% base substance mass frac- bread under predisposing conditions to 38 hours against 17 tion. hours without the additive, and to 139 hours against 42 hours Herein we present the results of analysis and definition of without the additive under normal conditions. the optimal parameters of new complex food additives from lac- The results of process trials have shown that additive intro- tic acid and its derivatives for improvement of microbiological duction in the dough making process inhibits successfully patho- safety and shelf life extension of food products. genic sporogenic bacteria causing rope spoilage of bread, delivers Performance trials of new complex food additives were con- the microbiological safety of the product, allows production of ducted in laboratory and, whenever possible, in industrial envi- non-perishable product to the applicable organoleptical, physical ronment. and chemical standards that features a more porous and delicate Experimental research was conducted in the laboratory fa- crumb structure and a homogeneous fine pore honeycomb pat- cilities of Russian Research Institute for Food Additives using tern compared to the reference samples (without the additive). the standard research methods and conforming to the applicable For enhancing the safety and storage stability of unsterilized regulations and specifications. canned products from dressed fish, trial development of a com- Testing parameters of complex food additives: titrated acid- plex food additive under Optima reference name (Table 2) was ity, active acidity, density, buffer capacity. conducted using various basic components which resulted in The testing parameters of complex food additives were quan- similar buffer capacity values. tified using the following methods: titrated acidity — ​by acid- base titration; active acidity — ​by potentiometric method using Table 2 Chemical analysis of trial samples of complex food pH‑150 MI analyzer; density — ​by densimetric method using additives for unsterilized canned fish production AON‑1 general purpose density meters with various measure- ment ranges; buffer capacity — ​by potentiometric method using Mass fraction,% pH‑150 MI analyzer. anion cation The counter-regulatory effect of lactate and acetate food Reference additives was evaluated by diffusion in beef extract agar inocu- name lated with testing cultures of rope spoilage of bread — ​Bacillus

pumilis and Bacillus subtilis sporogenic bacteria, and baking tests lactate acetate propionate sodium potassium water propylene glycol of wheat bread inoculated with spores of rope spoilage bacteria Optima‑1 42.9 3.3 1.5 11.8 — 40.5 — Bacillus cereus, including B. mesentericus and B. subtilis, under Optima‑2 27.4 16.1 3.5 12.2 1.9 38.9 — predisposing and normal conditions, following the applicable Optima‑3 32.7 9.4 3.5 11.0 1.9 41.5 — guidelines, in St. Petersburg branch of Breadmaking Industry Research and Development Institute. Optima‑4 22.6 13.8 3.5 10.0 1.9 33.1 15.0

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The importance of these properties of the additive for unsteril- Table 3 ized canned fish production was demonstrated on samples of fish Active acidity (pH) pickling brine. Table 3 shows that application of trial samples of of fish pickling brine samples with additives the additive in the amount of 1, 3 and 5% in the trial pickling brine Active acidity (pH) of fish brine Additive ensures achievement and stabilization of active acidity value from at mass fraction of the additive,% name 4.2 to 5.2 pH, which is optimal for the effect of the proteolytic en- 0 1 3 5 zymes contained in the fiber and activates the heterofermentative Brine (8% edible salt) lactic streptococci involved in biochemical fish maturation pro- Optima‑1 (pH 5.9) 4.8 4.8 4.8 cesses. Based on the ratio between the physic-chemical proper- ties and counter-regulatory effect of the trial samples, Optima‑1 Optima‑3 (pH 6.2) 6.8 5.0 5.0 5.0 additive was selected. The trials have shown that at the final salt Optima‑4 (pH 6.7) 5.2 5.2 5.2 concentration level in the fish within (5.0 ± 0.2)% range, the dura- Pickling brine (8% edible salt and 0.2% acetic acid) tion of herring fillet pickling is 36 hours. The analysis of fish pro- tein fraction variation during storage of unsterilized canned fish Optima‑1 (pH 5.9) 4.2 4.4 4.5 products at (2 ± 2) °C has demonstrated that the introduction of Optima‑3 (pH 6.2) 2.8 4.3 4.5 4.7 Optima‑1 complex food additive to the brine inhibits proteolytic Optima‑4 (pH 6.7) 4.4 4.7 4.9 processes compared to additive-free samples and delivers virtu- ally complete mold and yeast growth inactivation in the trial sam- Optima‑1 complex additive for enhancement of safety and ples of unsterilized canned fish samples. Eschericia coli, Staphylo‑ storage stability of unsterilized canned fish products features coccus aureus and sulphite reducing clostridia were not detected high buffer capacity delivering the achievement and stabiliza- in such samples either. The results of organoleptical analysis were tion of the optimal active acidity level for muscular tissue pro- in flavour of the samples of unsterilized canned fish products with teolytic fermentation. Optima‑1 introduction to pickling brine 2% fish weight additive in the pickling brine. The trial samples of has been found to deliver sustained consumer performance and unsterilized preserves from frozen Atlantic herring featured the at least 50% extension of the shelf life of unsterilized canned pleasant flavour, delicate and rich texture specific to matured fish products from dressed herring, subject to the recommended and conformed to the requirements of the regulation on produc- storage conditions. tion of unsterilized canned products from dressed fish. Optima‑1 additive was found to deliver at least 50% extension of the shelf 4. Conclusion life of unsterilized canned fish products subject to the recom- The optimal balance of the ingredients in the formula mended storage conditions. of AL‑1 complex additive delivers the optimal level of true Based on the findings of the research, new complex food ad- acidity combined with antimicrobial effect, and enhances ditives from lactate compounds, including acetate, propionate organoleptical performance and process parameters of food and propylene glycol containing components, were developed products. AL‑1 complex food additive features high counter- for enhancing microbiological safety and extension of shelf life regulatory effect on the pathogenic organisms of rope spoil- of wheat bread and unsterilized canned products from frozen At- age of wheat bread and can be used for production of non- lantic herring fillet. perishable products. AL‑1 complex food additive developed for non-perishable Optima‑1 complex additive delivers the achievement and wheat bread production features high counter-regulatory effect stabilization of the optimal active acidity level for muscular tis- on the agents of rope spoilage of bread. The experimental data sue proteolytic fermentation promoting the activation of het- obtained conforms to information contained in publication [10], erofermentative lactic streptococci involved in the biochemical according to which the formulations containing lactate and ac- processes of fish maturation. The application of this additive in etate additives can successfully substitute calcium propionate, a unsterilized canned herring fillet production processes delivers popular preservative agent in bread making industry. enhanced product quality and extended shelf life.

REFERENCES 1. Koo, O.-K., Eggleton, M., O’Bryan, C.A., Crandall, P.G., Ricke, S.C. sodium lactate. African Journal of Agricultural Research, 6(16), 3847–3852. (2012). Antimicrobial activity of lactic acid bacteria against Listeria DOI: 10.5897/AJAR11.714 monocytogenes on frankfurters formulated with and without lac- 7. Hussein, M.A., El-Ghareeb, W.R., Nasr, M.A.F. (2018). The effect of rose- tate/diacetate. Meat Science, 92 (4), 533–537. DOI:10.1016/j.meat- mary extract and lactic acid on the quality of refrigerated broiler fillets. sci.2012.05.023. Journal of Food Science and Technology, 55(12), 5025–5034. DOI: 10.1007/ 2. Wicklund, R., Paulson, D.D., Rojas, M.C., Brewer, M.S. (2007). The effects s13197–018–3441–2 of shelf-life enhancers on E. coli K12 survival in needle-injected, surface 8. Eveleva, V.V., Cherpalova, T.M., Shipovskaya, E.A. (2019). Technological contaminated beef strip steaks enhanced using recycled solutions. Meat innovations for treatment of casings. Theory and practice of meat process‑ Science, 75(2), 273–282. DOI:10.1016/j.meatsci.2006.07.012 ing, 4(2), 14–19. DOI: 10.21323/2414–438X‑2019–4–2–14–19 3. Mellefont, L. A., Ross, T. (2007). Effect of Potassium Lactate and a Po- 9. Mezenova, O. Ya., Leiumaa, E. A., Shenderyuk, V. M. (2012). Improvement tassium Lactate–Sodium Diacetate Blend on Listeria monocytogenes of the technology for producing the Baltic Sea herrings preserves. Fisher‑ Growth in Modified Atmosphere Packaged Sliced Ham. Journal of Food ies, 2, 109–111. (in Russian) Protection, 70 (10), 2297–2305. MS07–051 10. Pattison, T.-L., Lindsay, D., Von Holy, A. (2004). Natural antimicrobials 4. Smaoui, S., Hlima, H.B., Salah, R.B., Ghorbel, R. (2011). Effects of sodium as potential replacements for calcium propionate in bread. South African lactate and lactic acid on chemical, microbiological and sensory charac- Journal of Science, 100(7–8), 342–348. teristics of marinated chicken. African Journal of Biotechnology, 10(54), 11. Nevskaya, E.V. (2016). Development of technology of starter culture using 11317–11326. DOI: 10.5897/AJB11.1249 a strain, ladoumegue antagonistic activity against the causative agents of 5. Semenova, A. A., Lebedeva, L. I., Veretov, L. A. Motovilina, A. A., Verevki- microbial spoilage of bread. Znanie, 1–1(30), 65–69. (in Russian) na, M. I. (2011). New technology of semi-finished products with long 12. Sadeghi, A., Ebrahimi, M., Sadeghi, B., Mortazavi, S.A. (2019) Evaluation shelf life. Meat industry, 1, 22–24. (in Russian) of safety and antifungal activity of Lactobacillus reuteri and Pediococcus 6. Hecer, C., Ulusoy Sözen, B.H. (2011). Microbiological properties of me- diacetilactis isolates against aflatoxigenic Aspergillus flavus. Bulgarian chanically deboned poultry meat that applied lactic acid, acetic acid and Journal of Veterinary Medicine, 22(2), 190–199. DOI: 10.15547/bjvm.2046.

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AUTНOR INFORMATION Vera V. Eveleva — ​candidate of technical sciences, docent, Chief Researcher, Laboratory of technology and processing of biosynthesis products, Аll-Russian Research Institute for Food Additives — ​Branch of V. M. Gorbatov Federal Research Center for Food Systems of RAS, 191014, St.-Petersburg, Liteynу prospekt, 55, Tel: +7–812–273–75–24, e-mail: [email protected] *corresponding author Tatyana M. Cherpalova — ​candidate of technical sciences, Researcher, Laboratory of technology and processing of biosynthesis products, Аll-Russian Research Institute for Food Additives — ​Branch of V. M. Gorbatov Federal Research Center for Food Systems of RAS, 191014, St.-Petersburg, Liteynу prospekt, 55, Tel: +7–812–272–56–26, e-mail: [email protected] All authors bear responsibility for the work and presented data. All authors made an equal contribution to the work. The authors were equally involved in writing the manuscript and bear the equal responsibility for plagiarism. The authors declare no conflict of interest. Received 05.11.2019 Accepted in revised 02.12.2019 Accepted for publication 10.12.2019

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UDC 637.5:577.29:543.545 DOI: 10.21323/2618–9771–2019–2–4–18–24 Original scientific paper BIOCONVERSION OF SECONDARY PRODUCTS OF PROCESSING OF GRAIN CEREALS CROPS Irina S. Vitol *, Natalia A. Igoryanova, Elena P. Meleshkina All-Russian Scientific Research Institute of Grain and Products of its Processing — Branch of the V. M. Gorbatov Federal Research Center for Food Systems of RAS, Moscow, Russia

KEY WORDS: ABSTRACT grain, bran, bioconversion, enzyme A method has been developed for the production of organic ingredients from secondary products resulting preparations, dietary fiber, from the high-quality grinding of triticale and wheat into flour, which involves the enzymatic action of amy- biomodified bran, functional and lolytic enzymes to release starch polysaccharides while preserving the native properties of dietary fibers and technological properties biologically active substances associated with them. To a large extent, the features of the properties of the obtained ingredients are due to the number and composition of the components of dietary fiber of grain, as well as the morphological features of their structure. It is shown that the viscosity of aqueous colloidal sys- tems at a concentration of soluble dietary fiber of the ingredient 0.5 % increases 11 times; at a concentration of 1.0 % — 30 times, forming a viscous gel-like structure. This allows them to be used for gelling, thickening and stabilization of aquatic food systems. The use of ingredients with a high content of NLP in baking is pos- sible only taking into account their water absorption capacity. A method for the enzymatic modification of secondary products of processing of grain triticale was developed. On the basis of the study of the kinetics and efficiency of the effect of proteolytic and cellulolytic enzyme preparations (EP) and their compositions, optimal conditions for enzymatic modification (the EP dosage is 0.5…0.75 units of PA/g of bran, 0.3…0.4 units of CA/g of bran, the optimum temperature is 40–50 °С, pH is 5.0 and 3.5, the duration of reactions is 1.5 hours) have been determined. The use of cellulolytic EP allowed to increase the amount of reducing sub- stances and soluble protein by 1.5–2.5 times in comparison with the control sample. The biomodified bran obtained using the MEC «Shearzyme 500 L» + «Neutrase 1.5 MG» and «Viscoferm L» + «Distizym Protacid Ex- tra» has a high degree of hydrolysis of non-starch polysaccharides and proteins, is characterized by a certain ratio of high-, medium-, low-molecular peptides and amino acids, has different functional and technological properties. They can be used in the production of a wide range of general-purpose, functional and treatment- and-prophylactic food products.

1. Introduction special products [4,5,6]. They can be used to enrich and create new In recent years, there has been an increasing interest in sec- products of both general and special therapeutic and prophylactic ondary grain processing products as renewable raw materials — purposes [7,8,9,10,11]. promising sources of additional raw materials for obtaining food In the All-Russian Scientific Research Institute of Grain and for human consumption. The amount of secondary raw materials, Products of Its Processing, fundamental and applied research is formed during the processing of grain in flour mills, averages carried out to develop the basic methods for managing the techno- 21.3 % of the volume of flour production. When processing grain logical processes of preparing and grinding grain of various crops in cereals, the amount of secondary raw materials is much larger. in order to obtain products with a given chemical composition and For example, when processing wheat in cereals — up to 37 %, of properties; study the possibilities of biochemical transformation which 30 % falls on the flour, while the content of dietary fiber in of secondary products of grain processing (wheat, triticale, oats) these resources varies in a wide range and can reach 49–52 % [1]. into food ingredients containing dietary fibers, while preserving In addition, secondary products of grain processing are rich in the native structure of dietary fiber and biologically active sub- proteins 14–20 %, macro- and microelements, vitamins of group stances associated with them; develop methods for enzymatic B, vitamins A, D and E [2, 3]. modification of grain processing products (flour of various types, A promising direction in the development of the grain in- including high content of peripheral parts, bran) using multien- dustry, whose main task is the effective use of all components zyme compositions (MEС) based on proteolytic and cellulolytic of grain raw materials is deep processing. In the classical sense, enzyme preparations for the production of modified products this is the process of dividing the grain into constituents, as a (protein hydrolysate, structurally modified flour, biomodified result of which separate fractions of protein substances, for bran) with varying degrees and depth of hydrolysis of proteins example, gluten, can be isolated, concentrates and protein iso- and non-starch polysaccharides possessing different functional- lates obtained; various fractions of starch (A, B, C); soluble and technological specific properties. insoluble dietary fiber; some other components. The relatively The aim of the research is to develop a model for the biochemi- high cost of such drugs is compensated by a high content of the cal transformation of secondary products of grain processing, in main component, microbiological purity, a reduction in the costs particular triticale, into food ingredients with predicted tech- of their storage, and the convenience of using the drugs in the nological properties on the basis of studying the changes in the technological process. technological properties of products, depending on the nature Another approach is also possible in the rational use of grain of the feedstock, the chemical composition and the complex of processing products, namely: the use of flour from the peripheral insoluble dietary fibers, and the development of methods for parts of the endosperm of grain, including the aleurone layer enzymatic modification of tritical bran with the use of modern and the seed coat, as well as cereal bran, which may be limited to biotechnological methods. Realization of the set goal will allow traditional technologies (bakery, confectionery,) for further en- to design food products from grain with the specified composition zymatic modification and production of valuable components of and predicted properties.

FOR CITATION: Vitol I. S., Igoryanova N. A., Meleshkina E. P. Bioconversion of secondary products of processing of grain cereals crops. Food systems. 2019; 2(4): 18–24. DOI: 10.21323/2618–9771–2019–2–4–18–24

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2. Materials and methods the range of 20–70 °C and pH of 3.0–6.0. The hydrolysis efficiency The initial raw materials in our studies were the bran of the was estimated by RS accumulation using the Bertrand method. grinding and peeling systems formed by milling triticale into flour, Table 2 and secondary products (products of peeling) — in the production Characteristics of the enzymatic preparations of cereals. «Shearzyme 500 L» and «Viscoferm L» when effecting the The content of dietary fiber was determined on the device «GDE non-starch polysaccharides of triticale bran Enzymatic digester» of the company «Velp» (Italy) in accordance Indicator «Shearzyme 500 L» «Viscoferm L» with the enzymatic-gravimetric method 985.29, approved by Initial velocity, V0 (min) 30 30 AOAC in 1985; the fiber content — according to Genneberg and Optimum temperature, ºС 50 50 Shtoman [1]. Optimum рН 5.5 3.5 The soluble protein content was determined using the Lowry 0.4 units of Optimal amount of enzyme 0.3 units of xylanase method [12] and the protease activity — using the modified Anson cellulolytic preparation, units/g of bran ability/g of bran method [13], bovine serum albumin was used as the standard ability /g of bran substrate, amine nitrogen — using the formol titration method, and reducing substances (RS) — using the Bertrand method To estimate the efficiency of the studied enzyme preparations, [14]. The proteins and the products of proteolysis of triticale the enzymatic hydrolysis was carried out under the optimal condi- flour and bran were fractionated by molecular weight using the tions, which were selected experimentally. The incubation mixture gel chromatography method with a column with Toyopearl gel consisted of triticale bran, water (the hydromodule is 1:10), the HW‑55F [14]. appropriate buffer (20 % of volume) and an enzyme preparation The following were used as proteolytic and cellulolytic enzy- based on the final concentration of the corresponding optimum. matic preparations: «Neutrase 1.5 MG» — a bacterial metallopro- Sampling was carried out every 30 minutes for 2 hours, the samples teinase (Zn) produced by Bacillus amyloliquefaciens,); «Distizym were transferred to centrifugal glasses and centrifuged at 6000 Protacid Extra» — a fungal protease produced by Aspergillus niger rpm for 10 minutes. The supernatant was used to determine the (Döhler, Germany), «Shearzyme 500L» — a purified xylanase pro- reducing sugars (reducing substances) using the Bertrand method duced by Aspergillus oryzae and Aspergillus aculeatus, «Viscoferm and the amount of soluble protein using the Lowry method. L» — a balanced mixture of xylanase, β-glucanase, cellulase and The hydrolysis efficiency was estimated by the accumulation α-amylase produced by Aspergillus aculeatus (Novozymes, Den- of RS and soluble protein. The results are shown in Figures 3 and mark). All the preparations are recommended for the hydrolysis 4. It has been shown that the enzymatic preparation «Shearzyme of biopolymers of grain raw materials. 500 L» increases the amount of RS and soluble protein by 2 times; Table 1 presents the main kinetic characteristics of the en- and the preparation «Viscoferm L» increases the amount of RS zymatic reaction of hydrolysis of triticale brain proteins using by 1.5 times and the amount of soluble protein by 2.5 times. The bacterial and fungal proteolytic enzyme preparations. The hy- obtained data indirectly indicate the possibility of a significant drolysis was carried out at the optimum pH and temperature for increase in the nutritional value of secondary products of grain 30 minutes. It has been previously established that the reaction triticale processing. is zero order for 30 min. The enzyme preparations were added in The functional and technological properties were determined the amounts from 0.25 to 1.5 units of PA/g of flour, the substrate using the methods described in [7] and in [15, 16]. The water ab- concentration varied from 20 to 120 mg/ml. sorption capacity (WAC) was determined as the amount of water Table 1 adsorbed by the modified triticale bran after centrifugation. Characteristics of the enzyme preparations To determine the fat emulsifying capacity (FEC), 50 ml of dis- «Neutrase 1.5 MG» and «Distizym Protacid Extra» tilled water was added to the weighed amount of 1 g of modified when effecting triticale bran proteins triticale bran and suspended at 4000 rpm for 1 minute. Then 10 ml «Neutrase «Distizym Indicator of refined sunflower oil was added to the mixture and emulsified 1.5 MG» Protacid Extra» for 5 minutes at a rate of 8000 rpm. The obtained emulsion was Initial velocity, V (min) 30 30 0 centrifuged for 5 minutes at 2000 rpm. Optimum temperature, ºС 50 40 FEC was calculated as a ratio of the emulsion volume and the Optimum рН 5.5 3.5 overall system volume expressed as a percentage. The emulsion Optimal amount of enzyme preparation, stability (ES) was determined by heating the emulsion for 30 min at units of PA/g of bran 0.50 0.75 80 ºС, then cooled and centrifuged at 2000 rpm. ES was calculated Saturated substrate concentration, mg/cm3 100 100 as a ratio of the emulsion volume and the overall system volume expressed as a percentage. Taking into account the complex structure of the cell wall To determine the fat binding capacity (FBC), the weighed (the main component of bran), enzyme preparations with a whole amount was put into a pre-weighed centrifuge tube, 5 ml of re- complex of activities are required to degrade it and increase the fined sunflower oil was added and mixed for 1 minute at 1000 degree of protein extraction: cellulase, hemicellulase and pecto- rpm, then centrifuged for 15 minutes at 4000 rpm. The unad- lytic activity. sorbed oil was drained, the tubes were weighed and the FBC Table 2 presents the characteristics of the enzymatic reaction was calculated as a ratio of the weight of the bound oil to the of hydrolysis of non-starch polysaccharides of triticale bran when weighed amount. effected by the enzymatic preparations «Shearzyme 500 L» and The foaming capacity (FC) was determined by mixing a weighed «Viscoferm L». amount in 25 ml of distilled water in a graduated cylinder and The composition of the incubation mixture is the follow- thoroughly mixed, the volume was made up to 300 ml and shaken ing: milled triticale bran and water (the hydromodule is 1:10), a for 1 min. FC was expressed as a ratio of a foam height (mm) to phosphate-citrate buffer 0.1 M (20 % of volume) and an enzyme a liquid height (%). preparation with the activity from 0.1 to 0.5 activity units/g of The analyses were performed in triplicate, presenting the re- bran. It has been established that the reaction is zero order for sults as average arithmetic ones. The discrepancy between parallel 30 min. The optimum temperature and pH were revealed when assays did not exceed 3 % of the average arithmetic value with the studying the activity of the enzyme preparations under study in confidence probability P=0.95.

19 microns

WAC, g of water / g of ingredient 8.8 8.0 7.6 7.2

Studies have shown that the content of insoluble dietary fiber has a decisive effect on water absorption. A direct correlation was established between the water absorption capacity and the content of insoluble dietary fiber. The increase in the mass fraction of insoluble dietary fiber in the ingredients by 60-240% is accompanied by a corresponding increase in water absorption capacity by 60-270% (Figs. 1 and 2). ПИЩЕВЫЕ СИСТЕМЫ | Том 2 № 4 | 2019 FOOD SYSTEMS | Volume 2 № 4 | 2019 The mass fraction of insoluble dietary fiber in the ingredients, varying in the range of 63-81%, provides, as our studies show, a range of changes in water absorption capacity: from 3. Results and discussion microns The mass fraction of insoluble dietary fiber in the ingredients, According to the first line of research, the scientists7.2 to 8.8of the g ofInsti water- pervarying g of in ingredient the range (Figureof 63–811 %, and provides, Figure as 2). our studies show, a tute have developed a method for producing organicWAC ingredients, g of water range of changes in water absorption capacity: from 7.2 to 8.8 g from secondary products formed during the high-quality/ g of ingredient grinding of water8.8 per g of ingredient (Figure8.0 1 and Figure 2).7.6 7.2 of triticale and wheat into flour, which includes the enzymatic ac- tion of amylolytic enzymes to release starch polysaccharides while preserving the native properties of dietary fiber and Studiesbiologically have shown that the content of insoluble dietary fiber has a decisive effect on active substances associated with them. water absorption. A direct correlation was established between the water absorption capacity A sufficiently wide range of initial renewable raw materials and the technologies used make it possible to obtainand initialthe content fraction of insoluble dietary fiber. The increase in the mass fraction of insoluble products for further biochemical transformation, the composition of which varies in the total content of dietary fiber,dietary as wellfiber as infiber, the ingredients by 60-240% is accompanied by a corresponding increase in one of the components of dietary fiber (Tablewater 3). absorption capacity by 60-270% (Figs. 1 and 2). Table 3 Characterization of the complex of dietary fiberThe mass fraction of insoluble dietary fiber in the ingredients, varying in the range of of the initial fractions from the secondary products of triticale processing 63-81%, provides, as our studies show, a range of changes in water absorption capacity: from Figure 1. Mass fraction of insoluble dietary fiber (DF) The composition7.2 to 8.8 of g the of water per g ofin ingredientingredients from(Figure secondary1 and productsFigure 2). of processing complex of dietary fiber (DF), % Figure 1. Mass fractionof triticale: of insoluble 1, 2 — cuts dietary of grinding fiber and (DF) peeling in ingredients systems; from secondary Fractions 3, 4 — peeling products with a yield of 2.0 and 5.7 % from secondary products including the products of processing of triticale: total of triticale processing mass fraction of content 1, 2 – cuts of grinding and peeling systems; fiber from the of DF total DF content 3, 4 – peeling products with a yield of 2.0 and 5.7% when Bran from break systems 39.1 25.9 grinding triticale Bran from reduction systems 33.2 23.1 when bran output 2.0 % 60.6 25.2 peeling triticale bran output 5.7 % 43.4 24.9

The composition of the organic ingredient (concentrated dis- persions containing dietary fiber) obtained by biochemical trans- formation of the starting fractions is characterized by a higher mass fraction of the main component — dietary fiber, the content Figure 2. WBC ingredients derived from secondary of which is 66–81 %. The organic ingredient containing insoluble products of triticale processing: 1, 2 — cuts of grinding and dietary fiber is a light brown powder that is easily dispersibleFigure 1.in Mass fractionpeeling of systems; insoluble 3, 4 —dietary peeling fiber products (DF) within ingredients a yield from secondary water. The product yield, depending on the composition of the products of processingof 2.0 and 5.7 of %triticale: initial fraction of the feedstock, is 52–75 %. When1, using 2 – thesecuts of products grinding as foodand ingredients,peeling systems; it is important Considering possible solutions for the use of concentrated Figure 2. WBCto ingredientsknow3, 4 not– peeling only derived how products much from water withsecondary an a yieldingredient products of 2.0 can and of absorb, triticale5.7% but processing: dispersions containing dietary fiber as ingredients capable of also the amount of water that it can hold. The strength of moisture regulating the structure of food products, it is necessary, first 1, 2 – cuts of grinding and peeling systems; bonding by fibers is different for ingredients that differ both in the of all, to study one of the most important properties of dietary quantity and composition of the dietary fiber complex and in the fiber — their sorption ability. The properties of the resulting in- morphological features of the structure of the structures (Table 5). gredients are determined by the composition, structure and ratio of the components that form them. To a large extent, the features Table 5 of their properties are due to the number and composition of the Well-bonded moisture of ingredients containing insoluble dietary fiber from triticale recycled products components of dietary grain fibers entering the bran, as well as the morphological features of their structure [1]. So, with an in- Feedstock for ingredients containing dietary fiber crease in the particle size distribution of particles of ingredients triticale grinding triticale peeling containing dietary fiber, there is a steady growth trend in water Index products products absorption capacity (WAC) (Table 4). bran from bran from bran yield bran yield Table 4 break reduction 2.0 % 5.7 % Water absorption capacity of ingredients differing systems systems in particle size distribution Strongly bonded moisture 1.8 2.0 2.1 2.2 Particle size Pass 2.0 / Pass 0.95/ Pass 0.56/ Gathering g water / g ingredient distribution, gathering gathering gathering Figure 2. WBC ingredients derived from secondary products of triticale processing: 0.315 microns 0.355 0.63 0.157 1, 2 – cuts of grinding and peeling systems; WAC, g of water / g The ability of fiber to retain water is related to the degree 8.8 8.0 7.6 7.2 of ingredient of hydrophilicity and the amount of biopolymers, the nature of porosity and particle size. The surface of dietary fiber includes Studies have shown that the content of insoluble dietary fiber a system of pores whose sizes, depending on the morphological has a decisive effect on water absorption. A direct correlation structure of the source of dietary fiber, can vary over a fairly wide was established between the water absorption capacity and the range. This largely determines the strength of water retention by content of insoluble dietary fiber. The increase in the mass frac- dietary fiber. tion of insoluble dietary fiber in the ingredients by 60–240 % is Water held by insoluble dietary fiber is considered in three accompanied by a corresponding increase in water absorption forms: bonded to the hydrophilic groups of the polymer — strongly capacity by 60–270 % (Figs. 1 and 2). bonded; localized in the intercellular walls — easily accessible;

20 ПИЩЕВЫЕ СИСТЕМЫ | Том 2 № 4 | 2019 Figure 4. Absolute viscosityFOOD ofSYSTEMS aqueous colloidal| Volume systems 2 № 4 at| 2019 20°C We found that at an ingredient concentration of up to 0.5%, the temperature of the intermediate, the availability of which is determinedcolloidal by the medium size does Wenot foundsignificantly that at affectan ingredient the viscosity concentration of the colloidal of up to system. 0.5 %, However, of the matrix in which it is localized [1]. the temperature of the colloidal medium does not significantly Studies of the forms of binding of moisture absorbedat higher by dietary concentrations affect theof viscosityingredients of thecontaining colloidal system.soluble However,dietary fiber,at higher the effect of fiber showed the following. The amount of tightlytemperature bound moisture rises significantly.concentrations So, ofif ingredientsin a colloidal containing system at soluble an ingredient dietary concentrationfiber, of retained by hydration of the hydroxyl groups of cellulose and the effect of temperature rises significantly. So, if in a colloidal hydrophilic hemicellulose colloids varies slightly.1% The at a range temperature of system of 20 °C, at an the ingredient viscosity concentrationis 47.2 mPaS, ofthen 1 % at at 60 a temperature °C it is much of lower – 5.6 variation is 1.8–2.2 g of water / g of product (Table. 5). 20 °C, the viscosity is 47.2 mPaS, then at 60 °C it is much lower — mPaC (Figure 5). From the point of view of the technological properties of the 5.6 mPaC (Figure 5). ingredients, of most interest is easily removable moisture, not associated with fibers. Significant differences were found in the The ratio ofamount the tot ofal relativelyamount ofreadily water available absorbed moisture by an notingredient associated to the amount of its with fibers, caused by morphological features of the structure of easily accessible formthe dietary in each fiber particular complex ingredient (capillary containingmoisture capacity). insoluble For thedietary fiber largely studied ingredients, this indicator varies in the range — 4.8–6.3 g determines its propertiesof water during / g of the the product production (Figure process 3). and further storage of products. The ratio of the total amount of water absorbed by an ingredi- The developmentent to the of amount a method of its easilyof producing accessible ingredientsform in each particular(concentrated dispersions) containing solubleingredient dietary containingfiber with insoluble the properties dietary fiber of largelystabilizers determines of food systems was its properties during the production process and further storage accompanied by studiesof products. on the ability of the resulting ingredients to change the rheological The development of a method of producing ingredients (con- properties of aqueouscentrated systems. dispersions) The kinematic containing viscosity soluble of dietary solutions fiber ofwith various the concentrations Figure 5. Absolute viscosity of aqueous colloidal systems properties of stabilizers of food systems was accompanied by with different concentrations of the ingredient (1–0.2, was measured on astudies viscometer. on the ability of the resulting ingredients to change Figure 5.2–0.5, Absolute 3–1.0 viscosity %) and the of temperatureaqueous colloidal of the colloidalsystems systemwith different the rheological properties of aqueous systems. Theconcentrations kinematic of the ingredient (1 – 0.2, 2 – 0.5, 3 – 1.0%) and the temperature of the viscosity of solutions of various concentrations was measured Thus, for gelation,colloidal thickening system and stabilization of aqueous on a viscometer. food systems, it is possible to use ingredients containing soluble dietary fiber in concentrations of 0.5 % or higher, taking into ac- Thus, for gelation, thickening and stabilization of aqueous food systems, it is possible count the temperature of the colloidal system. to use ingredients containingStudies soluble on the dietary use of fiber ingredients in concentrations with a high of 0.5%content or higher,of taking dietary fiber in bakery have shown that the composition of the into account the temperaturefiber complex of the has colloidal a significant system. effect on the quality of bread. With Studies on thean useincrease of ingredients in the mass with fraction a high of content fiber in of the dietary composition fiber in bakeryof have the dietary fiber complex, the volume of bread and the quality shown that the compositioncharacteristics of the of fiber the crumb complex decrease has a when significant the water-absorbing effect on the quality of ability of the introduced ingredients is not taken into account [1]. Studies have shown that the presence of insoluble dietary fiber (NLF) in the dough does not adversely affect the qual- ity of the bread if there is enough water in the dough to swell all the hydrocolloids. So, when making ingredients containing insoluble dietary fiber in an amount of 2 % by weight of flour, Figure 3. Capillary moisture capacity of ingredients derived the optimal amount of water is 62–65 %. Ingredients contain- from secondary triticale products: 1 — bran of grinding systems; 2, 3 — peeling products with a yield of 2.0 and 5.7 % ing insoluble dietary fiber, when mixed with wheat flour dough Figure 3. Capillary moisture capacity of ingredients derived from secondaryduring triticale kneading, depending on the feedstock containing them Figure 4 presents dataproducts: on the absolute viscosity of col- and the morphological characteristics of the fiber structure, have 1 – bran loidalof grinding dispersed systems; systems 2, at3 –a peelingconcentration products of the with introduced a yield of 2.0a significantand 5.7% effect on the rheological properties of the dough ingredient of 0.05; 0.5 and 1.0 % at a temperature of a colloidal prepared with their participation. Most significantly, it manifests Figure 4 presesystemnts ofdata 20 ºС.on Itthe is shownabsolute that viscosity the viscosity of colloidalof aqueous dispersed col- itself systems in a change at a in the water-absorbing ability of the dough and loidal systems at a concentration of 0.5 % increases 11 times; its dilution, that is, indicators that determine the consistency concentration of theat a introducedconcentration ingredient of 1.0 % — of 30 0.05;times, forming0.5 and a 1.0%viscous at gel- a temperatuand propertiesre of a of the dough during the test and, as a result, the like structure. quality of the finished product (Tables 6, 7). colloidal system of 200 ºС. It is shown that the viscosity of aqueous colloidal systemsIt has been at a shown that ingredients containing insoluble di- etary fiber exhibit thickener properties in the product, moreover, concentration of 0.5% increases 11 times; at a concentration of 1.0% - 30 times,the morphologicalforming a structural features of the fiber structures of viscous gel-like structure. various ingredients can change the consistency of the product in different directions. This allows you to adjust the consistency of the product, ensuring its desired quality. Table 6 Rheological properties of the test with the ingredients Dough with an ingredient Liquefaction WAC of the Valorimetric containing insoluble of the test, dough, % rating, units dietary fiber min From bran break systems 76.2 80 82 From bran reduction systems 72.6 80 80 Figure 4. Absolute viscosity Without the ingredient 65.8 80 79 of aqueous colloidal systems at 20 °C Figure 4. Absolute viscosity of aqueous colloidal systems at 20°C 21 We found that at an ingredient concentration of up to 0.5%, the temperature of the colloidal medium does not significantly affect the viscosity of the colloidal system. However, at higher concentrations of ingredients containing soluble dietary fiber, the effect of temperature rises significantly. So, if in a colloidal system at an ingredient concentration of 1% at a temperature of 20 °C, the viscosity is 47.2 mPaS, then at 60 °C it is much lower – 5.6 mPaC (Figure 5).

Figure 5. Absolute viscosity of aqueous colloidal systems with different concentrations of the ingredient (1 – 0.2, 2 – 0.5, 3 – 1.0%) and the temperature of the colloidal system

Thus, for gelation, thickening and stabilization of aqueous food systems, it is possible to use ingredients containing soluble dietary fiber in concentrations of 0.5% or higher, taking into account the temperature of the colloidal system. Studies on the use of ingredients with a high content of dietary fiber in bakery have shown that the composition of the fiber complex has a significant effect on the quality of ПИЩЕВЫЕ СИСТЕМЫ | Том 2 № 4 | 2019 FOOD SYSTEMS | Volume 2 № 4 | 2019

Table 7 result, products of enzymatic modification of bran from triticale Quality characteristics of bread with ingredients grain with a different degree of hydrolysis of proteins and non- starch polysaccharides and various functional and technological properties have been obtained [5,9]. Dough with an The composition of 2 multi-enzyme compositions used for / 100 g ingredient containing 3 the enzymatic modification of triticale bran included: «Shear- insoluble dietary fiber zyme 500 L» + «Neutrase 1.5 MG» (MEC‑1) and «Viscoferm L» + «Dystizym Protacid Extra» (MEC‑2). The choice of enzyme Volumetric yield of Volumetric cm bread, of flour % Porosity, elasticity, Relative % organoleptic Total score assessment, preparations is caused by various specific effects and approxi- From bran break systems 781 88 29.7 10 mately the same effect optima: the optimum temperature is 50 °C; pH is 5.5–6.0 for MEC‑1 and 40 °C; pH is 3.5 for MEC‑2. Without the ingredient 725 86 32.2 10 The hydrolysis was carried out in 2 stages. At the first stage, a cellulolytic enzyme preparation was applied. At the second The use of organic ingredients containing insoluble dietary stage, a proteolytic enzyme preparation was applied. The dos- fiber in the manufacture of food products not only allows us to age of enzyme preparations, the substrate concentration and solve technological problems, but also does not create problems the duration of each stage were selected experimentally [6]. with a «clean label». In this case, the key role belongs to dietary Figures 6 а, b and Table 8 present the results of fractionation fiber, which is considered in the context of both their physiological of the products of proteolysis using the gel chromatography effects on the human body and their influence on the technologi- method on a column with Toyopearl gel HW‑55F. cal properties of food products containing them. Table 8 Our studies have shown that the use of ingredients containing Fractionation of the products of proteolysis of triticale insoluble dietary fiber does not adversely affect the volumetric bran proteins using MEC yield of bread when there is enough water in the dough to allow % of the total colloidal and biochemical processes to occur in the dough. This Fraction Molecular weight, Da allows you to get bread with a high volumetric yield without ad- Control MEC‑1 MEC‑2 ditional use of improvers. ≥ 700000 Peak I 6–13 35.81 23.67 19.55 The ratio of the amount of free and bound moisture absorbed (blue dextran yield) by each particular dispersion used for technological purposes Peak II 14–15 450000 ÷ 350000 13.26 14.79 12.62 largely determines the properties of the product, both during Peak III 16–19 300000 ÷ 100000 9.95 26.04 3.20 its production and subsequent storage. Dietary fiber is known to be an effective thickener in water systems. Our studies have Peak IV 20–22 100000 ÷ 50000 13.26 0 0 shown that significant changes in viscosity in aqueous colloidal Peak V 23–26 50000 ÷ 25000 10.08 5.02 1.77 systems occur when ingredients containing soluble dietary Peak VI 27–30 25000 ÷ 1500 5.31 2.54 0 fiber are added in an amount of 0.5 % or more. At concentra- tions above 2 %, it is possible to obtain gels of various textures. Peak VII 31–36 ≤ 1000 (tyrosine yield) 12.33 51.06 62.63 When deciding on the use of a particular ingredient containing soluble dietary fiber, it is important to consider the effect of The obtained experimental data on the kinetics of enzymatic temperature to thicken the food system, since its increase is reactions of hydrolysis of biopolymers of a grain substrate (different accompanied by a significant decrease in the viscosity of the types of flour and triticale bran); the degrees of hydrolysis and the colloidal system. ratio of fractions with different molecular weights using the gel The second direction of research was to develop methods for chromatography method have formed the basis for the develop- enzymatic modification of tritical bran. The enzymatic method for ment of biotechnological methods for modifying the products of the modification of plant proteins is preferable to physicochemi- triticale grain processing. cal modification, since its advantages are soft reaction modes, the The developed methods for modifying the products of triticale ability to regulate the degree of hydrolysis, a certain directivity grain processing include the following stages: and conservation of biological value [10,17,18,19]. ‰‰ the preparation of a suspension — bran: water (the hydro- In recent years, it has been found that, for the use of protein module is 1:4) hydrolysates, it is not necessary to obtain them with a high depth ‰‰ the preparation of solutions of enzyme preparations; the of hydrolysis, since peptides are also well absorbed by the human creation of MEC; organism. Protein hydrolysates are divided into 2 large groups: ‰‰ the enzymatic hydrolysis using MEC under the developed partially hydrolyzed proteins, fully hydrolyzed proteins. Each of conditions (the substrate concentration, the dosage of en- the hydrolysates has certain properties that determine the area of​​ zyme preparations, the optimum temperature and pH); the application. Completely hydrolyzed proteins have low antigenic inactivation of enzyme preparations; the product being ob- activity, which makes it possible to use them in hypoallergenic tained is hydrolyzed bran (an unclarified hydrolysate); children’s diets. Such hydrolysates contain free amino acids and ‰‰ centrifugation; the product being obtained is a hydrolysate short peptides. Partially hydrolyzed proteins include a wide range (a supernatant) and paste (a precipitate); of hydrolysis products. They include: fraction of free amino acids ‰‰ drying; the product being obtained is a dry hydrolysate and and short peptides; a sufficiently large number of oligopeptides; a hydrolyzed bran. significant amount of high molecular weight hydrolysis products. They are characterized as weakly and medium hydrolyzed proteins. To estimate the possibility of using the products obtained Hydrolysates belonging to this group do not differ significantly, in food branches, their functional and technological properties they are used as an easily digestible source of amino nitrogen in have been studied. specialized diets [8,9,10]. Tables 9 present the water binding capacity (WBC); the fat Bran with a high content of peripheral parts containing a large binding capacity (FBC); the fat emulsifying capacity (FEC); the number of non-starch polysaccharides, were modified using MEC emulsion stability (ES); the foam forming capacity (FFC) and the based on cellulolytic and proteolytic enzymatic preparations. As a foam stability (FS) of the modified triticale bran.

22 a different degree of hydrolysis of proteins and non-starch polysaccharides and various functional and technological properties have been obtained [5,9]. The composition of 2 multi-enzyme compositions used for the enzymatic modification of triticale bran included: "Shearzyme 500 L" + "Neutrase 1.5 MG" (MEC-1) and "Viscoferm L" + "Dystizym Protacid Extra" (MEC-2). The choice of enzyme preparations is caused by various specific effects and approximately the same effect optima: the optimum temperature is 50°C; pH is 5.5-6.0 for MEC-1 and 40°C; pH is 3.5 for MEC-2. The hydrolysis was carried out in 2 stages. At the first stage, a cellulolytic enzyme preparation was applied. At the second stage, a proteolytic enzyme preparation was applied. The dosage of enzyme preparations, the substrate concentration and the duration of each stage were selected experimentally [6]. Figures 6 а,b and Table 8 present the results of fractionation of the products of proteolysis

ПИЩЕВЫЕusing СИСТЕМЫ the gel chromatography | Том 2 № 4 | 2019 method on a column with Toyopearl FOODgel HW SYSTEMS-55F. | Volume 2 № 4 | 2019

a

b

Figure 6. Fractionation of the products of proteolysis of triticale bran proteins of MEC‑1 (a) and MEC‑2 (b) Figure 6. Fractionationusing the of gel the chromatography products of method proteolysis on a column of triticale with Toyopearl bran proteins gel HW‑55F of MEC-1 and MEC-2 using the gel chromatography method on a column with Toyopearl gel HW-55F Table 9 It is known that the functional properties of the products of Functional properties of the modified triticale bran enzymatic hydrolysis of protein raw materials depend on the Table 8. WBC,Fractionation FBC, of the products of proteolysisphysico-chemical of triticale properties bran proteins of the initialusing protein, MEC the specificity Sample* FAC, % ES, % FFC, % FS, % g/g g/g of the proteases used, the composition of MEC used, the condi- Fraction Molecular weight, Da % of the total Control — C1 1.56 1.32 52 58 50 32 tions for hydrolysis, the degree of hydrolysis and the ratio of the fractionsControl of proteolysisMEC products-1 with differentMEC molecular-2 weights Experiment 1 — E1 1.80 1.50 62 53 59 28 [11,18,19,20]. Experiment Peak2 — E2 I 1.20 61.40 – 13 56 46 ≥ 70000042 24 35.81 23.67 19.55 (blue dextran yield) The revealed differences in the functional properties in the ini- *) Control C1 — bran; Experiment 1 — bran + MEC1; Experiment 2 — bran + tial and modified products of triticale grain processing are related, MEC2 Peak II 14 – 15 450000 ÷ 350000 first of13.26 all, to the conditions14.79 for enzymatic12.62 modification (of the The functionalPeak III properties 16 – of19 bran from300000 triticale ÷grain 100000 and pH medium),9.95 the composition26.04 and specific3.20 effect of the enzymes the hydrolyzedPeak samples IV obtained20 – 22 using MEC1100000 and MEC2 ÷ 50000 dif- that are13.26 part of the composition0 of MEC; obtaining0 products of fer from each other. Thus, the water-binding capacity of the various degrees of hydrolysis, and the number of high-, medium- hydrolysedPeak bran Vin the first 23 option– 26 increases50000 by 16 ÷ %, 25000 in op- and low-molecular10.08 compounds;5.02 an increase1.77 or decrease in free tion 2 — onPeak the contrary,VI 27 it decreases– 30 by 12.625000 % with ÷ respect 1500 polar (charged)5.31 aggregations,2.54 hydrophilic and/or0 hydrophobic to the unhydrolysedPeak VII triticale 31 – bran.36 The similar≤ 1000 pattern (tyrosine can be groups,12.33 providing interactions51.06 with different62.63 types of substances. seen with respect to the foam forming capacity (Experimentyield) The obtained results indicate that the use of MEC on the basis 1: an increase of 18.0 %; Experiment 2: a decrease of 16.1 %). of cellulolytic and proteolytic enzyme preparations allows for an The fat binding and fat emulsifying capacity increases in both advanced destruction of proteins and non-starch polysaccharides experimental optionsThe obtained by 13.6 % experimental and 6.1 % and databy 19.2 on % the and kinetics of the of products enzymatic of triticale reactions grain processing;of hydrolysis to obtain of products 7.7 % respectively.biopolymers of a grain substrate (different types withof flour various and degrees triticale of hydrolysis bran); theand thedegrees ratio of of components The stability of the emulsion and foam of the modified triticale by molecular weight, which leads to a change in the functional bran is hydrolysisreduced: ES byand 8.7 %;the FS —ratio by 12.5 of % (Experimentfractions 1) andwith differentand technological molecular properties weights of the using initial brainthe andgel will allow to ES — by 20.7 %; FS — by 25.0 % (Experiment 2). find its new scopes in food products. chromatography method have formed the basis for the development of biotechnological 23 methods for modifying the products of triticale grain processing. The developed methods for modifying the products of triticale grain processing include the following stages: - the preparation of a suspension - bran: water (the hydromodule is 1:4) - the preparation of solutions of enzyme preparations; the creation of MEC; - the enzymatic hydrolysis using MEC under the developed conditions (the substrate concentration, the dosage of enzyme preparations, the optimum temperature and pH); the inactivation of enzyme preparations; the product being obtained is hydrolyzed bran (an unclarified hydrolysate); ПИЩЕВЫЕ СИСТЕМЫ | Том 2 № 4 | 2019 FOOD SYSTEMS | Volume 2 № 4 | 2019

4. Сonclusions native structure and properties of dietary fibers and associated bio- Two ways of increasing the efficiency of the use of secondary logically active substances and products of enzymatic modification products of grain processing in flour and cereals are suggested in of triticale bran with a certain ratio of high, medium, low molecular the example of triticale grain, namely: obtaining useful organic peptides and amino acids having certain functional and technologi- ingredients of functional purpose with technological properties cal properties. They can be used in the production of a wide range (thickening, structure formation) with maximum preservation of the of general, functional and therapeutic-prophylactic food products.

REFERENCES 1. Igoryanova, N.A., Meleshkina, E.P. (2017). Possibilities of using second- grain processing technology. Foods and Raw Materials, 5(2), 70–82. DOI: ary grain processing products to produce ingredients with food fibers. 10.21179/2308–4057–2017–2–70–82. Bakery products, 10, 41–44. (In Russian) 12. Lowry, O.H., Rosebrougt, N.J., Farr, A.L., Randall, R.J. (1951). Protein 2. Meleshkina, E.P., Vitol, I.S., Kandrokov, R. Kh. (2016). Products of triticale measurement with Folin phenol reagent. Journal of Biological Chemistry, grains as an object for enzymatic modification Storage and processing of 193(1), 265–275. farm products, 9, 14–18. (In Russian) 13. Anson, M. L. (1938). The estimation of pepsin, trypsin, papain, and ca- 3. Kazakov, E.D., Karpilenko G. P. (2005). Biochemistry of grain and bak- thepsin with hemoglobin. Journal of General Physiology, 22(1), 79–82. ery products. St. Petersburg: GIORD. — 512 p. ISBN: 5–901065–82–4 DOI: 10.1085/jgp.22.1.79 (In Russian) 14. Nechaev, A.P., Traubenberg, S.E., Kochetkova, A.A., Kolpakova, V.V., Vi- 4. Vitol, I.S., Karpilenko, G.P. (2015). Modification triticale flour using a pro- tol, I.S., Kobeleva, I.B. (2006). Food Chemistry. Laboratory practical work. teolytic Enzyme Preparations. Storage and processing of farm products, 9, St. Petersburg: GIORD. — 304 p. (in Russian) 17–22. (in Russian) 15. Toshev, A.D., Polyakova, N.V., Salomatov, A.S. (2012). The research of 5. Vitol, I.S., Meleshkina, E.P., Karpilenko, G.P. (2016). Bioconversion technological properties of № 2 puffed pearl barley grits. Food Process‑ of tritikale bran using enzyme preparations of cellulolytic and pro- ing: Techniques and Technology, 1(24), 77A‑81. (in Russian) teolytic action. Storage and processing of farm products, 10, 35–38. 16. Renzyaeva, T. V., Tuboltseva, A.S., Ponkratova, E. K., Lugovaya, A.V., Ka- (in Russian) zantseva, A. V. (2014). Functional and technological properties of pow- 6. Meleshkina, E.P., Vitol, I.S., Karpilenko, G.P. (2016). Modification of veg- dered raw materials and food additives for confectionary Food Processing: etable protein of triticale grain by means of biotechnological methods. Techniques and Technology, 4(35), 43–49. (in Russian) Bakery products, 5, 62–64. (in Russian) 17. Bezborodov, A.M., Zagustina, N.A., Popov, O.V. (2008). Enzymatic pro- 7. Zabodalova, L.A. (2015). Scientific foundations of functional products. cesses in biotechnology. Moscow: Nauka. — 335 p. ISBN: 978–5–02– St. Petersburg: Universitet ITMO. — 86 p. (in Russian) 035661–0 (in Russian) 8. Kolpakova, V.V., Zaitseva, L.V., Martynova, I.V., Osipov, Ye.A. (2007). Pro- 18. Dey, S.S., Dora, K.C. (2014). Optimization of the production of shrimp tein from wheaten bran: increase of output and functional properties. waste protein hydrolysate using microbial proteases adopting response Storage and processing of farm products, 2, 23–25. (in Russian) surface methodology. Journal of Food Science and Technology, 51(1), 16– 9. Vitol, I.S., Meleshkina, E.P., Karpilenko, G.P. (2017). Functional proper- 24. DOI: 10.1007/s13197–011–0455–4 ties of modified products of processing of triticale grain.Storage and pro‑ 19. Kaur, G., Sharma, S., Nagi, H.P.S., Dar, B.N. (2012). Functional properties cessing of farm products, 2, 27–29. (in Russian) of pasta enriched with variable cereal brans. Journal of Food Science and 10. Claver, I.P., Zhou, H.M. (2005). Enzymatic hydrolysis of defatted wheat Technology, 49(4), 467–474. DOI: 10.1007/s13197–011–0294–3 germ by proteases and the effect on the functional properties of result- 20. López-Sánchez, J., Ponce-Alquicira, E., Pedroza-Islas, R., de la Peña-Díaz, ing protein hydrolysates. Journal of Food Biochemistry, 29(1), 13–26. DOI: A., Soriano-Santos, J. (2016). Effects of heat and pH treatments and in 10.1111/j.1745–4514.2005.00004.x vitro digestion on the biological activity of protein hydrolysates of Am- 11. Meleshkina, E.P., Pankratov, G.N., Vitol, I.S., Kandrokov, R.H., Tulyakov, aranthus hypochondriacus L. grain. Journal of Food Science Technology, D.G. (2017). Innovative trends in the development of advanced triticale 53(12), 4298–4307. DOI: 10.1007/s13197–016–2428–0

AUTНOR INFORMATION Irina S. Vitol — candidate of biological sciences, docent, senior researcher, Laboratory of Biochemistry and Microbiology of Grain and Grain Products, All-Rus- sian Scientific Research Institute of Grain and Products of Its Processing — Branch of V. M. Gorbatov Federal Research Center for Food Systems of RAS, 127434, Moscow, Dmitrovskoe shosse, 11. Tel.: +7 926 709 02 07, e-mail: [email protected] *corresponding author Natalia A. Igoryanova — candidate of technical sciences, head microbiology sector, All-Russian Scientific Research Institute of Grain and Products of Its Pro- cessing — Branch of V. M. Gorbatov Federal Research Center for Food Systems of RAS, 127434, Moscow, Dmitrovskoe shosse, 11. Tel.: +7 499 976 43 02, e-mail: [email protected] Elena P. Meleshkina — doctor of technical sciences, acting director, All-Russian Scientific Research Institute of Grain and Products of Its Processing — Branch of V. M. Gorbatov Federal Research Center for Food Systems of RAS, 127434, Moscow, Dmitrovskoe shosse, 11. Теl.: +7 499 976 23 23, e-mail: [email protected] All authors bear responsibility for the work and presented data. All authors made an equal contribution to the work. The authors were equally involved in writing the manuscript and bear the equal responsibility for plagiarism. The authors declare no conflict of interest. Received 14.10.2019 Accepted in revised 22.11.2019 Accepted for publication 01.12.2019

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UDC 664.72 DOI: 10.21323/2618–9771–2019–2–4–25–30 Original scientific paper EQUIPMENT AND SCIENTIFIC STUDIES OF EXPERIMENTAL DATA ON STORAGE OF WHEAT GRAIN Yuri F. Markov*, Alexandra N. Buriak, Larisa G. Eresko Kuban Branch of V. M. Gorbatov’s Federal Research Center for Food Systems of Russian Academy of Sciences, Krasnodar, Russia

KEY WORDS: ABSTRACT experimental storage of wheat, A complex of technical solutions is presented which makes it possible to study the influence of external factors monitoring of parameters, on changes in the indicator of fat acidity value (FAV) and a range of other parameters characterizing the quality laboratory oil glazing of grain, of preservation of long-term stored wheat grains in South Russia. Storage conditions (natural climatic, model) control of the grain moisture, and also the effect of dust suppression treatment of grain by oil glazing that is carried out in some grain termi- water activity nals in the South of Russia were taken as controlled external factors. New data has been obtained on changes in parameters of food suitability of the wheat grains during storage under model conditions with varying degrees of severity of climatic conditions in South Russia. After 6 months of storage the most considerable changes in FAV value were found for 4th class of wheat stored in natural climatic conditions with intensive insolation, the increase was 1.7 mg KOH/1 g of fat (from 7.3 mg KOH/1 g of fat up to 9.0 mg KOH/ 1g of fat). Minimum changes in the same parameter for the same storage period were observed for the wheat of the 3rd class stored in a thermostat at a stable high temperature of 35 ºС (from 11.6 mg KOH/1 g of fat to 11.5 mg KOH/1 g of fat). For wheat of the 4th class, the changes were 0.7 mg KOH/1 g of fat (from 7.8 mg KOH/1 g fat to 8.5 mg KOH/1 g of fat). Analyzes of stored wheat grains subjected to dust suppression by the oil glazing showed similar results, which allows us to state the absence of a significant effect of oil glazing on changes in wheat properties during its storage. Based on the results of the experimental analysis and a generalization of the data obtained an assumption was made on the possible reasons for the lack of pronounced trends in the data for the expected increase of FAV value in food suitability of the wheat grain when it is stored under typical model conditions of South Russia. The likely reason for this is the corresponding moisture state of grain, the water activity of grain was about 0.45. Such a low value was due, in particular, to the fact that samples of model-stored grain had a limited volume and do not repro- duce the mass transfer processes that involve deep layers of the grain mass (because of the mass absence), which takes place during storage of grain in an industrial environment. The tools were proposed for the operational monitoring of the moisture state of wheat grain during storage. Moreover, a plan has been developed to expand the field of modelling storage processes in terms of varying the moisture state of the stored wheat grain samples with an assessment of the influence of moisture state on the dynamics of changes in the parameters to be controlled.

1. Introduction Such oil glazing is widely used for grain export shipment in the For wheat grains, as a raw material used for obtaining the Black Sea grain terminals in South Russia to increase the effi- corresponding grain products, standards of validity and fresh- ciency of transportation processes [2]. In particular, more than ness have not yet been defined. Also there are no systematized 10 million tons of Russian grain treated with such oil glazing data on the nature of FAV changes in wheat grain stored under has been shipped annually for more than six years. The indi- the various climatic conditions, which justifies the relevance cated technology of oil processing of grain came to Russia from of the current studies. The parameter of acid fat value is used the USA, where it has been generally used for more than 25 to assess the freshness and shelf life of grain products, in par- years [3]. ticular, wheat flour — here the relevant standards have been For the full implementation of the experimental part of these established [1]. studies the appropriate equipment is required. Partially, in this At the current period such studies were performed in labora- capacity we use well-known standard tools. In particular for the tory conditions. determination of FAV [4]. The methodological approaches during the research investi- The need for the laboratory modelling of storage conditions gations consist of the laboratory modelling of grain storage pro- of grain in South Russia including the operational monitoring of cesses and regular monitoring of the state of stored grain. This crucial parameters over long time intervals as well as the need laboratory modelling for climatic conditions of the South Russia to prepare wheat grain samples subjected to variable oil glazing is performed in special laboratory storage boxes. It is laboratory has resulted in the development and application of a range of conditions that allow deterministic sampling of the stored grain special technical solutions. Such solutions will find their appli- with appropriate analyzes and selective monitoring of the stor- cation also at the subsequent stages of research. In particular, age conditions. in case of large volumes of stored grain in far spread distances The data obtained at this stage in the climatic conditions delivery while monitoring of the grain condition. of South Russia will allow drawing of certain preliminary con- Thus, the aim of the research is to study the change of the clusions. fat acidity value (FAV) in the grain of food-grade wheat under The additional study requires an understanding of degree storage conditions of South Russia. The FAV characterizes the of FAV nature changes influence on the wheat grains treated lipid complex in terms of its content in unsaturated and free with the special dust-suppressing oil glazing during storage. fatty acids.

FOR CITATION: Markov Yu.F., Buriak A. G., Eresko L. G. Аппаратурное обеспечение и данные экспериментальных исследований опытного хранения зерна пшеницы. Food systems. 2019; 2(4): 25–30. DOI: 10.21323/2618–9771–2019–2–4–25–30

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2. Materials and methods There is a detailed description of the specified stand [5]. Storage was carried out in conditions as close as possible The overflow chamber 5, in which the studied sample of grain to the real climatic conditions of South Russia. The grain sam- was placed, is made in the form of a vertical section of pipe ples were placed into four containers. Two containers were with a diameter of 100 mm. In the upper and lower part of the installed in specially equipped boxes on the territory of the chamber 5 there are covers 3 and 15, through one of which a Kuban branch with slightly different conditions: in the shaded grain sample is loaded (and unloaded) into the chamber. The area under the influence of low-intensity natural insolation; overflow chamber 5 has a cross-shaped joint with a horizontal and the other was installed in the area exposed to intense in- section of pipe of the same diameter, which serves as the sup- solation. Two more containers were placed in a zone with ac- porting body 6, mounted on sliding bearings 16 and rotating tive thermostatic control: inside a thermostat with a stabilized around the longitudinal axis with the help of electric drive temperature of 35 ºС with temperature deviations within the 8. The electric drive itself is mounted on a fixed support 14. range of 0.5 ºС. In these thermostatically controlled containers On the same fixed support 14 is fixed the opposite the end two samples were placed: a sample treated with oil glazing, and face of the housing 6, connected to the rotating part of the an untreated sample. housing through an annular slip clutch 17. In order to prevent To ensure the most accurate temperature in the thermo- the exit of grain beyond the vertical cut of the pipe (chamber) statically controlled zone without dynamic fluctuations dur- 5 during its transverse rotation the restrictive diaphragms ing regulation the special technical solutions were applied. are installed in the interface between the housing 6 and the The inner surface of the thermostatically controlled volume chamber 5. was separated from the external environment by a jacket The dust sensor 10 is mounted on the bracket and is lo- filled with water coolant, inside of which there were thermal cated in the area of ​​the dust exit from the chamber 5. The pas- electric heating elements. The applied digital temperature sage through the measuring channel of the dust sensor has controller provided dual-circuit temperature stabilization: a vertical orientation for unimpeded passage of soaring dust stabilization of the coolant temperature in the jacket taking through it. into account the temperature of the thermostatically con- A solid-state laser emitter 9 and a digital video camera 1 trolled volume. are mounted on a fixed end of the housing. The laser beam 11 The general list of monitored parameters of wheat grain penetrates the entire horizontal space of the housing, passing samples: FAV, moisture, quantity and quality of gluten, vitres- through the overfill chamber and forming a light point on the cence, germination and sprouting energy. The determination of boundary partition — the screen. The viewing sector of the vid- these parameters is carried out according to the relevant domes- eo camera 2 covers the entire area of ​​the backlight of the laser tic standards that have foreign analogues. beam. The frequency of monitoring the state of the stored wheat With the help of the electric drive 8, the housing 6 rotates samples: FAV — 1 month, commodity classification parameters around its longitudinal axis. In this case the overflow chamber and additional parameters — 3 months. 5 rotates around its transverse axis. At the same time, the grain The dust-suppressing oil glazing of grain in industrial sample 13 located in the chamber 5 performs periodic recipro- conditions mentioned in the introduction was carried out by cating movements, permanently pouring from one end of the spraying of vegetable (sunflower) or mineral (food additive transversely rotating chamber to its opposite end. E905) oil into the grain stream in the beginning of grain trans- In the process of such pouring the dust present in the grain port line in the works. The concentration of oil in the grain sample is released off from the grain sample into the inner space during this treatment is less than 0.02 %. As a result of this (5, 6). This dust floating in the air of the working space (5.6) treatment, the volatility of grain dust was significantly re- passes through the restriction diaphragm 4 and enters the con- duced, which ensures a reduction in losses, improvement in trol passage of the dust sensor, which measures the concentra- the environmental and sanitary condition, increase of explo- tion of dust in the air. In parallel, the video camera 1 is used to sion safety level in the works, and reduction of power and waste register an image of the space section (5, 6) through which the disposal costs. The oil glazing agent sprayed in this way essen- laser beam 11 passes. tially belongs to the category of technological aids according Dust particles floating in the air are illuminated by a laser to TR TS029/2012 «Safety requirements for food additives, fla- beam, which makes it possible to observe the density of dust vors and technological aids». In the domestic interpretation, particles in the laser beam illuminated by a light cord and the only sunflower oil — refined, deodorized, frozen out — has so nature of the movement of these particles. Self-focusing the far been used as a glazing agent in this technology. This is op- laser beam, due to its coherence and monochromaticity gives timal item in its cost; it does not change the organoleptic and a clear visualization of the dust density distribution along the quality properties: grain safety, and allows processing at low entire length of the spatial coordinate of the camera viewing ambient temperatures. sector. Our studies were also intended to identify the possible im- The technique of grain sample processing at the described pact of sunflower oil introduced into the grain mass on the laboratory stand is as follows. A grain sample is loaded into the change of FAV index during storage of wheat grain in conditions overfill chamber. The drive is started, the dust sensor readings of South Russia. To ensure the introduction of a variable dose are recorded and the laser dust visualizer is video-recorded. The of sprayed oil into the grain sample, a special laboratory stand rotation is carried out until stationary values ​​of the measured for grain glazing was used, its scheme is shown in Figure 1. The parameters are reached. stand, on the one hand, provides atomizer spraying oil aerosol Further, a dust-suppressing reagent is introduced into the into the grain sample, and, on the other hand, has measurement grain sample with a spray nozzle-gun accompanying this rota- tools of the concentration of dust floating in the air overflow tion of the chamber and pouring of grain. As a result, a uniform chamber. Monitoring of the oil aerosol dosage was carried out by processing of the entire grain sample by dust-suppressing re- changing the oil level in the capacity of the atomizer. agent is achieved. Monitoring the dosage of oil aerosol was car- The main purpose of the stand was to assess the effective- ried out by changing the oil level in the atomizer capacity. ness of dust suppression by oil glazing of grains with different After that the measurements of dust within the control concentrations of sprayed oil and various types of glazers. interval were carried out. The experiments were performed in

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a) front view b) top view

c) side view

Figure 1. Diagram of a laboratory stand for grain glazing where: 1 — digital video camera; 2 — camera review sector; 3 — top cover of the overflow chamber; 4 — restrictive diaphragm; 5 — overfill chamber; 6 — carrying case; 7 — direction of rotation; 8 — rotation drive; 9 — solid state laser; 10 — dust sensor; 11 — focused laser beam and the axis of the spraying torch oil atomizer; 12 — restrictive screen; 13 — grain test zone; 14 — fixed support; 15 — bottom cover of the overflow chamber; 16 — sliding support; 17 — slip clutch several replications to verify reproducibility. As a dispenser sult, the complexity of their full formalized description. In this and oil atomizer the laboratory atomizer of the compression regard it is planned in the future to develop methods, prepare type was used. tools and conduct a similar kind of research on industrial vol- The SCADA system «MasterScada» was used as a data collec- umes of stored grain. tion and registration system. At the same time, a data exchange To enable remote monitoring of the parameters of grain protocol in ModBus format with data transfer via the RS‑485 se- mass during experimental storage, a special set of measuring rial interface line was implemented in the secondary electronic instruments has been prepared, which can be used both for unit of the dust sensor. Data transfer was carried out through the research purposes and for arrangement of monitoring of the OPC server «LectusOPC». grain state in industrial production conditions. The composi- Along with wheat grain samples that underwent laboratory tion of this set includes: thermic probes, hygro-thermic probes, oil glazing, a sample of grain that has passed oil dust glazing wireless and wired data transmission modules, data collectors, for suppression under industrial conditions at the Novorossiysk special software. Instruments for hygro-thermic control allow grain terminal was laid for experimental storage. determining the water activity of grain mass thus characteriz- Laboratory modelling of storage conditions cannot fully re- ing the stored grain mass from the point of view of its moisture produce storage conditions in an industrial volume grain ter- state [6]. minal. The reason of it is an insufficient knowledge of the dis- tributed and combined physical processes occurring in a large 3. Results and discussion volume grain terminal under the influence of variable external Table 1, Table 2, Table 3 and Table 4 show the data obtained conditions with heterogeneous initial conditions and, as a re- during the experimental storage of wheat grain samples.

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Table 1 The results of soft wheat grain storage of harvest of 2018 in an actively thermostatically controlled zone with a maintained temperature of 35 ºС (stated in June 2019) FAV, Gluten Period of storage, Sprouting mg КОН Moisture, % Vitrescence, % Germination, % month energy, % per 1 g of fat quantity, % quality, cu, IDC 3rd class Starting material 11.5 25.6 62.5 13.4 68 66 92

1 11.6 2 11.4 3 10.9 25.8 60 8.8 68 65 94 4 9.6 5 11.7 6 11.5 25.4 60 7.8 67 66 93 4th class Starting material 7.3 22.0 85 11.8 60 93 97

1 7.8 22.0 85 11.3 60 87 96 2 7.5 3 8.2 22.2 80 10.8 61 85 94 4 9.2 5 8.6 6 8.5 21.6 60 8.0 61 67 97 7 8.9 8 8.9 9 8.5 21.8 65 7.6 62 72 96 4th class (treated with oil glazing) Starting material 7.3 22.0 85 11.8 60 93 97

1 7.4 22.2 85 10.6 59 86 95 2 7.4 22.4 80 9.9 60 86 95 3 8.5 4 8.6 22.0 75 9.0 60 84 94 5 8.5 6 8.7 7 8.8 22.0 80 8.2 61 83 94

Table 2 The results of soft wheat grain storage of the harvest of 2018 under natural environmental conditions (in a box subjected to a regular exposure of direct sunlight, stated in June 2019) FAV, Gluten Period of storage, Sprouting mg КОН Moisture, % Vitrescence, % Germination, % month energy, % per 1 g of fat quantity, % quality, cu, IDC 3rd class Starting material 11.5 25.6 62.5 13.4 68 66 92

1 11.0 2 10.8 3 10.1 25.6 65 12.0 73 74 95 4 10.4 5 10.7 6 11.0 25.4 65 11.6 70 72 94 4th class Starting material 7.3 22.0 85 11.8 60 93 97

1 7.2 22.2 85 10.8 60 88 96 2 7.1 3 7.9 22.0 80 9.8 58 87 94 4 7.3 5 8.9 6 9.8 21.4 75 11.0 68 63 96 7 8.2 8 8.8 9 9.0 22.0 75 10.4 62 65 95

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Table 3 Storage results of soft wheat grain of harvest of 2018 in natural environmental conditions (in a metal container not subjected to regular exposure of direct sunlight, stated in June 2019) FAN, Gluten Period of storage, Sprouting mg КОН Moisture, % Vitrescence, % Germination, % month energy, % per 1 g of fat quantity, % quality, cu, IDC 3rd class Starting material 11.5 25.6 62.5 13.4 68 66 92

1 10.2 2 12.5 3 12.9 25.8 62.5 11.5 69 69 95 4 9.9 5 13.1 6 13.3 25.6 65.0 11.6 67 68 93 4th class Starting material 7.3 22.0 85 11.8 60 93 97

1 7.2 22.4 85 11.4 62 89 97 2 7.7 3 8.1 22.0 80 10.9 63 87 95 4 8.3 5 8.5 6 10.9 22.0 70 11.2 59 63 96 7 8.7 8 9.2 9 9.4 22.2 75 10.4 60 64 97

Table 4 The results of the analysis of a wheat grain sample subjected to dust suppression by oil glazing in an industrial environment FAV, Gluten Sprouting energy, mg КОН per Moisture, % Vitrescence, % Germination, % % 1 g of fat quantity, % quality, cu, IDC 8.9 23.0 75 12.8 62 87 93

An analysis of the data presented in the tables allows us to What can serve as explanations for these preliminary con- preliminarily state the following: it is not possible to identify clusions? From our point of view, there can be several reasons significant changes in the FAV parameter in experimental sam- for such results. In particular, one of the most probable reasons ples of wheat grain stored under conditions of South Russia for is that the grain is gradually dried (however it was initially in a both subjected to oil glazing and stored since 2016 at stable high dry state) under the conditions of increased temperature dur- (35 ºС) temperature. ing the experimental storage inherent to the conditions of South So, as shown in Table 1, there were no significant changes in Russia. As a result, moisture inside and on the surface of grain FAV value (11.6 mg KOH/1 g of fat) of 3rd class wheat stored for 6 acquires a more bound state leading to a substantially slowing months at a stably high temperature of 35 ºС. Other parameters down of biochemical and microbiological processes, which are of product classification also remained practically unchanged. the initiators of the start stages of grain self-heating and spoil- Wheat of the 4th class under the same conditions showed slight- age including the oxidative processes leading to the formation ly different but close results: the value of the FAV parameter of free fatty acids. during storage increased from 7.8 mg KOH/1 g of fat to 8.5 mg This situation significantly distinguishes the results of the KOH/1 g of fat, which increase can be considered insignificant. planned modelling of South storage conditions from the results The situation here is similar to the other parameters of product that could be obtained in real industrial grain storage conditions classification. in South Russia. Here it is understood that one of the main factor Table 2 and table 3 show the data obtained during storage which affects the potential deterioration in the quality preserva- of wheat grain samples in natural climatic conditions in un- tion of grain mass under real industrial conditions is the redis- shaded and shaded areas, respectively. Analysis of the data in tribution of moisture inside the grain mass. This results in local these tables shows that the FAV values have a slight tendency to drying of some zones and the concentration of moisture in other increase. At the same time changes of FAV of the 3rd class wheat zones of the grain terminal capacity. However, such a redistribu- stored in the zone with intensive insolation for 6 months were tion of moisture is caused by the moisture transfer under the in- not recorded with the basic value of 11 mg KOH/1 g of fat, where- fluence of external factors, mainly by the temperature gradient as such changes for wheat of the 4th class amounted as 2.6 mg and condensation effects that occur on the inner surface shells, KOH/1 g fat (from 9.8 mg KOH/1 g fat to 7.2 mg KOH/1 g of fat). especially for metallic hoppers. In the shaded zone the changes in FAV value were: 3.1 mg Moreover, it can be emphasized that the intensity of micro- KOH/1 g of fat (with 10.2 mg KOH/1 g of fat and 13.3 mg KOH/1 biological and biochemical processes in the grain is affected not g of fat) for the 3rd class, and 3.7 mg KOH/1 g of fat (c 7.2 mg by the mass fraction of moisture or moisture content in the grain KOH/1 g of fat to 10.9 mg KOH/1 g of fat) for the 4th class. Slight- as such, but by the amount of bound moisture or the moisture ly higher moisture values were​​ observed for the shaded zone. state of the grain. This parameter that uniquely characterizes

29 ПИЩЕВЫЕ СИСТЕМЫ | Том 2 № 4 | 2019 FOOD SYSTEMS | Volume 2 № 4 | 2019 the moisture binding of a material is the well-known as water velopment of microbiological and biochemical processes in the activity or Aw, which is widely used in many branches of the food products. These known threshold values ​​are: Aw < 0.65 — mois- and pharmaceutical industries of the developed countries [7]. For ture is completely inaccessible; 0.65 < Aw < 0.70 — moisture is grain, in particular, this parameter is numerically equivalent to partially available; Aw > 0.75 — moisture is widely available [9]. the relative humidity expressed in fractions of unit or the rela- With regard to grain, in particular wheat grain, such mois- tive humidity of intergranular air in thermodynamic equilibrium ture categories can be defined as «dry grain», «medium dry with grain. The practical implementation of this method does grain», «wet grain». not cause technical difficulties. The procedure for determination In order to confirm the relation between the critical values​​of of Aw actually comes down to measurement of the relative hu- the «grain moisture content» category and the critical values ​​of midity of intergranular air under conditions of thermodynamic the «grain moisture state» category, we can provide known data equilibrium. on the equilibrium moisture content in grain and intergranu- Aw = Pw / Po = Pvv / 100, (1) lar air in comparison with the data obtained from the normative documentation on the critical moisture values ​​of various types where: Pw is the pressure of water vapor in the food product system; of grain. Po is the vapor pressure of pure water; Rvv is a relative humidity in equilibrium in which the product does 4. Conclusion not absorb moisture and does not lose it into the atmosphere, %. The two-year experimental storage of samples of food-grade wheat including the grain samples treated with oil glazing under The indicated ratio is included in the basic thermodynamic modelling conditions of South Russia has not yet revealed the equation for determination the binding energy of moisture with expected significant changes in FAV value. In order to establish a material (Rebinder equation) [8]: the kinetic laws of this parameter change over a wide time range L = – R · T · Ln (Aw) (2) a continued experimental storage is required. Uniform oil glazing of a grain sample with an adjustable where: L is the work of separation of 1 mole of water from the dry base of the norm in laboratory conditions can be performed effectively due material (without changing of the composition); to the interaction of a cyclically pouring of grain sample and its T is the temperature, K; treating with an oil aerosol nozzled by an atomizer. R is the universal gas constant. The state of moisture in grain is potentially a factor that af- fects the kinetic processes course during storage. Defining the From the literature data the general threshold values of ​​Aw degree of moisture state influence on the kinetics changes in are known at which moisture is in a state accessible for the de- FAV value requires an additional cycle of research.

REFERENCES 1. Priezzheva, L.G., Meleshkina, E.P., Sorochinskii V. F., Verezhnikova, I.A., 6. Markov, Yu. F. (2019). Instruments for monitoring the state of quality Ignatova, L.G., Koval, A.I. (2017). Long-term storage of wheat flour in preservation of grain. Collection of materials of the 16th all-Russian scien‑ laboratory and production conditions. Bread products, 10, 44–47. (In Rus- tific and practical Conference, 16–22. (In Russian) sian) 7. Barbosa-Cánovas, G.V., Fontana Jr., A.J., Schmidt S. J., Labuza, T.P. (2007). Wa- 2. Markov, Yu.F., Buriak, A.G., Eresko, L.G. (2019). Modern methods, tools ter Activity in Foods. Fundamentals and Applications. Blackwell Publishing and standards in the field of assessment the quality of grain and grain and the Institute of Food Technologists. — 440 p. ISBN: 978–0–813–82408–6 products. Bread products, 7, 23–25. (In Russian) 8. Ermolaev, V.A., Shushpannikov, A.B. (2010). Investigation of water activ- 3. U. S. Congress, Office of Technology Assessment, Technology and Policy ity index of dry dairy products. Food processing: techniques and technol‑ for Suppressing Grain Dust Explosions in Storage Facilities, OTA-BP- ogy, 2, 20–25. (In Russian) ENV, Washington, DC, September 1995. 9. Safonova, Yu. A., Zharkova, I. M., Barinov. A. S. (2017). Influence of activ- 4. Priezzheva, L.G., Shuhnov, A.F. (2010). Method of definition of acid num- ity of water on properties of raw materials at storage. Bread products, 12, ber of fat in products of processing of grain. Food industry, 12, 61–63. 52–55. (In Russian) (In Russian) 10. Kemerbaev, A. Yu. (2001). The role of water in food and its functions. Al- 5. Markov Yu.F., Palladiev, A.A., Eresko L. G. (2016). Assessment of dust- maty: Mariya. —203 p. (In Russian) forming properties of the grain mass. The collection of materials of the 11. Zakladnoi, G.A., Dogadin, A.L., Abdiushev, М., Soskin, M., Markov, III all-Russian scientific-practical conference of young scientists and gradu‑ Yu. F. Metal silo has been turned into safe keeping grain facility. 9th ate students. All-Russian Research Institute of Tobacco, Shag and Tobacco Conference on Integrated Protection of Stored Products ISPS France, Products, 296–304. (In Russian) Talence: Agora — University Bordeaux 1, 20.

AUTНOR INFORMATION Yuri F. Markov — candidate of technical sciences, deputy director, Kuban branch of V. M. Gorbatov Federal Research Center for Food Systems of Russian Acad- emy of Sciences. 350042, Krasnodar, Kolhoznaya st., 3, Tel.: +7–861–255–30–02, E‑mail: [email protected] *corresponding author Alexandra N. Buriak — senior scientist, Kuban branch of V. M. Gorbatov Federal Research Center for Food Systems of Russian Academy of Sciences. 350042, Krasnodar, Kolhoznaya st., 3, Tel.: +7–861–255–30–02, E‑mail: [email protected] Larisa G. Eresko — senior scientist, Kuban branch of V. M. Gorbatov Federal Research Center for Food Systems of Russian Academy of Sciences. 350042, Kras- nodar, Kolhoznaya st., 3, Tel.: +7–861–255–30–02, E‑mail: [email protected] All authors bear responsibility for the work and presented data. All authors made an equal contribution to the work. The authors were equally involved in writing the manuscript and bear the equal responsibility for plagiarism. The authors declare no conflict of interest. Received 25.07.2019 Accepted in revised 30.10.2019 Accepted for publication 02.12.2019

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UDC 664:543.635: 57.084 DOI: 10.21323/2618–9771–2019–2–4–31–33 Original scientific paper DEVELOPING THE METHODS OF FOOD PRESERVATIVES EXTRACTION FROM COMPLEX MATRICES FOR BIOASSAY PURPOSE Artem V. Samoilov*, Natalia M. Suraeva, Sergey V. Koptsev, Sergey V. Glazkov All-Russian Scientific Research Institute of Technology of Preservation — Branch of the V. M. Gorbatov Federal Research Center for Food Systems of RAS, Vidnoye, Moscow Region, Russia

KEY WORDS: ABSTRACT sorbic acid, benzoic acid, The presence of food additives in food products may be associated with the risk of their toxic effects on hu- extraction, bioassay, man body. Therefore, the study of approaches to testing their safety seems to be a particularly urgent task. the Allium test, preserved The aim of this study was to determine the conditions for extracting food preservatives from the samples of vegetable products preserved pureed vegetables for further bioassay of the extract obtained in the Allium test. Onion roots were used as a test object in this method. Two extraction methods of benzoic and sorbic acids added to pureed vegetables have been developed. Distilled water and acetone were used as extracting solutions. The extrac- tion efficiency was evaluated on Shimadzu Prominence LC‑20 liquid chromatograph (Japan) in the ultraviolet range, wavelength 235 nm (benzoic acid), 285 nm (sorbic acid). According to the results of studies using both water and acetone as extractants, the degree of preservatives extraction was approximately the same and quite high. In the quantitative calculation of the preservatives content in pureed vegetables, the value of the correction factor was 0.8. However, due to certain production characteristics of this product, i. e. the stage of cauliflower homogenization, obtaining an extract with acetone seems to be more acceptable for the Allium test conditions.

1. Introduction also been published, in which onion roots were placed directly Focus on quality and safety of foods is currently increases, in food products such as drinks, juices, and milk [7,8,9]. How- including the approaches to healthy eating and minimizing ever, these experiments are not so unambiguous, because high the risks of diseases associated with the diet of various popu- concentration of carbohydrates in drinks and juices, and the lations. Particular attention is paid to food additives, as it is presence of emulsion in milk are undesirable factors in the Al- known that their use may cause toxic effects in the human lium test. It seems more appropriate to test extracts of food body. Long-term consumption of certain additives with food products that are acceptable for the Allium test, which allows is associated with the development of carcinogenic diseases, simultaneous and comprehensive assessment of various food pathological changes in reproductive system and digestive additives and their mixtures. Such tests are especially relevant tract [1,2,3]. However, these additives are often impossible to when developing new types of food products based on original replace in the mass production of foods, where they are used recipes or technological methods of processing and storage. as preservatives, sweeteners, colorants, emulsifiers, flavor en- In the literature, we didn’t find the data on extracts of food hancers and other components. Moreover, most products con- products to assess their toxicity in the Allium test, apart from tain a mixture of additives, the toxic effect of which may have the only work to detect pesticides in unwashed vegetables not only additive, but also a synergistic nature. It is important and grapes [10]. Therefore, the aim of the work was to develop to note that the products of these additives’ oxidation formed methods for extracting food preservatives from preserved pu- during the production and storage of foods are also chemically reed vegetables, which are acceptable for further bioassay of active compounds and show mutagenic activity [4]. these extracts in the Allium test. All these problems require the development of a new methodology for testing products, which will lead to further 2. Materials and methods revision of the toxicological hazard thresholds. Thus, it is pro- Benzoic acid (SIGMA-ALDRICH, lot.MKCG6487, Germany), posed to concentrate the main efforts on studying the mecha- sorbic acid (SIGMA-ALDRICH, lot.SLBW6722, Germany), and nisms of toxicant effects at the cellular and molecular level pureed cauliflower for infant nutrition purchased from a retail using modern methods of systems biology and bioinformat- network were used as the test objects. ics, which allow predicting toxicity at the whole-body level. Preservatives (benzoic and sorbic acids) were added by weight At the same time, the problem is being discussed of refusing to pureed vegetables individually and in a mixture, with a final animal experiments when possible [5]. Therefore, the use of mass fraction of 100 mg/kg. vegetable test systems may be preferable in solving these sci- entific problems. We believe that the Allium test, in which 3.1. Preparing the benzoic and sorbic acid stock solutions onion roots are used as a test object, meets the specified con- at a concentration of 1000 mg/dm3 ditions in the best way. This test is highly sensitive both to In a 100 cm3 volumetric flask, the corresponding acids were macro indicators (delayed root growth) and micro indicators weighed in amount of 0.1 g within the accuracy of 0.001 g, sepa- (cytotoxicity, genotoxicity, level of oxidative activity biomark- rately and together. 50 cm3 of water was added to the mixture ers, etc.) and is especially suitable for the analysis of complex of acids followed by dissolving in a water bath at a temperature mixtures of chemical compounds. Currently, the Allium test of 70 ± 5 °C for 30 minutes. The resulting solution was cooled to is widely and successfully used to study the toxic, cytotoxic a temperature of 20 ± 5 °C and diluted with water to the volume and genotoxic effects of food additives [6]. Several papers have of 100 cm3.

FOR CITATION: Samoilov A. V., Suraeva N. M., Koptsev S. V., Glazkov S. V. Developing the methods of food preservatives extraction from complex matrices for bioassay purpose. Food systems. 2019; 2(4): 31–33. DOI: 10.21323/2618–9771–2019–2–4–31–33

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3.2. Preparing a mixture of pureed vegetables benzoic acid extraction from pureed vegetables was high and with the preservatives amounted to 82.6 %. 90 g of pureed cauliflower was placed in three glasses. 10 cm3 Table 1 of the sorbic acid stock solution (prepared according to Section 3.1) Benzoic acid extraction results after aqueous extraction was added to the first glass to obtain a final concentration of The degree of Standard Recovery, % Average 3 benzoic acid deviation S, of the initial 100 mg/kg in pureed vegetables. 10 cm of the benzoic acid stock value, mg/kg extraction W , % mg/kg additive solution (prepared according to Section 3.1) was added to the i second glass to obtain a final concentration of 100 mg/kg in pu- 78.92 reed vegetables. 10 cm3 of the benzoic and sorbic acid mixture 85.00 82.60 3.23 82.6 stock solution (prepared according to Section 3.1) was added to 83.87 the third glass to obtain a final concentration of 100 mg/kg in pureed vegetables. The resulting mixtures were thoroughly mixed However, despite the high degree of benzoic acid extraction, on PE‑6500 laboratory shaker (Acroshim, Russia) for 20 min at the aqueous extract from pureed vegetables did not meet the above room temperature. Allium test parameters, due to the fact that it was a suspension. In bioassay, suspended particles could precipitate on onion roots 3.3. Extracting the preservatives from pureed vegetables and have a negative effect on their growth. We believe that the Mixtures of pureed vegetables with the preservatives (prepared reason for such stable aqueous suspensions is homogenization according to Section 3.2) of 10 g each were transferred to 100 cm3 of raw materials used in the manufacture of this type of pureed volumetric flasks within the accuracy of 0.001 g. 40 cm3 of acetone vegetables. All attempts to eliminate suspension and obtain a true (EKOS‑1, Russia) were added, mixed and placed in an ultrasonic solution, i. e. multiple dilution with water (4–8 times), centrifuga- bath (UNI0TRA UM‑4, Poland) for 15 minutes at room tempera- tion and filtering through filter paper were unsuccessful. On the ture. Then it was diluted with acetone to obtain 100 cm3, mixed other hand, it would be possible to dry pureed vegetables, and then and stored for 12 hours at 5 °C. After 12 hours, acetone mixtures obtain an aqueous extract of the dry residue, but this procedure were removed from the refrigerator, kept at room temperature for is long and laborious and it is necessary to apply high heating about 1 hour, and then filtered through a filter paper (Blue Rib- temperatures, which may lead to the formation of additional bon). 2 cm3 of each mixture filtrates were evaporated to dryness in toxicants in the extract. However, this extraction protocol can be Corning plastic tubes (Mexico) using Thermo Scientific evaporator used, for example, when testing tomato paste. It is known that in (USA) in nitrogen stream for 40 min at 60 °C. Then, 2 cm3 of HPLC the production of tomato-containing products, other technological water (Fisher Chemical, lot. 181206, Belgium) were added to the methods are usually used to achieve homogeneity, i. e. filtration evaporated residue in the same tube and mixed with vortex (IKA and evaporation. In this case, it is most likely that the use of an vortex genius 3, Germany) for 1 min. aqueous extract will be more appropriate. Similar extraction procedures were carried out under the same In order to eliminate the above disadvantages of pureed veg- conditions, but instead of acetone, distilled water was used in the etables aqueous extraction, acetone was used as an extractant in same amounts, excluding the evaporation and reconstruction the following experiments. Benzoic and sorbic acids are known procedures. to dissolve well in this solvent and its subsequent evaporation occurs under relatively mild temperature conditions (60 °C). As 3.4. Determining the preservatives concentration in the extracts shown in Tables 2 and 3, the percentage of extraction for benzoic The mass fraction of benzoic and sorbic acids was determined acid was only 6 % lower than in aqueous solution. Whereas, by high performance liquid chromatography (HPLC) method on the average degree of sorbic acid extraction was even higher Shimadzu Prominence LC‑20 liquid chromatograph (Japan) in the than for benzoic acid and amounted to 79.0 %. We believe that ultraviolet range at a wavelength of 235 nm (benzoic acid) and losses during extraction with acetone were associated with 285 nm (sorbic acid), respectively. insufficient solubility of benzoic and sorbic acids as low-polar A mixture of acetate buffer solution, pH 4.5 (Acros Organics, compounds (pKa 4.20 and 4.76, respectively). Therefore, it may Belgium) and methanol (Merck Millipore, Germany) was used be necessary to further dissolve the dry extract in water at a as the mobile phase. The components were separated using Hy- temperature of 70 °C. Based on the above, in the case of these persil BDS-C18 column (Thermo Scientific, USA), 150 × 4.6 mm, preservatives, when analyzing the toxic effects of extracts in in isocratic mode, with a volume ratio of 75:25, while the eluent the Allium test, the correction factor of 0.8 must be considered. velocity was 1 cm3 per minute. As far as we know, this is the first study on preparing the food The degree of extraction was estimated according to the con- extracts for bioassay, in which the effectiveness of the food ad- centration of the extracted preservatives. ditives extraction was studied. The quantification of extracted toxicants may be critical in understanding the mechanisms of 3. Results and discussion their biological effects. Preserved pureed cauliflower for infant food as a model Table 2 matrix of complex composition was chosen for studies of the Benzoic acid extraction results preservatives toxic effects in the Allium test. Extracts of such after extraction with acetone products are most likely do not contain any natural and syn- The degree of Standard Recovery, % Average benzoic acid deviation S, of the initial thetic toxicants, and therefore, the most accurate assessment value, mg/kg extraction W , % mg/kg additive of the toxic effects of food additives and their mixtures in this i vegetable matrix is ​​possible. According to the literature, in the 67.79 Allium test, onion roots are sprouted, as a rule, in distilled or 79.92 drinking water, in which the test compounds were dissolved; 73.92 and the pH of the solution should be not lower than 3.5 and 79.76 76.31 4.43 76.3 % not higher than 11.0 [11, 12]. Stock solutions of benzoic and 77.61 sorbic acids were within the indicated range. Therefore, in the 79.76 beginning, the simplest extraction option was studied using 75.47 distilled water. According to Table 1, the average degree of

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Table 3 Table 4 Sorbic acid extraction results Preservatives mixture extraction results after extraction with acetone after extraction with acetone The degree Standard Recovery, % The degree Average Standard Recovery, % Average Preser- of sorbic acid deviation S, of the initial of extraction value, deviation S, of the initial value, mg/kg vative extraction Wi, % mg/kg additive Wi, % mg/kg mg/kg additive 77.76 79.76 Benzoic 72.70 80.91 0.66 80.5 acid 80.53 73.77 80.91 71.64 79.0 6.57 79.0 % 78.76 Sorbic 85.00 77.92 78.53 0.54 78.7 acid 86.48 78.92 85.65 4. Conclusion To assess the interactions of preservatives and the specificity As a result of the research, two methods for obtaining extracts of their detection in pureed vegetables during extraction, experi- from preserved pureed vegetables acceptable for further bioassay ments were carried out with their mixture in a concentration of in the Allium test were first studied. For the first time, studies have 100 mg/kg of product. As shown in Table 4, the degree of extrac- been conducted to evaluate the effectiveness of extraction for food tion with acetone did not change significantly; the percentage preservatives added to the original product. The degree of extraction of benzoic and sorbic acids extraction remained within the same of these additives, benzoic and sorbic acids, was high and did not differ range. It should be noted that when both preservatives present between both methods. However, in relation to physical and chemical in the extract, evaluating the results in the Allium test require characteristics, only the acetone extract from this specialized infant the correction coefficient to be increased twofold, as not only an food met the requirements of the Allium test. We believe that the additive, but also a synergistic effect is possible or other, more aqueous extract may also be successfully used in this test for other complex interactions. vegetable products that are not homogenized during the production.

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AUTHOR INFORMATION Artem V. Samoilov — candidate of biological sciences, chief of laboratory, Laboratory of quality and safety of foodstuffs, All-Russian Scientific Research Insti- tute of Technology of Preservation — Branch of the V. M. Gorbatov Federal Research Center for Food Systems of RAS, 142703, Moscow region, Vidnoye, School str, 78, Tel.: +7–495–541–87–00, e-mail: [email protected] *corresponding author Natalia M. Suraeva — doctor of biological sciences, chief researcher, Laboratory of quality and safety of foodstuffs, All-Russian Scientific Research Institute of Technology of Preservation — Branch of the V. M. Gorbatov Federal Research Center for Food Systems of RAS, 142703, Moscow region, Vidnoye, School str, 78, Tel.: +7–495–668–70–37, e-mail: [email protected] Sergey V. Koptsev — researcher, Laboratory of quality and safety of foodstuffs, All-Russian Scientific Research Institute of Technology of Preservation — Branch of the V. M. Gorbatov Federal Research Center for Food Systems of RAS, 142703, Moscow region, Vidnoye, School str, 78, Tel.: +7–495–668–70–37, e-mail: kopt- [email protected] Sergey V. Glazkov — leading researcher, Laboratory of quality and safety of foodstuffs, All-Russian Scientific Research Institute of Technology of Preserva- tion — Branch of the V. M. Gorbatov Federal Research Center for Food Systems of RAS, 142703, Moscow region, Vidnoye, School str, 78, Tel.: +7–495–668–70–37, e-mail: [email protected] All authors bear responsibility for the work and presented data. All authors made an equal contribution to the work. The authors were equally involved in writing the manuscript and bear the equal responsibility for plagiarism. The authors declare no conflict of interest Received 18.10.2019 Accepted in revised 20.11.2019 Accepted for publication 05.12.2019

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UDC 637.14.04/0 DOI: 10.21323/2618–9771–2019–2–4–34–37 Original scientific paper STUDY OF FATTY ACID COMPOSITION OF MILK FOR CHEESE PRODUCTION Elena V. Topnikova*, Valentina A. Mordvinova, Galina M. Sviridenko, Ekaterina S. Danilova All-Russian Scientific Research Institute of Butter- and Cheesemaking — Branch of V. M. Gorbatov Federal Research Center for Food Systems of RAS, Uglich, Yaroslavl Region, Russia

KEY WORDS: ABSTRACT milk, cheese, milk suitability The article shows that the studied samples of raw milk in terms of composition (mass fraction of fat, protein, lac- for cheese, fat phase of milk, tose and milk solids-not-fat) met the criteria of suitability for cheese, and its physic-chemical properties (titrat- fatty acid composition able acidity, density, freezing point) were within acceptable limits. Moreover, rennet sample corresponded to the first and second classes. It was proved that the number of spores of lactating fermenting microorganisms met the requirements for milk for the production of any kind of cheese. It was established that the fatty acid composition of raw cow’s milk used to produce cheeses at various enterprises varied significantly. The data indicate the vari- ability of the content of all groups of fatty acids — low molecular weight, saturated, monounsaturated and poly- unsaturated. As a result of gas chromatography studies using the chromatographic complex «Chromos GX‑1000» with a flame ionization detector and a CP 88 quartz capillary column — Sil 88 for FAME100 m × 0.25 mm × 0.2 µm, it was found that the largest absolute fluctuations were for saturated (± 7.03 % of the average value) and monoun- saturated fatty acids (± 3.77 % of the average value). Absolute fluctuations in the group of low molecular weight fatty acids amounted to ± 2.62 %, and in the group of polyunsaturated — ± 1.02 % of the average value. The calcu- lation of the relative deviation showed that the most varied groups were the ones of low molecular weight fatty acids (± 28.40 rel.%) and polyunsaturated fatty acids (± 25.11 rel.%). At the same time, a relatively high content of certain fatty acids: myristoleic, palmitic, palmitoleic and low levels of stearic and oleic fatty acids, was revealed in individual milk samples.

1. Introduction Moreover, there is insufficient scientific data on how the The quality of any dairy product directly depends on the conditions of keeping and feeding animals affect the fat phase of composition and properties of the raw milk used, as well as on milk and its fatty acid composition. the features of technological methods and processing modes Currently, when milk is accepted at domestic enterprises, used in its manufacture. studies of the fat phase are extremely rare. In the European prac- In the cheese production, it is extremely important to use tice of cheesemaking, it is quite common to determine free fatty «milk suitable for cheese». This concept means a set of proper- acids in raw milk intended for making cheese, which can cause ties associated with the usefulness of the milk composition, not only a decrease in organoleptic characteristics and cheese its ability to coagulate under the action of rennet and to be spoilage, but also serve as inhibitors of the lactic acid process in a favorable environment for the development of starter mi- its manufacture [7]. croorganisms used in the cheese manufacture [1]. Consider- When identifying cheeses in circulation, the fatty acid com- ing this, during acceptance tests of milk at cheesemaking en- position is a very important indicator of this group of products terprises, its physical and chemical properties are evaluated: in terms of their naturalness. It is determined along with the or- density, titratable acidity, and freezing point. The composition ganoleptic characteristics of cheese and indicators of its physi- indicators are determined: the content of milk dry fat-free co-chemical composition. Many fatty acids and their derivatives substances, the mass fraction of fat, and protein. The rennet are involved in the formation of a characteristic cheese flavor test is determined and evaluated according to the indicators of and odor [8]. bacterial contamination, the absence of inhibitory substances, In this regard, the study of the fatty acid composition of milk including antibiotics. suitable for cheese is relevant. The data obtained will be the Moreover, for cheesemaking it is important to know not only starting point for studying changes in the fatty acid composition the total protein content in raw milk, but also the amount of true in the process of cheesemaking and substantiating the identi- protein, as well as the ratio of whey proteins and caseins. These fication indicators of cheeses made using various technologies milk quality criteria are not mandatory for milk acceptance, but related to their fat phase. directly affect the yield of cheese and the quality of the finished Based on the relevance of the issue, the aim of this work was product. Therefore, with a low yield of cheese from processed to study the fatty acid composition of milk used at different en- milk, it is advisable to determine the content of protein and non- terprises of the industry for the manufacture of cheeses of dif- protein nitrogen, calculating the content of the true protein, in- ferent species groups at different times of the year. cluding casein, as the main structural component of cheeses [2]. Scientific studies give great attention to the protein compo- 2. Materials and methods nent of cow’s milk. In addition, the content of various fractions Raw cow’s milk obtained in different regions and in different of casein and whey proteins is studied depending on the breeds seasons of the year served as research objects. 25 samples of raw of animals and their feeding diets [3,4]. The presence of genes cow’s milk were examined. that contribute to a simultaneous increase in milk yield and its To assess its suitability for cheese, standardized methods protein content is established [5,6]. For improving these particu- were used to determine the composition (mass fraction of fat, lar indicators, corresponding diets for feeding lactating animals protein, lactose and milk solids-not-fat) and physico-chemical are being developed. properties of raw milk (titratable acidity, density, freezing point).

FOR CITATION: Topnikova E. V., Mordvinova V. A., Sviridenko G. M., Danilova E. N. Study of fatty acid composition of milk for cheese production. Food systems. 2019; 2(4): 34–37. DOI: 10.21323/2618–9771–2019–2–4–34–37

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The rennet test, the presence / absence of antibiotics and inhibi- sample 1 mm3; injector temperature 220 °C; temperature pro- tory substances were determined. The number of somatic cells, gram of the thermostat: 1) 100 °C — 4 min, then a temperature the number of mesophilic aerobic and facultative anaerobic mi- increase of 5 °C for 20 min; 2) 170 °C — 20 min, a temperature croorganisms, and the number of spores of lactic-fermenting increase of 5 °C for 9 min; 3) 215 °C — 30 min (analysis dura- microorganisms determined the evaluation of the microbiologi- tion — 77 min); carrier gas — nitrogen (pressure in front of the cal suitability of milk for the cheese manufacture. Its organolep- column — 2.7 kgf/cm2). A standard mixture of Supelko 37 Com- tic characteristics were evaluated as well. ponent FAME Mix fatty acid methyl esters (Supelko, USA) was To determine the fatty acid composition of raw milk, a fat used as an identification mixture. Processing of the obtained phase was isolated from it. With this purpose, the analyzed sam- data was carried out by the method of internal normalization ples were placed in two centrifuge tubes of 50 cm3, centrifuged using the «Chromos» program. at 10,000 min‑1 for 15 min, and the upper fat fraction was taken after centrifugation. The resulting fat fraction was placed in a 3. Results and discussion glass with a capacity of 250 cm3, 150 cm3 of hexane were added Quality indicators of raw cow’s milk used to make cheese are (Vekton JSC, Russia), carefully mixed with a blender at maximum shown in Table 1. speed for 1 min, and the hexane layer with the dissolved fat was According to the data of the table, all the milk used for the separated. cheeses manufacture met the criteria of suitability for cheese in The resulting solutions of milk fat fractions were transferred terms of organoleptic, physico-chemical and microbiological in- to round-bottom flasks with a capacity of 250 cm3 for subsequent dicators. Rennet, as a specific indicator of suitability for cheese, removal of the solution, which was carried out at a temperature corresponded to the first and second classes. The number of of 70 °C using a rotary evaporator. spores of lactate-fermenting clostridia met the requirements for To obtain methyl esters of fatty acids, 0.1 g of fat was trans- milk for the production of any kind of cheese. ferred to a 10 ml centrifuge tube, 2 cm3 of hexane and 100 μl of a The results of evaluating the fatty acid composition of the 2 M solution of sodium methylate in methanol (Sigma-Aldrich, studied milk are shown in Table 2. According to the data ob- USA) were added; the tube was closed with a stopper, vigorously tained, the fatty acid composition of raw cow’s milk used to mixed for 2 min, it was settled for 5 min to separate the trans- produce cheeses at various enterprises varied significantly. The parent upper phase containing methyl esters, which was then data indicate the variability of the content of all groups of fatty transferred to the vial and used to determine the fatty acid com- acids — low molecular weight, saturated, monounsaturated and position. polyunsaturated. Fatty acid composition tests were performed on a Chromos The largest absolute fluctuations were found for saturat- GX‑1000 chromatographic complex with a flame ionization ed (± 7.03 % of the average) and monounsaturated fatty acids detector (Chromos LLC, Russia) and a CP 88 silica capillary (± 3.77 % of the average) groups. Absolute fluctuations in the column Sil 88 for FAME100m × 0.25mm × 0.2µm (Agilent Tech- group of low molecular weight fatty acids amounted to ± 2.62 %, nologies, USA). During the research, the following chromato- and in the group of polyunsaturated — ± 1.02 % of the average graphic modes were established: the volume of the introduced value. The calculation of the relative deviation showed that the Table 1 Indicators of the investigated raw milk for the cheese manufacture № (item Name of indicator Indicator value Value of the admissible indicator number) 1. Organoleptic indicators: Clean, free from foreign odors and flavors, not Clean, free from foreign odors and flavors, —— flavor and odor peculiar to the fresh natural milk. Mild weedy not peculiar to the fresh natural milk flavor and odor are allowed —— color From white to light cream From white to light cream Homogeneous liquid without sediment and Homogeneous liquid without sediment and flakes. —— consistence flakes Freezing is not allowed. 2. Mass fraction, %: —— solid-not-fat 8.3–9.2 Not lower than 8.2 —— fat 3.2–4.6 Minimum 3.2 —— protein 3.0–3.4 Not lower than 3.0 3. Density, kg/m3 1027–1028 Not lower than 1027 4. Titratable acidity, оТ 16.0–19.0 From 16 to 18 5. Freezing point, °С 0.520–0.528 Not lower than 0.520 6. Rennet, class I–II Not lower than I–II Number of mesophilic aerobic and facultative 7. 3.3 · 104–5,0 · 105 Maximum 5 · 105 anaerobic microorganisms, CFU/см3 For cheeses with a low temperature of the second Number of spores of lactic-fermenting heating — not more than 13000 spores/dm3; 8. 11–600 microorganisms, spore/dm3 For cheeses with a high temperature of the second heating — no more than 2500 spores/dm3. 9. Number of somatic cells, cells / cm3 5 · 104–4 · 105 Maximum 5 · 105 10. Inhibitory substances Not found Not allowed 11. Antibiotics Not found Not allowed

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Table 2 Fatty acid composition of milk used in different regions for cheese production

Indicators for the fatty acid composition of the investigated milk (n** = 25) Name of fatty acid ± Standard deviation at P = 0.95 Range Average value (for n determination results) С4:0 2.58–3.78 3.05 0.25 С6:0 1.73–2.71 2.08 0.21 С8:0 1.06–1,66 1.25 0.13 С10:0 2.45–3.69 2.83 0.32 С10:1 0.21–0.37 0.25 0.04 С12:0 2.78–4.14 3.30 0.37 С14:0 9.84–12.71 11.06 0.89 С14:1* 1.27–2.28 1.64 0.21 С16:0 24.80–35.93 29.13 3.01 С16:1* 1.85–2.84 2.36 0.29 С18:0 7.69–12.86 10.07 1.39 С18:1* 18.78–27.89 24.26 2.87 С18:2* 2.60–4.55 3.50 0.55 С18:3* 0.22–0.77 0.54 0.16 С20:0 0.15–0.31 0.18 0.03 С22:0 0.05–0.15 0.08 0.02 Others 3.56–5.74 4.43 0.48 Low molecular weight 7.88–11.84 9.22 0.23 Saturated 59.01–70.06 63.03 3.46 Monounsaturated 22.41–32.03 28.26 2.96 Polyunsaturated 2.92–5.06 4.04 0.63 Notes: * the calculation was made by the sum of the isomers; ** n is the number of determination results. most varied groups were the ones of low molecular weight fatty other meal, glycerin, palm fat and other energy supplements de- acids (± 28.40 rel.%) and polyunsaturated fatty acids (± 25.11 scribed in the literature [17,22,23,24]. rel.%). Fluctuations in the fatty acid composition of the studied All identified features of the fatty acid composition of milk milk is consistent with data from other scientists, whose publica- can affect the nutritional value of the cheeses made from it and tions indicate the dependence of this indicator on many factors: affect the formation of organoleptic characteristics of cheese. season of the year and climatic conditions [9,10,11,12,13], breed of animals [14,15,16] and their feeding diets [17,18,19,20,21]. 4. Conclusion An increase in the content of polyunsaturated fatty acids Significant fluctuations were found in the fatty acid compo- was characteristic of summer milk. This is consistent with simi- sition of cow’s milk used to make cheese. The greatest relative lar ideas of foreign and domestic researchers, who attribute the fluctuations were registered at the levels of polyunsaturated increase in this indicator to the consuming of green food by lac- fatty acids and low molecular weight fatty acids. tating animals in the summer period [10,12]. Studies have also As there is no possibility to determine the fatty acid profile found that individual milk samples were characterized by an in- in each batch of milk at the acceptance control, it is advisable to creased content of the following fatty acids: capric, myristoleic, adjust the standard values ​​of the fatty acid profile for cheese, in- palmitic and palmitoleic in relation to the values ​​of these indi- cluding variations associated with a change in the fatty acid pro- cators specified for milk fat in national regulatory documents. file of milk depending on the conditions of keeping and feeding In some samples, a reduced content of stearic and oleic acids animals. It requires the expansion of research in the collection was found in relation to these documents. Such deviations may of more extensive statistical material on the fatty acid profile of be associated with special diets for feeding lactating animals, milk suitable for cheese and cheeses made from it of different including the use of highly concentrated feeds, sunflower and species groups.

REFERENCES 1. Sviridenko, G.M. (2009). Microbiological risks in the production of milk 5. Bigaeva A. V. (2019). Comparative analysis of cow milk technological prop- and dairy Moscow: Russian Academy of Agricultural Sciences. — 246 p. erties with different genes of ϰ-casein. XIII International Scientific and ISBN: 978–5–85941–338–6 (in Russian) Practical Conference of Young Scientists and Specialists of Organizations in 2. Myagkonosov, D.S., Mordvinova, V.A., Abramov, D.V., Delitskaya, I.N. the Field of Agricultural Sciences «Advanced research and new approaches (2014). Special features of proteolysis in different groups of cheese types. to the production and processing of agricultural raw materials and food Cheesemaking and buttermaking, 2, 24–27. (in Russian) products.» Uglich: VNIIMS, 52–57. ISBN978–5–6043305–1–7 (in Russian) 3. Ignat’eva, N.L. (2019). Protein content and structure in milk of cows- 6. Tyulkin S.V. (2018). The effect of cows genotype on their productivity and daughters of bulls of domestic and import selection. All-Russian scientif- Milk quality. Food systems, 1(3), 38–43. DOI: 10.21323/2618-9771-2018- ic-practical conference «Advanced achievements of science in the dairy 1-3-38-43 (in Russian) industry». Vologda: GMKHA, 216–221. (in Russian) 7. Kukhtyn, M.D., Pokotylo, O.S., Karpyk, G.V., Kravchenyik, K.J., Shynka- 4. Borovitskiy, M.V. (2012). Study of the influence of the breed of cows on ruk, O.J. (2015). Сhanges in the content of free fatty acids composition of the composition and properties of milk and cheese production. Author’s the milk under the influence of psychotropic microorganisms. Scientific abstract of the dissertation for the scientific degree of Candidate of Tech- Messenger of LNU of Veterinary Medicine and Biotechnologies. Series: nical Sciences. Kemerovo: КеmТIPP. — 16 p. (in Russian) Veterinary Sciences, 17(1–4 (61), 50–55. (in Ukrainian)

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8. Lopez, Ch., Maillard, M.-B., Briard-Bion, B., Camier, B., Hannon, J. A. 16. Khastayeva, A.Z., Smagulov, А.К., Zhamurova, V.S., Topnikova, E.V., (2006). Lipolysis during Ripening of Emmental Cheese Considering Umirzakov, B.U., Nurgalieva, M.T. (2019). Technological properties of Organization of Fat and Preferential Localization of Bacteria. Journal milk from the cows of black-motley and alatau breeds in various seasons of Agricultural and Food Chemistry, 54 (16), 5855–5867. DOI: 10.1021/ of lactation. International Journal of Innovative Technology and Exploring jf060214l Engineering, 8(8), 2299–2303 9. Appalonova, I.V., Smirnova, E.A., Nikonorova, N.P. (2012). Study of fatty 17. Chouinard, P.Y., Girard, V., Brisson, G.J. (1998). Fatty Acid Profile and acid composition of milk lipids. Food industry, 11, 72–75. (in Russian) Physical Properties of Milk Fat from Cows fed Calcium Salts of Fatty Ac- 10. Samoylov, A.V., Kolpakov, E. Yu., Suraeva, N.M., Petrov, A.N., Volodarska- ids with Varying Unsaturation. Journal of Dairy Science, 81(2), 471–481. ya, T.K., Goreva, T.A (2017). Seasonal variations in fatty acids composi- DOI: 10.3168/jds.S0022–0302(98)75599–7 tion of cow’s milk. Bulletin of KrasGAU, 9, 35–40. (in Russian) 18. Collomb, M., Sollberger, H., Bütikofer, U., Sieber, R., Stoll, W., Schaeren, 11. Abilleira, E., Collomb, M., Schlichtherle-Cerny, H., Virto, M, De Renobales, W. (2004). Impact of a basal diet of hay and fodder beet supplemented M., Barron, L.J.R. (2009). Winter/spring changes in fatty acid composition with rapeseed, linseed and sunflowerseed on the fatty acid composition of farmhouse Idiazabal cheese due to different flock management sys- of milk fat. International Dairy Journal, 14(6), 549–559. DOI: 10.1016/j. tems. Journal of Agricultural and Food Chemistry, 57(11), 4746–4753. DOI: idairyj.2003.11.004 10.1021/jf900460u 19. Postavneva, E.V. (2010). Chemical composition of milk of Black-and- 12. Heck, J.M.L., Van Valenberg, H.J.F., Dijkstra, J., van Hooijdonk, A.C.M. White cows of various genotypes. Zootechniya, 1, 30–31. (in Russian) (2009). Seasonal variation-in the Dutch bovine raw milk composition. 20. Kharitonov, E.L., Panyushkin D. E. (2016). Feed and metabolic factors of Journal of Dairy Science, 92(10), 4745–4755. DOI: 10.3168/jds.2009–2146 the milk fatty acid composition in cows. Problems of Biology producing 13. Larsen, M.K., Nielsen, J.H., Butler, G., Leifert, C., Slots, T., Kristiansen, animals, 2, 76–106. (in Russian) G.H., Gustafsson, A.H. (2010). Milk quality as affected by feeding regi- 21. Buryakov, N.P. (2009). Feeding high producing dairy cattle. Мoscow: mens in a country with climatic variation. Journal of Dairy Science, 93(7), Prospect. — 416 p. ISBN: 978–5–98597–148–4 (in Russian) 2863–2873. DOI: 10.3168/jds.2009–2953 22. Kasatkin, I.A., Serkova, A.N. (2019). Energy supplement for highly productive 14. Soyeurt, H., Dardenne, P., Gillon, A., Croquet, C., Vanderick, S., Mayeres, cows. All-Russian scientific-practical conference «Advanced achievements P., Bertozzi, C., Gengler, N. (2006). Variation in fatty acid contents of milk of science in the dairy industry.» Vologda: GMKHA, 271–221. (in Russian) and milk fat within and across breeds. Journal of Dairy Science, 89(12), 23. Yurin, D.A., Yurina, N.A., Esaulenko, N.N. (2017). Effective approaches 4858–4865. DOI: 10.3168/jds.S0022–0302(06)72534–6 to feeding highly productive cows. Effective Livestock, 1(131), 16–18. 15. DePeters, E.J., German, J.B., Taylor, S.J., Essex, S.T., Perez-Monti, H. (in Russian) (2001). Fatty acid and triglyceride composition of milk fat from lactating 24. Gusarov, I.V., Fomenko, P.A., Bogatyrev, E.V. (2019). Bulk feed in the diet Holstein cows in response to supplemental canola oil. Journal of Dairy of dairy cows in the Vologda region. Cheesemaking and buttermaking, 5, Science, 84(4), 929–936. DOI: 10.3168/jds.S0022–0302(01)74550-X 54–56. (in Russian)

AUTНOR INFORMATION Elena V. Topnikova — doctor of technical sciences, director, All-Russian Scientific Research Institute of Butter- and Cheesemaking — Branch of V. M. Gor- batov Federal Research Center for Food Systems of RAS. 152613, Yaroslavl Region, Uglich, Krasnoarmeysky Boulevard, 19, Tel .: + 7–910–666–93–93, E‑mail: [email protected] *corresponding author Valentina A. Mordvinova — candidate of technical sciences, head of research department of cheesemaking technology and serum processing, All-Russian Sci- entific Research Institute of Butter- and Cheesemaking — Branch of V. M. Gorbatov Federal Research Center for Food Systems of RAS. 152613, Yaroslavl Region, Uglich, Krasnoarmeysky Boulevard, 19, Tel.: +7–915–970–36–38, E‑mail: [email protected] Galina M. Sviridenko — doctor of technical sciences, chief research scientist, head of microbiological research department of milk and dairy products, All-Rus- sian Scientific Research Institute of Butter- and Cheesemaking — Branch of V. M. Gorbatov Federal Research Center for Food Systems of RAS. 152613, Yaroslavl Region, Uglich, Krasnoarmeysky Boulevard, 19, Tel .: + 7–903–823–56–88, E‑mail: [email protected] Ekaterina N. Danilova — junior researcher of research department in butter technology, All-Russian Scientific Research Institute of Butter- and Cheesemak- ing — Branch of V. M. Gorbatov Federal Research Center for Food Systems of RAS. 152613, Yaroslavl Region, Uglich, Krasnoarmeysky Boulevard, 19, Tel.: + 7–960–527–61–48, E‑mail: [email protected] All authors bear responsibility for the work and presented data. All authors made an equal contribution to the work. The authors were equally involved in writing the manuscript and bear the equal responsibility for plagiarism. The authors declare no conflict of interest. Received 21.11.2019 Accepted in revised 05.12.2019 Accepted for publication 17.12.2019

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UDC62/66 DOI: 10.21323/2618–9771–2019–2–4–38–41 Original scientific paper MECHANISMS OF DESTRUCTION AND SYNTHESIS OF LIQUID MEDIA, USED IN THE FOOD INDUSTRY UNDER NON-EQUILIBRIUM CONDITIONS Tatyana V. Savenkova1*, Alexander R. Karimov2,3, Mikhail A. Taleysnik1, Timofei V. Gerasimov1, Nikolai B. Kondratev1 1 All-Russian Scientific Research Institute of Confectionery Industry — Branch of V. M. Gorbatov’s Federal Research Center for Food Systems of RAS, Moscow, Russia 2 National Research Nuclear University «MEPhI» Moscow, Russia 3 Institute for High Temperatures of RAS, Moscow, Russia

KEY WORDS: ABSTRACT liquid medium, invert syrup, The formation of food liquid medium structures containing at least 70 % of disperse particles with high disper- cavitation, compressibility, siveness has been considered. The possible formation mechanisms of food liquid medium structure when slow selectivity, microscopic level, (hydrodynamic) and quick (acoustic) processes create favorable conditions for cavitation have been studied. The modified properties possibility to control these processes for initiation of mechanical and kinetic reactions that change the structure of the medium has been demonstrated. The invert syrup has been selected as the study object. The change in the invert syrup structure before and after such cavitation treatment has been recorded with the use of metallograph- ic microscope Nikon Eclipse MA100. The decrease in disperse phase sizes from 2–3 µm to 0.1–0.4 µm along with establishing the high uniformity of component distribution as compared to the syrup without cavitation process treatment has been detected.

1. Introduction of the liquid disperse media (LDM) widely used in confectionery In the existing food production technologies practically all used industry in the form of solutions and meltdowns [5]. media, for example, confectionery masses, are produced in large The possibility to control selective microscopic processes from volumes [1] for which the principle of local equilibrium is applied the macrolevel, in particular, change in macroscopic parameters when the formation processes of the treated media structure hap- is related to initiation of mechanical and chemical processes of pen so slowly so that they can be considered as quasi-equilibrium the LDM spatial structure formation that occur at the microscopic media [2]. And the system is exposed to external impacts in its level (Figure 1) [3,6]. entire volume in all spatial scales from molecular to macroscopic When these processes run at the microlevel we hope to get for a considerable period of time, so the kinetic reactions between response in the form of change in physical and chemical properties the medium components take place in the entire volume until the of the media at the macroscopic level. Such mechanisms can be relevant locally equilibrium values are achieved. These features expected to become the basis for manufacturing of new products lead to a range of natural restrictions that prevent obtaining the with specified properties. products with the specified structural and physical & chemical properties [3,4]. 2. Materials and methods The purpose of the work is the formation of food liquid medium The chosen object of study is the invert syrup with the amount structures with large amount of dry substances in the conditions of dry and reducing agents about 78–80 %, i. e. with 100 % de- of the combined hydrodynamic and acoustic impacts. composition of saсcharose into glucose and fructose, what is To obtain such products it appears that it is necessary to widely used in the production of pastry confectionery products go beyond the scope of quasi-equilibrium approach, start us- and marshmallow gummy products. The invert syrup produced ing mechanisms of non-equilibrium and unsteady structure under the conditions of cavitation impact on 20 samples and formation. monitoring without cavitation impact on 5 samples have been This article describes one of the possible approaches to solving considered. the issue, the approach that is based on the combined effect of In the studies the principle of the combined hydrodynamic hydrodynamic and acoustic processes impacts on the structure and acoustic impacts has been applied in the laboratory cavitation machine with ultrasonic transducer fed by the power of 250 W and oscillation frequency of 21–24 Hz. For the operational control of the process by gravimetric method, the moisture analyzer MB23 (Ohaus, USA) has been used. The structure and sizes of the disperse particles have been determined using the microscope observation on the inverted metallographic microscope Nikon Eclipse MA100 (with resolution of ×100, ×500 in the reflected light) with the DS-L2 control device for the DS camera head (Nikon, Japan). The reducing agents have been determined in accordance Figure 1. Hierarchic levels of mechanical and chemical with GOST 5903–89 «Confectionery. Methods for determination changes of sugar».

FOR CITATION: Savenkova T. V., Karimov A. R., Taleysnik M. A., Gerasimov T. V., Kondratev N. B. Mechanisms of destruction and synthesis of liquid media, used in the food industry, under non-equilibrium conditions. Food systems. 2019; 2(4): 38–41. DOI: 10.21323/2618–9771–2019–2–4–38–41

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Figure 2. The mechanism of interaction of particles of the disperse phase

The sizes and quantity of particles in the visualization field of As a result of this process the specific surface area reduces, the the optical microscopy with the same magnification by x500 lens density decreases, and simultaneously the cavities of different sizes and the set microscope grade scale in µm have been determined form. Such cavities with gas phase content are, by their physical in accordance with GOST R8.774–2011 «State system for ensuring nature, the cavitation nuclei which physical characteristics may the uniformity of measurements. Disperse composition of liquid change with time. All this indicates that the syrup can have the media. Determination of particle size by dynamic light scattering». property of compressibility [6,9].

3. Results and discussion 3.2. Cavitation dynamics and kinetics of macromolecules 3.1. Structural features of macromolecular media The reviewed microscopic properties of the LDM are diversely The used confectionery media, for example, emulsions, invert represented at the macrolevel and can be used for solving various and concentrated sugar syrups that differ by content of disperse process tasks. Particularly, the cavitation nuclei dynamics can particles of at least 70 % with high dispersiveness and a total be controlled in a specified way by duly stimulating their growth absence of translational motion, have the peculiar features of and further collapse with the help of the combination of quick gel [4, 7, 8, 9]. These features can include compressibility, sig- and slow hydrodynamic processes [1,11,12]. For example, these nificant amount of solid phase high-molecular particles that are processes can be organized in a given manner in the LDM flow in coupled and interlink together by molecular forces forming the the laboratory cavitation machine (Figure 4). macromolecular structures of various configurations and sizes [9]. We will try to demonstrate the occurrence of these properties on the example of production of the invert syrup — the medium that belongs to one of the simplest macromolecular media where the structure formation processes can occur in the simplest way. In the syrup production process, the absorptive bound mois- ture envelope is formed around the solid phase particles due to molecular force field that always appears as a result of non- compensated molecular forces in the interphase surface layer [9,10]. The saccharose inversion in the syrup is associated with the increase in the number of molecules, the favorable conditions for particles approach to the distance of 2H (disperse medium thickness) are created, and then while the distance between the particles reduces, the deformation processes increase and the free Figure 4. General view (a) and schematic circuit diagram surface energy is accumulated (Figure 2) that results in forma- (b) of the laboratory cavitation machine tion of intermolecular bindings. The strength of these contacts is determined by molecular forces. In this case, the quick process means the acoustic impact with The inversion of saccharose with its high concentration of frequency of 21–24 kHz, and the slow process means the param- about 80 % is associated with the increased amount of fructose eters change of the medium that flows with the given geometric and glucose to ensure favorable conditions for formation of ag- configuration for specified timeτ ( g). In the machine reactor, during gregates, chains and clusters of different spatial sizes, and their the sharp speed increase and pressure drop (from Р1 to Р2, Р3), the configuration can change for multiple times under the influence configuration of aggregates changes and the number of bubbles of both internal and external factors (Figure 3). significantly increases (Figure 5). The pressure increases at the reactor outlet that can lead to bubble collapse with the saved energy release practically in the point, and this, in its turn, can lead to breaking of disperse phase contacts. It should be expected that such selective impact is capable to initiate the various kinetic reactions changing the structure of the considered media. For example, during the bubble collapse, the released energy can appear to be sufficient for water molecule excitation, ionization and dissociation [1,12,13]. That is why it can be assumed that these water particles colliding with the Figure 3. Schematic illustration of the fructose and fragments of the broken macromolecules will join them to form glucose molecules aggregates configuration new kinds of materials.

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–1 Then the condition τg . ω will determine the possibility of the bubble dynamics control due to change in hydrodynamic parameters of the flow. It is convenient to reformulate this ratio in the following form: Lω . 1. (3) V3 Therefore the ratios (1)-(3) determine the allowable technical parameters of the system when the energy excitation of cavita- tion nuclei with acoustic waves and control of their dynamics are possible.

3.4. Practical results The conducted experiments with invert syrup in the machine illustrated in Figure 4 and Fig. 5 have demonstrated the dispersive- ness change with the combination of hydrodynamic and acoustic cavitation. It has been established that the sizes of the produced glucose and fructose aggregates decreased: the particles with practical sizes of about 0.1–0.4 µm were observed (Figure 7). Figure 5. Schematic circuit diagram of the cavitation And the produced medium is characterized by high uniformity machine reactor of component distribution of 92–94 % as compared to the syrup 3.3. Prerequisites for structure formation without cavitation treatment. In order to determine the conditions for implementing such structure formation processes, we will assess in stages the charac- teristic values of time and spatial scales of the induced wave move- ments of the LDM and oscillation properties of separate bubbles. Depending on the ratio between the practical values, the different ways of the cavitation nuclei evolution will be implemented. In the linear approximation, the natural frequency of bubbles oscillation for adiabatic process is determined by the ratio [14]:

ωb = υs / b , where b is the practical radius of the bubble, υs = √3γp/ρ, here γ — the adiabatic exponent for the gas in the bubble, p — gas pressure in the bubble, ρ — density of the liquid surrounding Figure 7. Structural change of the invert syrup produced without the bubble. As it was mentioned above, the quick dynamic pro- cavitation impact (a) and with cavitation impact (b) cess means the acoustic impact with practical frequency ω (see Fig. 4) that is responsible for the bubble energy excitation that is implemented by 4. Conclusions It has been demonstrated that the combination of quick acous- ω . ω . (1) b tic and slow hydrodynamic impacts makes it possible to control Furthermore, the length of this acoustic excitation wave λ the structure formation processes in the food media. One of pos- must be less than the practical size of the bubble (or, at least, sible mechanisms is related to the gas bubble formation in the commensurately), i. e. the following ratio must be observed studied medium with its further collapse when the bubble energy λ n b. (2) is released in the amount that is commeasurable to the practical In this case, the acoustic dispersion and resonance absorption molecular scale (see Figure 1). This selective impact is capable to of the acoustic waves by the bubbles are possible. In the opposite initiate various kinetic reactions changing the structure of the case which is likely to be implemented in the considered experi- considered media [1,12,13]. ment, the oscillating mode will be implemented in the entire area of That is why processes of this type can become the basis of the acoustic impact. And the excitation can transfer from acoustic advanced technologies in obtaining products with given properties vibrations to bubbles in the entire impacted medium in general and structure. For example the technology of making the pastry and during the time less than ω–1. confectionery with the use of invert syrup to produce emulsion In the case being considered, the slow process is the change also in the conditions of the combined cavitation effects is associ- in macroscopic properties of the liquid in the given geometric ated with the decrease in saccharose particle size from 25 to 6 µm configuration, controlling the growth and collapse of bubbles. As it and increase in their quantity almost by 75 times. This provides can be seen from Fig. 4 and Fig. 5, the main change in flow rate and the output of finished products with a new changed coagulation pressure takes place in the waveguide area due to change in the tube and crystallization structure with modified properties: increased absorptivity, high strength along with crumbliness and significantly cross-section. Let’s assume the maximum rate V3 in the waveguide area as the practical flow rate, and the length of the waveguide L reduced recyclable waste. In the future it is supposed to develop L the technology of «green» fruit jelly on the basis on the invert τ as the practical spatial scale, then the value g = can be taken syrup with the use of fresh fruits and vegetables to preserve na- V3 as the measure of the slow process. tive vitamins and minerals.

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REFERENCES 1. Aksenova, L.M., Kochetov, V.K., Lisitsyn, A.B., Nikolsky, K.N., Panfilov, Osipov, M.V. (2015). Theoretical and practical aspects of development of V.A., Podkorutov, N.V., Semenova, A.A., Taleysnik, M.A. (2015). Food biscuits with a modified carbohydrate profile for patients with type 2 dia- technology and nano transformations of biopolymers. Krasnodar: Diapa- betes. Voprosy pitaniia, 84(6), 122–131. (in Russian) zon-V. — 304 p. ISBN: 978–5–91050–168–7 (in Russian) 9. Karimov, A.R., Taleysnik, M.A., Savenkova, T.V., Aksenova, L.M., Gera- 2. Prigogin, I., Kondepudi, D.(2002). Modern thermodynamics. From heat simov, T.V. (2018). Physical and chemical features of dynamic of poly- endints to dissipative structures. Мoscow: Mir. —460 p. ISBN: 978–5–03– meric fluid. Food systems, 1(3), 44–54. DOI: 10.21323/2618–9771–2018– 003538–3 1–3–44–54 (in Russian) 3. Karimov, A.R., Taleisnik, M.A., Savenkova, T.V., Aksenova, L.M. (2019). 10. Uriev, N.B., Taleysnik, M.A. (1985). Food disperse systems. М: Agro- The influence of velocity field on simple chemical reactions in viscous promizdat. —296 p. (in Russian) flow. Physica Scripta, 94(4), 045002 DOI: 10.1088/1402–4896/aafc15 11. Margulies, M.A. (2000). Sonoluminescence. Advances In Physical Sciences, 4. Rebinder P. A. (1979). Surface effect in disperse system. Physicochemical 170(3), 285–287. mechanics. Selected works. М: Nauka. — 384 p. (in Russian) 12. Novitskiy, B.G. (1983). The use of acoustic vibrations in chemical-techno- 5. Khmelnitskiy, R.A. (1988). Physical and colloid chemistry. Мoscow: Vys- logical processes. Moscow: Chemistry. —192 p. (in Russian) shaya shkola. —400 p. ISBN: 5–06–001257–3 (in Russian) 13. Pokrovskiy, V.N. (1994). Low-frequency dynamics of dilute solutions of 6. Caruso, M.M., Davis, D.A., Shen, Q., Odom, S.A., Sottos, N.R., White, linear polymers. Advances In Physical Sciences, 164(4), 375–391. S.R., Moore, J.S. (2009). Mechanically-induced chemical changes in poly- 14. Naugolnykh, K.A., Ostrovskii, L.A. (1990). Nonlinear wave processes in meric materials. Chemical Reviews, 109(11), 5755–5798. DOI: 10.1021/ acoustics. Мoscow: Nauka. —236 p. ISBN: 5–02–000682–3 (in Russian) cr9001353 15. Savenkova, T.V., Soldatova, E.A., Kiseleva, T.L., Glazkova, I.V., Zhilin- 7. Savenkova, T.V., Osipov, M.V., Kazantsev, E.V., Kochetkova, A.A, Voro- skaya, N.V. (2015). The role of the food industry in dietetic therapy of bieva, I.S., Kiseleva, T.L. (2016). The production technology of diabetic the population. Specialized confectionery diabetic food. Voprosy Pitaniia, confection with modified carbohydrate profile. Research Journal of Phar‑ 84(6), 107–115. (in Russian) maceutical, Biological and Chemical Sciences, 7(6), 3123–3130. 16. Dorn, G.A., Savenkova, T.V., Sidorova, O.S., Golub, O.V. (2015). 8. Kochetkova, A.A., Vorobyova, V.M., Vorobyova, I.S., Sharafetdinov, Сonfectionery goods for healthy diet. Foods and raw materials, 3(1), 70– Kh. Kh., Sarkisyan, V.A., Semin, M.O., Savenkova, T.V., Soldatova, E.A., 76. DOI: 10.12737/11240

AUTНOR INFORMATION Tatyana V. Savenkova — doctor of technical sciences, professor, Director, All-Russian Scientific Research Institute of Confectionery Industry — Branch of V. M. Gorbatov Federal Research Center for Food Systems of RAS, 107023, Moscow, Electrozavodskaya Str, 20, bld. 3, Tel.: +7–495–963–64–09, e-mail: conditer- [email protected] *corresponding author Alexander R. Karimov — doctor of physical and mathematical sciences, professor, professor, department of Electrophysical installations, National Research Nuclear University «MEPhI» 115093, Moscow, Kashirskoe av., 31 E‑mail: Mikhail A. Taleysnik — candidate of technical sciences, leading researcher, laboratory technology of production of flour confectionery products, All-Russian Scientific Research Institute of Confectionery Industry — Branch of V. M. Gorbatov Federal Research Center for Food Systems of RAS, 107023, Moscow, Electro- zavodskaya Str, 20, bld. 3, Tel.: +7–495–962–17–35, e-mail: [email protected] Timofei V. Gerasimov — candidate of technical science, leading researcher, laboratory technology of production of flour confectionery products, All-Russian Scientific Research Institute of Confectionery Industry — Branch of V. M. Gorbatov Federal Research Center for Food Systems of RAS,107023, Moscow, Ekektro- zavodskaya str., 20, Tеl.: +7–495–962–17–39, E‑mail: [email protected] Nikolai B. Kondratev — doctor of technical sciences, chief researcher, department of modern quality assessment methods, All-Russian Scientific Research Institute of Confectionery Industry — Branch of V. M. Gorbatov Federal Research Center for Food Systems of RAS,107023, Moscow, Ekektrozavodskaya str., 20, Tеl.: +7–495–962–17–39, E‑mail: [email protected] All authors bear responsibility for the work and presented data. All authors made an equal contribution to the work. The authors were equally involved in writing the manuscript and bear the equal responsibility for plagiarism. The authors declare no conflict of interest. Received 30.10.2019 Accepted in revised 20.11.2019 Accepted for publication 13.12.2019

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UDC 664.046.3 DOI: 10.21323/2618–9771–2019–2–4–42–47 Original scientific paper AN EFFECT OF STORAGE AND TRANSPORTATION TEMPERATURE ON QUANTITATIVE AND QUALITATIVE COMPOSITION OF MICROFLORA OF PLANT PRODUCTS Svetlana V. Avilova1, Vladimir N. Kornienko1*, Aleksey A. Gryzunov1, Anna A. Vankova2 1 All-Russian Scientific Research Institute of Refrigeration Industry — Branch of V. M. Gorbatov Federal Research Center for Food Systems of RAS, Moscow, Russia 2 Russian State Agrarian University — Moscow Timiryazev Agricultural Academy, Moscow, Russia

KEY WORDS: ABSTRACT storage, transportation, The results of the study on changes in the composition and quantity of epiphytic and endophytic microorganisms temperature regime, plant of plant products during storage and transportation are presented. For the investigation, the authors took apple products, microorganisms, fruits and leafy spicy green products that had biological peculiarities and allowed investigating processes of the microbiological spoilage long-term and short-term main stages (cold storage, transportation by refrigerated transport, presales storage) of the continuous cold chain on the way to a consumer. Apple fruits were placed in storage in cold chambers with the temperature regimes of plus (2–3) °C and minus (1–2) °C, where they were stored for 90 days. The vegetative organs of dill and parsley were transported during 8 hours by a refrigerated truck and placed in the commercial refrigeration equipment at two temperature regimes (4–5) °C and (0–1) °C for 72 hours for presales storage. The results of the microbiological analysis showed that the number of endophytic microorganisms (bacteria, yeasts and molds) was lower by 1–3 orders of magnitude in apple fruits and by 2–3 times in green vegetables compared to the number of epiphytic microorganisms. It was established that the regime of storage at negative temperatures completely inhibited the development of epiphytic bacteria on fruits, significantly delayed the multiplication of epiphytic yeasts and molds; while at a positive temperature the number of bacteria increased approximately by 10–17 times, yeasts by 180 times and molds by 3 times. The dynamics of changes in the number of endophytic microorganisms during storage showed the same trend that was observed for epiphytic microorganisms. Analysis of the microbial quantity after transportation of green products showed an increase in abundance of the revealed groups of epiphytes and endophytes by 1.5–3 times upon absolute prevalence of bacteria. After short-term stor- age, a significant growth of the revealed microbial groups was found; with that, their quantity was 1.5–6.5 times higher at (4–5) °C than at (0–1) °C. The authors experimentally confirmed the conclusion that with respect to reduction of losses due to microbiological spoilage and extension of shelf life, the cold storage regime of the stud- ied plant products at near-zero temperatures is preferable compared to the regimes of storage at higher positive temperatures.

1. Introduction possible to significantly retard life processes in microorganisms Products of plant production are necessary for the human body increasing therewith storability of plant products, their resistance as a source of vitamins, carbohydrates, microelements, mineral to diseases in the process of distribution from a manufacturer to salts , amino acids and other substances. Fruits and vegetables are a consumer [2,8]. consumed mainly in the fresh state; therefore, to ensure healthy Artificial cold that underlies all modern technologies for indi- and safe nutrition, it is necessary to carry out not only chemical- vidual links of the continuous cold chain (CCC) allows a substantial toxicological but also microbiological control of products [1]. reduction of losses due to a decrease in microbiological spoilage, Spoilage of plant products occurs due to the natural physi- maximum retardation of physiological and biochemical processes ological processes and microbial activity [2]. According to some and consequently prolongation of shelf-life of fruits and vegetables estimates, a proportion of registered losses caused by the microbial without significant deterioration of their quality [9]. The CCC as activity reaches (30–40)%. Spoilage occurs throughout the food an inter-branch organizational technological complex is intended chain — during harvest, processing and storage of crops, during for food safety and storability assurance at a high-level through- transportation, wholesale and retail sales up to consumption by out the way from a producer to a consumer (refrigerated storage, population [3]. transportation by refrigerated transport, presales storage) due One of the solutions to the strategic task of reducing food losses to continuity of the specified thermal condition of foods [10,11]. is regulation of microbiological spoilage, which requires knowl- Violation of required thermal regimes, emergence of cyclic and edge of the composition of the microbial community including variable temperature effects of the environment during storage both microorganisms typical for a certain type of plant products and transportation lead to a sharp acceleration of the processes of and spoilage microorganisms. The use of knowledge about micro- bacterial spoilage in products, which negatively affect their quality bial behavior and their habitat for regulation of microbiological indicators and safety in a sales process [12, 13,14,15]. parameters will provide producers and suppliers of fresh fruit A quantity and composition of microorganisms on the sur- and vegetable products with an opportunity to improve existing face and in internal tissues of fruits and vegetables at different technologies of storage and transportation [4,5,6]. ambient temperatures are studied insufficiently. Investigations Preservation of fruits and vegetables in the fresh state is in this direction are of great practical importance and create a achieved due to a reduction of the vital processes both directly basis for improving parameters of storage and transportation of in plant products and in microbial community [7]. By regulating plant products. environmental conditions (temperature, humidity, gas compo- Fruits and vegetables intended for storage are the living veg- sition, quantity of available nutrients, acidity and others), it is etative and reproductive organs of plants, which surface is abun-

FOR CITATION: Avilova S. V., Kornienko V. N., Gryzunov A. A., Vankova A. A. An effect of storage and transportation temperature on quantitative and qualitative composition of microflora of plant products. Food systems. 2019; 2(4): 42–47. DOI: 10.21323/2618–9771–2019–2–4–42–47

42 ПИЩЕВЫЕ СИСТЕМЫ | Том 2 № 4 | 2019 FOOD SYSTEMS | Volume 2 № 4 | 2019 dantly inhabited by microorganisms that were given the name apples of the «Champion» variety; yeasts, which are the typical «epiphytes» [16,17]. Recently, more and more information have inhabitants of the phyllosphere, were practically absent on the appeared showing that plant internal tissues are also inhabited apples of both varieties, and molds were present in low numbers by microorganisms called endophytes [18,19,20,21,22]. on the surface of the «Royal Gala» fruits (Figure 1 a, b, e). The aim of the investigation was to study the composition The number of endophytic yeasts was extremely negligible and number of epiphytic and endophytic microorganisms of dif- on the apple fruits of both varieties and endophytic molds were ferent types of fruit and vegetable products during storage and absent (Figure 1 d, f). In the «Champion» variety, endophytic transportation at different temperature regimes. bacteria were found in small numbers (Figure 1b). During storage, a quantity of the studied microbial groups 2. Materials and methods changed depending on a variety and temperature conditions. At Apple fruits and leafy spicy green products were taken for the positive temperature regime of plus (2–3) °C, the maximum the investigation. This choice was based on differences in their number of epiphytic bacteria was revealed on the apples of the biological peculiarities that allowed investigating processes of «Champion» variety. During storage, their number increased by the long-term and short-term main stages of the CCC on the way approximately 15 times (Fig. 1a). The storage regime at negative to a consumer. temperatures of minus (1–2) °C completely inhibited the develop- Storability of apple fruits is largely determined by duration of a ment of epiphytic bacteria on fruits of both varieties (Figure 1a). period of post-harvest ripening at corresponding temperature and The number of yeasts increased during storage with more humidity regimes (storage in a cold chamber). Due to low duration intensive multiplication of yeasts at a positive temperature than of transportation compared to the storage period, a temperature at a negative temperature. For example, at the temperature re- of transportation practically does not influence apple storability. gime of (2–3) °C, the number of yeasts increased by 180 times Only mechanical damages during transportation can be the main on the «Royal Gala» variety and by 10 times on the «Champion» causes of subsequent quality deterioration. variety compared to the initial values before storage; while at a The objects of the investigation of seed fruit crops were the temperature of minus (1–2) °C, it increased by approximately 95 apple varieties «Royal Gala» and «Champion», which were placed times on the «Royal Gala» fruits (Fig. 1c). Therefore, a negative in storage in cold chambers (with a positive temperature of plus storage temperature did not completely inhibit the development (2–3) °C and with a negative temperature of minus (1–2) °C) and of epiphytic yeasts on the «Royal Gala» variety but retarded their stored for 90 days. multiplication. An increase in the number of yeasts on the fruits Storability of leafy vegetables is low; their preservation almost of the «Champion» variety was observed only at a positive storage entirely depends on external conditions upon supply to a consumer. temperature and they were not revealed at a negative temperature Leafy vegetables are characterized by slight loss of water due to (Figure 1c). the highly developed leaf surface, thin cell walls and cover tissues, The composition of epiphytic microorganisms usually contains weak water holding capacity of colloids. Therefore, spoilage and molds, including phytopathogenic that contaminate fruits during quality deterioration of fresh green products mainly occur during harvest, transportation and storage, and quickly develop upon the short periods of their transportation and sale [23]. favorable temperature and humidity conditions. However, molds The objects of investigation of green cultures were dill variety were not found on the fruits upon placement of the apples of the «Alligator» and parsley variety «Kharkovchanka». Freshly harvested studied varieties in storage (Figure 1 e). green products were delivered (for 8 hours by refrigerated trucks) During storage at the temperature regime of (2–3) °C, molds and stored before sale (for 72 hours in the retail refrigerated equip- developed on the «Champion» fruits in a quantity that was three ment) at two temperature regimes: (4–5) °C and (0–1) °C. times higher than the values obtained after storage at the tem- The microbial quantity and composition in apple fruits were perature regime of minus (1–2) °C (Figure 1 e). In the «Royal Gala» analyzed by the standard method of plating on the glucose pep- variety, molds were not revealed on the fruit surface after storage tone medium (GPM) [24]. To detect yeasts, 40 % lactic acid was both at positive and negative temperatures (Figure 1 e). additionally introduced into the medium. To detect epiphytic The results of the microbiological analysis showed that the microorganisms, the surface of the preliminarily weighted apples number of endophytic microorganisms was by 1–3 orders of mag- was swabbed with sterile water. Then, the fruit surface was steril- nitude lower than epiphytic (Figure 1 b, d, f). In the structure of ized with 96 % ethanol, the cover tissue was removed with a sterile the microbial community, endophytic bacteria were absent in the scalpel and an average sample of internal storage tissues was taken «Royal Gala» fruits; individual bacterial cells were found only inside for detection of endophytic microorganisms. The consecutive serial apple tissues of the «Champion» variety (Figure 1b). dilutions were prepared and pour plating on the GPM was carried The dynamics of changes in the number of endophytic yeasts out in duplicate by the conventional methods [25]. during storage showed the same trend that was observed for epi- The microbial complex of green cultures upon transportation phytic yeasts. At a temperature of (2–3) °C, the multiplication of was also analyzed by the standard method of plating on the GPM. endophytic yeasts was more intensive: an increase by 25 times To detect epiphytic microorganisms, a specimen of an average was found in the fruits of the «Royal Gala» variety. In the fruits sample was washed in sterile water in the process of agitation in of the «Champion» variety, yeasts were not revealed after stor- a shaker for 10 min. To detect endophytes, the washed samples age at a negative temperature, while at a positive temperature, of green vegetables were transferred to 96 % ethanol, agitated for a decrease in their initial number by approximately 2 times was 1 min., rinsed several times with sterile water and mechanically established (Figure 1 d). chopped until obtaining homogeneous mass. During storage of apples of the «Champion» variety at the temperature regime of (2–3) °C, a large number of molds developed 3. Results and discussion in internal tissues (Figure 1f). At the same time, the development Apple fruits. As the results of the investigation showed, micro- of molds was completely inhibited in the fruits of this variety at organisms (bacteria, yeasts and molds) were found both on the the temperature regime of minus (1–2) °C. In the fruits of the surface and in internal tissues. «Royal Gala» variety, the development of mold cells was not re- Microbial community of epiphytic microorganisms was differ- corded during storage. Therefore, a negative ambient temperature ent on the apples of the analyzed varieties before placement in completely stopped the growth of endophytic molds on the fruits storage (Figure 1). For example, bacteria were found only on the of both varieties (Figure 1f).

43 temperaturetemperature regime of regime (2 – 3) of ° С(2, the– 3) number °С, the of number yeasts ofincreased yeasts increased by 180 times by 180 on timesthe “Royal on the “Royal Gala” varietyGala” and variety by 10and times by 10on timesthe “Champion” on the “Champion” variety compared variety compared to the initial to the values initial values before storage;before whilestorage; at awhile temperature at a temperature of minus of(1 minus– 2) ° С(1, it– increased2) °С, it increased by approximately by approximately 95 95 times on timesthe “Royal on the Gala” “Royal fruits Gala” (Fig. fruits 1c). (Fig. Therefore, 1c). Therefore, a negative a negativestorage temperature storage temperature did not did not completelycompletely inhibit the inhibit development the development of epiphytic of epiphytic yeasts on yeasts the “Royalon the Gala”“Royal variety Gala” butvariety but retarded retardedtheir multiplication. their multiplication. An increase An increasein the number in the ofnumber yeasts of on yeasts the fruitson the of fruitsthe of the “Champion”“Champion” variety was variety observed was observedonly at a onlypositive at a storagepositive temperature storage temperature and they andwere they not were not revealed atrevealed a negativeat a temperaturenegative temperature (Figure 1c). (Figure 1c). The compositionThe composition of epiphytic of epiphytic microorganisms microorganisms usually containsusually containsmolds, includingmolds, including phytopathogenicphytopathogenic that contaminate that contaminate fruits during fruits harvest,during harvest,transportation transportation and storage, and storage,and and quickly developquickly upondevelop favorable upon favorable temperature temperature and humidity and humidityconditions. conditions. However, However, molds were molds were not foundnot on foundthe fruits on theupon fruits placement upon placement of the apples of the of athepples studied of the varieties studied invarieties storage in (Figure storage (Figure 1 e). 1 e). During storageDuring atstorage the temperature at the temperature regime of regime (2 – 3)of °(2С, –molds3) °С ,developed molds developed on the on the “Champion”“Champion” fruits in fruitsa quantity in a thatquantity was thatthree was times three higher times than higher the thanvalues the obtained values obtainedafter after storage atstorage the temperature at the temperature regime of re minusgime of (1 minus – 2) °(1С (Figure– 2) °С1(Figure e). In the1 e). “Royal In the Gala” “Royal Gala” variety, moldsvariety, were molds not were revealed not revealedon the fruiton thesurface fruit aftersurface storage after bothstorage at positiveboth at andpositive and negative temperaturesnegative temperatures (Figure 1 (Figure e). 1 e). The resultsThe of results the microbiological of the microbiological analysis showedanalysis thatshowed the numberthat the ofnumber endophytic of en dophytic microorganismsmicroorganisms was by 1 was− 3 byorders 1 − of3 ordersmagnitude of magn loweritude than lower epiphytic than epiphytic (Figure 1 (Figure b, d, f).1 In b, d, f). In the structurethe structureof the microbial of the microbial community, community, endophytic endophytic bacteria werebacteria absent were in absent the “Royal in the “Royal ПИЩЕВЫЕGala” СИСТЕМЫfruits;Gala” individual fruits; | Том 2individual №bacterial 4 | 2019 cellsbacterial were cells found were only found inside only app insidele tissuesFOOD apple ofSYSTEMS tissues the “Champion” of| Volume the “Champion” 2 № 4 | 2019 variety (Figurevariety1b). (Figure 1b).

endophytic bacteria epiphyticepiphytic bacteria bacteria endophytic bacteria 16 16 0,025 0,025 14 14 g g

0,02 g g 0,02 12 12 10

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CFU / 0,015 3 3 3 8 3 8 ·10 ·10 ·10 6 ·10 6 0,01 0,01 4 4 0,005 0,005 2 2 Number of microorganisms Number Number of microorganisms Number Number of microorganisms Number 0 of microorganisms Number 0 0 0 1 1 1 1 epiphytic epiphyticyeasts 2 yeasts2 endophyticendophytic yeasts2 yeasts2 а) b) 1,8 1,8 epiphytic epiphyticyeasts yeasts 0,25 endophytic0,25 endophytic yeasts yeasts 1,6 1,6 а ) а ) b ) b) g g g 1,4 g 1,4 0,2 0,2 1,8 1,8 1,2 1,2 0,25 0,25 CFU /

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3 1

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0,8 ·10 ·10 ·10 ·10 0,2 of microorganisms Number Number of microorganisms Number 0,2 of microorganisms Number Number of microorganisms Number 0,1 0,1 0,6 0,6 0 0 0 0 0,4 0,4 0,05 0,05 1 1 1 1 Number of microorganisms Number 0,2 of microorganisms Number

Number of microorganisms Number 0,2 Number of microorganisms Number 2 2 2 2 0 0 0 0 1 1 1 1 c) 2 2 2 d) 2 c) c) 4 4 d) d) epiphytic epiphyticmolds molds endophyticendophytic molds molds c ) c ) d ) d) 0,12 0,12 2 2 epiphytic epiphyticmolds molds endophyticendophytic molds molds

0,1 g g 0,1 g g 1,5 1,5 0,12 0,12 2 2 0,08 0,08 CFU / CFU / CFU / CFU / 3 3 3 3 g 0,1 0,1 1 g g g 1 0,06 0,06 1,5 1,5 ·10 ·10 ·10 0,080,04 ·10 0,08 CFU / 0,04 CFU / CFU / CFU / 0,5 3 3 0,5 3 3 1 1 0,060,02 0,06 ·10

0,02 ·10 Number of microorganisms Number ·10 ·10 Number of microorganisms Number Number of microorganisms Number 0,04 of microorganisms Number 0,04 0 0 0 0 0,5 0,5 1 1 0,02 0,021 1 2 Number of microorganisms Number 2 of microorganisms Number 2 Number of microorganisms Number Number of microorganisms Number 2 0 0 0 0 e) f) Figure 1. An effect of storage temperature conditions on a quantity of ep1iphytic and1 endophytic microorganisms 1 1 2 2 e ) 2 e ) (thousands 2 CFU/g) in apple fruits: f ) f) 1 — «Royal Gala» variety; 2 — «Champion» variety.  — after storage (regime: (2–3) °C;  — before placement in storage;  — after storage (regime: minus (1–2) °C Figure 1. An effect of e ) storage e temperature ) conditions on a quantity of f ) epiphytic f) and Spicy leafy greens.Figure The 1. results An ofeffect the microbiological of storage analysis temperature and yeasts, conditions and those onof the a endophyticquantity microbialof epiphytic complex onlyand by of freshendophytic green productsendophytic microorganisms during microorganisms transportation (thousands and (thousandsthe CF followingU/g) in CF appleU/g)bacteria. fruits: in appleThe vegetative fruits: organs of parsley separated from plants short-term1 ‒ “Royal storage Gala”before salevariety; showed 2 that‒ “Champion” microorganisms variety. (bac- were slightly different from dill regarding the microbial composi- Figure 1.1 ‒An“Royal effect Gala” of storage variety; temperature2 ‒ “Champion” conditions variety. on a quantity of epiphytic and teria, yeasts‒ before andFigure molds) placement were1. presentAn in effectstorage; both onof the storage surface andtemperature in tion — conditions bacteria and filamentouson a quantity fungi wereof foundepiphytic on the andsurface endophytic microorganisms‒ before placement (thousands in storage; CF U/g) in apple fruits: internal ‒tissuesafter endophyticof storage dill and (regime: parsley. microorganisms The (2 total– 3) microbial°С; (thousands number CFU/g)and in in internal apple tissues fruits: (Figure 2). changed1 ‒ depending“Royal Gala” ‒onafter a species variety; storage of the 2 (regime: ‒green“Champion” culture. (2 – 3) variety.°С; The absolute prevalence of bacteria was common for all types ‒ after1 storage‒ “Royal (regime: Gala” minusvariety; (1 2 – ‒2)“Champion” °С variety. The structure‒ before of placement‒ thebeforeafter epiphytic storage placement in microbial storage; (regime: in complex storage; minus of the(1 –dill2) °Сof green crops. Their number was a thousand times higher than the leafy apparatus before transportation was presented by bacteria number of yeasts and molds (Figure 2 a, c, e). The initial number ‒ after storage‒ after (regime: storage (2(regime: – 3) °С (2; – 3) °С; The dynamicsThe dynamics of changes of inchanges the number in the of number endophytic of endophytic yeasts during yeasts storage during showed storage showed ‒ after storage‒ after (regime: storage minus(regime: (1 minus– 2) °С (1 – 2)44 °С the same thetrend same that trend was thatobserved was observedfor epiphytic for epiphyticyeasts. At yeasts. a temperature At a temperature of (2 – 3) of ° С(2, –the3) °С, the multiplicationThemultiplication dynamics of endophyticThe dynamics ofof changesendophytic yeasts of in changeswas theyeasts more number in was intensive: the moreof number endophytic intensive: an ofincrease endophytic yeasts an byincrease during25 yeasts times bystorage duringwas25 times found showed storage was in found showed in the fruitssame theoftrend thefruitssame that“Royal oftrend was the Gala” that“Royalobserved wasvariety. Gala” observedfor In epiphyticvariety. the forfruits In epiphyticyeasts. theof thefruits “Champion”At yeasts. ofa temperaturethe “Champion”At avariety, temperature of (2yeasts variety, – 3) ofwere ° С(2yeasts, not–the3) were °С, notthe multiplicationrevealed aftermultiplicationrevealed storageof endophytic after at storageof a negativeendophytic yeasts at a temperature,negativewas yeasts more temperature,was intensive: while more atintensive: an awhile positiveincrease at an a temperature, bypositiveincrease 25 times temperature,by awas25 decrease times found awas indecrease found in thetheir fruits initial thetheirof number thefruits initial “Royal of by number the approximate Gala” “Royal by variety. approximate Gala”ly 2In variety.times the lyfruits was 2In times esttheof ablished thefruits was “Champion” estof (Figure ablishedthe “Champion” 1variety, (Figured). yeasts 1variety, d). were yeasts not were not revealed Duringafterrevealed storage storage Duringafter at storage ofa storage negativeapples at ofaof temperature, negative applesthe “Champion” of temperature, the while “Champion” variety at awhile positive at variety the at a temperaturetemperature, positive at the temperaturetemperature, regime a decrease of regime (2a indecrease – of (2 in – their3) °С initial, a largetheir3) number°С numberinitial, a large by number of approximatenumber molds by of devapproximate moldselopedly 2 times dev in elopedlyinternal was 2 times est in ablishedtissues internal was est (Figure (Figure ablishedtissues1f). 1(Figure (Figured). At the1f). 1same d). At thetime, same the time, the During storageDuring of storage apples ofof applesthe “Champion” of the “Champion” variety at variety the temperature at the temperature regime of regime (2 – of (2 – 3) °С, a large3) °С number, a large of number molds ofdev moldseloped dev in elopedinternal5 in tissues internal5 (Figure tissues1f). (Figure At the1f). same At thetime, same the time, the

5 5 developmentdevelopment of molds ofwas molds completely was completely inhibited inhibitedin the fruits in theof thisfruits variety of this at variety the temperature at the temperature regime ofregime minus of(1 minus– 2) °С (1. In – 2)the ° Сfruits. In theof thefruits “Royal of the Gala” “Royal variety, Gala” the variety, development the development of mold of mold cells wascells not recordedwas not recordedduring storage. during Therefore,storage. Therefore, a negative a negativeambient temperatureambient temperature completely completely stopped thestopped growth the of growth endophytic of endo moldsphytic on molds the fruits on thofe bothfruits varieties of both (Figurevarieties1f). (Figure 1f). Spicy leafySpicy greens. leafy Thegreens. results The ofresults the microbiologicalof the microbiological analysis analysisof fresh ofgreen fresh green products productsduring transportation during transportation and the followingand the following short-term short storage-term before storage sale before showed sale thatshowed that microorganismsmicroorganisms (bacteria, (bacteria, yeasts and yeasts molds) and were molds) present were both present on the both surface on the and surface in internal and in internal tissues oftissues dill and of parsley. dill and Theparsley. total Themicrobial total microbial number changed number dependingchanged depending on a species on aof species the of the green culture.green culture. The structureThe structureof the epiphytic of the epiphytic microbial microbial complex complexof the dill of leafy the dill apparatus leafy apparatus before before transportationtransportation was presented was presented by bacteria by andbacteria yeasts, and and yeasts, those and of thosethe endophytic of the endophytic microbial microbial complex complexonly by onlybacteria. by bacteria.The vegetative The vegetative organs of organs parsley of separatedparsley separated from plants from were plants were slightly differentslightly differentfrom dill from regarding dill regarding the microbial the microbial composition composition - bacteria - andbacteria filamentous and filamentous fungi werefungi found were on foundthe surfa on cethe and surfa in ceinternal and in tissues internal (Figure tissues2). (Figure 2). The absoluteThe prevalenceabsolute prevalence of bacteria of wasbacteria common was commonfor all types for allof greentypes crops.of green Their crops. Their number wasnumber a thousand was a thousandtimes higher times than higher the numberthan the ofnumber yeasts ofand yeasts molds and (Figure molds2 (Figure a, c, e).2 a, c, e). The initialThe number initial ofnumber epiphytic of epiphytic bacteria onbacteria freshly on harvested freshly harvested parsley beforeparsley transportation before transportation was aboutwas 1.5 abouttimes 1.5 highertimes than higher on dillthan (Figure on dill 2a).(Figure A quantity2a). A ofquantity epiphytic of epiphytic bacteria wasbacteria was several timesseveral higher times than higher those than of endophyticthose of endophytic bacteria: bacteria:the number the ofnumber epiphytic of epiphytic bacteria onbacteria on dill was dill2.5 wastimes 2.5 higher times and higher on parsleyand on 3parsley times higher3 times than higher the thannumber the ofnumber endophy of ticendophy tic bacteria, respectivelybacteria, respectively (Figure 2 (Figure a, b). 2 a, b). Analysis Analysisof microbial of microbial quantity quantityafter transportation after transportation of green ofproducts green productsshowed anshowed an increase inincrease abundance in abundance of the revealed of the revealedgroups of groups epiphytes of epiphytes and endophytes and endophytes (Figure 2). (Figure In our 2). In our ПИЩЕВЫЕopinion, СИСТЕМЫ opinion,this effect | Тthisом was 2 №effect 4associated | 2019was associated with changes with changesin temperature in temperature-FOODhumidity SYSTEMS- humidityconditions | Volume conditions of 2 № 4 | 2019of transportationtransportation [23]. [23].

epiphyticepiphyticbacteria bacteria endophyticendophytic bacteria bacteria

200 200 120 120 100 100 g g g g 150 150 80 80 CFU / CFU / CFU / CFU / 3 3 60 3 100 60 3 100 ·10 ·10 ·10 ·10 40 40 50 50 20 20

0 of microorganisms Number 0 0 of microorganisms Number 0 Number of microorganisms Number Number of microorganisms Number 1 1 1 1 epiphyticepiphytic 2yeasts yeasts2 3 2 2 3 3 4 4 endophyticendophytic yeasts3 4yeasts 4 а) b) 0,2 0,2 0,12 0,12 epiphytic yeasts

epiphyticg yeasts 0,1 g 0,1 endophytic yeasts g 0,15 0,15 g endophytic yeasts a ) a ) 0,08 b ) b) 0,2 0,08 0,12 CFU / CFU / 0,2 0,12

0,1 CFU / 3

0,1 CFU / 3 0,06 0,06 3 3

g 0,1 g ·10 g 0,15 0,1 ·10 ·10

g 0,04 0,15 ·10 0,04 0,05 0,05 0,08 0,08 CFU / 0,02 0,02

0,1 CFU / CFU / 3 0,06

0,1 CFU / 3 3 0 0 0,06 3

Number of microorganisms Number 0 Number of microorganisms Number 0 ·10 Number of microorganisms Number Number of microorganisms Number ·10 ·10 10,05 1 0,04 0,05 2 2 ·10 0,04 1 1 3 4 3 4 2 2 0,02 3 4 3 4 0 0,02 0 Number of microorganisms Number 0 Number of microorganisms Number Number of microorganisms Number 1 6 60 1 2 of microorganisms Number 1 2 3 4 1 2 3 4 2 3 4 c ) c ) 3 d ) 4 d) c) d) epiphyticepiphytic molds molds endophyticendophytic molds molds c ) c ) d ) d) 0,4 0,4 0,12 0,12 0,1 0,1 g g g

epiphytic molds g endophytic molds 0,3 0,3epiphytic molds endophytic molds 0,08 0,08 0,4 0,12 CFU / CFU / CFU / 0,4 CFU / 0,12 3 3

0,2 3 0,2 3 0,06 0,06 0,1 g 0,1 g ·10 g ·10 ·10 g 0,3 ·10 0,04 0,04 0,30,1 0,1 0,08 0,080,02 0,02 CFU / CFU / CFU / 3 CFU / 0,2 3 0,06 3 0,20 0 3 0,060 0 Number of microorganisms Number Number of microorganisms Number Number of microorganisms Number Number of microorganisms Number ·10 ·10 0,04 ·10 ·10 0,04 1 0,1 1 2 1 1 2 0,1 2 3 3 0,02 2 3 3 4 4 0,02 4 4 0 0 Number of microorganisms Number

0 e) 0 of microorganisms Number f) Number of microorganisms Number 1 of microorganisms Number 1 Figure 2. 1An effect of storage 2temperature conditions on a quantity of epiphytic1 and endophytic2 microorganisms 2 3 4 2 3 4 3 (thousands4 CFU/g) in spicy green vegetables: 3 4 1 — dill at the regime of (4–5) °C; e ) e )  — before transportation; f ) f) 2 — dill at the regime of (0–1) °C;  — after transportation; 3 — parsley at the regime of (4–5) °C;  — after presales storage 4 — parsley at the regime of (0–1) °C e) f) Figure 2. Figure An effect 2. An of effecte )storage of temperature storage temperature conditions conditions on a quantity on a quantity off) epiphytic of epiphytic and and of epiphytic bacteria on freshly harvested parsley before trans- of epiphytes and endophytes (Figure 2). In our opinion, this effect portationendophytic was aboutendophytic microorganisms 1.5 times microorganisms higher (thousandsthan on dill (thousands (Figure CFU/g) 2a). in CFU/g) spicywas associated greenin spicy vegetables: withgreen changes vegetables: in temperature-humidity conditions A quantity1 ‒ dill of epiphytic at1 ‒thedill regime bacteria at the of wasregime (4 several - 5) of times° С(4; -2 higher 5)‒ dill°С than; at2 ‒thedill ofregime transportationat the of regime (0 -[23].1) of ° С(0; -3 1)‒ parsley°С; 3 ‒ atparsley the at the thoseFigureregime of endophytic of2.regimeFigure (4An –bacteria: 5) effectof 2.°С (4;An the 2– of‒ 5) numbereffect parsley °storageС; 2 of of‒ at epiphyticparsley temperaturethestorage regime atbacteria temperaturethe of regimeconditions (0 - 1) Figureof °Сconditions (0 on -3 presents1) a ° Сquantity onthe charactera quantityof epiphytic of changes of epiphytic inand the air tempera and - on dill was 2.5 times higher and on parsley 3 times higher than ture in the body of a refrigerated truck and on the surface of fresh endophytic‒ beforeendophytic microorganismstransportation;‒ before transportation;microorganisms (thousands (thousands CFU/g) in CFU/g) spicy green in spicy vegetables: green vegetables: the number of endophytic1 ‒ dill at bacteria, the regime respectively of (4 (Figure - 5) °2С a,; b).2 ‒ dillgreen at vegetablesthe regime at theof transportation(0 - 1) °С; 3 temperature ‒ parsley regimeat the of Analysis1 ‒‒dillafter of atmicrobial transportation; the‒ after regime quantity transportation; of after (4 -transportation5) °С; 2 ‒ ofdill green at the (0–1) °C.regime Asof a (0result - 1) of the°С ;impact 3 ‒ parsleyof the external at the peak incoming productsregime ‒showedafter ofregime anpresales(4‒ increase–after5) of ° Сstorage presales (4in; 2 abundance– ‒5)parsley ° storageС; 2of ‒theatparsley the revealed regime at groups the of regime (0 -heat1) of° loadС (0 upon- 1) °doorС opening during (7–10) min. for unloading ‒ before transportation;‒ before transportation; ‒ afterFigure transportation;‒ after3 Figurepresents transportation;3 presentsthe character the character of changes 45of changesin the air in temperature the air temperature in the body in the of bodya of a refrigerated‒ afterrefrigerated presales ‒truckafter storagepresalesand truck on storageandthe surfaceon the ofsurface fresh ofgreen fresh vegetables green vegetables at the transportationat the transportation temperaturetemperature regime of regime (0 - 1) of °С (0. As - 1) a °resultС. As of a resultthe impact of the of impact the external of the peakexternal incoming peak incoming heat heat load uponFigureload door upon3 opening Figurepresents door 3duringopening presentsthe character (7 during - the10) character min.(7of -changes 10)for unloadingmin.of inchanges for the unloading partair in temperature ofthe products partair temperatureof productsatin differentthe body atin different points theof abody points of a refrigeratedof sale, theofrefrigerated sale,requiredtruck the and requiredtemperaturetruck on and thetemperature regimesurfaceon the of regimeofsurface transportation fresh of of greentransportation fresh ofvegetables greengreen ofcropsvegetables greenat wasthe crops not transportationat maintained wasthe nottransportation maintained temperatureduring mostduringtemperature regimepart most (80 of- 90%)partregime (0 -(80 1)of -of°the90%)С (0. transportationAs - of1) a the°resultС. transportationAs of a theperiod;result impact of with period;the of that,impact the with externala temperature of that, the peakaexternal temperature incomingon thepeak surface onincoming hea thet surface heat loadof green upon vegetablesofload door green upon opening vegetables doorvaried duringopening in varieda range (7 during - in10) of a (5rangemin. (7 - -7) 10)for of °С (5unloadingmin.. - 7) for °С unloading. part of products part of productsat different at differentpoints points of sale, theof requiredsale, the temperaturerequired temperature regime of regime transportation of transportation of green cropsof green was crops not maintainedwas not maintained during mostduring part most (80- 90%)part (80 of -the90%) transportation of the transportation period; with period; that, witha temperature that, a temperature on the surface on the surface 7 7 of green vegetablesof green vegetables varied in varieda range in of a (5range - 7) of°С (5. - 7) °С.

7 7 ПИЩЕВЫЕ СИСТЕМЫ | Том 2 № 4 | 2019 FOOD SYSTEMS | Volume 2 № 4 | 2019

ТемператураTemperature onповерхности the product surface продукции t, °С ТемператураAir temperature воздуха in the body в кузове of a truck 20

15

10

5

0 τ, min 0 25 50 75 100 125 150 175 200 225 250 275 300 325 350 Figure 3. Changes in the air temperature in the body of a refrigerated truck and on the surface of fresh green vegetables at the transportation temperature regime of (0–1) °C and an ambient temperature of (21–23) °C part of productsFigure at 3. different Changes points in ofthe sale, air the temperature required tempera in the- bodyetative of aorgans refrigerated of green productstruck and were on slowed the surface down. For example, ture regimeof offresh transportation green vegetables of green crops at the was transportation not maintained temperature the number regime of epiphytic of (0 -and1) endophytic°С and an bacteria increased by during mostambient part (80–90 temperature %) of the of transportation (21 - 23) °С period;. with approximately 3.6 and 3.8 times in dill, by 4.4 and 7.7 times in that, a temperature on the surface of green vegetables varied in parsley, respectively, compared to the initial data (before trans- a range of (5–7) °C. portation). The number of epiphytic yeasts increased by 4.5 times During transportation,During moisture transportation, of warm ambientmoisture air ofentered warm ambienton dill, the air number entered of theepiphytic body and of endophytica refrigerated molds by 3 times the body oftruck a refrigerated upon opening truck upon the opening door, thewhich door, led which to ledcondensate and 2 times dropout in parsley, and respectivelyformation (Figureof condensed 2 c, e, f). to condensatemoisture dropout on and the formation leaf surfaceof condensed of moisturespicy green on vegetables. The favorable conditions for the leaf surfacepenetration of spicy greenof microorganisms vegetables. The favorable into the conditions lamina were4. Conclusioncreated. Moreover, substances from the for penetration of microorganisms into the lamina were created. Based on the performed research, it can be concluded that the Moreover,surface substances of fromtissues the surfacedissolved of tissues in the dissolved formed in the water. quantitative These substances and species compositionserved as ofnutrition the microbial for community formed water.microorganisms These substances and served stimulants as nutrition for thefor microor development,- of plant which products accelerate and thed character their penetration of its changes into depend on the ganisms andinternal stimulants tissues for ofthe a development, leaf. which accelerated temperature parameters of storage and transportation. their penetration intoAs internala result tissues of the of aconducted leaf. experiments, itFor was apple found fruits thatof the a studied sharp varietiesgrowth that in werethe stored at a As a resultmicrobial of the conductedcommunity experiments, of green it productswas found thatwas a not positiveobserved temperature, immediately the number after of the the revealedend of microbialthe groups sharp growth in the microbial community of green products was increased: bacteria by approximately 10–17 times, yeasts by 95 to not observedtransportation immediately process.after the endFor of theexamp transportationle, the number 180 times, of moldsepiphytic by 3 times.bacteria At the increasedsame time, theby temperature process. Forapproximately example, the number 1.5 times of epiphytic on dill bacteria and parsley increased at theregime temperature of minus (1–2) °Cregime completely of (0 - 1) inhibited °С (Figure the activity of epi- by approximately2a). With 1.5 timesthat, onthe dill number and parsley of epiphyticat the temperature yeasts on phyticdill and bacteria epiphytic (the «Champion» molds on variety), parsley considerably did not delayed the regime of change(0–1) °C (Figure(Figure 2a).2 Withc, e). that, An the increase number ofin epiphyticendophytic multiplication microorganisms of bacteria at the and temperatureyeasts (the «Royal regime Gala» variety), and yeasts onof dill (0 and - 1) epiphytic °С was molds found: on byparsley approximately did not change 1.6 timesalso significantlyfor bacteria slowed on dill down and or parsley;completely by stopped 3.5 the growth (Figure 2 c, e). An increase in endophytic microorganisms at the of endophytic microorganisms. Epiphytic bacteria and endophytic temperaturetimes regime for ofmolds (0–1) °C on wasparsley found: (Figure by approximately2 b, f). 1.6 molds, which cause spoilage of fruits of these varieties at positive times for bacteria Aton dillthe andsame parsley; time, by the 3.5 number times for of molds epiphytic on temperaturesbacteria increased of storage, by wereapproximately not revealed 3 in times the conditions of parsley (Figureon dill 2 b, andf). parsley at the temperature regime ofnegative (4 - 5)temperatures. °С (Figure 2a). The number of At theepiphytic same time, yeaststhe number on dill of epiphytic and epiphytic bacteria moldsincreased on parsleyWithalso respect increased to reduction by 3of times losses from(Figure microbiological 2 c, spoil- by approximately 3 times on dill and parsley at the temperature age, the regime of refrigerated storage of apple fruits at negative regime ofe). (4–5) °C With (Figure that, 2a).the Thenumber number of of endophyepiphytic ticyeasts microorganisms temperatures isincreased preferable comparedas well: to bacteriathe recommended by regimes on dill andapproximately epiphytic molds 2.5 on parsleytimes alsoon dill increased and by by 3more times than of 5 storage times aton positive parsley; temperatures. molds by 3.5 times on (Figure 2 c,parsley e). With (Figure that, the2 number b, f). Endophytic of endophytic yeasts microorgan in both- typesIt is of generally green vegetablesagreed that short-term were absent violation (Fig.2 of temperature- isms increasedd). as well: bacteria by approximately 2.5 times on humidity regimes has no effect on food quality indicators [26]. At dill and by more thanHowever, 5 times onafter parsley; 3 days molds of by presales 3.5 times storage on the in same the time,retail variations refrigerated in ambient equipment temperature upon and humidity, as parsley (Figure 2 b, f). Endophytic yeasts in both types of green in the case of transportation of spicy green vegetables, can cause vegetablesmaintaining were absent (Fig.2adequate d). temperature regimes of storage,the formation certain of condensedmicrobiological moisture onindicators the surface of of plant prod- However,spicy after green 3 days vegetables of presales storagesignificantly in the retail deteriorated. refriger- ucts, an increase in the intensity of its breath, the development ated equipment uponAt maintainingthe temperature adequate regime temperature of (4regimes - 5) °Сand, ther multiplicatione was an ofincrease epiphytic in and the endophytic number microorganisms, of of storage,epiphytic certain microbiological bacteria by indicators approximately of spicy green6 times veg- on which,dill and finally, by more can lead than to loss7 times of quality on parsley,and product spoilage etables significantlyepiphytic yeastsdeteriorated. by approximately 10 times on dillduring and epiphytic following storage.molds by 20 times of parsley At the temperature regime of (4–5) °C, there was an increase Analysis of microbial quantity after transportation of green in the number(Figure of epiphytic2 a, c, bacteriae) compared by approximately to the in6 timesitial dataproducts (before showed transportation). an increase in abundanceThe number of the revealedof groups on dill andendophytic by more than microorganisms 7 times on parsley, also epiphytic increased: yeasts by bacteriaof epiphytes by more and endophytesthan 7 times by 1.5–3 in timesdill andupon byabsolute preva- approximately 10 times on dill and epiphytic molds by 20 times lence of bacteria. After short-term storage, a significant growth of of parsley (Figure 2 a, c, e) compared to the initial data (before 8 the revealed groups of microorganisms took place; with that, their transportation). The number of endophytic microorganisms also in- quantity was 1.5–6.5 times higher at (4–5) °C than at (0–1) °C. creased: bacteria by more than 7 times in dill and by approximately As the performed investigations show, the multiplication 13 times in parsley; molds by 5.5 times in parsley (Figure 2 b, f). and development of microorganisms during transportation re- At the temperature regime of storage of (0–1) °C, the develop- duced resistance of green crops to further damage of tissues by ment and multiplication of the microbial community of the veg- microbiota.

46 ПИЩЕВЫЕ СИСТЕМЫ | Том 2 № 4 | 2019 FOOD SYSTEMS | Volume 2 № 4 | 2019

Near-zero temperatures of transportation and storage reduced plant products significantly depends on the temperature regimes the growth of the microbial community during following short-term and their variation during storage and transportation. storage of green products by several times. Therefore, to maintain Detection of microbial contamination of fruits and vegetables high quality of green crops and reduce losses, it is necessary to before their movement along the main stages of the CCC will store and transport them at a temperature of 0 °C — 0.5 °C. When permit improving (regulating) temperature-humidity parameters adhering to these conditions, shelf-life of green products can be of the processes of storage, transportation and presales storage significantly prolonged. of plant products, increasing their quality, reducing losses and It was established that the process of the development and extending shelf-life. multiplication of epiphytic and endophytic microorganisms of

REFERENCES 1. Shirokov, E.P., Polegaev, V.I. (1989). Storage and processing of fruits and 14. Robert, D. (2010). Guide to Refrigerated Transport. Paris: International vegetables. Moscow: Agropromizdat. — 302 p. ISBN: 5–10–001284–6 Institute of Refrigeration. — 182 p. (in Russian) 15. Leif BØgh-Sorensen .(2006). Recommendations for Processing and Han- 2. Vankova, A.A. (2012). Microbiological processes in storage and process- dling of Frozen Foods. Paris: International Institute of Refrigeration. — ing of vegetable products. Textbook. Moscow: State Agrarian Universi- 174 p. ty — Moscow Timiryazev Agricultural Academy. — 58 p. ISBN: 978–5– 16. Bab’eva, I.P., Chernov, I. Yu. (2004) Biology of yeasts. Мoscow: KMK Sci- 9675–0708–3 (in Russian) entific Press Ltd. — 221 p. ISBN: 5873171793 (in Russian) 3. Kudryashova, A.A. (1986) Microbiological principles of preservation of 17. Glushakova, A.M., Chernov, I.Y. (2010). Seasonal dynamics of the struc- fruits and vegetables. Moscow: Agropromizdat. —189 p. (in Russian) ture of epiphytic yeast communities. Microbiology (Mikrobiologiya), 79(6), 4. Emtsev, V.T., Mishustin, E.N. (2005).). Microbiology: textbook for higher 830–839. DOI: 10.1134/S0026261710060160 educational institutions. Moscow: Drofa. —445 p. (in Russian) 18. Chernov, I. Yu. (2013). Yeasts in nature. Moscow: KMK Scientific Press 5. Filtenborg, O., Frisvad, J.C., Thrane, U. (1996). Moulds in food spoil- Ltd. —221 p. ISBN: 978–5–87317–927–5 (in Russian) age. International Journal of Food Microbiology, 33(1), 85–102. DOI: 19. Blagoveschenskaya, E. Yu., Dyakov, Yu. T. (2005). Fungal endophytes 10.1016/0168–1605(96)01153–1 of cereal grasses. Mycology and Phytopathology, 39(3), 1–15. (in Rus- 6. Blackburn, C.de W., McСlure, P.J. (2009). Foodborne Pathogens: Hazards, sian) Risk Analysis and Control. CRC press. —521 p. ISBN: 0–8493–1213–2 20. Mathews, J.F., Clay, K. (2001). Influence of fungal endophyte infection on 7. Blackburn, C.de W., (2008). Микробиологическая порча пищевых plantsoil feedback and coomunity interactions. Ecology, 82(2), 500–509. продуктов. St. Petersburg: Profession. — 784 p. ISBN: 978–5–93913– DOI: 10.1890/0012–9658(2001)082[0500: IOFEIO]2.0.CO;2 146–9 (in Russian) 21. Müller, C.B., Krauss, J. (2005). Symbiosis between grasses and asexual 8. Gould, G.W. (1989). Heat-induced injury and inactivation. In book: Mech- fungal endophytes. Current Opinion in Plant Biology, 8(4), 450–456. DOI: anisms of Action оf Food Preservation Procedures. London; New York: 10.1016/j.pbi.2005.05.007 Elsevier Applied Science. pp. 11–42. ISBN: 1–85166–293–6 22. Rosenblueth, M., Martínez-Romero, E. (2006). Bacterial endophytes and 9. Herbert, R.A. (1989). Microbial growth at low temperatures. Mechanisms their interactions with hosts. Molecular plant-microbe interactions, 19(8), of Action оf Food Preservation Procedures. London; New York: Elsevier 827–837. DOI: 10.1094/MPMI–19–0827 Applied Science. pp. 71–96. ISBN: 1–85166–293–6 23. Gryzunov, A.A., Kornienko, V.N., Avilova, S.V. (2018). Calculation of Re- 10. Scheglov, N.G.(2003). Refrigeration technology of food products. Pyatig- frigeration Capacity of Refrigerated Trucks for Intracity Transportation orsk: KZ. — 208 p. (in Russian) of Food Products. Storage and processing of farm products, 2, 100–104. 11. Encyclopedia «Food technologies». Vol. 16 «Technologies of refriger- (in Russian) ated processing and storage of food products», book 2. Uglich: ID Uglich, 24. Netrusov, A.I., Egorova, M.A., Zakharchuk, L.M. (2005). Practical 2019. — 298 p. (in Russian) course on microbiology. Мoscow: Academy. —608 p. ISBN: 576951809X 12. Gryzunov, A.A., Kornienko, V.N. (2014). Structural analysis of refrigerated (in Russian) vehicles — refrigerators for intercity transportation of perishables food- 25. Tepper, E.Z., Shilnikova, V.K., Pereverzeva, G.I. (2004). Practical course on stuffs. Kholodilnaya Tekhnika, 12, 45–48. (in Russian) microbiology. Moscow: Drofa. — 256 p. (in Russian) 13. Gryzunov, A.A., Kornienko, V.N. (2015). About the test of compliance of 26. Gryzunov, A.A., Ivatsevich, B.P., Yanovskiy, R.B. (2005). Special vehicles heat engineering characteristics of special vehicles for perishables with for perishables. Kholodilnaya Tekhnika, 2, 28–31. (in Russian) international standards. Kholodilnaya Tekhnika, 5, 47–50. (in Russian)

AUTНOR INFORMATION Svetlana V. Avilova — candidate of agricultural sciences, docent, leading research scientist, Laboratory of refrigeration processing and storage of food products, All-Russian Scientific Research Institute of Refrigeration Industry — Branch of V. M. Gorbatov Federal Research Center for Food Systems of the RAS, 127422, Moscow, Kostyakova str., 12. Теl.: +7–495–610–80–92, e-mail: [email protected] Vladimir N. Kornienko — candidate of technical sciences, docent, leading research scientist, Laboratory of refrigeration processing and storage of food prod- ucts, All-Russian Scientific Research Institute of Refrigeration Industry — Branch of V. M. Gorbatov Federal Research Center for Food Systems of the RAS, 127422, Moscow, Kostyakova str., 12. Теl.: +7–495–610–80–92, e-mail: [email protected] *corresponding author Aleksey A. Gryzunov — senior research scientist, Laboratory of refrigeration processing and storage of food products, All-Russian Scientific Research Institute of Refrigeration Industry — Branch of V. M. Gorbatov Federal Research Center for Food Systems of the RAS, 127422, Moscow, Kostyakova str., 12. Теl.: +7–495– 610–80–92, e-mail: [email protected] Anna A. Vankova — candidate of biological sciences, docent, Department of microbiology and immunology, State Agrarian University — Moscow Timiryazev Agricultural Academy, 127550, Moscow, Timiryazevskaya str., 49. Tel.: +7–499–976–09–66, e-mail: [email protected] All authors bear responsibility for the work and presented data. All authors made an equal contribution to the work. The authors were equally involved in writing the manuscript and bear the equal responsibility for plagiarism. The authors declare no conflict of interest. Received 22.10.2019 Accepted in revised 25.11.2019 Accepted for publication 10.12.2019

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UDC 664.8.036.5 DOI: 10.21323/2618–9771–2019–2–4–48–52 Original scientific paper DEVELOPMENT OF PASTERIZATION MODES FOR HIGH-SUGAR CANS IN CONTINUOUS ACTING PASTEURIZERS Galina P. Pokudina*, Marina V. Trishkaneva, Raisa A. Volkova Russian Research Institute of Canning Technology — Branch of V. M. Gorbatov’s Federal Research Center for Food Systems of RAS, Vidnoe, Moscow region, Russia

KEY WORDS: ABSTRACT continuous-acting pasteurizer High-sugar canned fruits, such as jam, marmalade, confiture, are characterized by a high content of soluble dry (CAP), pasteurization modes, solids and high acidity, which allows to apply to them pasteurization processes as heat treatment. In comparison high-sugar products, packing with sterilization processes, heat treatment of hermetically sealed canned food during pasteurization is imple- temperature, required and actual mented at temperatures less than 100 °C. lethality The article presents the results of the development of scientifically-based pasteurization modes for high-sugar canned food for industrial continuous-acting pasteurizers (CAP — continuous-acting pasteurizers) of open type with a temperature in the heating area less than 100 °C. For today, such pasteurizers are widely spread due to the simplicity of their design, high productivity, with the option to reduce consumption of water, steam, electricity and to use it for the various types of packaging. So, the actual task of this article is the establishment of canned fruit pasteurization modes for such pasteurizers. The development of pasteurization modes for high-sugar canned food using strawberry jam as a sample was car- ried out in the experimental equipment simulating the operation of a continuous-acting pasteurizer. The studies were implemented to confirm that the developed modes of heat treatment in the CAP as a matter of fact provide industrial sterility of the produced high-sugar canned food. Optimal pasteurization modes have the following parameters: heat treatment in an autoclave sterilizer at a heating temperature of the heating medium 97 °C and stage-by-stage cooling of the products with water at temperatures 70 °C, 50 °C and 30 °C.

1. Introduction The required lethality of heat treatment for canned food is The pasteurization process is preferable in comparison with referred to normative value equal to duration of heating in the the sterilization process in the subsequent manufacture of canned least heated point of the product with the base temperature, food for a range of products made of processed fruit. These prod- which ensures the destruction of vegetative forms and spores of ucts include the group of high acidic products with pH less than microorganisms, that cause the spoilage of the product [6, 9,10,11]. 4.2 and high-sugar canned food with a content of soluble dry Meanwhile choosing the new pasteurization modes it is necessary solids within the range from 57 to 72 %, such as marmalade, to take into account their impact on organoleptic parameters and jam, confiture and etc. [1]. High-sugar canned food is obtained nutritional value of canned products. The developed mode must by boiling of fruits and berries with sugar till the desired con- guarantee the absence of the most heat-resistant microorganisms centration of soluble dry solids (sugars). Such canned food is in the canned food, that can cause the spoilage of the product also characterized by the low content of active water αw — less during storage, and also provide the organoleptic properties of than 0.75. Canned food with a low value of aw (less than 0.85) the product [12]. are susceptible for spoiling mainly due to development of fungi The choice of the optimal pasteurization mode also depends on and yeast in them, but the bacteria, do not develop when aw in the type of packaging of the finished product, the design features the products is less than 0.85 [2, 3]. of the pasteurizers used (immersive, steaming, irrigational), as Thus the high concentration of sugars and organic acids in well as the type and method of supplying a of the thermal agent combination with the low content of active water in high-sugar (steam, water, water streams) to heat the canned products in the canned fruit lead to destruction of yeast, fungi and vegetative pasteurization zone. forms of bacteria during heat treatment with temperatures be- Continuous-acting pasteurizers (CAP) of immersive and ir- low 100 °C, which is typical for the processes of the canned food rigational type are usually used for pasteurization of high-sugar pasteurization. products, packaged in glass jars or in heat-resistant polymer Thermal pasteurization for food preservation is based on packaging [13, 14]. In the industrial version this pasteurizer is a the concept of lethal effect of heat treatment on microorgan- closed chamber with moving mesh transporter belt inside. The isms (heating) [4, 5]. Choosing the parameters of pasteuriza- chamber consists of several zones: hot water pasteurization zone tion modes — temperature and duration of products heating in (heating zone) and 1–3 cooling zones. The cooling process is car- autoclaves and pasteurizers, first of all the fact matters that the ried out by water or combined air and water method. In this case heat treatment mode provides industrial sterility of the produced there are two types of cooling zones: air cooling zones and water canned food. In industrial-sterile canned food the microbiological cooling zones. The product remains in continuous motion during stability and safety of the product are preserved throughout the the entire pasteurization cycle in the CAP. The loading and the shelf life defined for it. Canned food, heated at the temperature unloading processes are mechanized; the temperatures of the below 100 °C, conforms to the requirements of industrial sterility heating and cooling zones are set in accordance with the selected in case if the actual lethality of the heat treatment process Afact pasteurization mode. The temperature is maintained and recorded is equal or slightly exceeds the value of the required lethality automatically, which ensures uniformity of the temperature area Z Z AT: Аfact ≥ AT [6, 7, 8]. in the CAP zones [15, 16, 17].

FOR CITATION: Pokudina G. P., Trishkaneva M. V., Volkova R. A. Development of high-sugar-sufficient pasterization modes canning in continuously-acting paster- izers. Food systems. 2019; 2(4): 48–52. DOI: 10.21323/2618–9771–2019–2–4–48–52

48 ПИЩЕВЫЕ СИСТЕМЫ | Том 2 № 4 | 2019 FOOD SYSTEMS | Volume 2 № 4 | 2019

The pasteurization of the canned food with using CAP has a ment 1, packaged in glass jars 2 with screw lids, the diameter of range of advantages: the neck 68 mm and 350 cm3 volume (Jar 1) and the diameter ‰‰ no time intervals between packing and pasteurization of the of the jar neck is 82 mm and the volume is 650 cm3 (Jar 2), the canned food; temperature of the canned food packing before its immer- ‰‰ decreased duration of the pasteurization process; sion in an autoclave at 70 °C.

‰‰ reduced labor costs due to complete mechanization and au- ‰‰ device «Citronics», Swiss, for measuring of aw tomation of loading and unloading of canned food; ‰‰ sample test-culture of fungi-strains of Penicillium glaucum ‰‰ reduced consumption of water, steam and electricity per unit fungi spores of production; ‰‰ agarised nutrient medium DG 18 with dichlorane and glyc- ‰‰ option to produce many types of canned food. erin according to ISO16000–17:2008 3. Thus the aim of the research is development of heat treatment There was assembled the experimental stand for develop- mode for the high-sugar products (marmalade, jam, confiture, ment of pasteurization modes, on this stand the simulation of berries pureed with sugar, etc.), providing industrial sterility of the operation of a continuous immersive pasteurizer was car- the produced cans for continuous-acting pasteurizers with tem- ried out. Initially the canned strawberry marmalade was heated perature below 100 °C without backpressure. up to a temperature inside the product at 70 °C and immersed in the experimental vertical autoclave stand. The product in 2. Materials and methods the autoclave was carried out heated up at temperatures of the The following equipment, materials and methods were used heating area: 97 °C, 95 °C, 90 °C (simulation of the CAP heating to perform the work: zone). Cooling was carried out in 3 thermostats with maintaining ‰‰ temperature recorder with automatic data processing (calcu- the temperature of the cooling area, respectively, 70 °C, 50 °C lation of the actual lethality according to Bigelow) of brand and 30 °C (cooling zone of the pasteurizer). The heating and «Ellab» CTF 9008 (Denmark) with a set of thermocouples for cooling medium was water. Temperature of the canned food, measuring of the temperature from 100 to 350 °C with an ac- heating and cooling medium was measured by the temperature curacy of 0.1 °C. It was used to measure the temperature of recorder «Ellab». The schematic drawing of the experimental the heating area in the autoclave and the temperature at the stand is shown in Figure 1. least heated point of the product; ‰‰ laboratory electric vertical autoclave VEE‑2–1–0.08–0.3–3 3. Results and discussion UHL (Russia) with automatic setting of heat treatment The required lethality for strawberry marmalade pasteuriza- modes, with working pressure‑3 MPa and temperature of tion modes with a pH value = 4,2 4(in foreign practice a control pH the heating area 135 °C. The autoclave simulates the heating value = 4.6 is used in accordance with document4) is selected from zone of a continuous pasteurizer, in the experiment it was the conditions that ensure the death of mold spores of Penicillium used without backpressure; Z = 15 °C glaucum, and is equal to AT = 80 °C = 40 ÷ 80 conditional minutes. ‰‰ three WCH‑12 high-temperature circulation thermostats 1 GOST 34113–2017 Gams. General specifications (Korea) with constant maintenance of water temperature 2 GOST 5717.2–2003 Glass jars for canned food. Basic parameters and di- 70 °C, 50 °C and 30 °C (the simulation of pasteurizer cooling mensions zones). 3 ISO 16000–17–2008 Indoor air — Part 17: Detection and enumeration ‰‰ strawberry marmalade with the content of soluble sol- of moulds — Culture-based method 4 ids‑68 %, manufactured according to the normative docu- CAC/RCP 23–1979 (Rev.2–1993) «Recommended international code of hygienic practice for low and acidified low acid canned foods».

Water

Tubular electric heater (TEH)

Power 220W

Figure 1. The scheme of the experimental stand, simulating the work of CAP 1 — can with thermocouple, 2 — autoclave, 3 — cooling section (thermostats), 4 — temperature recorder, 5 — temperature sensor, 6 — water valve, 7 — heating control unit Вода - Water Tubular electric heater (TEH) Power 220W Figure 1. The scheme of the experimental stand,49 simulating the work of CAP 1 - can with thermocouple, 2 - autoclave, 3 - cooling section (thermostats), 4 - temperature recorder, 5 - temperature sensor, 6 - water valve, 7 - heating control unit.

. eslts and disssion The required lethality for strawberry marmalade pasteurization modes with a pH value = 4,2 1(in foreign practice a control pH value = 4.6 is used in accordance with document4) is selected from the conditions that ensure the death of mold spores of enicillium glaucum, and

is equal to =1 =40 ÷ 80 conditional minutes. =0𝑜𝑜 𝑧𝑧 𝑜𝑜𝐶𝐶 The𝐴𝐴 heating𝑇𝑇 𝐶𝐶 capacity of canned food during pasteurization depends mainly on its con- texture determined by the recipe, the size and material of the package, as well as the initial temperature of the product and the pasteurization temperature. The greatest efficiency of the pasteurization mode can be achieved by increasing the pasteurization temperature or the product packing temperature. The rate of heat transfer in the jars with the product was determined by heating of the canned food in an open autoclave (which simulated the heating zone with water at a tempera- ture of 97 °C, 95 °C, 90 °C), and cooling in 3 tanks with water at a temperature of 70 °C, 50 °C and 30 °C turn-by-turn (cooling zone of the pasteurizer). The temperature of the heating

1 CAC/RCP 23-1979 (Rev.2-1993) «Recommended international code of hygienic practice for low and acidified low acid canned foods». 5

9

9

9

ПИЩЕВЫЕ СИСТЕМЫ | Том 2 № 4 | 2019 FOOD SYSTEMS | Volume 2 № 4 | 2019

The heating capacity of canned food during pasteurization without cooling. This product is packed in 2 stages: first — the

depends mainly on its texture determined by the recipe, the size berries are packed in a jar, and then they are poured by syrup. 9 and material of the package, as well as the initial temperature of Pasteurization modes were developed for high-sugar products the product and the pasteurization temperature. The greatest effi- (marmalade), the packing temperature of which cannot be raised ciency of the pasteurization mode can be achieved by increasing the to the temperature of the pasteurization zone, according to the pasteurization temperature or the product packing temperature. production technology. 9 The rate of heat transfer in the jars with the product was deter- ProductsПро дwithолж aи lowтел packingьность temperatureобработки, (70 °C)мин - mustProce stayssing 9time, min mined by heating of the canned food in an open autoclave (which longer in the pasteurization zone. For other types of high-sugar simulated the heating zone with water at a temperature of 97 °C, canned foodЛета (jams,льнос confituresть - Letha liand,ty, cberriesonditi onpureedal mi n.with sugar), 95 °C, 90 °C), and cooling in 3 tanks with water at a tempera- which packingПродолж temperatureительность can об рbeаб raisedотки, мhigher,ин - P theyrocess willing meettime ,themi n Продолжительность обработки, мин - Processing time, min ture of 70 °C, 50 °C and 30 °C turn-by-turn (cooling zone of the requirements of industrial sterility even higher, if pasteurization pasteurizer). The temperature of the heating and cooling water is carriedЛеF тoutiаgлuь reaccordingно с3.ть Th - Lee ttohcah theliartyt, modes ocondf thitie developedonteaml miperan. t uforre marmalade.changes of the heating area in the autoclave (at as well as the temperature of the product, was measured by the AccordingЛетальн toос тtheь - Lexperimentalethality, cond itidataona lobtained min. at the stand, thermocouples and «Ellab» potentiometer. The hot junction of the charts95 )of a temperaturend strawber changesry marm curvealad ein w thehe nheating heate darea in aand gl athess jar 2 Figure 3. The chart of the temperature changes of the heating area in the autoclave (at thermocouple was placed into the least heated area of the product. temperatureFigure in 3. the Th leaste char heatedt of th pointe tem pofera thetu rproducte chang withes of indication the heatin g area in the autoclave (at 95 ) and strawberry marmalade when heated in a glass jar 2 In high-sugar products heat transfer occurs only due to thermal 95of the) actualandB eslt raolethalityww btehrreyre valuem ararem (Figure aplardees ew n2ht eandend h etFigurehatee dd ian 3) aret a golfa ssconstructed.p rjaroc 2essing of experimentally obtained heating conductivity, so the hot junction of the thermocouple was placed The heating of canned food for recording of temperature change in the geometric center of the product packed in a glass jar [17]. curvescurves Bwas aelndo wcarried p tahsetree uoutarreiz inaptir ethreeonsen tme dreplications.od thees doaft ah oigf hp-rsoucegssarin cga onfs efxoprer cimonetintnuousally ob-tacaintiendg heaimmting ersive pasteur- Below there are presented the data of processing of experimentally obtained heating The pasteurization modes were calculated basing on the fact Below there are presented the data of processing of experi- curves and pasteurization modes of high-sugar cans for continuous-acting immersive pasteur- that the about 1/3 value of the actual lethality is achieved by the cmentallyiuzrervess aatnd h obtained eapatistneugr iarz aheatingetiaon te mmod pcurveseraes otfu h randiegsh -97 spasteurizationug ar c, a95ns for caon ndmodesti nuous90 of- ac. ting immersive pasteur- izers at heating area temperatures 97 , 95 and 90 . heat-conducting products during their heating up, and 2/3 are ihigh-sugarzers at heaFoti rcansn gh iarg foreha- s tcontinuous-actingeumgpaerar ptruodures 97cts (s, timmersive95ra wb eandrry 90 pasteurizersm arm. alade) at, packed in a glass jar 1 on the stand, achieved during cooling down. heating area temperatures 97 °C, 95 °C and 90 °C. For high-sugar products (strawberry marmalade), packed in a glass jar 1 on the stand, According to the obtained experimental data on heating rate wasFor prF high-sugaroocr ehssigehd-s uign productsa rt hperodu follc (strawberrytos w(sitnragw bmeodrr marmalade),ye ms aromf apladste)e packed,u priaczaktie dinon i na wa itglhass th jear ca1 olcnu tlhaeti sontand o,f the actual le- was processed in the following modes of pasteurization with the calculation of the actual le- of the canned food calculated the value of the actual lethality. To wglassas p rjaroc e1ss oned the in tstand,he foll owaswin processedg modes o fin p theaste followingurization w modesith the of ca lculation of the actual le- calculate the lethality rate, the temperature in the product was tpasteurizationthhaalilityt:y: with the calculation of the actual lethality: thality: read at regular intervals (every 5 minutes). The corresponding 2 10 10 1 =1 2 10 10 1 =1 lethality coefficients for these product temperature readings were 2 10 10 1 =01 =0001= 001 00 0 0 30 30 0 = 001 0 0 30 0 added and the resulting sum was multiplied by the accepted time о о о о 22оо 101о0о10о10о1о1о =1 2оС − 10оС − 10оС − 1оС =1 = =1 interval and in this way the actual lethality value was obtained. СС−−0 С −С −0 С −С30−С С 0= 1 = = С − 0С0 − 0 С0− 30 С 3 0 0 =0 о 0о 0о 30о The lethality value is the value of lethality per 1 minute at a 30о 10о 10о 1о =1 3300оСо− 101о0Со−10о1С0−о1оС1. о =1 = 1 certain temperature in the least heated point of the product. The 0 СС− 0 С −С 0 С −С30 С . С =0 1 = =1 1 0 −−0 − −0 − 30− . . 0 =01= 1 00о 0о 0 0о 0 30о 30 lethality value was calculated by the formula: оС оС оС оС оСо − оС о− оС −о оС о СThe− obtainedС − Сvalues− Сof the actual lethality were compared Tproduct – Tbas С − С − С − С l = 10 , The obtained values of the actual lethalitZy = w15e °Cre compared with the value of the re- z with theTh evalue obta iofne dthe va lrequiredues of th elethality — actual leth a liAtTy = w80e °Cre =c o40m p÷are 80.d with the value of the re- where The obta=in1ed values of the actual lethality were compared with the value of the re- quThe choicetionired mode lettionha liof t isymode the- determined optimal= is10 determined = pasteurization0 by the by0 durationthe modeduration isof determined ofthe. theTh pasteurizatione pasteurizationcho ibyce of the op process.timal pas tFor eForur imarmalade,z amarmalade,- packed packed Z — is the constant of thermal stability of the test-microorganism, °C. quired lethality - =0 = 0 0 . The choice of the optimal pasteuriza- qutheire durationd lethali tofy -the =pasteurization0 =1 process. For marmalade,. The packed choice of the optimal pasteuriza- quired linet htheali tglyass- jar= 1,0 this = mode0 was 0 the following: . The choice of the optimal pasteuriza- In this case z = 15 °C, and the base temperature (Tbas) is 80 °C. inin thethe glass glass jar jar 1, this1, this mode mode was the was following: the following: 2 210 1010 1011 During the tests on the stand, the marmalade was heated up 0 0 30 withwith the thevalue value of the of theactual actual lethality 0 0 30 with the value of the actual lethality to a packing temperature 70 °C, which is 3–5 °C below the tem- о о о о о =о1 о о perature used in production conditions. In production conditions lethality=1 0С −= 0С01− С − С 0С −= 0С01− С − С the packaged product immediately enters the pasteurizer almost 𝑧𝑧 𝑧𝑧 For high-sugar products (strawberry marmalade), packed in a glass jar 2, the pasteuri- For𝐴𝐴 high-sugar𝑐𝑐𝑐𝑐 products𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛 (strawberry𝑚𝑚𝑚𝑚𝑛𝑛 marmalade), packed in a glass jar 2, the pasteuri- 𝐴𝐴 𝑐𝑐𝑐𝑐𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛 𝑚𝑚𝑚𝑚𝑛𝑛 zation modes are following: zation modes are following: Pasteurizer 40 min 10 min 10 min 20 min =1 − − − 0 = 2 . Lethality о о о о 40 min 9710Сmin70 10С min50 С20 min30 С =𝑧𝑧1 − − − 0 = 2 . 9о о о о 𝐴𝐴 𝑐𝑐𝑐𝑐𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛 𝑚𝑚𝑚𝑚𝑛𝑛 97 С 45 min70 С10 min50 С10 min30 20Сmin 𝑧𝑧 =1 − − − 0 = 0 95о С 70о С 50о С 30о С 𝐴𝐴 𝑐𝑐𝑐𝑐𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛 𝑚𝑚𝑚𝑚𝑛𝑛 Product 45 min 10 min 10 min 20 min =𝑧𝑧1 − − − 0 = 0 о о о о 𝐴𝐴 𝑐𝑐𝑐𝑐𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛 𝑚𝑚𝑚𝑚𝑛𝑛 95 С 50 min70 С10 min50 С10 min30 20С min =1 𝑧𝑧 − − − = 1 о о о о 0 90 С 70 С 50 С 30 С 𝑧𝑧 50 min 10 min 10 min 20 min 𝐴𝐴 =1 𝑐𝑐𝑐𝑐𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛 𝑚𝑚𝑚𝑚𝑛𝑛 − − − 𝐴𝐴0 = 1 𝑐𝑐𝑐𝑐𝑛𝑛𝑛𝑛𝑛𝑛𝑡𝑡𝑡𝑡𝑜𝑜𝑜𝑜𝑜𝑜𝑜𝑜 𝑚𝑚 𝑚𝑚𝑛𝑛 о о о о 90 С 70 С 50 С 30 С 𝑧𝑧 Lethality, conditional min.

The comparison of the obtained values of the actual lethality with the value of the re- 𝐴𝐴 𝑐𝑐𝑐𝑐𝑛𝑛𝑛𝑛𝑛𝑛𝑡𝑡𝑡𝑡𝑜𝑜𝑜𝑜𝑜𝑜𝑜𝑜 𝑚𝑚𝑚𝑚𝑛𝑛 quired lethality - =40 ÷ 80 . shows that the optimal pasteurization The comparison of the𝑜𝑜 obtained values of the actual lethality with the value of the re- 𝑧𝑧=15 𝑜𝑜𝐶𝐶 mode for marmalade,𝑇𝑇= 8packed0 𝐶𝐶 in a glass jar 2, was the following: 𝐴𝐴 𝑐𝑐𝑐𝑐𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛 𝑚𝑚𝑚𝑚𝑛𝑛 quired lethality 0- 10 =4010 ÷ 8020 9 . shows that the optimal pasteurization 𝑜𝑜 with the value of the actual lethality 𝑧𝑧=150 𝑜𝑜𝐶𝐶 0 30 Processing time, min mode for marmalade,𝑚𝑚𝑖𝑖𝑛𝑛 𝑇𝑇= 8packed𝑚𝑚0𝑚𝑚𝑛𝑛𝐶𝐶 𝑚𝑚in𝑚𝑚𝑛𝑛 a glass𝑚𝑚𝑚𝑚𝑛𝑛 jar 2, was the following: =1 о 𝐴𝐴 о о 𝑐𝑐𝑐𝑐о 𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛 𝑚𝑚𝑚𝑚𝑛𝑛 Figure 3.= The Сchart2 of theС temperatureС changesС of the Figure 2. The chart of the temperature changes of the 0 − − − Продолжheatingи0тель areaност 1ьin0 о theбра бautoclaveот1ки0, мин (at 95 °C) - P2r0ocess iandng tim strawberrye, min heating area in the autoclave (at 97 °C) and strawberry 𝑧𝑧 with the value of the actual lethality marmaladeSo, 0for whenthe established0 heated in30 a glassmodes jar 2of pasteurization the optimal modes are: the heating marmalade when heated in a glass jar 1 𝐴𝐴 𝑚𝑚𝑖𝑖𝑛𝑛 𝑚𝑚𝑚𝑚𝑛𝑛 𝑐𝑐𝑐𝑐𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑚𝑚𝑚𝑚𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛 𝑚𝑚𝑚𝑚𝑚𝑚𝑚𝑚𝑛𝑛𝑛𝑛 Лет=а1льностьо - Lethalityо, conditionоal min. о 0 =(heatС treatment)2− С − in anС autoclave − С at a temperature of the heating area 97 °C and step-by-step 50 Fi𝑧𝑧gure 3. coolingThe char tof o f thethe tproductemperatu rbye c hwateranges withof the temperaheating artureea i n70 th e°C, aut o50cla v°Ce (a andt 30 °C. The time of food 𝐴𝐴 So, for 𝑐𝑐𝑐𝑐the𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛 established𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛 𝑚𝑚 𝑚𝑚𝑛𝑛modes of pasteurization the optimal modes are: the heating 95 ) and strawexposureberry marminal athede w heatinghen heat ezoned in a is g l25ass jminutesar 2 (for jar 1) and 40 minutes (for jar 2), in 1-st and 2- (heat treatment) in an autoclave at a temperature of the heating area 97 °C and step-by-step Below thndere coolingare prese nzonested thethe da tjarsa of areproc exposedessing of eforxp er10im eminutes,ntally ob tainin ethed h lasteatin coolingg zone for the product, cooling of the product by water with temperature 70 °C, 50 °C and 30 °C. The time of food curves and pasteupackagedrization m odine glasss of higjarh- s1ugisar cooledcans for forconti15nuous minutes,-acting andimm theersi veproduct,pasteur -packed into the glass jar 2 is exposure in the heating zone is 25 minutes (for jar 1) and 40 minutes (for jar 2), in 1-st and 2- izers at heating arcooledea temp forera t20ure sminutes 97 , 95. and 90 . For high-sugar products (strawberry marmalade), packed in a glass jar 1 on the stand, nd cooling zonesThe temperaturethe jars are ofexposed the productfor 10conveyed minutes,to the in CAPthe last pasteurization cooling zone zone, for mustthe not product, be was processed in the following modes of pasteurization with the calculation of the actual le- packagedlower in thanglass 70jar °C. 1 Increasingis cooled forthe product15 minutes, packing and temperature the product, on 5packed °C will intoreduce the the glass pasteu jarr- 2 is thality: 2 10 10ization 1 mode, and =that1 will reduce the time of exposure in the heating zone for 7-10 minutes. cooled for 20 minutes. = 001 0 0 30 0 2о 10о 10о 1Theо canned food, produced according to the obtained pasteurization modes, meet the С С TheС temperatureС = 1of = the product conveyed to the CAP pasteurization zone, must not be − 0 − 0 − 30 0 30о 10о 10requirementsо 1о of industrial sterility. When determining the compliance of canned food with the С −lowerС − thanС − 70 °C.С Increasing =1 = the1 product packing temperature on 5 °C will reduce the pasteur- 0 0 0 30 . 0 о о requirementsо о of industrial sterility, the absence of mold fungi (including enicillium glaucum) С −izationС − mode,С − andС that will reduce the time of exposure in the heating zone for 7-10 minutes. and yeast was confirmed by plating of DG 18 with dichlorane and glycerin into agarized nu- The obtaiThened v acannedlues of t hfood,e actua lproduced lethality w eaccordingre compared wtoit hthe the obtainedvalue of th epasteurization re- modes, meet the trient=1 medium. quired lrequirementsethality - =0 of = industrial0 0 sterility. When. The c hdeterminingoice of the optimtheal p complianceasteuriza- of canned food with the

requirements of industrial sterility, the absence of mold fungi (including enicillium glaucum9 ) and yeast was confirmed by plating of DG 18 with dichlorane and glycerin into agarized nu- trient medium.

9 tion mode is determined by the duration of the pasteurization process. For marmalade, packed in the glass jar 1, this mode was the following: 2 10 10 1 0 0 30 with the value of the actual lethality tion mode is determined by the duration of the pasteurization process. For marmalade, packed о=1 о о о С С С С 0 −= 001−in the gl−ass jar 1, this mode was the following: tion𝑧𝑧 modetion is mode determined is determined by the durationby the duration of the pasteurizationof the pasteurization process. process. For marmalade, For marmalade, packed packed 2 10 10 1 For high-sugar products (strawberrywith the marmalade), value of the actual packed lethality in a glass jar 2, the pasteuri- 𝐴𝐴in the glinass the jar gl 1,ass this jar𝑐𝑐𝑐𝑐 mode 1,𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛 this was0 mode𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛 the was following:0𝑚𝑚 the𝑚𝑚𝑛𝑛 following:30 о о о о zation2 modes120 1 0are following:=110 1 0С −= with0С01− the valueС − of Сthe actual lethality ПИЩЕВЫЕ0 0 СИСТЕМЫ300 30 |with Том the 2 value№ 4 of | the2019 actual lethality FOOD SYSTEMS | Volume 2 № 4 | 2019 о о о 𝑧𝑧 о =1 о=1 о оFor highо -sugar products (strawberry marmalade), packed in a glass jar 2, the pasteuri- 40 minС −= 0С1001− min СС − 10ССmin С 20 min =1 0 −0 −= 0𝐴𝐴01−− − −𝑐𝑐𝑐𝑐 𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛 𝑚𝑚 𝑚𝑚 𝑛𝑛 0 = 2 . 𝑧𝑧 о 𝑧𝑧 о о о 97 СFor high70For-sugarСzation high products-sugar 50modes Сproducts (strawberryare following:30 (strawberryС marmalade), marmalade), packed𝑧𝑧 packed in a glass in a jarglass 2, jarthe 2,pasteur the pasteuri- i- 𝐴𝐴 𝐴𝐴 For 𝑐𝑐𝑐𝑐high-sugar𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑐𝑐𝑐𝑐𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛 products𝑚𝑚𝑛𝑛𝑚𝑚𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛 𝑚𝑚 𝑚𝑚(strawberry𝑛𝑛 marmalade), packed in a of industrial sterility, the absence of mold fungi (including Penicil‑ zation modeszationglass arejarmodes following:2,40 the aremin pasteurizationfollowing:10 min 10 modesmin are20 min following:𝐴𝐴 =1 𝑐𝑐𝑐𝑐𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛lium𝑛𝑛 glaucum𝑛𝑛𝑛𝑛𝑛𝑛 𝑚𝑚𝑚𝑚)𝑛𝑛 and yeast was confirmed by plating of DG 18 with 45 min 10 min 10−min 20− min − = 1 0 = 2 . − 97−о С 70о С− 50о С 30 о С 0 = 0 dichlorane and glycerin into agarized nutrient medium. 40 minо 4010minminо 1010minmin 10о20minmin 20 minо =1 =1 𝑧𝑧 95 С − 70− С− 50− − С − 30 С 0 =02= 2 . . о о о о о о о о 𝑧𝑧 𝐴𝐴 𝑐𝑐𝑐𝑐𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛It is𝑛𝑛 𝑛𝑛𝑛𝑛𝑛𝑛necessary𝑚𝑚𝑚𝑚𝑛𝑛 to take into account the characteristics of the 97 С 9770СС 704550minС С 501030СminС 3010Сmin 𝑧𝑧 20 min𝑧𝑧 =1 − − − 𝐴𝐴 0 = 𝑐𝑐𝑐𝑐0𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛 equipment𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛 𝑚𝑚 available𝑚𝑚𝑛𝑛 at the enterprises in order to optimize the 50 min 10 min о 10 minо 20 minо 𝐴𝐴 𝐴𝐴о 𝑐𝑐𝑐𝑐=1𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛 𝑐𝑐𝑐𝑐𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛 𝑚𝑚𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑚𝑚𝑛𝑛 𝑚𝑚𝑚𝑚𝑛𝑛 45 min 4510minmin 109510minminС 102070minminС 20 min50 С = 1 30 С = 1 =𝑧𝑧 1 pasteurization modes, then to produce experimental batches of −− − − − − − −− =00= 00 о о о о о о о о о о о 0 9095 С 9570С70С С705050minС С505010С30СminС 303010СminС 𝑧𝑧 20 min𝑧𝑧 𝑧𝑧 𝐴𝐴 =1 𝑐𝑐𝑐𝑐canned𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛 𝑛𝑛𝑛𝑛𝑛𝑛food,𝑚𝑚 𝑚𝑚put𝑛𝑛 them for 3-month storage, further open the − − − = 1 о о о 𝐴𝐴о 𝑐𝑐𝑐𝑐0𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛 𝑚𝑚𝑚𝑚𝑛𝑛can and conduct microbiological studies in accordance with the 50 min 5010minmin 109010minminС 102070minminС 20 min50 С𝐴𝐴 =130 С=1𝐴𝐴 𝑐𝑐𝑐𝑐𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛 𝑚𝑚𝑚𝑚𝑛𝑛𝑐𝑐𝑐𝑐𝑛𝑛𝑛𝑛𝑛𝑛𝑡𝑡𝑡𝑡𝑜𝑜𝑜𝑜𝑜𝑜𝑜𝑜 𝑚𝑚𝑚𝑚𝑛𝑛 − − − − − − =01= 1 𝑧𝑧 о о о о о о о о 0 requirements of industrial sterility. 90 С 9070СС 7050С С 5030С С 30 С 𝑧𝑧 𝑧𝑧 𝐴𝐴 𝑐𝑐𝑐𝑐𝑛𝑛𝑛𝑛𝑛𝑛𝑡𝑡𝑡𝑡𝑜𝑜𝑜𝑜𝑜𝑜𝑜𝑜 𝑚𝑚𝑚𝑚𝑛𝑛 The comparison of the obtained𝐴𝐴 values𝐴𝐴 of𝑐𝑐𝑐𝑐 𝑛𝑛𝑛𝑛𝑛𝑛the𝑡𝑡𝑡𝑡𝑐𝑐𝑐𝑐 actual𝑜𝑜𝑜𝑜𝑜𝑜𝑜𝑜𝑛𝑛𝑛𝑛𝑛𝑛𝑡𝑡𝑡𝑡𝑚𝑚𝑜𝑜𝑜𝑜𝑜𝑜𝑜𝑜 𝑚𝑚𝑛𝑛lethality𝑚𝑚𝑚𝑚𝑛𝑛 with the value of the re- The comparisonThe ofcomparison the obtained of the values obtained of the values actual of lethalthe actual- lethality4. Conclusionwith the value of the re- ity withThe the comparison value of theof the required obtained lethality values of — the A actualZ = 15 °C lethality = 40 ÷ with80 the valueUpon of the the r e-results of the work: quired Thelethality comparison- of the obtained=40 ÷ values 80 of the actual lethalityT = 80 °C .withshows the value that of the the roptimale- pasteurization conditionalquired min, lethalityshows𝑜𝑜 that- the optimal=40 ÷ pasteurization 80 mode for. shows 1. thatThere the optimal have beenpasteurization developed the optimal pasteurization quired lethality 𝑧𝑧- =15 𝐶𝐶 =40 ÷ 80 𝑜𝑜 . shows that the optimal pasteurization quired lethality - =40𝑜𝑜 𝑜𝑜÷ 80 𝑧𝑧=15 𝑜𝑜𝐶𝐶 . shows that the optimal pasteurizationmodes of high-sugar canned food for continuous-acting marmalade, packed𝑜𝑜 in a glass jar 2, was the following: mode for marmalade,mode𝑇𝑇 for= 𝑧𝑧8packed= 0marmalade,15𝐶𝐶𝑜𝑜𝐶𝐶 in𝑇𝑇= a 8packed 0glass𝐶𝐶 injar a 2,glass was jar the2, was following: the following: 𝑧𝑧=𝐴𝐴15 𝑜𝑜𝐶𝐶 𝐴𝐴 𝑐𝑐𝑐𝑐𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑐𝑐𝑐𝑐𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑚𝑚𝑛𝑛𝑚𝑚𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛 𝑚𝑚𝑚𝑚𝑛𝑛 pasteurizers of immersive type with the temperature of the mode formode marmalade, for marmalade,𝑇𝑇= 8packed0 𝐶𝐶𝐴𝐴𝑇𝑇= 8 packedin0 𝐶𝐶a glass in ajar glass 2,𝑐𝑐𝑐𝑐 was 𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛jar 2,the𝑛𝑛 was𝑛𝑛𝑛𝑛𝑛𝑛 following: the𝑚𝑚𝑚𝑚 𝑛𝑛following: 0 𝐴𝐴10 0 10 10 𝑐𝑐𝑐𝑐2𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛0 10𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛 𝑚𝑚20𝑚𝑚𝑛𝑛 0 10 10 20 with the value of the actual lethalityheating area below 100 °C and for various types of jars. 0 10 10 20 0 with0 withwith the3 value0the value valueof the ofactual of the the lethalityactual actual lethality 0 0 𝑚𝑚𝑖𝑖𝑛𝑛00 𝑚𝑚with𝑚𝑚𝑛𝑛3030 the𝑚𝑚 value𝑚𝑚𝑛𝑛 of𝑚𝑚 the𝑚𝑚𝑛𝑛 actual lethality 2. The developed modes of pasteurization of high-sugar canned 0𝑚𝑚𝑖𝑖𝑛𝑛 𝑚𝑚0 𝑚𝑚𝑛𝑛 о 3𝑚𝑚0𝑚𝑚𝑛𝑛 о 𝑚𝑚𝑚𝑚𝑛𝑛 о о 𝑚𝑚𝑚𝑚𝑖𝑖𝑛𝑛 𝑖𝑖𝑛𝑛 𝑚𝑚о 𝑚𝑚𝑛𝑛 =𝑚𝑚1𝑚𝑚𝑛𝑛𝑚𝑚о 𝑚𝑚𝑛𝑛 𝑚𝑚о𝑚𝑚𝑛𝑛𝑚𝑚𝑛𝑛 о𝑚𝑚𝑚𝑚𝑛𝑛 lethalityо=1о о о =о С2о о С о С С food can be recommended for continuous-acting pasteur- ==1 0 = С2−0 С − −С − −С − 0 ==С2С−2 С −С С − СС С 0 𝑧𝑧 − 𝑧𝑧 − − izers of irrigational type. In this case, preference should be 𝑧𝑧 So, for the establishedSo, for the modes established of pasteurization modes of pasteurization the optimal the optimal modes are: the heating 𝑧𝑧 So,𝐴𝐴 for So,the 𝐴𝐴 forestablished 𝑐𝑐𝑐𝑐the𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛 established𝑛𝑛 modes𝑛𝑛𝑛𝑛𝑛𝑛𝑐𝑐𝑐𝑐𝑚𝑚𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛 𝑚𝑚 𝑛𝑛modesof pasteurization𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛 of 𝑚𝑚pasteurization𝑚𝑚𝑛𝑛 the optimalthe optimal modes modes are: the are: heatinggiven the heating to pasteurizers with a temperature in the heating zone 𝐴𝐴 modesSo, for𝑐𝑐𝑐𝑐 are: 𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛the the 𝑛𝑛established𝑛𝑛𝑛𝑛𝑛𝑛 heating𝑚𝑚𝑚𝑚𝑛𝑛 (heat modes treatment) of pasteurization in an autoclavethe at a optimal modes are: the heating (heat treatment)(heat intreatment) an autoclave in an at autoclavea temperature at a of temperature the heating areaof the 97 heating °C and notsteparea -less by97-step than°C and 95 °C. step-by-step 𝐴𝐴(heat treatment)temperature in𝑐𝑐𝑐𝑐 an 𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛ofautoclave the𝑛𝑛 heating𝑛𝑛𝑛𝑛𝑛𝑛 at a𝑚𝑚 temperature 𝑚𝑚area𝑛𝑛 97 °C ofand the step-by-step heating area cooling97 °C and step-by-step 3. For achieve the industrial sterility of high-sugar cans and (heat treatment)coolingof the product of coolingthe in product anby of waterautoclave theby waterproductwith temperaturewith at by temperaa water temperature turewith 70 °C, 70 tempera °C,50 °C of50 tureand°Cthe and 7030 °C.heating 30°C, °C. 50 The°Carea andtime 97 30of °Cfood°C. andThe timestep -ofby food-step cooling of the product by water with temperature 70 °C, 50 °C and 30 °C. The time ofto food reduce the duration of the pasteurization process, it is The time of food exposure in the heating zone is 25 minutes (for exposure inexposure the heatingin zonethe heating is 25 minutes zone is (for 25 jar minutes 1) and (for40 minutes jar 1) and (for 40jar minutes2), innecessary: 1-st (for and jar 2- 2), in 1-st and 2- coolingexposurejar ofin 1) and thethe heating product 40 minutes zone byis 25 (forwater minutes jar 2),with (for in 1-stjartempera 1) and and 2-nd 40ture minutes cooling 70 °C, (for zones jar50 2),°C in and1-st and30 2°C.- The time of food —— to ensure the product packing temperature from 70 °C and ndthecooling jars are ndzones exposedcoolingthe jars zones for are 10 exposedthe minutes, jars forare in10 exposed theminutes, lastfor coolingin 10 the minutes, last zone cooling forin the zone last for coolingthe product, zone for the product, exposurend cooling inzones thethe heating jars are exposedzone isfor 25 10 minutes minutes, in(for the jarlast 1)cooling and zone40 minutesfor the product, (forhigher jar till2), toin the 1- stmaximum and 2- permissible values (85–90 °C); packagedthe product, in glass packagedjar 1 is cooledin glass for jar15 1 minutes, is cooled and for the 15 product, minutes, packed and into the glass jar 2 is packaged in glass packagedjar 1 is cooled in glass for 15jar minutes, 1 is cooled and thefor product,15 minutes, packed and into the theproduct, glass jar—packed— 2to is ensure into the minimum glass jar 2 distance is between the capping ma- the product, packed into the glass jar 2 is cooled for 20 minutes. nd coolingcooled zones for 20 minutesthe jars. are exposed for 10 minutes, in the last coolingchine zone and for thethe entry product, of the product to the pasteurizer in cooled for 20The minutes temperaturecooled. for 20 of minutesthe product. conveyed to the CAP pasteuri- order to minimize the loss of product temperature in the zation Thezone, temperature must not ofbe the lower product than conveyed 70 °C. Increasingto the CAP the pasteurization product zone, must not be packagedThe intemperature glass jar of The1 theis temperatureproduct cooled conveyed for of15 the tominutes, product the CAP conveyed pasteurizationand theto product, the zone, CAP must pasteurizationpacked not package;be into zone,the glass must notjar be2 is lowerpacking than temperature 70 °C. Increasing on 5 °C the willproduct reduce packing the pasteurizationtemperature on 5mode, °C will reduce the pasteur- —— to provide automatic maintenance of the set temperatures cooledlower than andfor 70 20that °C. minutes willlowerIncreasing reduce than. the 70the product°C. time Increasing of packing exposure thetemperature product in the heating packingon 5 °C zone willtemperature reducefor the on pasteu 5 °C r-will reduce the pasteur- ization mode, and that will reduce the time of exposure in the heating zone for 7-10 minutes.on zones of heat treatment (pasteurization) and cooling. 7–10 minutes. ization mode, and izationthat will mode, reduce and the thattimewill of exposure reduce the in thetime heating of exposure zone for in 7 the-10 4.heatingminutes. Continuous-acting zone for 7-10 minutes. pasteurizers can be recommended for TheThe temperatureThe canned canned food, food, ofproduced produced the product according according conveyed to to the the obtainedobtainedto the pasteurizationpasteuri CAP -pasteurization modes, meet the zone, must not be The canned food, produced according to the obtained pasteurization modes, meetthe the production of other types of high-sugar and high-acid zation modes, meetThe the canned requirements food, produced of industrial according sterility. to the When obtained pasteurization modes, meet the lower thanrequirements 70 °C. of Increasing industrial sterility. the product When determining packingthe temperature compliance of cannedon 5 ° foodCcanned will with reducefood:the marmalade, the pasteu jams,r- concentrated juices and to- requirementsdetermining of industrial the compliancesterility. When of canneddetermining food thewith compliance the requirements of canned food with the requirementsrequirements of industrial of sterility, industrial the absencesterility. of When mold fungidetermining (includingthe enicillium compliancemato glaucum of products, canned) food etc. with the izationrequirements mode, of industrial and that sterility,will reduce the absence the oftime mold of fungi exposure (including in enicillium the heating glaucum zone) for 7-10 minutes. and yeast wasrequirements confirmed ofby industrialplating of sterility,DG 18 with the dichloraneabsence of and mold glycerin fungi in (includingto agarized enicillium nu- glaucum) and yeast was confirmed by plating of DG 18 with dichlorane and glycerin into agarized nu- trientThe mediumcannedand. yeast food,was produced confirmed accordingby plating of to DG the 18 obtainedwith dichlorane pasteurization and glycerin modes,into agarized meet n u-the trient medium. requirements oftrient industrial medium sterility.. 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51 ПИЩЕВЫЕ СИСТЕМЫ | Том 2 № 4 | 2019 FOOD SYSTEMS | Volume 2 № 4 | 2019

INFORMATION ON THE AUTНORS Galina P. Pokudina — senior Researcher, Laboratory of canning production processes and equipment, Russian Research Institute of Canning Technology — Branch of V. M. Gorbatov Federal Research Center for Food Systems of RAS, 142703, Moscow region, Vidnoye, School str, 78, Tel.: +7–495–668–70–31 e-mail: [email protected]. *corresponding author Marina V. Trishkaneva — candidate of chemical sciences, leading researcher, Laboratory of scientific and technical analysis, Russian Research Institute of Canning Technology — Branch of V. M. Gorbatov Federal Research Center for Food Systems of RAS, 142703, Moscow region, Vidnoye, School str, 78, +7–495– 548–51–22, e-mail: [email protected] Raisa A. Volkova — leading research scientist, Laboratory of quality and food safety, Russian Research Institute of Canning Technology — Branch of V. M. Gor- batov Federal Research Center for Food Systems of RAS, 142703, Moscow region, Vidnoye, School str, 78, +7–495–668–70–37, e-mail: raj [email protected]. All authors bear responsibility for the work and presented data. All authors made an equal contribution to the work. The authors were equally involved in writing the manuscript and bear the equal responsibility for plagiarism. The authors declare no conflict of interest.

Received 02.08.2019 Accepted in revised 25.11.2019 Accepted for publication on 16.12.2019

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