Prosiding II International Symposium “Innovations in Life Sciences” (ILS 2020)

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The technology of curd cake with sucralose: a infrared spectroscopy analysis Olena Aksonova1* , Sergey Gubsky1 , Dmitry Torianik1 and Victoria Evlash1 1Educational and Research Institute of Food Technology and Business, Kharkiv State University of food technology and trade, Kharkiv, Ukraine Abstract. The article presents the results of developing technology for the production of curd cake using a low-calorie sweetener sucralose. The implementation of this direction in the form of a partial replacement in the sugar formulation for effective sweeteners sucralose is proposed. As a prototype, a sugar-based recipe was used, in which a partial replacement (at the level of 50%) of sugar with sucrose was carried out. Sucralose was formulated as the commercial sweetener TM Splenda, which contains maltodextrin and sucralose. All samples, including the control on sugar, were subjected to organoleptic evaluation, which showed the absence of extraneous flavors in all samples and their similarity in terms of sweetness compared to control. IR spectra of the sweetener, curd cake with sugar, as well as curd cakes with 50% and 100% sugar substitution for sweetener were obtained and analyzed. The analysis showed the presence of identical characteristic bands on the spectra of the sweetener and samples of sucralose cakes, which suggests that the sweetener TM Splenda does not undergo thermal degradation when baking curd cake. However, the literature analysis indicates the danger of the formation of toxic gaseous substances that are obtained during the thermal treatment of products with sucralose. 1 Introduction The glоbal trend оf an inсrease in the inсidenсe оf diabetes amоng the рорulatiоn and the emergenсe оf a high рerсentage оf оverweight рeорle are рrоmрting the fооd industry tо develор reсiрes fоr lоw-sugar flоur соnfeсtiоnery оr tо соmрletely reрlaсe it with highly effeсtive sweeteners [1]. The wide range of sweeteners and the growing attention to them are explained by their significant economic advantages in relation to sugar, since they all have a sweetness coefficient much higher than that of sucrose and are easy to use. [2]. In general, they can be conditionally divided into two groups in terms of use in food technologies. Substances with almost "zero calories" and very intense sweet taste, which are used in small quantities to replace the sweetness of much higher amounts of sugar and are approved for use in the food industry, can be classified in the first group. The second group includes caloric substances that can replace both physical mass and the sweetness of sugar. Products of this type, sometimes referred to as “sugar replacers”or “bulk sweeteners,” include the sugar alcohols (also called “polyols”). Sucralose belongs to the sugar substitutes of the first group. From a chemical point of view is a chloroderivative of sucrose and is known as the food additive E955. It is abоut 600 times sweeter than sugar but, this faсtоr varies deрending оn the level оf suсralоse being used. Like sugar, suсralоse is a white, сrystalline, nоnhygrоsсорiс, free-flоwing роwder and freely sоluble in water. This sweetener has a zerо glyсemiс index and zerо сalоriс соntent. Although sucralose is made from sugar, the human body does not recognize it as a sugar and does not metabolize it; therefore, it provides sweetness with no calories. Sucralose was approved as a food additive in USA, Canada, UK and European countries, followed by others. It is оn sale as a рrоduсt under the trademark SРLENDA® (Brand Sweetener оf Tate & Lyle РLС, UK), whiсh alsо inсludes оther related соmроnents. Flоur соnfeсtiоnery, and in рartiсular сheese сake, are in demand amоng соnsumers. And the develорment оf a teсhnоlоgy fоr the рrоduсtiоn оf сheese сake with a рartial reрlaсement оf sugar with suсralоse is relevant fоr sоlving the рrоblems оf оverweight and рreventing tyрe 2 diabetes. Earlier [3] we offered technology of curd cake with suclarose as sweetener. The organoleptic analysis of the obtained samples showed that the sweetness levels for sugar curd cake with and sucralose curd cake areidentical. This fact indirectly indicates the thermal stability of the sweetener used in the technology. This technology was based on an approach with partial replacement of sugar with sucralose. In the process of optimizing this technology, two problems were identified. The first is related to the need for compensation of effect of added sugar on food quality. It is known that sugar is a structure-forming component [4]. This problem was solved by adding more formulation ingredients (curd). This procedure increased the amount of solids, the amount of which was reduced by partial replacement of sugar. The second problem was related to the safety of using sucralose in this technology, which requires a baking temperature of 170 ° C to prepare the *Corresponding author: [email protected] © The Authors, published by EDP Sciences. This is an open access article distributed under the terms of the Creative Commons Attribution License 4.0 (http://creativecommons.org/licenses/by/4.0/). BIO Web of Conferences 30, 01001 (2021) https://doi.org/10.1051/bioconf/20213001001 ILS 2020

product. This sweetener is known to be thermally stable up to temperatures 120 С [1]. With regard to the behavior of sucralose as the temperature rises further, many questions arise about its ability to form chlorinated by-products. This issue has been studied in detail in numerous original research [5-9] and review papers [10, 11]. Table 1 provides an overview of some of the research on the thermal decomposition of sucralose. In most publications, the object of these studies is pure sucralose. Only in work [5] real samples of food with sucralose were investigated. However according [8], these results lack credibility due to inefficient separation technique with thin-layer chromatography used in analysis of degradation products. Most research suggests that thermal decomposition of Sucralose is associated with the release of chlorides, leading to the formation of possibly harmful compounds [11]. The most detailed analysis on this issue was carried out on the basis of literature data in [10]. According to this review, available data indicate that harmful compounds, some of which are carcinogenic, can occur when sucralose, and especially foods containing sucralose such as canned vegetables or baked goods, are heattreated. When sucralose is heated to temperatures above 120 ° C, the sweetener gradually decomposes and dechlorinates. Temperatures of 120-150 ° C are possible during industrial food production and processing, and are also reached in private households during the preparation and baking of foods containing sucralose. This can lead to the formation of potentially hazardous chlorinated organic compounds such as polychlorinated dibenzo-pdioxins (PCDD), dibenzofurans (PCDF) and chloropropanols. However, there are currently insufficient data for final conclusions. The data in Table 1 indicate that the spectroscopic method in the infrared region was used as one of the most common experimental physicochemical methods for studying the decomposition products of sucralose. Fourier transform infrared spectroscopy is a powerful as a primary tool that facilitates the study of materials in matrix of foods. The technique allows monitoring the changes in various chemical bonding compounds. On the other hand, FTIR spectroscopy can be used as the technique of complementary measurement to help interpret the results along with other techniques. This determined the choice of the method in this work, the purpose of which was to study the possibility of the safe use of sucralose as a sweetener in the production of baked goods using the example of curd cake production. Table 1. Study of thermal degradation of sucralose Samples Method Results References Baked goods (yellow cake, cookies, graham crackers) with radioactively 14C- marked Sucralose Radiolabeling techniques (by liquid scintillation), thin-layer chromatography a. Sucralose minimally degrades in baked goods in range 180–300 °C. b. The marked Sucralose did not react with any other ingredients in the baked goods. c. The authors concluded that Sucralose is "heat-stable" [5] Sucralose in buffered solutions at pH 3, 7, and 11 High performance liquid chromatography, Ionmetry using a chloride selective electrode a. The biggest change appeared to occur after 120°C and at temperatures up to 140°C, there was a greater amount of chloride released at all three pH levels b. The largest class of compounds produced was furans. c. The compound produced in the highest amounts was levulinic acid. [6] Sucralose with assay > 98.0% (HPLC - grade) Differential scanning calorimetry, thermogravimetric analysis, infrared (IR) spectroscopy a. The sucralose is thermally stable up to 119 C and above this temperature the thermal decomposition takes place in three steps up to 550 C and without melting. b. The endothermic peak at 131°C (thermogravimetric analysis) and 128°C (differential scanning calorimetry) is due to the thermal decomposition with release of constitution water and hydrogen chloride. c. IR spectra also confirm that the thermal decomposition occurs above 119 C [7] Sucralose Pyrolysis-gas chromatography–mass spectrometry a. Degradation of sucralose and glucose may generate a levoglucosenone a well documented degradation product. b. Caution should be exercised in the use of sucralose as a sweetening agent during baking of food products containing lipids due to the potential formation of toxic chloropropanols. [8] Technicalgrade sucralose Differential scanning calorimetry, thermogravimetric analysis, IR spectroscopy, hot-stage microscopy, high-resolution mass spectrometry a. Decomposition of sucralose happens around 125 C in form of carbon dioxide along with the formation of hydrogen chloride and other minor compounds. b. Chlorinated derivatives, including polychlorinated aromatic hydrocarbons were confirmed. c. These findings not only corroborate the suspected instability of sucralose to high temperatures, but also indicate that even exposed to mild conditions the formation of hazardous polychlorinated compounds is observed. [9] 2 BIO Web of Conferences 30, 01001 (2021) https://doi.org/10.1051/bioconf/20213001001 ILS 2020

2 Experimental 2.1 Materials The ingredients used in this study obtained from local stores in city Kharkiv, Ukraine. sucralose as SPLENDA TM was purchased from tate&lyle plc, uk and used was food grade. according to the manufacturer's information on the packaging, the sweetener contains maltodextrin (ìd) and sucralose (according to information available on the internet, the content of sucralose is 1%). potassium bromide (reachim, russia) used was analytical grade. 2.2 Sampling A curdcake was manufactured according to [3]. The product included wheat flour, butter, milk cheese 9%, melange and baking powder. In the production of curdcake was used sucralose in the form of sweetening SPLENDA TM. The manufacturer of this brand states that the product contains not only sucralose but also maltodextrins. The full chemical composition in percentage on the package is not indicated. However, according to literature data, this product contains 1% sucralose. Therefore, a sweetener based in sucralose was added based in a partial (50%) and full (100%) sugar substitute in and amounted to 0.10 and 0.05 g, respectively. Cheese cakes samples with a mass of 100 g were baked in silicone forms at a temperature of 170° C for 30 minutes. The samples obtained using this technology were designated as S50 and S100, respectively, for curd cake with 50% and 100% sugar substitution. The prototype on sucrose sample was considered in the study as Control. 2.3 Methods Fourier transform infrared coupled to attenuated total reflectance (ATR–FTIR) spectra of curd cake samples were obtained using Nicolet iS5 FT-IR spectrometer with the versatile iD5 ATR Accessory (Thermo Scientific, USA) as the result of the accumulation of 32 scans with a resolution of 4 cm-1 in the range of 4000-600 cm-1 . FTIR spectra of SÐLENDA were recorded between 4000 and 400 using a the same spectrometer with Thermo Scientific Base Adapter in a potassium bromide matrix. Discs of matrix were prepared by first mixing 0.5 mg of dried sample with 500 mg of KBr in an agate mortar and then pressing the resulting mixture. A pellet containing only potassium bromide was prepared and used as the reference sample for transmission measurements. Each spectrum was converted to ASCII format to be further processed. Processing of spectroscopic data obtained including spectral pretreatment (correction of baseline, ATR correction and correction for ambient carbon dioxide, smooth of spectra) were performed with the Know It All Informatics System 2020, Academic Edition (John Wiley&Sons Inc., USA) software. 3 Results and discussion Obtained in the FTIR spectra of the sweetener TM SPLENDA is given on Fig. 1. As already mentioned, the sweetener contains maltodextrin and sucralose, and the content of maltodextrin is approximately one hundred times higher than that of sucralose. Thus, it was maltodextrin that made the main contribution to the obtained results. Based on literature data [12], the spectrum for MD exhibited a strong broad absorption band centered at 3300 cm-1 and a weak band at ~1635 cm-1 which arise from OH-stretching and OH-bending (in plane) modes, respectively. An absorption band of CH stretching vibrations was found at 2900 cm-1 . The spectrum of the sweetener shows that the broad intense band observed for maltodextrin at 3300 cm-1 is shifted to 3420 cm-1 . An upward shift is also observed for the band corresponding to CH-stretching absorption band 2930 cm-1 . The shift of these bands can be explained by the presence of sucralose, which contains chlorine atoms, which, possibly, affect the redistribution of electron density within the molecule and reduce the possibility of the formation of intra- and intermolecular bonds for hydroxyl groups. A weak band at 1635 cm-1 is observed on the spectrum of the sweetener as well as on the spectrum of pure MD. The results of studies indicate the existence in the spectrum of characteristic bands of 635 cm-1 , which correspond to strength vibrations of the C-Cl bond in the sucralose molecule [13]. It is this band that makes it possible to identify sucralose in the composition of the sweetener. The spectra of samples Control, S50 and S100 are shown in Fig. 2. The contribution of water to the formation of the OH-stretching absorption band in carbohydrate-water systems is very strong. In these systems clusters of water molecules could be categorized into three families according to their hydrogen bond organization. The first, originating from tetrahedrally bonded water molecules, namely networking water, locates in the region of around 3300cm-1 to 3314 cm-1 . The second is intermediate water, which is a weakly or distorted hydrogen bonded family locates at around 3441 to 3470 cm-1 . The last family is free or multimer water, which originates at around 3570 to 3610 cm-1 from water monomers and dimers and locates [14-16]. The spectra of cake samples have bands with peaks at 3275 cm-1 - Control, 3281 cm-1 - S50 and 3355 cm-1 for S100, which indicates that water in the product, is mainly in the form of networking water (Fig.3). It should also be noted the shift of this band to the region of large values with increasing concentration of the sweetener. For sample S100, this value is already 3355 cm-1 , which is very close to the indicator of this band in the spectrum of pure sweetener 34203420 cm-1 . This again speaks in favor of the fact that the sweetener is preserved during heat treatment. In addition, the water begins to change to the state intermediate water weakly or distorted hydrogen bonded. 3 BIO Web of Conferences 30, 01001 (2021) https://doi.org/10.1051/bioconf/20213001001 ILS 2020

Fig. 1. FTIR spectra of SPLENDA. Fig. 2. Representative ATR-FTIR spectra in the fingerprint region of the curd cake samples: control (red), S50 (blue) and S100 (black) The spectra next from the fingerprint region, in the range of 1800 to 1600 cm-1 , which usually arise from HO-H-bending of water molecules [17]. Thus, the bands in the range of 16501630 cm-1 can be attributed to water of crystallization, and the spectra clearly show the dependence between the sugar content and the amount of water of crystallization: in a sample with 100% sugar replacement by sweetener, this band is the most intense, cm-1 cm-1 4 BIO Web of Conferences 30, 01001 (2021) https://doi.org/10.1051/bioconf/20213001001 ILS 2020

3270 3280 3290 3300 3310 3320 3330 3340 3350 3360 0 20 40 60 80 100 (OH) band position (1/cm) % sugar peplacement Fig. 3. Band positions of OH-stretching vibration for curd cake samples and in the sample with sugar it is practically absent (Fig 2). This region of 1800 to 800 cm-1 the so called a “fingerprint” region. Vibrations in this region are mainly attributed to in-ring CO-stretching, interring COC-, COH- and CCH- bending, and symmetrical deformations of CH2 groups. Vibrations of 10351030 cm-1 are identified as aromatic C-H planar bending vibrations. This band is clearly visible on the sweetener spectrum. It is absent in the spectrum of the sample with sugar, and in the spectra of samples on sucralose this band is shifted to a lower region - 1021 cm-1 and 1023 cm-1 for samples S50 and S100, respectively (Fig. 2). This fact may also be a reason to think that the sucralose molecule has not undergone thermal degradation. However, one should take into account the risks of the appearance of substances hazardous to health during the heat treatment, which can also be identified using IR spectra. Most studies suggest that thermal decomposition of Sucralose is associated with the release of chlorides, resulting in the formation of possibly harmful compounds [11,18]. It should be noted, that in the obtained spectra of samples of curd cake with sucralose, peaks at 1743 cm-1 are observed. These peaks, as in [7], can be attributed to the carbonyl stretching frequency appeared after heated the sample is up to 130ºC. However, these peaks may not necessarily be associated with the degradation of sucralose, most likely the thermal transformation of other nutrients present in the formulation also contributes to their appearance. This fact is also confirmed by the fact that the same band is present in the spectrum of the "control" sample, in the production of which sucralose was not used. Therefore, rather, this band can be explained by the occurrence of the Maillard reaction. The results of FTIR analysis presented in [9] show that at the point of decomposition of sucralose one can observe characteristic profiles of water (ranges from 40003200 cm-1 and 20001200 cm-1 ), carbon dioxide (the main peak at 24002300 cm-1 ), hydrogen chloride (range from 31002600 cm-1 ) and chloroacetaldehyde (main peak at 18501700 cm-1 ). However, such a complex system as curd cake requires water. As for the bands that can be attributed to carbon dioxide, they were not found in the finished product. The latter suggests that the identification of CO2 is relevant when studying the thermal decomposition of pure sucralose. In such complex systems as food systems, it is not correct to conclude that sucralose decomposes by the presence of peaks that can be attributed to CO2. The peak attributed to chloroacetaldehyde may also be associated with a carbonyl group, which does not have to be part of chloroacetaldehyde. For example, the Maillard reaction leads to the formation of numerous pore products with a rather complex and often unknown structure, among which there are compounds containing a carbonyl group. In addition, the 18501700 cm-1 region can be associated with a number of carbonyl compound group frequencies [19]. Among the compounds hazardous to health that can appear in food during the thermal decomposition of sucralose, there may also be: chloropropanols, PCDD and PCDF congeners and dioxin-like polychlorinated biphenyls (dl-PCBs), Polychlorinated naphthalene (PCN) congeners. However, it should be noted that chloropropanols in the study [8] were found in the released gas phase. The authors [20] concluded that the heating (roasting) of Sucralose in the presence of soybean oil and beef at 250°C led to the formation of various toxic PCDF and PCDD congeners which were transferred from the solid to the gas phase. Moreover, they emphasized that it is important to ensure effective ventilation when cooking with Sucralose in order to reduce the exposure risk of consumers to the occurring volatile PCDFs and PCDDs via inhalation. The authors concluded that, due to thermal decomposition during heating (roasting) to 160°C in the presence of soybean oil and beef, Sucralose became a source of chloride, which in turn promoted the formation of dl-PCB compounds, which were finally found in the oil vapors. They also hypothesised that an “appropriate” use of chloride-containing additives during cooking could therefore help to reduce exposure of consumers to dlPCBs [21]. In [22] it is said that that heating of Sucralose together with peanut or olive oil free of PCN, can lead to the formation of various PCN congeners as well as PCDD, PCDF and PCB compounds, which were released with the vapors generated during the further course of the process. It should be noted separately that most of the studies in which the thermal degradation of sucralose was studied were carried out at temperatures above 200°C. These temperatures are significantly higher than those used in baked goods technologies. You should also pay attention to studies on the study of the temperature gradient in baked goods during their production [23-25]. So the author of [23] showed that the temperature of the central layers of biscuit crumb is 9798°C at the end of baking. Similar studies for muffins [24] indicate a temperature at 5 BIO Web of Conferences 30, 01001 (2021) https://doi.org/10.1051/bioconf/20213001001 ILS 2020

the end of baking of 103104 °C. The crumb temperature at the end of baking for muffins was 100101°C, which was reached for muffins on sucrose in 25 minutes, and on fructose in 23 minutes [25]. In the same work, the authors noted that the temperature in the samples during baking does not exceed the decomposition temperature of sucralose. But on the surface, the temperature rises to 120°C for 12 minutes of baking for muffins on sucrose and from 12 to 24 minutes of baking - up to 160°C at a working chamber temperature of 180°C. Thus, we can conclude that the thermal decomposition of sucralose during the manufacture of curd cake does not occur, although it cannot be ruled out that part of sucralose is involved in thermal degradation. It should also be noted that the issue of products generated during this process remains open and requires integrated approaches. 4 Сonclusion An analysis of the literature showed that Sucralose is dechlorinated and this may lead to the formation of chlorinated organic compounds with possibly harmful potential (e.g. polychlorinated dibenzo-para-dioxins or dibenzofurans or chlorpropanols). The literary analysis also showed that, it can be concluded that substances hazardous to humans were detected in the gas phase formed during the heat treatment of the food product, and were not identified in the finished product using the analysis of IR spectra. Moreover, most of the research has been conducted at temperatures higher than those used to make curd cake. Based on the currently available data, it is impossible to judge the degree of formation of toxicologically significant congeners and to attribute them directly to the presence of sucralose. Since the ongoing Maillard reactions lead to the formation of not only dioxins, but also other potentially toxic compounds. The spectra of cake samples have bands with peaks in range 32753355 cm-1 , which indicates that water in the product is mainly in the form of networking water. A shift of this band to the region of large values is observed with an increase in the concentration of the sweetener. For sample S100, this value is already 3355 cm-1 , which is very close to the indicator of this band in the spectrum of pure sweetener - 3420 cm-1 . This once again speaks in favor of the fact that the sweetener is preserved during heat treatment. The spectra of samples of curd cakes with sucralose have peaks in the range of 10351030 cm-1 , which are identified as aromatic C–H planar bending vibrations. This band is clearly visible on the sweetener spectrum. It is absent in the spectrum of the sample on sugar, but in the spectra of samples on sucralose this band is shifted to a lower region. This also suggests that the sweetener has not undergone thermal degradation. This also indicates that chlorine is in a bound state, and this indicates the impossibility of forming substances hazardous to human health. Analysis of IR spectra cannot be proposed as a 100% reliable method for identifying potentially hazardous substances in food (chloropropanols, polychlorinated dibenzo-para-dioxins, polychlorinated dibenzofurans congeners, dioxin-like polychlorinated biphenyls and polychlorinated naphthalene congeners). The presented results indicate that further studies of the thermal behavior of sucralose in food products are needed using a combination of physicochemical methods. Moreover, both the final product and the products that are obtained at different stages of heat treatment should be investigated. References 1. K. O’Donnell, M.W. 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BfR - Bundesinstitut für Risikobewertung. 2019. “Harmful Compounds Might Be Formed When Foods Containing the Sweetener Sucralose Are Heated,” BFR (No 012/2019):1–22. 11. S.S. Schiffman, K. I. Rother, Journal of Toxicology and Environmental Health - Part B: Critical Reviews, 6, pp. 399 (2013). 12. E. Sritham, S. Gunasekaran, Food Hydrocolloids, 70, pp. 371 (2017). 13. B. Guven, S. D. Velioglu, I. H. Boyaci, Gida, The Journal of Food, 44, pp. 274 (2019). 14. M.E. Gallina, P. Sassi, M. Paolantoni, A. Morresi, and R. S. Cataliotti, The Journal of Physical Chemistry B, 110, pp. 8856 (2006). 15. A. Lerbret, P. Bordat, F. Affouard, Y. Guinet, A. Hédoux, L. Paccou, D. Prévost, and M. Descamps, Carbohydrate Research, 340, pp. 881 (2005). 16. J. Malsam, A. Alptekin, The Journal of Physical Chemistry B, 113, pp. 6792 (2009). 6 BIO Web of Conferences 30, 01001 (2021) https://doi.org/10.1051/bioconf/20213001001 ILS 2020

17. A. Gharsallaoui, B. Rogé, J. Génotelle, M. Mathlouthi, Food Chemistry, 106, pp. 1443 (2008). 18. S. S. Schiffman, Chemical Senses , 37, pp. 671 (2012). 19. J. Coates, Encyclopedia of Analytical Chemistry. Chichester (UK: John Wiley & Sons, Ltd., 2006) 20. J. Wu, S. Dong, G. Liu, B. Zhang, M. Zheng, Journal of Agricultural and Food Chemistry, 59, pp. 5444 (2011). 21. S. Dong, J. Wu, G. Liu, B. Zhang, M. Zheng, Food Control, 22, pp. 1797 (2011). 22. A. Fernandes, M. Rose, J. Falandysz, Environment International, 104, pp. 1 (2017). 23. E. Bondarenko, Studies of the Kinetics of the Biscuit Baking Process Semi-Finished Products (Kiyv: KTIFI, 1979). 24. V. V. Dorokhovich, Development of rational technologies of diabetic flour confectionery products based on fructose (Kiyv: NUFT, 2000). 25. A. Dorokhovich, N. Lazarenko, Progressive Technique and Technologies of Food Production Enterprises, Catering Business and Trade, 1, pp. 414 (2012). 7 BIO Web of Conferences 30, 01001 (2021) https://doi.org/10.1051/bioconf/20213001001 ILS 2020

Study of the impact of the food biopolymer system on the clinical-biochemical status of white inbred mice in the invivo experiment Tatyana Alekseeva1* , Mihail Korystin1 , Ekaterina Klimova1 , Yuliya Kalgina1 , Lidiya Vitruk2 , and Lyudmila Malakova1 1Department of Service and Restaurant Business, Voronezh State University of Engineering, Voronezh, Russia 2Department of Foreign Languages, Voronezh State University of Engineering, Voronezh, Russia Abstract. Studies have been conducted on the effect of the food biopolymer composition on the clinicalbiochemical status of white inbred mice. The experiment showed that the development of laboratory animals was carried out without abnormalities with the complete safety of all individuals. Clinicalbiochemical dynamics, glucose monitoring and the results of experiments in the exchange of macro and microelements of plasma of mouse blood indicated the normal flow of metabolic processes. In addition, the animals that consumed food additives had a higher rate of absorption of feed carbohydrates and a predominance of osteosynthesis processes in animal bone tissue. The studies conducted allow us to conclude that enrichment and trace elements increase the availability of trace elements, as well as activate the formation and strengthening of the bone tissue of the body. 1 Introduction One of the main tasks of the food industry is the creation of commercial food lines to correct the nutritional status of the human body. We have developed the component composition and the technology of biopolymer composition on the basis of domestic raw materials of deep processing for food enrichment. The enrichment composition included albumin of animal origin, bagasse wheat germ and white bean seeds. The purpose of the work was to conduct studies on the influence of the developed food system on the clinical and biochemical status of laboratory animals. Similarity in the structure and functioning of the digestive system (stomach, small and large intestines, liver) and the white mouse suggests that the morphofunctional and physiological changes (or absence thereof) obtained in the experiment Intestinal tract organs under investigation in mice may also occur in humans under different environmental conditions [1-6]. 2 Experimental The experiment was carried out on white inbred mice of the BALB/c line, the age of the animals 15 days. Throughout the experiment, mice were kept in standard vivar conditions, under natural light, without limiting access to food and water, in standard polycarbonate cages of five individuals each. Prior to the start of the study, all animals underwent a two-week adaptation period. For the experiment, the animals were randomized and divided into two groups of 15 individuals. Intact animals (group 1 – control) received standard granulated feed PC-120-1 (LLC “Laboratornas”, Russia); animals of group 2 (experiment) were kept on the diet with the addition of food additive. In an experiment to study the effect of an enriching additive on the clinical and biochemical status of laboratory animals, the body weight of mice in each group was determined daily, and the growth rate relative to the primary data was determined on the basis of the results obtained [7-10]. The blood taken from the tail vein before the beginning, on the 21st day and after the end of the experiment was examined on a number of clinicalbiochemical indicators by sets of test-systems «Vital Development Corporation» (Russia, St. Petersburg). The general protein was determined using the biurettic method according to the instructions for the diagnostic set [11-17]. In order to carry out quantitative analysis, finished reagents and test samples were applied according to the recommended scheme. The samples were mixed, incubated for 15 minutes at 18-25 °C. Photometration was performed against the idle sample at a wavelength of 540 nm. Cholesterol level was controlled using enzymatic colorimetric method in accordance with instructions [11- 17]. Ready reagents and test samples were entered in accordance with the schematic in the instructions during the experiment. After mixing, the samples were incubated for 5 minutes at 18-25 °C. Photometration was performed against the idle sample at a wavelength of 500 nm. Colour stability was maintained for at least 1 hour while being protected from direct sunlight. The glucose content was determined using the glucose-oxidation method without deproteinization. In accordance with the instruction on the proposed scheme, quantitative analysis was performed [10-17]. The samples *Corresponding author; [email protected] © The Authors, published by EDP Sciences. This is an open access article distributed under the terms of the Creative Commons Attribution License 4.0 (http://creativecommons.org/licenses/by/4.0/). BIO Web of Conferences 30, 01002 (2021) https://doi.org/10.1051/bioconf/20213001002 ILS 2020

were mixed, incubated for 15 minutes at 18-25 °C. Photometration was performed against the idle sample at a wavelength of 505 nm. Colour stability was maintained for at least 1 hour while being protected from direct sunlight. Low-density lipoproteins (LDL) were defined by an enzymatic colorimetric method with selective (no deposition) protection on the use of a diagnostic set [11- 17]. The test samples were incubated for 10 minutes at 18-25 °C. Photometration was performed against the idle sample at a wavelength of 500 nm. Next, the samples were centrifuged for 10 minutes at 4000 g. Solutions for blank and calibration samples were not centrifuged. Transparent supernatant was used to determine the concentration of LDL. LDL was determined in all samples within an hour. High density lipoproteins (HDL) were determined by an enzymatic method with immuno-inhibition without deposition in accordance with the procedure [11-17]. The samples were mixed and incubated for 5 minutes at 37 °C, measuring adsorption A1, a sample of the idle sample. The samples were then re-mixed, incubated for 5 minutes at 37 °C, adsorption A2 was measured. The difference of adsorptions ΔA = A2 - A1 was then calculated. The LDP concentration in the sample (E) was then calculated with the formula: E = · ∕ (1) Where: is calibrator concentration; is adsorption difference of the calibrator; is the difference in the adsorption of the experimental sample. The phosphorus level was determined by UV method without deproteinization according to the instructions [11-17]. Experimental samples after mixing were incubated for 5 minutes at 18-25 °C. Photometry was performed against an idle sample at a wavelength of 340 nm. The color stability was maintained for at least 24 hours while being protected from direct sunlight. The calcium quantity was determined by the colorimetric method with Arsenazo III in accordance with the instructions for the application of the diagnostic set [11-17]. Complete reagents and test samples were added according to the recommended scheme. After mixing the samples were incubated for 5 minutes at 18- 25 °C. Photometration was performed against the idle sample at a wavelength of 340 nm. Color stability was maintained for at least 1 hour while being protected from direct sunlight. Alkaline phosphatase activity in serum and blood plasma was determined by an optimized kinetic method using the instruction [11-17]. A certain scheme was used for the application of finished reagents and test samples. This was followed by an incubation of experimental samples at a 37 °C water bath for 30 minutes. After the sample was cooled, appropriate reagents were introduced into the test tubes. The samples were then mixed and incubated for 5 minutes at 37 °C. Photometry was performed against the idle sample at a wavelength of 405 nm. The colour was stable for at least 8 hours in the absence of direct sunlight. Statistical processing of the results of experimental studies was carried out using Statistic 6.0 software package for Windows using methods of descriptive statistics (mean error, standard deviation). The results are presented as М ± SЕ. To determine the reliability of the obtained values, the Student T-test was used. The differences were considered significant when the value of the criterion is p ≤ 0,05 [18-19]. 3 Results and discussion The study of the influence of the food biopolymer system on the clinical and biochemical status of laboratory animals was carried out for 36 days. An in vivo experiment on white mice showed that the development of animals in the early stages took place without significant deviations in both groups. The safety of animals (experience and control) for the entire observation period was 100 %. This indicates the absence of toxic effects of feed on the organs and tissues of laboratory animals. During the first 21 days of the experiment, the increase in body weight in groups 1 and 2 was 32.8 %. It was recorded that by day 36 the weight gain in animals of the experimental group was 17,6 % higher than in animals of the intact group. This indicates a higher digestibility of feed containing an enriching additive compared to standard feed. Clinical-biochemical indicators showed the normal flow of metabolic processes (Table 1). Table 1. Results of research on exchange of macro and microelements of blood plasma clinical-biochemical status of laboratory animals in the invivo experiment Number of trial days Trial group 1 (Control) 2 (Experiment) Glucose, mmol/l 0 4,5±0,21 21 4,4±0,29 36 4,4±0,26 4,5±0,82 Total Protein, g/l 0 54,0±1,8 21 55,0±2,0 36 54,0±1,3 55,0±1,1 Cholesterol level, mmol/l 0 2,2±0,10 21 2,1±0,09 36 2,3±0,27 2,2±0,28 LDL, mmol/l: 0 0,44±0,05 21 0,51±0,06 36 2,46±0,12 1,27±0,11 HDL, mmol/l 0 0,71±0,14 21 0,84±0,12 36 3,61±0,16 2,96±0,15 Phosphorus level, mmol/l 0 0,8±0,05 21 0,7±0,04 36 0,8±0,04 0,9±0,04 Calcium level, mmol/l 0 1,9±0,13 21 1,7±0,15 36 1,9±0,14 2,1±0,16 Alkaline Phosphatase level, mcmol/l 0 113,0±5,0 21 116,0±7,0 36 115,0±3,5 115±3,5 2 BIO Web of Conferences 30, 01002 (2021) https://doi.org/10.1051/bioconf/20213001002 ILS 2020

The common protein of the experimental and control animal groups in all blood samples tested did not exceed the physiological standard and averaged 54-55 g/l for all groups. This indicates the normal flow of exchange processes in both pilot groups. Blood plasma cholesterol at the end of the experiment in the target animals was 4,5 % above the physiological norm, while the corresponding figure for group 2 was within the physiological norm. This shows that there is no negative effect of the feed with the bioactive additive on the lipid exchange of living organisms. The LDL level was also within the physiological norm (no more than 3 mmol/L), and the HDL rate increased by an average of 4,6 times in all groups. This fact is not evidence of any negative processes, but primarily shows the presence of an active level of parietal digestion and lipid intake in the body [20-22]. The control of glucose content in both groups showed that its amount fluctuated slightly within the physiological norm. A slightly higher glucose content (4,5±0,82 mmol/L) was observed in blood plasma in experimental animal group 2, this is 2,3 % higher than the blood glucose level of the animal control group at the end of the experiment. These data indicate a higher rate of assimilation of feed carbohydrates of groups of animals with enriched food systems [20-22]. The studies also monitored the exchange of some microelements. Results of experiments to exchange macro and microelements of plasma in mice in an in vivo experiment to study the effect of feedstuff enrichment on the clinical-biochemical status of animals in the experimental group showed that calcium and phosphorus were increased (by 12,5 % and 10,5 % respectively) compared to mice of the intact group (Table I). This confirms the higher availability and digestibility of microelements in feed containing a biopolymeric composition. Studies have found the same level of alkaline phosphatase against a background of stable microelements (calcium, phosphorus), indicating normal synthesis of collagen by osteoblasts under moderate activity of osteoclasts. This fact indicates the predominance of osteosynthesis in the bone tissue of animals that consumed food with bioadditive. 4 Сonclusion Studies conducted using bioassay in vivo techniques allow concluding that the developed food enrichment additive does not show any signs of toxicity and is biologically safe for the health of living organisms. It was found that a more significant increase in body weight of mice was recorded in the experimental group compared to the control group, which indicates a high digestibility of feed containing an enriching food additive. The dynamics of clinical and biochemical indicators shows not only the normal course of metabolic processes in laboratory animals, but also a higher level of carbohydrate absorption in the feed of the experimental group. In this group, calcium and phosphorus levels were higher than those of animals in the intact group, indicating the most frequent high availability and assimilation of micronutrients in the feed. The same level of alkaline phosphatase against the background of a stable content of trace elements confirms the normal synthesis of collagen by osteoblasts against the background of moderate activity of osteoclasts. This fact indicates the predominance of osteosynthesis processes in the bone tissue of experimental animals. The results obtained allow us to speak about the feasibility of introducing a developed biopolymer enriching additive into the composition of food products [23-27]. References 1. L.V. Antipova, N.S. Rodionov, and Е.S. Popov, Journal of Higher Educational Institutions. Food Technology, 1, 8–11, (2018) 2. S.V. Belokurov, N.S. Rodionova, and E.V. BelokurovaJournal of physics, 1015, 32–107, (2018) 3. Т.V. Alexeeva, Yu.О. Kaigina, and А.P. Fursova, Commodity expert of food products, 9, 69–77, (2019) 4. N.A. Zherebtsov, T.V. Zyablova, and T.N. Popova, Applied Biochemistry and Microbiology, 2, 145– 149, (2001) 5. D. Pass and G. Freeth, ANZCCART News, 4, 1–4, (1993) 6. M. Fitzgerald and J. Nolan P., J. Anat, 11, 67–71, (1971) 7. I.P. Zapadnuk, Laboratory animals. Breeding, maintenance, experimentation (Vysha shkola, Kiev, 1983) 8. E. Satinoff and I. Whishalow, The behavior of the laboratory rat. A handbook with tests (Oxford University Press, NY, 2005) 9. C. Gordon, Temperature Regulation in Laboratory Rodents (Cambridge University Press, N-Y, 1993) 10. V. Zutphen, V. Baumans, and A. Beynen, Principles of Laboratory Animal Science( Elsevier, Amsterdam, 2001) 11. Reference book. Biochemestry. 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19. Е.Yu. Ludupova, N.V. Rinchinova, and R.V. Dugarova, Healthcare, 12, 80–88, (2015) 20. N.N. Karkishchenko and S.V. Grachev, Guidance on Laboratory and Alternative Models in Biomedical Technologies (Profile, Moscow, 2010) 21. Reference book. Physiological, biochemical and biometric standards of experimental animals (“LEMA” Publishing, Saint Petersburg, 2013) 22. Reference book. Farm animals. Physiological and biochemical parameters of the body (VNIIFBiP, Borovsk, 2002) 23. V.D. Bogdanov, О.V. Sazarova, and Т.G. Saharova, Scientific papers of Dalrybvuz, 37, 93–98, (2016) 24. Т.V. Alexeeva, А.А. Rodionov, and А.А. Vesnina “Economics. Innovations. Quality management, 3, 127–131, (2015) 25. L.Yu. Lazhentseva, University Journal. Food Technology, 1, 108–110, (2009) 26. E.V. Belokurova, N.S. Rodionova, and S.A. Solohin, Journal of Advanced research in technical science, 1, 2474–5901, (2016) 27. L.V. Golubeva, E.A. Pozhidaeva, and E.S. Popov, Indian Journal of scienct and technology, 42, 104– 224, (2016) 28. N.N. Alekhina, E.I. Ponomareva, and S.I. Lukina, Journal of engineering and applied sciences, 12, 2623–2627, (2016) 29. Е.I. Ponomareva, V.I. Popov, and I.E. Esaulenko, Nutrition issues, 5, 75–81, (2017) 30. L.V. Golubeva, Е.А. Pozhidaeva, and G.М. Smolsky, Food Industry, 1, 60–64, (2020) 4 BIO Web of Conferences 30, 01002 (2021) https://doi.org/10.1051/bioconf/20213001002 ILS 2020

Study of properties of jelly-fruit marmalade with herbal additives Maya Artamonova1* , Inna Piliugina2 , and Olena Aksonova2 1Department of Technology of Bread, Confectionary, Pasta and Food Concentrates, Kharkiv State University of Food Technology and Trade, Kharkiv, Ukraine 2Department of Chemistry, Microbiology and Hygiene of Food, Kharkiv State University of Food Technology and Trade, Kharkiv, Ukraine Abstract. The results of studies of the properties of jelly-fruit marmalade with plant additives from Sudanese rose, rose hip and pumpkin during storage for 3 months are presented. It is shown that during the shelf life organoleptic, physicochemical quality indicators of marmalade with plant additives meet the requirements of current regulations, the color intensity is maintained. New products have a high content of β-carotene, anthocyanins, minerals, tannins and pectin. 1 Introduction Confectionery is a large group of high-calorie foods that are in great demand among the population. In particular, there is an increase in production and demand for jelly products. This is due to their attractive appearance, pleasant taste, low price. However, the main significant disadvantage of confectionery is their low physiological value due to the almost complete absence of important biologically active substances, such as vitamins, dietary fiber, minerals and the like. Therefore, in today's market competition, it is important to improve existing technologies and develop new types of jelly products. This is possible through the development of innovative technologies, the creation of products of high nutritional and biological value, special purpose, as well as the use of new raw ingredients. The search for new non-traditional types of raw materials for the production of jelly-fruit marmalade remains topical. Technologies for the manufacture of jelly products allow the introduction of plant supplements in various forms: in natural form, products of processing of plant raw materials, products of processing of nontraditional plant raw materials and their combinations. Scientists are actively developing new technologies for jelly-fruit marmalade using plant raw materials or products of its processing. Thus, the technologies of fruit and jelly marmalade for health purposes with the use of rhubarb, elderberry and water-alcohol extracts of thyme, violet, oregano are known. The introduction of these additives allows to introduce into the finished product catechins, flavonols and anthocyanins. The finished product has good taste and appearance, increased nutritional value and pharmacological effect 1.The technology of jelly-fruit marmalade with alcohol extract of Dunaliella salina microalgae and concentrated apple juice has been developed. It is shown that new types of marmalade have increased antioxidant activity 2. There is a method of obtaining jelly-fruit marmalade on agar and fructose with the addition of sea buckthorn juice. Replacement of apple puree with sea buckthorn juice in the marmalade formulation has significantly increased the antioxidant properties of products 3. The technology of jelly-fruit marmalade using hydrated pectin, which is obtained from medicinal dandelion, has been developed. New technology allows to obtain products with increased nutritional value 4.Scientists have developed a technology of jelly-fruit marmalade on pectin with the replacement of granulated sugar with fructose and 30% of apple puree on pureed fruits chokeberry, which allowed to obtain a diabetic and functional product 5.The use of chokeberry puree in the recipe of jelly-fruit marmalade on agar allowed to obtain products with high content of anthocyanins 6.The expediency of using the pulp of the fruits of wild garlic in the production of jelly-fruit marmalade on agar of high nutritional value of functional orientation is proved 7.The possibility of introducing beet juice, sea buckthorn and cranberry puree into the recipe of jellyfruit marmalade on gelatin to increase its nutritional value has been established, namely the introduction of bioflavonoids and betaine into the composition of products8.The use of semi-finished products from Jerusalem artichoke in the technology of fruit-jelly marmalade was proposed, which made it possible to increase the nutritional value of products and reduce the calorie content 9. A number of publications are devoted to the use in the technology of jelly-fruit marmalade on pectin plant additives obtained by cryogenic technologies – cryopowders and cryopaste 1016.The use of cryopowders from grapes, rose hips, sea buckthorn and *Corresponding author: [email protected] © The Authors, published by EDP Sciences. This is an open access article distributed under the terms of the Creative Commons Attribution License 4.0 (http://creativecommons.org/licenses/by/4.0/). BIO Web of Conferences 30, 01003 (2021) https://doi.org/10.1051/bioconf/20213001003 ILS 2020

cryopaste from quince, apples, carrots, pumpkins, grapes in the production of jelly-fruit-marmalade to increase its nutritional value, complete exclusion from the prescription composition of synthetic dyes and flavors 1012. In addition, the introduction of these additives to the prescription composition of marmalade helped to increase the antioxidant properties of products that are stable during the guaranteed shelf life.13, 14.It is shown that new types of jelly-fruit marmalade on pectin had high quality indicators during storage 15, 16. In works 17, 18 the expediency of using in the technology of jelly-fruit marmalade on gelatin cryopaste and cryopowder from grapes is shown. The innovative solution allows to obtain high quality products with maximum preservation of biologically active substances and reduced consumption of apple puree by 10-15% and citric acid by 20-25%, as well as the complete exclusion of dyes and flavors. However, the task of increasing the nutritional value of jelly-fruit marmalade on gelatin through the use of plant supplements from Sudanese rose, rose hip and pumpkin in these works was not solved. The authors of this publication are devoted to the solution of the problem described above. New types of jelly-fruit marmalade based on gelatin with herbal additives have been developed, namely Sudanese rose powder, rosehip cryopowder and pumpkin cryopaste. Studies of the properties of new types of marmalade with plant additives during the shelf life were not performed. The study of organoleptic and physicochemical properties of new products are necessary to justify their storage conditions and type of packaging. Thus, to solve the problem of maintaining over time the quality, color fastness of new types of jelly-fruit marmalade on gelatin with plant additives from Sudan rose, rose hip and pumpkin, it is important to timely investigate their properties during storage. 2 Experimental The purpose of this work was to study the organoleptic and physicochemical properties, chemical composition of jelly-fruit marmalade on gelatin with plant additives during storage, justification of optimal storage conditions for products and type of packaging. Object of research organoleptic, physicochemical properties and chemical composition of the product during its storage for 3 months. Subject of research jelly-fruit marmalade on gelatin with plant additives. Fruit jelly differed in the type of herbal additive Sudanese rose powder; cryopowder from rose hips and cryopaste from pumpkin (Fig. 1). Marmalade made on gelatin according to traditional recipes was used as a control one. Plant additives obtained by low-temperature technologies, due to fine grinding are a concentrate of biologically active substances. They contain a significant amount of low- and high-molecular phenolic compounds, dietary fiber, vitamins, glycosides, organic acids, macroand micronutrients and have antioxidant, immunomodulatory properties, as well as high coloring ability, good taste and aromatic characteristics. The marmalade was stored at a temperature of 18 ± 2 ˚С, relative humidity 75-80%, without access to light for three months, packed in a plastic wrap and a cardboard box. Control points during the experiment were the 1st, 2nd, 3rd months. Fig. 1. Plant additives of: a Sudanese rose; b – rose hip; c pumpkin. Quantitative characterization of the quality of marmalade samples and its study during the shelf life was carried out based on the main physicochemical parameters, namely mass fraction of moisture, mass fraction of reducing substances, total acidity. These indicators were determined using standard methods 19, 20. Color stability and color intensity of products with powder from Sudanese rose at certain time intervals were determined by spectrophotometric method. To do this, the optical density of the marmalade solution was determined on an SF-46 spectrophotometer at a wavelength of 510 nm, using cuvettes with a layer thickness of 10-2 m. The change in optical density was expressed as a percentage, while the initial optical density was taken as 100%. The content of β-carotene was determined by colorimetric method. The method is based on the ability of β-carotene to dissolve in petroleum ether, as a result of which the solution turns yellow. The color intensity of the solution is proportional to the β-carotene content in it. Determination of the content of anthocyanin substances was carried out by the method of pH differential spectrophotometry. The content of pectin substances in new types of jelly-fruit marmalade was determined by the calcium pectate method, which is based on the precipitation of pectic acids in the form of calcium salts. The method of determination of tannins was based on their easy oxidation by potassium tetraoxomanganate in an acidic environment in the presence of indigosulfonic acid. 3 Results and discussion Organoleptic and physicochemical quality indicators of jelly-fruit marmalade on gelatin with plant additives are given in tables 1, 2. It was found that at the end of the shelf life, the visual perception of the color of new types of marmalade remained unchanged, the taste, appearance and consistency of the samples changed somewhat. Products decrease in volume, the consistency becomes gummier, 2 BIO Web of Conferences 30, 01003 (2021) https://doi.org/10.1051/bioconf/20213001003 ILS 2020

the sour taste becomes more pronounced, which is confirmed by the data of physical and chemical studies. It is proved that the color intensity of jelly-fruit marmalade with Sudanese rose powder after 3 months of storage was 100%. Thus, the color of the new type of marmalade remains stable during the guaranteed shelf life. The study of physicochemical parameters during storage for three months showed that at the end of the storage period the moisture content in new types of marmalade decreases to 11.9-13.4%; the total acidity is 20.4-22.0 degrees; the content of reducing substances is 4.8-5.1%. The change in properties during storage is explained by a whole complex of physicochemical and microbiological processes. The transformations are associated with the hydrolysis of the carbohydrate component, redox reactions, condensation, and polymerization. However, all indicators were within the limits corresponding to the requirements of regulatory documents for this type of product. The nutritional value of the developed types of marmalade was assessed. In jelly-fruit marmalade on gelatin with the addition of these plant additives, the content of minerals significantly increases (Table 3). It is noted that the new types of jelly-fruit marmalade on gelatin in comparison with the sample made according to the traditional formulation, contain 1.2-2.3 times more phosphorus, 1.2-1.7 times more iron, 1.1-1.6 times more calcium. The content of potassium and magnesium also increases in the products. In general, with the introduction of herbal supplements, the mineral content in marmalade increases by 1.1-1.3 times. It is determined that new types of marmalade with plant additives are characterized by a higher content of biologically active substances compared to jelly-fruit marmalade made according to traditional formulation (Table 4). The β-carotene (mg / 100 g) was found in freshly prepared marmalade: control contains (1.30 ± 0.04); marmalade with Sudanese rose powder - (2.25 ± 0.09); marmalade with rosehip cryopowder and pumpkin cryopaste - (2.77 ± 0.25). That is, the consumption of 100 g of freshly made marmalade provides 45...55% of βcarotene from the daily value of an adult. After a month of storage, the content of β-carotene in the marmalade with pumpkin cryopaste and rosehip cryopowder was 0.15 ± 0.03 mg per 100 g of product. In jujube with powder from the Sudanese rose, β-carotene was destroyed, which is explained by its instability in an acidic condition and upon contact with atmospheric oxygen. According to the results obtained (Table 4), the content of anthocyanins, pectin and tannins at the end of the shelf life remained almost unchanged. So, the possibility of storing jelly-fruit marmalade on gelatin with plant additives from Sudanese rose, rose hips and pumpkin has been proven during the guaranteed shelf life of 3 months, provided that it is packed in plastic wrap and a cardboard box. It has been established that the consumption of 100 g of new types of marmalade will satisfy the daily need of healthy residents of Ukraine from 18 years of age and older in pectin substances by 3.7-5.7%. Consuming 50 g of Sudanese rose marmalade will satisfy the daily need for anthocyanins by 120%. Table 1. Organoleptic quality indicators of freshly made jelly-fruit marmalade on gelatine with plant additiveseriment. Indicators Characteristics of jelly-fruit marmalade on gelatin control with Sudanese rose cryopowder with rose hips cryopowder and pumpkin cryopaste Form Correct, with a clear contour, without deformation Surface Smooth, sprinkled with sugar Color Yellow Dark pink Light orange Taste and smell Pleasant, without foreign taste and smell Pleasant, with a slight smack of Sudanese rose without any foreign taste and smell Pleasant, with a taste of pumpkin and rose hips without foreign taste and smell Consistence Jellylike Table 2. Physicochemical indicators of the quality of jelly-fruit marmalade on gelatin with plant additives Sample of marmalade Shelf life, months Mass fraction of moisture, % Total acidity, degrees Mass fraction of reducing substances, % Control 0 23.0±1.0 11.6±0.9 12.3±0.4 3 12.9±0.9 16.2±0.7 4.2±0.7 Marmalade with Sudanese rose cryopowder 0 22.2±1.2 16.4±0.5 14.3±0.4 3 13.4±0.9 22.0±0.3 5.1±0.7 Marmalade with rose hips cryopowder and pumpkin cryopaste 0 22.5±1.2 13.2±0.5 13.6±0.6 3 11.9±0.8 20.4±0.4 4.8±0.8 According to normative documents (DSTU 4333: 2004) not more than 24 7.522.5 not more than 28 3 BIO Web of Conferences 30, 01003 (2021) https://doi.org/10.1051/bioconf/20213001003 ILS 2020

Table 3. The content of minerals in the studied sample of innovative marmalade Mineral substance Daily intake for an adult, mg Content in marmalade, mg / 100 g control with Sudanese rose cryopowder with rose hips cryopowder and pumpkin cryopaste К 2000 94.1 96.7 110.6 Ca 1000 99.54 159.24 105.67 Mg 400 24.34 25.02 26.07 Fe 15 1.55 2.56 1.9 Р 800 3.90 4.63 8.98 Total 223.43 282.86 253.22 Table 4. Chemical composition of jelly-fruit marmalade on gelatine with plant additives Sample of marmalade Shelf life, months Name of substances Anthocyanins, mg / 100 g Pectic substances, mg / 100 g Tannins (by tannin), mg / 100 g Control 0 182±2 324±9 3 188±2 320±9 Marmalade with Sudanese rose cryopowder 0 124.0±6.2 227±6 352±10 3 121.0±6.0 230±7 350±10 Marmalade with rose hips cryopowder and pumpkin cryopaste 0 1.2±0.1 187±5 330±9 3 1.1±0.1 187±5 330±9 4 Conclusion Thus, the results of studies of the properties of jelly-fruit marmalade with plant additives from Sudanese rose, rose hips and pumpkin during storage for 3 months showed that during the storage period the organoleptic, physicochemical indicators of the quality of new products comply with the requirements of the current regulatory documentation, intensity color is preserved. New products have an increased content of β-carotene, anthocyanins, mineral, tannins and pectin substances. Based on the results of research on new products, formulations and technology were developed, draft regulatory documents in the form of technical specifications and technological instructions, and the safety of the technology was assessed using the HACCP system. References 1. A. Bashta, T. Leshchynska, Scientific Works of NUFT, 53, 63-70 (2013) 2. E. Kuznetsova, Ya. Brindza, E. Klimova, A. Borovkov, I. Gudvilovich, Ya. Jondareva, E. Kuznetsova, Food Industry, 4, pp. 14-19 (2019) 3. G. Magomedov, I. Lobosova., S. Zhurakhova, Technique and technology of food production, 46, 50- 54 (2017) 4. N. Tiguntseva, S. Evstafyev, News of higher educational institutions. Food technology, 4, 36-39 (2015) 5. L. Lobosova, M. Magomedov, S. Zhurakhova, Bulletin of the Voronezh State University of Engineering Technology, 4, 256-260 (2016) 6. A. Tabarovich, E. Stepanova, V. Bakaytis, Food industry. Raw materials and additives, 7, 53-57 (2017) 7. S. Glazyrin, N. Tipsina, Bulletin KrasGAU, 10, 218- 220 (2014) 8. E. Pankova, S. Agafonova, Bulletin of Youth Science, 14, 1-6 (2018) 9. G. Magomedov [et al.], Confectionery production, 4, 38-39 (2011) 10. M. Artamonova, N. Shmatchenko, Scientific works of the Odessa National Academy of Food Technologies, 46, 177-180, 11. M. Artamonova, N. Shmatchenko, Baker, 6, 36-37 (2015) 12. M. Artamonova, I. Piliugina, N. Shmatchenko, Nutrition: innovative aspects of technology, energyefficient production, storage and marketing, рр. 229- 256 (2015) 13. M. Artamonova, I. Piliugina, N. Shmatchenko, S. Gubsky, Nutrition: innovative aspects of technology, energy-efficient production, storage and marketing, 117-142 (2015) 14. S. Gubsky, M. Artamonova, N. Shmatchenko, I. Piliugina, E. Aksenova, Eastern-European Journal of Enterprise Technologies. Technology and equipment of food production, 4, 43-50 (2016) 15. N. Shmatchenko, M. Artamonova, O. Aksonova, S. Oliinyk, Food Science and Technology, 12, 87-94 (2018) 16. M. Artamonova, N. Shmatchenko, Study of the influence of plant cryoadditives on the quality of jelly-fruit marmalade during storage in Innovative aspects of food and hotel industry equipment 4 BIO Web of Conferences 30, 01003 (2021) https://doi.org/10.1051/bioconf/20213001003 ILS 2020

development in modern conditions: materials of the second international scientific-practical conference, 2017, KhSUFTT, Kharkiv,Ukraine, pp. 203-204 17. M. Artamonova, N. Shmatchenko, I. Piliugina, Technology of jelly-fruit marmalade with the use of cryoadditives from grapes in International scientific and practical conference Technical sciences: history, the present time, the future, EU experience, 27-28 December 2019, Wloclawek, Poland 18. M. Artamonova., I. Piliugina, N. Shmatchenko, Nutrition: trends in energy efficient production, storage and marketing, 210-232 (2020) 19. Y. Picó, Chemical Analysis of Food: Techniques and Applications (Academic Press,Elsevier, 2012) 20. S.S. Nielsen, Food Analysis (Springer, Science+Business Media, LLC,2010) 5 BIO Web of Conferences 30, 01003 (2021) https://doi.org/10.1051/bioconf/20213001003 ILS 2020

An innovative concept for the technology of jelly-fruit marmalade using vegetable cryopastes Maiia Artamonova1* , Natalia Shmatchenko1 , Tetyana Gavrysh2 ,and Liudmyla Pikh2 1Department of technology of bread, confectionary, pasta and food concentrates, Kharkiv State University of Food Technology and Trade, Kharkiv, Ukraine 2Department of food technologies and processing industries, Kharkiv Petro Vasylenko National Technical University of Agriculture, Kharkiv, Ukraine Abstract. The results of research on the innovative concept of the technology of jelly-fruit marmalade with vegetable cryoadditives from carrots and pumpkin are presented. The use of innovative developments in marmalade makes it possible to increase productivity and improve the quality of products. It is shown that organoleptic, physicochemical quality indicators of marmalade with plant additives meet the requirements of current regulations, the colour intensity is maintained. The new products have an increased content of vitamin C, β-carotene and pectin. 1 Introduction In modern economic conditions, food companies need to constantly improve existing products, production technologies, methods of their implementation, implement technical innovations, develop new products and bring them to market [1]. A sign of innovation is the requirement that the product, process, method, etc. must be new or significantly improved for the company. Innovations include products, processes or methods that the company first created. Another sign of innovation is that a product, process, marketing method or organization must be implemented. A new or improved product is introduced when it begins to be marketed [2]. Sugar confectionery due to the content of a large number of easily digestible sugars stimulates mental activity and provides energy to the human body. Also, the consumption of sugary confectionery promotes the body's production of the pleasure hormone - serotonin. However, today it is very difficult to choose confectionery without synthetic additives, because, unfortunately, modern confectionery production uses dyes and flavours, synthetic and identical to natural. Many of these supplements are not just harmful but dangerous. One of the most common products containing synthetic additives are jelly products, so it is important today to create products using natural ingredients with high nutritional and biological value. The peculiarity of jelly products is their composition, which includes gelforming components: pectin, gelatine, agar-agar, as well as sugar-molasses syrup, fruit puree, fruit juices, sugar, molasses. Thanks to pectin, a soluble dietary fiber, marmalade is a useful food product. The current state and prospects of improving the quality and nutritional value of jelly products by adding plant supplements are based on an analytical review of literature and patent sources [3-7]. To significantly reduce the risk of chronic diseases and meet the needs for micronutrients, daily consumption of fresh fruits and vegetables is recommended. However, fruits and vegetables have a very short shelf life due to their high moisture content. In addition, they have seasonal and regional availability, which limits the longer use of these products. To overcome these negative aspects, they are widely used to produce processed products such as jams, jellies and marmalades. [8]. It is established that the use of various additives of plant origin in marmalade technologies is one of the most promising technological methods to provide them with health and therapeutic properties, but in many cases the additives require additional mechanical or heat treatment, which leads to loss of biologically active substances. Therefore, today alternative methods of processing and storage of fruit and vegetables are processing by lowtemperature technologies. Today in the food industry it is promising to use plant additives with increased amounts of biologically active substances obtained by modern technologies, such as cryogenic freezing and grinding [9, 10]. Highly frozen vegetable supplements contain a significant amount of vitamins, carotenoids, pectin, which strengthen the immune system, as well as have a detoxifying effect on the human body. Grinding of raw materials as a technological technique is widely used in various branches of the food industry and largely ensures the quality of the finished product. In particular, as the particle size of the product decreases during grinding, its surface area increases. This allows more complete extraction of biologically active nutrients, aromatic substances with solvent, and also leads to increased *Corresponding author: [email protected] © The Authors, published by EDP Sciences. This is an open access article distributed under the terms of the Creative Commons Attribution License 4.0 (http://creativecommons.org/licenses/by/4.0/). BIO Web of Conferences 30, 01004 (2021) https://doi.org/10.1051/bioconf/20213001004 ILS 2020

digestibility of crushed products by the human body. The feasibility of using cryogenic grinding is closely related to the cost-effectiveness of the process. The cost of cooling is offset by the advantages of cryogenic technology, which for food products are determined by their quality, namely: the preservation of vitamins, aromatic and other biologically active substances (BAS), increasing the bioavailability and physiological efficiency of products. And this is equivalent to creating an additional amount of finished product of higher quality [11, 12]. Therefore, to implement the innovative idea of the technology of jelly-fruit marmalade and increase its nutritional value, we have proposed the use of vegetable cryopastes from carrots and pumpkin. 2 Experimental The aim of this work was to study the organoleptic and physicochemical properties of jelly-fruit marmalade on pectine with vegetable additives. The object of research is organoleptic, physicochemical properties of the product. Subject of research – jelly-fruit marmalade on pectin with vegetable additives. The resulting marmalade differed in the type of herbal supplement – cryopastes from pumpkin and carrots. Due to the low-temperature technology of their production, cryoadditives are a concentrate of biologically active substances, contain a significant amount of low- and high-molecular phenolic compounds, dietary fiber, vitamins, organic acids, macro- and microelements and have antioxidant, immunomodulatory properties and high colouring ability, taste and odour characteristics [11, 13]. Sampling for physicochemical studies of experimental samples was performed according to DSTU 4619: 2006. Determination of organoleptic characteristics of finished products was performed according to DSTU 4683: 2006. The moisture content of the finished products was determined refractometrically according to DSTU 4910: 2008, for this purpose a 50% solution was prepared: to a portion of 5 g of marmalade was added 5 ml of distilled water and dissolved in a water bath to a homogeneous solution. The titratable acidity of the finished products was determined according to DSTU 5024: 2008. To express the acidity in% of citric acid, the acidity in degrees was multiplied by the milliequivalent (0.07). The mass fraction of vitamin C was determined using the titrometric method according to GOST 24556-89. The method is based on the redox reaction between ascorbic acid and the indicator 2,6-dichlorophenolindophenol (Tillmans reagent). The content of carotenoids was determined by colorimetric method according to GOST 13496.17-95. The method is based on the ability of carotene to dissolve in petroleum ether or gasoline, giving a yellow colour, the intensity of which is proportional to the carotene content. The content of pectic substances in plant raw materials and products of its processing was determined by one of the most accurate methods: calcium-pectate, which is based on the precipitation of pectic acids in the form of calcium salts. Determination of strength was performed on a penetrometer "Labor", recording the destructive weight. 3 Results and discussion The Today, the main tasks of the food industry are: the creation of health products, new technologies for food production, recovery and use of food waste, ensuring the biosafety of packaging materials and more. Manufacturers are faced with the task of paying more attention to foods that meet the needs of consumers in a healthy lifestyle. As a result, there is a need to innovate in food technology and raw material processing, to withstand competition among manufacturers [14]. To create safe, high-quality, health-promoting products on the market, non-heat treatment technology is available. Innovative cryogenic food processing is often perceived as an alternative to heat treatment. The research conducted in the work pays attention to improving the functionality of food products due to the minimal processing of fresh vegetable raw materials, its quality, safety and ease of use. Of particular interest are studies on the "path to commercialization" for selected new technologies using plant additives obtained by cryotechnology [15]. The innovative strategy of this study provides: ‒improvement of jelly-fruit marmalade technology with the use of vegetable cryopastes, which has certain competitive advantages over traditional technologies in this segment; ‒offer products to expand the range of jelly products without the use of synthetic additives (flavours, essences, dyes); ‒creation of new types of marmalade on pectin with high content of biologically active substances. Taking into account the above information, a description of the innovative product was compiled (Table 1). The use of innovative developments in the production of marmalade makes it possible to increase productivity and improve the quality of products. The technological process for the production of jellyfruit marmalade contains the following stages: preparation of raw materials, preparation of the marmalade mass, the formation and gelation of the marmalade mass, removal of the marmalade from the molds and sprinkling with sugar, proofing (drying) of the marmalade, packing, packaging products. To establish rational dosages of pumpkin and carrot cryopastes in marmalade technology, experimental samples with different numbers of cryopastes were made. Jelly-fruit marmalade on pectin with apple puree was chosen for control [16]. Cryopaste was used as a fruit additive in the samples studied and synthetic essences were completely excluded from the formulation. To do this, the marmalade mass was added thawed vegetable cryopaste at the end of the boiling stage in the amount of 5.0-25.0% of the total weight of the system. 2 BIO Web of Conferences 30, 01004 (2021) https://doi.org/10.1051/bioconf/20213001004 ILS 2020

Table 1. An innovative concept of a new product - jelly-fruit marmalade with theuse of vegetable cryopastes Indicator Characteristic Sources of implementation Product appearance Fruit jelly marmalade has the correct form with a clear drawing and accurate contours, without deformation. The surface is evenly sprinkled with white sugar Achieved by complying with the rational parameters of the technological process and the use of gelling agents and cryopastes Target segment The product is intended for consumption by the general population В2С: general population through wholesale and retail trade enterprises Competitive advantages Fruit jelly marmalade with vegetable cryopastes and cryopowders is characterized by high content of vitamin C, β-carotene, pectin and reducing the prescription amount of the gelling agent Achieved through the implementation of technological properties of plant cryoadditives Packaging Packing in paper-cardboard, polymeric and polyethylene materials is provided Product weight from 200 g to 7 kg depending on the form of sale Assortment The range of marmalade depends on the type of cryopaste that is added The assortment is formed taking into account: technological purpose, form of sale and demand Shelf life 3 months Storage conditions for marmalade: in dry, clean, wellventilated warehouses at an air temperature of 15 to 18 ° C and a relative humidity of no more than 75% Estimated wholesale price per 1 kg up to UAH 75 Achieved through the use of cryopastes and depends on the cost of raw materials The first step was to determine the strength of the obtained samples of marmalade with cryopastes for all dosages. The strength of the control sample was (τ0 = 12.5±0.63 kPa). It is established that the addition of cryopastes increases the strength of marmalade, namely, with the introduction of cryopastes from carrots in the amount of 5.0-20.0%, the strength increases by 8-34%, from pumpkin - by 17-42%. The increase in strength is due to a significant amount of pectin in plant cryopastes, so we can assume the possibility of reducing the amount of pectin according to the recipe. With a further increase in the prescription amount of cryopastes to 25%, the strength index decreases and the structure of marmalade becomes viscous and brittle, which does not meet the requirements. In the course of organoleptic studies, it was found that at dosages of cryopastes in the amount of 5.0-20.0% of the total weight of the marmalade system has the correct shape, with a clear contour, without deformation, consistency - gel-like, not gummy. The taste and smell of the obtained products are pronounced due to the presence of cryopastes, without foreign taste and smell, but with an increase in the number of cryopastes there is a sour taste. Colour - from light orange to dark orange. When breaking the sample, the inner surfaces are transparent, without the inclusion of additives. The outer surface of the marmalade is evenly sprinkled with white sugar, elastic. In the case of adding cryopastes in the amount of 25.0% of the total weight of the system and above, the marmalade has a brittle not gel-like consistency, but more like jam, this is due to a violation of the pectin-sugar-acid balance. Physico-chemical indicators of quality of samples of marmalade with a dosage of cryopastes in the amount of 5.0-20.0% were studied It is established that marmalade for all dosages meets the requirements of regulatory documentation [17] for physicochemical parameters (mass fraction of moisture in the range of 15.0-24.0%; total acidity 7.5-22.5 degrees; mass fraction of reducing substances is not more than 28.0%). The acidity of the samples and the content of reducing substances in them increases according to the amount of additive. The strength index of marmalade when adding cryopastes increases compared to the control (τ0 = 12.5±0.63 kPa) The strength of marmalade with the addition of cryopastes increases compared to the control by 8…58%, which gives grounds to reduce the prescription amount of gelling agent. According to the results of studies of strength indicators with a decrease in the prescription amount of pectin by 5.0-25.0%, it was found that it is advisable to reduce the prescription amount of pectin by 20.0% for all tested samples of marmalade. When reducing the amount of pectin by more than 20.0% of the prescribed amount, the strength of marmalade becomes lower than the control. Thus, in the course of the research, the most rational dosages of cryopastes and pectin were selected, and formulations of new types of marmalade were developed: ‒with carrot cryopaste in the amount of 10.0% of the total weight of the system with a reduced amount of gelling agent by 20.0%; ‒with pumpkin cryopaste in the amount of 10.0% of the total weight of the system with a reduced amount of gelling agent by 20.0%; ‒with carrot paste in the amount of 15.0% of the total weight of the system with a reduced amount of gelling agent by 20.0%; ‒with cryopaste from pumpkin in the amount of 15.0% of the total weight of the system with a reduced amount of gelling agent by 20.0%; Plant cryopastes contain a significant amount of biologically active substances, so it was important to determine the content of vitamin C, β-carotene and pectin in new samples of marmalade (Table 2). 3 BIO Web of Conferences 30, 01004 (2021) https://doi.org/10.1051/bioconf/20213001004 ILS 2020

Table 2. Content of vitamin c, β -carotin, and pectin substances in marmelad with addition of cryopastes Marmalade Content of biologically active substances Vitamin C, mg / 100 g β-carotene, mg / 100 g Pectin substances, mg / 100 g according to the traditional formulation 0.40±0.02 trace 1.55±0.08 with carrot cryopaste in quantity (from the total weight of the system): 10.0% 0.56±0.03 3.50±0.18 1.50±0.08 15.0% 0,90±0.03 5.10±0.26 3.45±0.18 with pumpkin cryopaste in quantity (from the total weight of the system): 10.0% 1.20±0.06 3.00±0.15 2.70±0.14 15.0% 1,80±0.09 4.50±0.23 3.90±0.20 As can be seen from the table, the content of vitamin C in marmalade with the addition of vegetable cryopastes in the amount of 10.0 and 15.0% increases approximately 1.4-4.5 times according to the type and number of cryopastes compared to the control. The control sample of marmalade does not contain β-carotene, and 100 g of marmalade with cryopaste from carrots and pumpkin in the amount of 10.0-15.0% contains 3.1-5.0 mg / 100 g of β-carotene depending on the type and number of cryopastes. The content of pectin substances in marmalade with the introduction of fruit and vegetable cryopastes increases up to 2.5 times. Based on the physiological needs of a person, the daily amount of vitamin C is 75 mg, carotene - 5 mg, pectin substances - 5 g, and anthocyanin substances - 80 mg. To provide marmalade with the status of a product of health-improving action, the content of these physiologically functional ingredients should be 20-50% of the daily requirement of a person [16, 17]. Therefore, it is important to calculate the content of these biologically active substances in relation to the average daily human need for 100 g of new products (Table 3). Data are given in % in relation to the average daily requirement of an adult. The table shows that the content of β-carotene in marmalade with the addition of cryopaste from carrots and pumpkin in the amount of 10-15% is 60.0-102.0% of the average daily human need. The content of pectin substances in new samples of marmalade is 30.0-78.0% of the average daily human need. Therefore, these samples can be considered products of health effects. 4 Сonclusion An innovative idea to expand the range of jelly-fruit marmalade with increased nutritional value was identified. Formulations and technologies of marmalade with the addition of vegetable cryopastes have been developed. The use of cryopastes made from carrots and pumpkin has reduced the prescription amount of pectin by 20%. Organoleptic and physicochemical parameters of new products meet the requirements of current regulations. The resulting marmalade products have a high content of vitamin C, β-carotene and pectin. Based on the results of research on new products, draft regulatory documents in the form of technical conditions and technological instructions were developed, the safety of technology was assessed using the HACCP system. Table 3. Content of vitamin c, β-carotin and pectin substances in marmalade with addition of cryopastes in relation to the average daily need of adults Marmalade The content of substances relative to the average daily requirement when consuming 100 g of marmalade, % Vitamin C β-carotene Pectin substances According to the traditional formulation 0.5 - 31.0 with carrot cryopaste in quantity (from the total weight of the system): 10.0% 0.6 70.0 30.0 15.0% 1.2 102.0 69.0 with pumpkin cryopaste in quantity (from the total weight of the system): 10.0% 1.6 60.0 54.0 15.0% 2.4 90.0 78.0 References 1. N. Skopenko, Herald of Lviv Polytechnic National University, 684, 66-71(2010) 2. N. Chorna, Innovative development of food production and food security risks ( monograph, Lviv, 2012) 3. A. Bashta, T. Leshchynska, Scientific Works of NUFT, 53, 63-70 (2013) 4. N. Sanzharovskaia, O. Khrapko, “Technical sciences, 64, 95-98 (2017) 5. N. Ershova, N. Tarasenko, “Composition for the production of marmalade,” Patent RF, no. 2650549, (2018) 6. G. Tsyibikova, S. Batumunkeeva, ‘Vegetable marmalade,” Patent RF, no. 2631307, (2017) 7. Magomedov G. O., Method for producing jelly marmalade using concentrated pumpkin paste. Patent RF, no. 2603895 (2016). 8. T. Swer, S. Rani, Kh. Bashir, Processing of Fruits and Vegetables From Farm to Fork,366 (2019) 9. I. Syazin, G. Kasyanov, Food technology, 4, 123- 124 (2012) 10. M. Ahmedov, G. Kasyanov, A. Ramazanov, Z. Yaralieva, “Innovative technologies for the production of fruit and vegetable cryopowders,” Abstract journal “Food and processing industry”, No. 3, pp. 135-149 11. R. Pavlyuk, V. Pogarskaya, V. Pavlyuk, A. Berestova, N. Maksimova, Cryomechanochemistry in food nanotechnology, monograph, (2015) 12. R. Pavliuk, “New about carotenoids and oxidative enzymes of carotenoid vegetables during cryogenic "shock" freezing and grinding,” Advanced 4 BIO Web of Conferences 30, 01004 (2021) https://doi.org/10.1051/bioconf/20213001004 ILS 2020

techniques and technologies of food production, restaurant business and trade, No. 1, pp. 52-60 13. V. Poharska, “Study of cryomechanical destruction and mechanochemistry processes in the development of nanotechnology of frozen carotenoid plant additives,” Eastern-European Journal of Enterprise Technologies, 84, pp. 39-46, 2016) 14. Ch. Galanakis, “Innovation Strategies in the Food Industry,” 1st ed., Academic Press, (2016) 15. O. Tokusoglu, “Introduction to Innovative Food Processing and Technology,” Natural Science and Discovery, 4, pp. 85–87, (2015) 16. L. Kuznetsova, M. Sidanova, Production of marmalade-pastille products, Moscow: DeLi plus, (2012) 17. State Standart 4333-2004, Marmalade. General technical conditions, Kyiv, The State Committee of Ukraine for Technical regulation and Consumer policy, (2004) 5 BIO Web of Conferences 30, 01004 (2021) https://doi.org/10.1051/bioconf/20213001004 ILS 2020

Structuring meat systems using natural biopolymers Boris Baranov1* , Aleksandr Sokolov2 , and Yuri Boltenko3 1Plekhanov Russian University of Economics, Moscow, Russia 2Plekhanov Russian University of Economics, Moscow, Russia. Moscow State University of Food Production, Moscow, Russia 3Belgorod State National Research University, Belgorod, Russia. Abstract. The features of the structure and properties of raw materials, which are a source of biopolymers of natural origin are presented. The main focus of the theoretical part is paid to the study of the process of structuring of biopolymers (proteins and polysaccharides). Practical approbation was carried out on the example of meat-containing meat and meat chopped products and semi-finished products. 1 Introduction Important areas of research are the stabilization of various foods environments, including the study of the properties of colloids [1-5], structuring of food systems with the disclosure of biopolymer potential (proteins, polysaccharides, etc.), technology development in various industries, including food recipes based on natural raw materials. The main tasks of work are to compare the features of structuring the biopolymers of different groups and their influence on the technological and consumer properties of culinary products on a meat basis. 2 Experimental In the new laws and regulatory documents of Russia, the principle is presented - the rational use of raw materials of industries, including the food and catering industry. Therefore, the development of the theory and practice of the formation of the structure, properties of biopolymers and food, as well as the scientific substantiation of the methods of their optimal processing is the most important scientific problem. The goal of the work is a scientific substantiation of the possibilities of using various natural sources of raw materials to obtain gelling agents (in the comparative aspect - proteins, polysaccharides) and their rational use in the technology of food and culinary products. Waste of meat processing production were used as raw materials of animal origin - pork skins (for example, canned freezing), edge sections of the skins. The use of pork ears, beef lips, etc. is allowed. The chemical composition of these types of raw materials is presented in Table 1. Analysis of data on the chemical composition of raw material sources allowed to choose raw material rich in protein biopolymers, in particular the skin of pork, ears of cattle. However, the first type of raw material is more preferable due to a more homogeneous histological structure (absence of cartilage tissue) and a greater amount of resources of its blanks, what is more practically. Since the raw material was selected according to the criterion of biopolymers content, the next stage of the work was aimed at developing an optimal scheme for transforming the structure and disclosing the functional and technological potential of this type of protein raw material. This method is not considered in detail, but it is necessary to note the principle of partial hydrolysis with solutions of inorganic and organic compounds (allowed to use as food additives) in a wide range of pH changes. This method allowed us to regulate the structure and properties in a directed manner, on the other hand, it does not require fundamental changes in the design of the equipment, it can be performed at the temperature of the workshop. Combined hydrolysis of raw materials allowed the development of a semi-finished product, conditionally named protein structuring agent (PSA), which can be dried using a sublimation unit. At the same time, the focus has been on structuring food systems using this semi-finished product (PSA). One of the most scientifically interesting processes is the thermotropic gelation, but its rationale requires the study of fractions of food disperse systems. Loosening of tissue after hydrolytic effects was detected according to the light and scanning electron microscopy (SEM). In particular, there is an increase in interfiber spaces between bunches of fibers (PV) and their disorientation are noted. The hydrolytic effect led to the fact that the swelling BF and primary fibers became insufficiently distinguishable (Fig. 1). The fibrillar structures were more homogeneous. Loosening and longitudinal splitting of layers and bundles of fibers and cells due to separation of fibers or thinning of layers of loose connective tissue were revealed. . *Corresponding author: [email protected] © The Authors, published by EDP Sciences. This is an open access article distributed under the terms of the Creative Commons Attribution License 4.0 (http://creativecommons.org/licenses/by/4.0/). BIO Web of Conferences 30, 01005 (2021) https://doi.org/10.1051/bioconf/20213001005 ILS 2020

Table 1. Chemical composition of animal raw materials Name of raw materials Moisture Fats Proteins Ash Water soluble Total Complete substances Collagen groups Pork raw Pork skin (saline) 62.0± 0.4 10.1± 0.5 27.8± 0.8 1.9± 0.6 25.9±0.6 0.60± 0.03 1.20± 0.07 Skin (inter-nipple part) 42.6 42.5 13.7 3.4 10.3 0.14 1.06 Skin (marginal sections) 54.0 18.5 28.4 ̶ 9.6 0.25 1.10 Ears 60.9 14.1 21.0 8.4 12.6 0.70 3.30 Cattle raw Lungs 77.5 4.7 15.2 7.3 7.9 1.00 1.60 Ears 69.8 2.3 25.2 10.5 14.7 0.70 2.00 Lips 71.0 4.9 20.3 9.3 11.0 0.90 2.90 Rumen 76.0 3.9 18.0 10.4 7.6 1.40 0.70 The influence of the thermal process on the change in the microstructures of the PSA was studied. The features of the microstructure were established at the level of light microscopy to identify general changes in tissues and SEM — to determine the features of the architectonics of collagen fibers found throughout the entire thickness of the samples with a large depth of field. Pores of various diameters found in almost all layers of the samples (Fig. 1), recorded in PSA samples, probably influenced by grinding, thermal gelation, etc. There are other features of the ultrastructure, but in detail in terms of technology they are not significant. Professor G.P. Andrianova and co-authors reported that the modification changes the properties of functional groups of biopolymers, i.e. free amino, carboxyl and -OH groups appear and increase their hydration. The flooding of collagen structures was also initiated by electrostatic interaction with water dipoles, which, apparently, increased the content of tightly bound moisture and, as a result, increased the moisture binding ability. In particular, PSA exceeded the feedstock by this indicator by about 4%. Apparently this is due to the activation of biopolymer centers, which increases the effect of interaction between proteins and water dipoles. A definite role here was played by the "composite" structure of raw materials ‒ complex biopolymers of connective tissue (collagen, elastin, glycoproteins, etc.). Based on the previously stated assumption that after heat treatment, the compound-type systems are formed that not only positively affect the functional and technological properties of minced meat products, but also contribute, a priori, to a decrease in losses of broth or nutrient solution. Consequently, the production efficiency of meat products increases. The molecular weight of the protein (viscometric method) in the colloidal solution was determined after a model heat treatment. It is known that the molecular weight of tropocolagen is 300-500 kDa, and of individual polypeptide chains of collagen group proteins is from 80 to 125 kDa. α-, β- and γ-chains with molecular weight of 100, 200 and 300 kDa were isolated from the products of thermal denaturation of collagen. According to the data the molecular weight of dissolved protein of the PSB was approximately 200 kDa. Therefore, it can be assumed that the presence of mainly high molecular structures similar to doublestranded β-polypeptides capable of thermotropicaly structuring the systems of the "protein protein" group, "protein-water"group, of participating in gelation. Fig. 1. Ultrastructure of the protein structuring agent (×600). Further research was aimed at obtaining a standard jelly based on category ΙΙ by-products ‒ sample 1 (according to the Recipe book) and model jelly (sample 2) using PSA. During the histological examination of sample 1, it was found that it is a homogeneous basophilic colored mass of biopolymers, in which particles of connective tissue are detected, which are relatively preserved by the structural organization (cell elements are not detected, the fibers are highly loosened), as well as fibers fragments, mainly elastic, turned red, fig. 2; microstructure in fig. 3 seems more homogeneous. The summation of all data (chemical composition; sensory; histological; molecular weight) made it possible to obtain the most complete and detailed histological picture of fibrous collagen products, positive changes in its architectonics (relative homogeneity of systems, cohesion, etc.). Jellies were prepared according to the standard recipe and experimental recipe (Table 2), according to the Recipe book. Based on the data in Table 2, the prototype with the introduction of PSA has a slight advantage in protein content and rheological quality indicator, which confirms 2 BIO Web of Conferences 30, 01005 (2021) https://doi.org/10.1051/bioconf/20213001005 ILS 2020

the effect of structure formation in these protein systems and improves the "tenderness" of meat-based products, which is important. Table 2. Composition and some properties of jellies Indicator Sample control prototype Moisture, % 78 74 Protein, % 9.6 11.3 Fat, % 10.2 10.6 The maximum share stress, kPA 1.2 1.5 Jelly temperature, С 15.7 16.5 Fig. 2. Ultrastructure of the control jelly (×≈200). Fig. 3. Ultrastructure of a jelly with a protein structuring agent (×≈200). In addition, technological processes for the production of gelled products, the hydrolysis of biopolymers and, in addition, the introduction of the necessary prescription ingredients provide high organoleptic characteristics, can be «fat replacers», etc. [4]. Calculation of the energy value of the finished jelly data showed that it is for the control sample 130.2 (kcal / 100 g), for prototype 140.6 (kcal / 100 g), which is relatively small. In addition, we can expect the enrichment of these products with protein biopolymers, which are a source of essential substances and and dietary fiber analogues. The next stage of the work is devoted to the study of natural biopolymers using alginates as an example in the technology of meat products and semi-finished products. Sodium alginate is produced from natural sources; it is soluble in water. When calcium ions (gluconate, calcium lactate, etc.) are introduced into sodium alginate solutions, water-insoluble sodium-calcium polyguluronates are formed. Calcium ion connects the polysaccharide molecules with each other, their aggregation and structuring occurs. Since the interaction with calcium ions occurs quickly, calcium salts are poorly soluble in water and weakly dissociating acids are used to slow down the process, which contributes to the gradual release of calcium ions sufficient to form a gel formation. It is very important that calcium alginate forms thermostable jellies, insoluble in water, which makes it possible to use it for structuring products subjected to thermal culinary processing. At this stage, the object of research was chopped meat and fish products. "Chopped Steak" was prepared without the addition of sowbelly to eliminate the ots unwanted influence in determining the structural and mechanical characteristics. "Natural Fish Schnitzel" was produced according to traditional technology. As a structuring additive, we used the dry mixture developed by us, consisting of food sodium alginate, calcium carbonate and food citric acid. The amount of added mixture ranged from 0.5 to 1.5% by weight of minced meat (Table 3), which corresponds to the source [6]. Tests of chopped products were performed comprehensively by physicochemical, including rheological, and organoleptic standard methods. Therefore, changes in indicators the moisture-holding capacity (MHC), the moisture releasing capacity (MRC), as well as water activity, have a positive effect on the value of losses during heat treatment and product yield (Table 4) and other consumer properties. Table 3. The influence of the structuring mixture on MHC, MRC and Aw minced meat and fish The amount of the added mixture,% MHC,% MRC, % Moisture, % Water activity (aw) to the mass of minced to total moisture Minced meat 0 (control) 57.1+0.7 83.9+0.7 20.8+0.4 68.1+0.8 0.989+0.001 0.5 59.7+0.7 89.1+0.8 15.4+0.3 67.5+0.8 0.988+0.002 1.0 62.1+0.8 94.2+0.9 11.6+0.3 67.2+0.9 0.987+0.002 1.5 63.0+1.4 97.1+1.5 9.4+0.3 66.5+1.3 0.986+0.003 Minced fish 0 (control) 48.1+0.6 60.4+0.7 16.5+0.2 75.7+0.9 0.994+0.001 0.5 55.5+0.6 69.3+0.7 11.0+0.2 75.1+0.9 0.993+0.002 1.0 65.3+0.7 83.7+0.9 6.3+0.3 74.6+1.1 0.992+0.002 1.5 78.9+1.3 96.6+1.2 3.2+0.2 73.9+1.5 0.991+0.002 3 BIO Web of Conferences 30, 01005 (2021) https://doi.org/10.1051/bioconf/20213001005 ILS 2020

Table 4. The dependence of the yield of the finished product on the amount of structuring mixture The amount of additive, in% by the mass of minced meat The yield of the finished product, g Mass loss with heat treatment, % “Steak” “Schnitzel” “Steak” “Schnitzel” 0 (control) 53.0+0.5 75.4+0.5 29.3+0.5 20.1+0.5 0.5 57.5+0.6 76.5+0.7 23.4+0.6 18.0+0.7 1.0 59.8+0.8 78.8+0,8 20.3+0.7 16.3+0.8 1.5 63.2+1.0 81.7+1.1 15.7+1.0 12.6+1.2 A generalization of the research results made it possible to confirm the concept of the formation of structures based on natural biopolymers, some salts, etc., to give a fairly complete and detailed histological picture of fibrous protein systems and food products, positively assess the physicochemical changes, including architectonics (relative homogeneity of system, cohesion, etc.), therefore, the texture and other organoleptic properties of food products. The performed developments correspond to the modern level of research on biopolymers, their functional and technological features and applications [5, 6], including the production of meat semi-finished products and products, such as hamburgers, cutlets, etc. [7]. As a result of the research, it was found that the mass loss during the frying of the semi-finished product "Chopped Steak" and "Natural Fish Schnitzel" is reduced in proportion to the amount of additive and is reduced by almost 2 times when adding 1.5% of the test mixture. The introduction of a structuring mixture leads to a sharp increase in the moisture-holding capacity (MHC) of minced meat (up to 97% to the total moisture) and the at the same time a decrease in the moisture releasing capacity (MRC) of minced meat by half, and fish by 2,5 times (Table 1). 3 Сonclusion As a result of comprehensive research, it was found that the introduction of biopolymers of various classes, including gelling mixtures, significantly increases the rheological and technological properties of meat systems, in particular, jellies, which optimizes quality. Therefore, in general, there was a tendency to increase their quality. A comparative study of structures using alginates as an example showed the positive dynamics of moisture binding, mass loss and yield; in addition, the modulus of elasticity of the meat is increased, as well as the adhesion to stainless steel and fluoroplastic, which obviously occurs as a result of the pronounced functional and technological features of the calcium alginate gel. For example, the stickiness of minced meat with the addition of 1.5% of the mixture is almost two times higher than that of the control sample; the maximum share stress of the finished product is reduced, i.e. products become more tender and juicy, which correlates and with organoleptic assessment. References 1. E.I. Titov, A.Yu. Sokolov, E.V. Litvinova, S. Kidyaev, D.I. Shishkina, and B.A. Baranov, Foods and Raw Materials. 2, 7(2), 387–395 (2019) 2. A.YU. Sokolov, E.I. Titov, D.I. SHishkina, and E.V. Litvinova, Commodity expert on food products, 2, 63–66. (2019) 3. M.S. Brewer, Meat Science, 91 (4), 385–395 (2012) 4. J.-H. Choe, H.-Y. Kim, J.-M. Lee, et. al. Meat Science, 93 (4), 849–854 (2013) 5. M.C. Gomez-Guillen, B. Gimenez, M.E. LopezCaballero, and M.P. Montero, Review Food Hydrocolloids, 25 (8), 1813–1827 (2011) 6. Rodrigo Tarte, Ingredients in Meat Products. Properties, Functionality and Applications.(Professiyaб Sankt-Peterburg, 2015) 7. E.M. Desmond, D.J. Troy, and D.J. Buckley, Journal of Muscle Foods, 9 (3), 221–241 (1998) 4 BIO Web of Conferences 30, 01005 (2021) https://doi.org/10.1051/bioconf/20213001005 ILS 2020

Promising unconventional plant raw materials for food production Elena Belokurova1* , Anna Derkanosova1 , Yana Dombrovskaya1 , and Tatyana Malyutina1 1Voronezh State University of Engineering Technologies, Voronezh, Russia Abstract. Article contains information about new technologies of flour and bakery products on the basis of multicomponent structure products or the flour composite mixes executed from a flour of pumpkin and a thistle, a dry leaf of a stevia and dry lactulose, and also mixes from different types of grain, such as: corn, buckwheat, rice, millet. 1 Introduction Due to the fact that flour confectionery products are popular among the population, due to their attractive appearance and high taste qualities, for domestic producers the scientific and technical problem is the creation of highly effective technologies, expanding the range of products, the development of original formulas, creating functional products, improving consumer properties and nutritional value, improving their structure. The use of ready-made concentrates with the introduction of biologically active raw materials, products of multicomponent composition, which were named flour composite mixtures (FCM), is one of the priority directions for solving these problems [1]. Powder components such as pumpkin and thistle flour, dry stevia leaf and dry lactulose were considered to create a new generation of products based on FCM. Although additives in a dry form remain in the natural biological environment for a long time, when enriching FCM it is necessary to maintain stability in the quality of the components until the system needs to be converted to an aquatic composition. The FCM under consideration are thermophiles, and pumpkin meal is a rich source of complete and easily digestible vegetable protein, its content reaches 40%. The high biological and nutritional value of pumpkin meal is largely due to its unique mineral composition [1]. 2 Experimental Thistle flour is useful to eat regularly for prevention and as part of the complex treatment of atherosclerosis, varicose veins, coronary heart disease, arterial hypertension, inflammatory heart disease and blood vessels, as it is present in its composition: proteins, mono- and dysaccharides, flavonoids and flavolignans (silybin, silichristine, silidianine, taxifolin, neohydrocarpine, quercetin, etc.), carotenoids, vitamins E, K, D, vitamins of group B, chlorophyll, essential oil (0.08 %), resins, mucus, biogenic amines (histamine, tyramine), enzymes, alkaloids, bitterness, saponins, as well as various macro- and microelements (magnesium, potassium, manganese, calcium, iron, zinc, selenium, chrome, copper, aluminum, boron, vanadium, etc.). It was found that lactulose both separately and in combination with bifidobacteria promotes the absorption of calcium and increases the strength of bones in osteoporosis, it is classified as oligosaccharides. Lactulose is a white crystalline substance, odourless, sweet to the taste and well soluble in water. Stevia leaves contain mineral compounds, organic substances, vitamins A, C, E, P; flavonoids, essential oils, amino acids, pectins, sterebins. This valuable set of chemical compounds helps to rationalize the process of hormone synthesis in the human body, which allows: provide tissue respiration, normalize the work of enzyme systems, reduce cholesterol, restore carbohydrate-proteinlipid metabolism, stabilize blood pressure, stimulate digestion and urinary system [2]. The results of research of products manufactured on the basis of FCM with introduction of biologically active raw materials are presented in Tables 1, 2. The mass fraction of the moisture of the raw materials used is shown in Table 3. Table 1. Comparative assessment of sample viscosity Indicator Control Lactulose content of samples in relation to to the FCM, % Pumpkin seed flour content in samples at to to the FCM, % Contents thistle flours in reference to to the FCM, % 0.8 1.1 13.5 15.0 11.0 13.0 Viscosity, Ра·s 4.5 4.0 3.3 4.5 6.0 3.9 6.8 *Corresponding author: [email protected] © The Authors, published by EDP Sciences. This is an open access article distributed under the terms of the Creative Commons Attribution License 4.0 (http://creativecommons.org/licenses/by/4.0/). BIO Web of Conferences 30, 01006 (2021) https://doi.org/10.1051/bioconf/20213001006 ILS 2020

Table 2. Comparative moisture assessment of samples Indicators Control Lactulose content of samples in relation to to the FCM, % Pumpkin seed flour content of samples in relation to to the FCM, % Thistle flour content of samples in relation to to the FCM, % 0.8 1.1 13.5 15.0 11.0 13.0 Humidity (dough),% 28.7 28.9 36.4 28.0 33.6 27.0 32.6 Humidity (crumb), % 26.1 22.3 29.2 25.4 28.4 25.4 28.7 Table 3. Mass fraction of moisture of applied vegetable raw materials 3 Results and discussion The proposed types of plant raw materials have high antioxidant activity, which will give the products based on them functional properties (Table 4). As a result of the conducted researches technical and technological cards for flour products with the content of dry leaf stevia and biologically active raw materials were developed and introduced into production: TTK on “Lactulose sponge” with the content of dry lactulose 0.8 % to FCM mass; TTK on “Pumpkin sponge” with the content of pumpkin flour 13.5 % to FCM mass; TTK on “Spiky sponge” with the content of 11 % to FCM mass. The next stage of the study was the development of new bakery products, the formulation of which is introduced flour from different cereals to replace part of wheat flour, which allows to balance products on the qualitative and quantitative composition of protein, as well as increase the content of dietary fiber, vitamins and trace elements to the recommended consumption rates. This allows you to manage the chemical composition and biological value of products, while changing their organoleptic characteristics: taste, aroma, appearance, which allows you to expand the range of products. Table 4. Antioxidant activity of the investigated samples № sample Sample name Content of antioxidants, mg/g 1 Biscuit based on FCM 0.008 2 Biscuit to traditional technologies (control) 0.013 3 Stevia leaf 0.110 4 Biscuit with lactulose content, % 0.8 0.038 5 11 0.045 6 Biscuit with pumpkin flour, % 135 0.033 7 15.0 0.043 8 Pumpkin flour 0.026 9 Biscuit containing thistle flour, % 11.0 0.130 10 13.0 0.142 11 Thistle flour 0.191 When assessing the quality, bakery products cooked according to a classical recipe were taken for control; prototypes were prepared according to a similar recipe with the replacement of a part of wheat flour of the highest grade by a selected mixture; the recipe and technological modes of dough testing are given in Table 6. In the process of fermentation of the dough every 30 minutes physical-chemical and structural-mechanical parameters were determined, as a result of a number of studies the best samples were recognized with the introduction of mixtures 2, 4, 5 into the recipe. For these products there was carried out the qualification evaluation of quality, which provides the evaluation of the totality of organoleptic, physicochemical and structural-mechanical parameters of products. The evaluation of product quality levels provides the definition of values of quality indicators of the evaluated products and their comparison with the basic ones [4]. Thus the concept of relative value of the indicator of quality of production qi which is defined by a differential method is used. Its essence consists in separate comparison of unit quality indicators of the considered product with similar basic indicators. For this purpose the relative quality index was defined by the formula: qi = рi / ра (1) where рi, ра – is the value of the i-th quality index for the evaluated and base samples respectively. The comprehensive quality assessment involves the use of comprehensive property set indicators, which must take into account the significance of each property, i.e. assess the extent to which individual property values influence the final quality indicator. Each individual quality index is characterised by two parameters – the relative index and the weighting. When determining the weighting factors, the main criteria are identified, which most fully reflect the ability of the product to fulfil its primary purpose. In this case, the traditional point scales used in the current standards are taken into account [5]. The quality indicators of bakery products, on the basis of which the qualification evaluation was carried out, are given in Table 7. The values of complex quality indicators of the examined samples of bakery products are given in Table 8. From the obtained data, we determined a generalized complex quality indicator, which was: for control – 1.0; sample 2 – 1.2; sample 4 – 1.18; sample 5 – 1.12. The appearance of samples of received bakery products is shown in Fig. 1. Product name Mass fraction of moisture, % Verona Concentrate 5.3 Pumpkin flour 5.5 Thistle flour 4.7 Lactulose 6.8 Stevia leaf (variety in vintro) 7.2 2 BIO Web of Conferences 30, 01006 (2021) https://doi.org/10.1051/bioconf/20213001006 ILS 2020

Table 5. Ratio of components of selected mixtures, % Blend number Ratio of mixture components, % Corn Buckwheat flour Rice flour Wheat flour 1 39 33 20 8 2 40 32 20 8 3 41 31 20 8 4 42 30 20 8 5 47 25 20 8 Table 6. Recipe and modes of dough testing process Name of raw materials, semi-finished goods and process indicators Raw material consumption and process parameters Wheat flour bakery top grade, kg 88-94* Bakery yeast extruded, kg 1.5 Food salt, kg 1.3 Composite blend, kg 6-12** Water, kg by calculation Initial temperature, °C 27.0 Duration fermentation, min 180.0 The acidity is final, hail, no more than 4.0 Note: * – intervals of prescription composition are given depending on the type and dosage of mixtures; ** – batching intervals of mixtures depending on their composition. Table 7. Quality indicators of bakery products Indicator The value of the quality indicator Control Sample 2 Sample 4 Sample 5 Organoleptic Appearance, score 4 5 5 4 Form, score 4 5 5 4 Crust color, score 4 5 5 5 Bakery, point 5 5 5 5 Promise, point 5 5 5 5 Taste, score 3 5 5 4 Smell, score 3 5 5 4 Physico-chemical Humidity, % 42,8 42,2 42,5 44,2 Acidity, hail 3,0 3,2 3,2 3,0 Porosity, % 73,0 76,0 74,0 72,0 Specific volume cm3 /100 g 324,0 350,0 342,0 330,0 Structural and mechanical 60,0 65,5 65,0 60,0 General deformation of crumb compression, etc 60,0 65,5 65,0 60,0 Table 8. Complex indicators of product quality by studied properties Sample product Value of complex quality parameters Organoleptic Physicochemical structuralmechanical Control 1.00 1.00 1.00 Sample 2 1.31 1.20 1.09 Sample 4 1.28 1.19 1.08 Sample 5 1.23 1.12 1.00 Sample 2 Sample 4 Sample 5 Fig. 1. Samples of bakery products 4 Сonclusion Thus, the use of such valuable plant raw materials as pumpkin and thistle flour, dry stevia leaf, dry lactulose, as well as flour from mixtures of corn, buckwheat, rice, millet in the formulation composition of flour confectionery and bakery products will expand the range of confectionery and bakery products, enrich them with useful functional ingredients, give functional orientation. Products on the proposed technology will be in demand among the population, as they have high taste qualities, improved nutritional value. These products can be recommended for feeding different groups of the population, including children of primary and school age. References 1. E.V. Belokurova and A.A. Derkanosova, Bulletin of VSUET, 2, 119–124 (2013) 2. Ya.P. Kolomnikova, A.A. Derkanosova, and E.V. Litvinova, Economics. Innovation Quality control, 2 (11), 139–143 (2015) 3. E.V. Belokurova, T.N. Malyutina, and N.M. Derkanosova, “Influence of the hop extract on the infecting microflora of rye starches”. Collection of Articles of the VIII International Scientific and Technical Conference dedicated to the 90th anniversary of the Faculty of Technology VGUIT, 2019, Voronezh. 4. E.V. Belokurova, S.A. Solokhin, and A.A. Rodionov, Technologies of food and processing industry of agro-industrial complex - Healthy food products, 3 (11), 51–55, (2016) 5. S.V. Belokurov, N.S. Rodionova, E.V Belokurova., and T.V. Alexeeva, Journal of Physics: Conference Series (see books), 1015, 1-4 (2018) 3 BIO Web of Conferences 30, 01006 (2021) https://doi.org/10.1051/bioconf/20213001006 ILS 2020

Comparative characteristics of white and black ceylon tea based on morphological features of the leaf Natalya Durnova1* , Margarita Simakova1 , Daniil Isaev1 , Alexander Simakov2 ,and Inna Simakova2 1Saratov State Medical University named after V.I. Razumovsky, Saratov, Russia 2Saratov State Agrarian University named after N.I. Vavilov, Saratov Russia Abstract. The central question of the research was addressed to comparative characteristics of white and black Ceylon tea based on morphological features of the leaf. The relevance of the study is due significant increasing falsifications of black and white tea on markets in Russia and all around the world. It should be noted that tea is still incomplete and insufficiently studied crop, despite its centuries-old use in human nutrition. Thus, the purpose of this work was to study the microstructure of black and white tea to identify differences in morphological characteristics of the leaf and develop markers for identifying its variety. The objects of study of this work were white and black varieties OP1 teas of Nandana Tea Factory (Sri Lanka). In our research were used research methods includes organoleptic and microscopical methods. According to the results micro- and macroscopic studies of black and white tea samples showed that there are significant morphological differences in the structure of leaves of different varieties, such as the size and density of hairs, the density of stomata, the location of druses and sclereids, and outgrowths of the leaf edge. These signs can serve as reliable markers for identifying the grade of tea. 1 Introduction Tea is the oldest and one of the most common drinks in the world, the use of which is related to authentic cuisine and traditions of many nations. For various reasons, many nations consider tea not only as a drink, but also as a first necessity product, a strategic component of the country's food security basket. In accordance with this, tea consumption should be based on modern scientific ideas about the product, knowledge about its benefits, confidence in its quality and safety. It should be noted that tea is still incomplete and insufficiently studied crop, despite its centuries-old use in human nutrition. And it has been known for more than 5,000 years. For example, today there are contradictions in scientific researches related to the chemical composition of various types of tea and its impact on the human body. Some scientists have pointed out that highquality Chinese green teas have the most valuable mineral composition and the highest protein content in comparison with black and white teas [1]. Results of scientist’s works Unachukwu UJ, Ahmed S from Lehman College, Bronx, NY, USA shown that the polyphenols content in green teas is similar to some varieties of white tea; some white teas have comparable amounts of common catechins with some green teas, but less antioxidant capacity. It is assumed to be connected with the less non-catechin antioxidants content than in green teas. [2] While in the research of Santana-Rios G, Orner GA the powerful antimutagenic activity of white tea in comparison with green tea has been revealed. Nine major components found in green tea have also been found in white tea, including epigallocatechin-3-gallate (EGCG) and some other polyphenols. [3] The inconsistency in the conclusions can be probably explained both by a discrepancy in the quality of the studied types of tea to their classification, and by other reasons that affect the chemical composition of the final product and its impact on the human body. In the opinion of the authors of this work, in order to remove contradictions and increase the share of truth, you need to be sure that the tea to be studied belongs to the variety indicated on the packaging in accordance with the international classification of tea varieties. One of the simplest and most accessible tools as markers of authenticity and compliance of tea indicated on the label variety, in addition to the generally accepted organoleptic methods, can be the morphological characteristics of the tea leaf. Unfortunately, there are not enough works in scientific periodicals devoted to the study of morphological features of tea raw materials that identify its grade. There are works devoted to leaf morphology as an identifier of the most important hereditary traits that determine the essential difference between Thea sinensis and Thea assamica varieties. For example, in the book of W. Weixin «Encyclopedia of healing tea» [4] is indicated that when studying the tissues and cells of the tea leaf, it has been found that on the upper and lower sides it consists of single-layer cells of the epidermis, under which, depending on the type of tea bush, there are one, two or three layers of palisade tissue, and under them there are cells of the spongy parenchyma. On the lower surface of the leaf there are stomata and hairs. Most of the hairs are found on young leaves and stems. Scientists Xiaofang Zhu, Yi Zhang *Corresponding author: [email protected] © The Authors, published by EDP Sciences. This is an open access article distributed under the terms of the Creative Commons Attribution License 4.0 (http://creativecommons.org/licenses/by/4.0/). BIO Web of Conferences 30, 01007 (2021) https://doi.org/10.1051/bioconf/20213001007 ILS 2020

found that the quality of the aroma of black tea and oolong tea depends on the cuticle wax. However, there is still no comprehensive documentation on the cuticle of the tea leaf [5]. Thus, the purpose of this work was to study the microstructure of black and white tea to identify differences in morphological characteristics of the leaf and develop markers for identifying its variety. 2 Experimental Objects of study. The objects of study of this work were white and black teas of Nandana Tea Factory (Sri Lanka). The share of Ceylon tea in the world production is about 9-10%. In Sri Lanka, different types of tea are produced, only the Assamese variety of the tea plant is cultivated. The rationale of the research objects is the varying degree of their fermentation during processing and the growth phase of shoots during harvesting. White tea varieties are non-fermented (oxidation up to 12 % of the total amount of tanning substances of the raw material), black tea is fermented – oxidation within the limit of 35- 45 % of the total content of tanning substances. Black tea varieties OP1 in accordance with the international classification. Orange Pekoe, category 1 – a variety of high-quality large-leaf tea with an infusion of medium strength. The variety is made from the first and second flushes of fresh leaves which grow on the highest point of the tea tree. White tea is a drink of elite quality. The composition contains tips and the first upper leaf. It should be noted that white and black teas belong to baihao teas. Black tea is dominant on the world market, especially prevalent in temperate, cool, humid (wet) climate countries. To get such a tea, they should undergo the leaf three stages of processing: withering, twisting and fermentation. White tea is a very rare, exceptional in quality tea, consisting entirely of tips and first leaves. Its almost colorless infusion combines the advantages of all types of tea: it has a very delicate and light aroma, rich and strong (especially when re-brewing) taste, has a wide range of healing properties. All five types of tea existing in the world (black, green, yellow, red, white) in all the variety of its types can be made from leaves of the same bush, what indicates how important is the quality of raw materials and method of its processing. Research methods. The studies were carried out on the basis of the laboratories of the department "Food Technology" of the Federal State Budgetary Educational University of Higher Education “Saratov State Agrarian University named after N.I. Vavilov”, and the Department of General Biology, Pharmacognosy and Botany of the Federal State Budgetary Educational Institution of Higher Education “V.I. Razumovsky Saratov State Medical University of the Ministry of Healthcare of the Russian Federation”. The study of raw materials was carried out in accordance with the requirements of GF XIV OFS 1.5.1.0003.15 “Leaves” and OFS 1.5.3.0003.15 “Technique for microscopic and microchemical studies of medicinal plant materials and herbal medicines”. For the analysis of whole leaves, whole leaves or pieces of a leaf plate with an edge and vein, pieces of a leaf from the base and apex, pieces of a petiole (if the leaf has a petiole) were taken. The raw material was enlightened in the following way: a few slices of the raw materials were placed in a flask or vial, then the sodium hydroxide 5 % solution, diluted with water (1:1) was added and the raw material was boiled for 2 − 5 minutes depending on the thickness and density of objects, avoiding strong softening. Then the contents were poured into the glass, the liquid was drained through 2 to 4 layers of gauze, which closed the glass, and the raw material is washed thoroughly with water, each time pouring water through the same gauze. The beaker content was transferred to a small amount of water in a Petry dish. Particles of the raw material, remaining on the gauze, were washed in the same Petry dish. From the water the pieces were removed with a scalpel or spatula and placed on a glass slide in a drop of the glycerin 33 % solution. Pieces of raw, enlightened and placed on a glass slide, were separated with a scalpel or dissecting needle into two parts, one of them gently turned. A piece of stem was placed on a glass slide. Thin stalks were crushed with a scalpel or reverse end of a dissecting needle to release the epidermis. From the thick stalks the epidermis was removed with the help of dissecting needles or razor, the rough inner parts of the petiole which prevent getting a good micropreparation of the epidermis were also removed. The object was covered with a cover glass, if necessary, lightly fell on the net reverse end of the dissecting needle and gently warmed to remove air bubbles, after cooling, was considered the sheet on both sides and the epidermis of the petiole under a microscope, first at low then at high magnification. At different magnifications, using the macro sheet and screws, the upper and lower epidermis and the deep structure of the sheet located beneath the epidermal (inclusion, vessels, etc.) were explored. Preparation of cross sections of petioles: slices were made from soaked petioles by clamping pieces of petiole into a bottle stopper (cork plug) or elderberry core. A piece of elderberry or bottle cork was cut in half and a piece of leaf was clamped between the two halves. For the preparation of transverse slices, the surface of the piece should be prepared so that it is strictly perpendicular to the axis of the petiole or vein of the sheet. Prepared sections were placed in a Petry dish with water, from where the sections were removed and viewed under a microscope. Organoleptic evaluation of white and black tea was carried out in accordance with GOST 32573-2013 Black tea. Specifications (Edition as amended) and GOST 33481-2015 Tea partially fermented. Technical conditions. 3 Results and discussion At the first stage of work, an organoleptic evaluation of tea raw materials was carried out. 2 BIO Web of Conferences 30, 01007 (2021) https://doi.org/10.1051/bioconf/20213001007 ILS 2020

A photograph of black tea is shown in Fig. 1. Leaves are whole and well twisted, homogeneous. Kidneys do not occur. The sizes of the leaves are from 0.5 to 2.0 cm in length. Sometimes there are pieces of the stem, they are more straight, stiff, black, about 1.0-1.5 cm in length. Tea infusion is homogeneous, brown-red. It has a wonderful aroma by brewing and tart flavour. In appearance, black tea, grade OP1, meets the requirements of GOST 32573-2013 and the international classification of tea varieties. The dried raw materials of white tea (Fig. 2) are solid leaves of silver-green colour with a petiole, with copious pubescence on the underside of the leaf. Fig. 1. Photo of raw black tea. Fig. 2. Photo of raw white tea. A leaf blade from the edges is turned inward through the ventral side. The sheet’s size is 1.2-3.0 cm in length and up to 0.5 cm in width has a crescent-shaped bend to the dorsal side. The main vein is visible with the naked eye on the underside of the leaf. Fragmented parts of leaves are occasionally found. Inside some leaves, tips are found that are morphologically similar to the rest of the leaves, but smaller and folded into a tube. Tea infusion is clear, taste and smell are floral, delicate, well perceptible. Organoleptic characteristics of white tea brand Nandana tea factory (Sri Lanka) corresponds to GOST 33481-2015 Tea is partially fermented. Specifications and international classification of tea varieties and refers to white types of tea. After brewing the dried raw black tea (Fig. 3), the leaves unfold, parts of leaf blades and central veins become well distinguishable. Unfolded raw materials are usually pieces of stems, less often the central vein without a leaf blade. The leaf blade is heterogeneous in color, from dark to light green. Dimensions usually do not exceed 11 cm. The edge of the leaf remaining on some parts of the leaf blades is smooth, slightly serrated. The top of the leaf is sharp; the base is wedge-shape. The leaf blade is thin. The raw material of white tea does not change shape after brewing (Fig. 4), but the color of the leaves changes from silver-green to dark green, almost black. Tips are clearly visible. The leaf is quite thick, brittle, when trying to take a sample for a micropreparation, the upper and lower epidermis peeled off easily, and the sheet itself broke. Fig. 3. Raw black tea after brewing. Fig. 4. Raw white tea after brewing. Microphotographs of tea preparations are shown in Fig. 5 and 6. Fig. 5 shows a micrograph of a leaf of black tea. One can see a large number of stomata per surface unit, rare hairs, clusters of druses, small veins that are not in focus, can be also seen. Fig. 6 shows a micrograph of white tea, where a large number of hairs and outgrowths of the leaf blade, which are not visible to the naked eye, can be seen at a glance. On the lower epidermis of white tea (Fig. 7-8), such a high density of hairs is observed that they completely overlap the overview of the underlying structures. The places of attachment of hairs to the epidermis are visible. The epidermal cells with some cellular structures and the attachment sites of hairs are clearly visible. It can be 3 BIO Web of Conferences 30, 01007 (2021) https://doi.org/10.1051/bioconf/20213001007 ILS 2020

seen that between adjacent hairs from 1 to 3 epidermal cells are located. Fig. 5. A leaf blade of black tea. Magnification 1010. Fig. 6. Leaf plate of white tea. Magnification 104. Black tea hairs are rarer and shorter than white tea hairs (Fig. 9). Hairs are unicellular. Figures 10–11 clearly show the stomata as well as the cuticle folds. Presumably these are stomata of paracytic type. The guard cells and their thickened inner wall are visible. Sometimes stomata are found whose size exceeds the size of the surrounding stomata. Stomata on white tea (Fig. 12) are extremely difficult to find and examine because they are rare, and hairs hinder the review. Only two guard cells are poorly distinguishable. Fig. 7. Hairs of white tea. Magnification 1010. Fig. 8. Hairs of white tea. Magnification 1040. Fig. 9. Hairs of black tea. Magnification 1010. In conducting bundles of black tea (Fig. 13-14), spiral thickenings of the vessel walls are clearly visible. Thicker vessels are located in the center of the conducting bundle; to the periphery, the diameter of the vessels decreases. In some areas, the conducting bundles are surrounded by several (2-3) even rows of druses. In conducting bundles of white tea (Fig. 15), spiral thickenings of the vessel wall are clearly visible. Thicker vessels are located in the center of the conducting bundle; to the periphery, the diameter of the vessels decreases. Fig. 10. Stomata of black tea. Magnification 1040. 4 BIO Web of Conferences 30, 01007 (2021) https://doi.org/10.1051/bioconf/20213001007 ILS 2020

Fig. 11. Stomata of white tea. Magnification 1040. Fig. 12 Stomata of white tea. Magnification 1040. Reduced structures representing the tops of the serrated edges of the leaf are visible in some areas of the leaf blade of black tea (Fig. 16-17). It can be seen that, as in white tea, in black tea, the conducting bundles are approaching to these outgrowths. The outgrowths apparently do not play a significant role at this stage of the leaf development. Fig. 17 shows a dead, partially damaged, lignified, but preserved almost entirely on its original place outgrowth. Along the entire edge of the white tea leaf in the middle of the leaf blade, such outgrowths with vessels, which are approaching to them, are observed (Fig. 18– 19). Morphologically, these outgrowths are similar to the top of a leaf plate (Fig. 20). It is visibly that the outgrowths of the leaf edge can be heterogeneous in size and shape, but have the same general structure. Fig. 13. Conducting bundles of black tea. Magnification 1040. Fig. 14. Conducting bundles of black tea. Magnification 10100. Fig. 15. Conducting bundles of white tea. Magnification 1040. Fig. 16. Outgrowths of the leaf edge of black tea. Magnification 1040. Fig. 17. Outgrowths of the leaf edge of black tea. Magnification 1040. 5 BIO Web of Conferences 30, 01007 (2021) https://doi.org/10.1051/bioconf/20213001007 ILS 2020

Fig. 18. Outgrowths of the edge of a leaf of white tea. Magnification 1010. Fig. 19. Outgrowths of the edge of a leaf of white tea. Magnification 10100. Fig. 20. Outgrowths of the edge of a leaf of white tea. Magnification 1010. Astrosclereids in the leaves of black tea (Fig. 21) are very common anywhere in the leaf blade and have the most diverse form. Druses in the leaves of black tea are found everywhere. Often they are concentrated in even rows along the periphery of the conducting bundles (Fig. 14). In a section of the central vein near the base of a leaf of black tea (Fig. 22), two very large, almost rectangular, sclereids are located at both ends of the conducting bundle on the border with the leaf blade. Sclereids in white tea leaves (Fig. 23) are found near the top of the leaf; they were not found in the rest of the leaf blade. The edge of the leaf near the apex is whole. Druses are found near the apex and are concentrated at the edge of the leaf. Druses, like sclereids, were not found in other parts of the white tea leaf. The upper epidermis of the leaf of black tea is represented by highly sinuous cells (Fig. 24). Fig. 21. Sclereids in a leaf plate of black tea. Magnification 1010. Fig. 22. Large sclereid on a slice of black tea leaf. Magnification 104. Fig. 23. Druses and sclereids of a leaf of white tea. Magnification 104. Fig. 24. The upper epidermis of a leaf of black tea. Magnification 1040. 6 BIO Web of Conferences 30, 01007 (2021) https://doi.org/10.1051/bioconf/20213001007 ILS 2020

4 Сonclusion According to the results of a macroscopic research, the raw material of white tea is a flush of 1-2 leaves and a leaf bud of silver-green color, curled up, not too dry, of medium size. Black tea – black, dry, angular, wrinkled parts of the leaf blade and pieces of the stem. Microscopic examination of white tea showed that the density of hairs on its leaves is extremely high, the hairs are unicellular, long. Stomata are rare; they are difficult to detect and examine. Root buds of denticles are found along the entire edge of the leaf. Sclereids and druses are found only at the top of the leaf blade. The density of hairs on the leaves of black tea is much lower, and the hairs themselves are shorter. The density of stomata, on the contrary, is much higher. Around the conductive beams are several even rows of druses. The ends of the cloves on the edge of the leaf of black tea are lignified and fall. Sclereids and druses are distributed throughout the leaf blade. Thus, micro- and macroscopic studies of black and white tea samples showed that there are significant morphological differences in the structure of leaves of different varieties, such as the size and density of hairs, the density of stomata, the location of druses and sclereids, and outgrowths of the leaf edge. These signs can serve as reliable markers for identifying the grade of tea. Acknowledgment The authors thank Nandana Tea Factory and personally Mr. Gunasoma Wanigasekara for providing tea samples for research. References 1. M. Czernicka, G. Zaguła, M. Bajcar, B. Saletnik, and C. Puchalski, Rocz Panstw Zakl Hig., 68(3), 237–245 (2017) 2. U.J. Unachukwu, S. Ahmed, A. Kavalier, J.T. Lyles, and E.J. Kennelly, J.Food Sci., 75(6), 541–548, (2010) 3. G. Santana-Rios and G.A. Orner, “Potent antimutagenic activity of white tea in comparison with green tea in the Salmonella assay”, 495(1-2), 61–74, (2001) 4. Electronic document: access mode https://med.wikireading.ru/76027 5. X. Zhu, Y. Zhang, Z. Du, X. Chen, X. Zhou, X. Kong, W. Sun, Z. Chen, C. Chen, and M. Chen, Reports, 8(1), 14944 (2018) 7 BIO Web of Conferences 30, 01007 (2021) https://doi.org/10.1051/bioconf/20213001007 ILS 2020

Chemical composition and nutritional value of the new citrus pomace feed additive Oleksii Karunsky1 , Іgor Nikolenko1* , Maria Madani2 ,and Oleksii Danchuk1 1Odessa State Agrarian University, Odessa, Ukraine 2Odessa National Academy of Food Technologies, Odessa, Ukraine Abstract. Research aimed at studying the chemical composition and nutritional value of a new feed additive from citrus pomace was conducted. The article presents the results of research on the technological line of granulation of citrus waste, studies their chemical composition, determines the gross content of proteins, fats, assimilated carbohydrates, including simple sugars and identifies the effectiveness of use in the diets of farm animals. The positive influence of the technological line of citrus waste granulation has been established, which allows to obtain high quality granules. Chemical analysis of citrus granules showed that 100 g contains: proteins - 6.15 g, fats - 0.99 g, carbohydrates - 21.8 g, of which simple sugars 14 g, the energy value of this feed additive is Kcal (kJ) - 120.7 (505.7). We believe that the data on the qualitative characteristics and nutritional value of feed additives from citrus pomace suggest that this additive can be used in cattle feeding. 1 Introduction Domestic and world experience of increasing the production of livestock products and reducing their cost shows that in recent years an increase in animal productivity by 65% has been achieved by improving their feeding system and advanced housing technologies. It follows that the organization of rational full-fledged feeding of farm animals is one of the main conditions for further increase of their productivity [3, 5, 9]. As the knowledge of the animal's need for nutrients increases, so does the concept of the level of a complete diet. Detailed norms and rations of feeding farm animals reflect modern knowledge, on the basis of which a scientifically sound and well-founded feeding system has been developed [4]. Full supply of animals with all nutrients, minerals and biologically active substances, determines not only the level of productivity, but also the amount of feed costs for production [5]. With the transfer of animal husbandry to an industrial basis, it is important to develop a system of complete feeding of animals, which ensures their high productivity. It is established that the animal body needs the optimal amount of nutrients and biological substances [4]. Every year the science of feeding develops tools for the use of various biological and chemical feed additives. High productivity of animals, support of reproductive functions of an organism, effective use of forages is impossible without inclusion in a diet of forage additives which provide necessary level of biologically complete feeding. They are a mandatory ingredient in any diet for farm animals [6,7]. Data on the biological role of individual feed additives, their interaction with other nutrients, dosage taking into account the species and age of animals, methods of administration, the impact on productivity and product quality are necessary for qualified use [8]. The use of supplements also helps to save feed, as a scientifically sound combination of all nutrients and biologically active substances in the diet provides the fullest digestion and assimilation by the body, compared with the components of the diet, which are used separately, fed separately. The production of feed additives allows to increase the high-energy properties of the diet, to effectively use the ingredients of the diet. The possibility of preparing feed additives in the form of granules avoids self-sorting of components and overdose of micronutrients and improve the consumption of feed ingredients. In this regard, it is important to study the chemical composition and nutritional value of citrus pomace and the effectiveness of their use for feed purposes in feeding farm animals. The main raw materials for the production of new feed additives are citrus pomace. The use of new feed additives in the diets of farm animals is possible only after studying their effectiveness in feeding animals, the impact on health and productivity and their registration in Ukraine. The purpose of our work was to develop a scheme of the technological line of granulation of citrus waste, to establish their chemical composition. To achieve this goal, the task of our research was: *Corresponding author: [email protected] © The Authors, published by EDP Sciences. This is an open access article distributed under the terms of the Creative Commons Attribution License 4.0 (http://creativecommons.org/licenses/by/4.0/). BIO Web of Conferences 30, 01008 (2021) https://doi.org/10.1051/bioconf/20213001008 ILS 2020

- determination of the gross content of proteins, fats, assimilated carbohydrates, BAS in particular simple sugars; - identification of the effectiveness of use in the diets of farm animals. 2 Materials and methods The experimental part of the work on the development of the technological line of granulation of citrus waste was performed on the basis of LLC "LAMPOCHKA" of the city of Chernomorsk, Odessa region. Laboratory studies to study the chemical composition of citrus granules were conducted in the Research and Service Company "OTAVA" in 2019. The fat content in the sample №1157 / 3 was determined according to the method of GOST 32905- 2014 (ISO 6492: 1999) "Feed, compound feedstuff, feedstock. Method for determination of crude fat content", by extracting crude fat from the product with a solvent, subsequent removal of the solvent, drying and weighing the extracted fat [9, 10]. The protein content in sample 1157/3 was determined according to the method of GOST 13496.4-93 " Feed, compound feedstuff, feedstock. Methods for determining the content of nitrogen and crude protein" by measuring the nitrogen content and converting the result to the protein content. The total content of digestible carbohydrates was defined as the sum of simple sugars and carbohydrates that are easily hydrolyzed ("inverted"). Prior to hydrolysis, the content of simple sugars is determined, followed by the determination of the total invert sugar, which consists of the original simple sugars contained in the sample and the simple sugars formed by hydrolysis. The content of simple sugars in sample 1157/3 was determined according to the method of GOST 5903-89 "Confectionery". Methods for determination of sugar by measuring the amount of monovalent copper oxide formed by the iodometric method due to the reaction with reducing substances (simple sugars). Accordingly, the gross content of digestible carbohydrates was determined in a similar manner, after acid hydrolysis of dissolved poly- and oligosaccharides to glucose. BAS, mineral and vitamin composition of citrus pomace was determined by generally accepted methods [9, 10]. Methods for determining the above indicators involve working with crushed dry sample. The sample was pre-dried to constant weight at 65°C and ground to a powder. To convert the measurement results to the native state of the sample, the content of volatile substances and water in it was determined. The sample was dried on an automatic moisture meter to a constant weight at a temperature of 65oC. The moisture and volatile matter content was 2,55 ± 0,35% by weight of the sample. 3 Results and discussion Taking into account the useful properties of citrus pomace, a method of processing citrus pomace into feed additives was developed. When citrus was extruded, up to 55-60% of moisture evaporated from the extrudate, which ensured the moisture content in the feed additive was not more than 12-13%, which allows to store the feed additive for a long time. The humidity of the mixture before extrusion was not more than 17-19%. The introduction into the mixture of 20% sunflower meal with a moisture content of 10%, 3- 7% molasses with a moisture content of 20%, citrus extrudate 75% with a moisture content of 15%, made it possible to obtain a mixture with a moisture content of 15%. Given the results of studies of citrus feed additives, the most effective is the inclusion in the feed additive: sunflower meal - 20%, molasses - 5% and 75% of citrus fruits. Citrus pomace is separately prepared for processing, crushed on a grater to a particle size of 2-3 mm, homogenized and dosed, after which sunflower meal, molasses is mixed in a mincer to form a homogeneous mixture. The granular process is the compression of a homogeneous mass by pressing rollers and the surface of the matrix with round holes. Compression takes place under the action of moisture, heat and pressure, at this time the homogeneous mixture acquires thermoplastic properties and through the holes of the matrix takes the form of granules. The temperature of the obtained granules was + 60- 80oС, after that they are sent to the cooler, where they are cooled to a temperature of 18-20 oС. The humidity of the cooled granules did not exceed 6-7%. The floury fraction is obtained after sieving the granules, sent for further granulation. A sample of cylindrical granules with a diameter of 5-6 mm was obtained in Fig. 1. Fig. 1. Photo of the obtained sample. To use the obtained citrus granules for feed purposes, we conducted a study of chemical composition and nutrition. The results of determining the content of proteins, fats and carbohydrates are listed on the native state of the sample №1157/3 and the result of calculating the energy value per 100 g of product are shown in table 1. Chemical analysis of citrus granules showed that 100 g contains: protein - 6.15 g, fat - 0.99 g, carbohydrates - 2 BIO Web of Conferences 30, 01008 (2021) https://doi.org/10.1051/bioconf/20213001008 ILS 2020

21.8 g, of which simple sugars 14 g, the energy value of this feed additive is Kcal (kJ) - 120.7 (505.7). The mineral and vitamin composition of citrus per 100 grams of product are shown in table 2. Scientific and economic experiment to study the effectiveness of the use of citrus granules in the diets of dairy cows was carried out for 105 days by the group method according to the scheme shown in table 3. Table 1. Chemical composition and energy value of the sample №1157 / 3 per 100 g of product (for the native state of the sample) Indicators, units of measurement Value (per 100 g) Protein, g 6,15±0,03 Fat, g 0 The content of digestible carbohydrates ** (of them simple sugars), g 21,8±0,04 (14,0±0,1) Energy value, kcal (kJ) 120,7 (505,7) ** - the sum of simple sugars and hydrolyzed (digestible) carbohydrates. Feeding of cows was double, carried out individually, taking into account the specified feed and their residues. The total nutritional value of the rations was determined by detailed norms, the milk yield of cows was carried out individually once every 10 days. The average daily milk yield for the study was taken twice in the transition period and in the main and in the final at the end of each period in proportion to the amount of milk. Food consumption during the experiment was quite high. The composition and nutritional value of the average daily rations of experimental animals are shown in table 4. The data in table 3 indicate that when using a feed additive from citrus pomace, the nutritional value of the diet is improved by the sugar - protein ratio (0,62:1 against 0,8:1,2). Table 2. Mineral and vitamin composition of citrus sample №1157/3 per 100 grams of product Vitamins Macronutrients Trace elements А – 0,05 mg K – 197,0 mg Fe – 0,3 mg В1 – 0,04 mg Ca – 34,0 mg Na – 13,0 mg В2 – 0,03 mg Mg – 13,0 mg Cl – 3,0 mg В3 – 0,3 mg P – 23,0 mg B – 180 mcg В6 – 0,006 mg S – 9,0 mg J – 2,0 mcg В9 – 5,0 mcg - Co – 1,0 mcg С – 60,0 mg - Mn – 30,0 mcg Е – 0,2 mg - Cu – 67,0 mcg Н – 1,0 mcg - F– 17,0 mcg РР – 1,0 mcg - Zn– 200,0 mcg Dairy productivity of cows is shown in table 5. As can be seen from table 5, the average daily milk yield was higher in cows of the research group by 1.8 kg or 15.5%. Therefore, the effectiveness of feeding dairy cows with a feed additive from citrus feed for 105 days of scientific and economic experiment had a positive effect on milk productivity. The cost of feed for milk production of 4% fat was almost the same 0.97-0.98 feed units. Table 3. The scheme of the experiment Group Number of animal heads Equalization period (15 days) Main period (60 days) Final period of 30 days) Control 10 the basic ration the basic ration the basic ration Experimental 10 the basic ration the basic ration + 1,5 kg citrus pomace the basic ration Table 4. The composition and nutritional value of the average daily rations of experimental animals FEED GROUPS Control Experimental corn silage, kg 30 30 wheat straw, kg 2 2 sainfoin hay, kg 3 3 compound feed, kg 3 2 citrus supplement, kg - 1.5 salt, g 100 100 The ration contains: feed units, kg 16,5 16,2 digestible protein, g 1250 1185 dry substances, kg 17,2 17,8 Ca, g 152,4 148,3 F, g 78,4 81,2 fiber, % 16,4 16,7 Sugar-protein ratio 0,62- 1 0,8-1,2 Table 5. Dairy productivity of cows Indexes Groups 1 Control 2 Experimental Average daily hopes, kg 11,6 13,4 The fat content in milk,% 3,8 3,8 The protein content in milk,% 3,64 3,61 Feed costs per 1 kg of milk 4% fat, feed. units 0,98 0,97 4 Сonclusion 1. Studies indicate a positive impact of the technological line of granulation of citrus waste, which allows to obtain high quality granules. 100 g contains: proteins - 6.15 g, fats - 0.99 g, carbohydrates - 21.8 g, of which simple sugars 14 g, the energy value of this feed additive is Kcal (kJ) - 120.7 (505.7). 2. When using a feed additive from citrus pomace, the nutritional value of the diet is improved by sugar - protein ratio (0.62: 1 vs. 0.8: 1.2). 3. Introduction to the diet of dairy cows feed additives from citrus pomace during stable lactation increases the average daily hope by 1.8 kg or 15.5%, feed costs for 3 BIO Web of Conferences 30, 01008 (2021) https://doi.org/10.1051/bioconf/20213001008 ILS 2020

milk production of 4% fat was almost the same 0.97-0.98 feed units. References 1. V.M. Kandyba, I.I. Ibatul, V.I. Kostenka, Theory and practice of normalized feeding of cattle, (PP Ruta, Zhytomir, 2012) 2. A. Kotlyar, Pig breeding, 4, 20-23 (1994) 3. V.Yu. Chumachenko, S.V. Stoyanovskyy, P.Z. Lahodyuk [et al.], Guide to the use of biologically active substances in animal husbandry ( Harvest, Kyiv, 1989) 4. V.Y. Hnoevoy, A.K. Tryshen, Y.V. Hnoyevoy, Biomorphological organization and nutritional value of feed ( Kharkiv: FLP Borovyn A.V., (2017) 5. V.V. Pan’ko, The use of a complex and feed additive from non-traditional raw materials in the diets of young pigs ( Dissertation, Simferopol, 1995) 6. H. Humenyuk, Feed industry, 3, 28-29, (1997) 7. B.V. Yehorov, O.I. Shapovalenko, A,V, Makaryns’ka, Premix production technology. Handbook (Center for Educational Literature, Kyiv, 2007) 8. F.Y. Svezhentsov, V.N. Korobko, Unconventional feed additives ( Art-Press, Dnepropetrovsk 2004) 9. V.K. Kononenko, I.I. Ibatulin, V.S. Patrov, Workshop on the basics of scientific research in animal husbandry ( Higher Education, Kyiv 2003). 10. V.M. Sheyko, N.M. Kushnarenko, Organization and methods of research: Textbook ( Knowledge, Kyiv, 2004) 4 BIO Web of Conferences 30, 01008 (2021) https://doi.org/10.1051/bioconf/20213001008 ILS 2020

Recipe modeling and the study of the chemical composition of functional fish culinary products Olga Kutina1 ,and Irina Korotkyh2* 1All-Russian Research Institute, Fisheries and Oceanography, Moscow, Russia 2 Belgorod State National Research University, Belgorod, Russia Abstract. The biochemical composition of the new fish culinary product is considered. The results make it possible to attribute the product to functional products. Compositions with a given set of nutritional value indicators were simulated by optimizing multicomponent formulations. Products are characterized by preset properties: functionality, balance of amino acid, mineral and vitamin compositions, high organoleptic characteristics due to the inclusion of original plant materials (spinach, green peas, cauliflower, eggplant, etc.) and individual methods for their preliminary preparation. The technology and the recipe are protected by a patent [1], technical standards [2], have been tested in production conditions. Product samples participated in the nomination “Innovative Product” at the international exhibition of food products, beverages and raw materials for their production PRODEXPO in 2015-2018. Development “Fish loaf” received a silver medal in 2015. 1 Introduction “Fundamentals of state power of the Russian Federation for the period up to 2020” [3], “Fundamentals of state power of the Russian Federation in the field of healthy nutrition for the period up to 2020” [4], “Doctrine of food security of the Russian Federation” [5], “Strategy improving the quality of food products in the Russian Federation until 2030" (approved by Order of the Government of the Russian Federation of June 29, 2016 No. 1364-p) [7] and the Action Plan for the implementation of the Strategy for improving the quality of food in the Russian Federation until 2030 (approved by Order of the Government of the Russian Federation of April 19, 2017 No. 738 -p) [8]. All of these documents address issues related to the creation of conditions that ensure the satisfaction of the needs of various population groups for healthy nutrition in accordance with the requirements of medical science, as well as taking into account established traditions, habits and economic conditions. Raw fish is the most important component of a healthy diet due to the uniqueness of the source of raw materials: - balanced amino acid composition; - the presence of essential polyunsaturated fatty acids of the omega-3 family. But the development of products based on fish with desired properties is difficult, because the variability of the chemical composition of fish is approximately 40%, which is a fairly high indicator. In order to expand the range of fish products, for the systematic use in the composition of food rations by all age groups of a healthy population, taking into account the rational use of fish raw materials and scientifically based nutritional standards, innovative products have been developed. The chemical composition of this product, justified from the point of view of evidencebased medicine, allows us to attribute it to a functional product. Modeling of compositions with a given set of nutritional value indicators was carried out by optimizing multicomponent formulations. 2 Experimental The following research methods were used in the analysis of the developed products: - for laboratory research of fish products, sampling and preparation of samples was carried out according to a single technique in accordance with the requirements of GOST 7636-85 [9], GOST 31339-2006 [10]; - determination of protein content - Kjeldahl method according to GOST 25011-2017 [11]; - the amino acid composition of the proteins was determined on an AAA-835 amino acid analyzer ("Hitachi", Japan) by liquid chromatography; - tryptophan content was determined according to GOST 32201-2013 (ISO 13904: 2005) [12]; - the vitamin content was determined by gas-liquid chromatography on a device Crystalux-4000M (Russia) on the reverse phase; - the potassium and magnesium content was determined by the photometric method according to GOST 26449.1-85 [13]; - determination of phosphorus - according to GOST 26657-97 [14]. To solve optimization problem, we applied the computer program of the Kuban State Technological University "Generik", intended for aided design and *Corresponding author: [email protected] © The Authors, published by EDP Sciences. This is an open access article distributed under the terms of the Creative Commons Attribution License 4.0 (http://creativecommons.org/licenses/by/4.0/). BIO Web of Conferences 30, 01009 (2021) https://doi.org/10.1051/bioconf/20213001009 ILS 2020

calculation of multicomponent recipes of functional food products 15. Statistical processing of individual indicators was carried out according to the Microsoft Excel-7.0 program with the determination of the average value, standard deviation and significance of differences by Student. Experimental studies were carried out in 3-5 replicates. 3 Results and discussion To obtain estimated information on the nutrient content in the composition of the modeled recipe, the material balance was used: (1) where Сi – is a mass fraction of a particular macro or microcating substance in the recipe; aij – is the value of the mass fraction of the i-th nutrient in the j-th ingredient (component); xj – mass fraction of the j-th component in the recipe. In the calculation process, the amino acid composition of prescription ingredients was used. From the obtained materials, data were selected that are characterized by the highest rates of desirability function (DF), which reflects the degree of balance of amino acids in product proteins. Taking into account the restrictions imposed on the quantitative content of components set during the development of food products (their sum is accepted as a unit) and permissible deviations from the values of the mass fractions of nutrients from reference ones, it was proposed to use the Lagrange function and the equation system in the form of conditions of the Kuhn-Tucker theorem for the convex programming problem to model recipes. Its solution allows us to obtain the vector x mass fractions of the recipe, the most balanced nutrient composition. Modeling of the most balanced recipe does not always determine the highest quality of the finished food product, therefore, in most cases, not one, but a sufficiently wide range of options for the composition of the prescription composition is required for its development. For this purpose, it is proposed to divide the process of its optimization into two stages. The first is the modeling of the recipe as the definition of all possible options for the quantitative ratio of the ingredients included in it. The second is a qualitative assessment and selection of several of its most optimal options. Harrington's desirability function was used as a generalized criterion for assessing the quality of the modeled recipe, which ensures the independence of the properties of particular indicators with different dimensions and a range of variable values, and at the same time it allows you to combine relative complex and simple unit quality criteria into one formula: (2) where Y – is a comprehensive quality criterion; pi − particular criteria (functions) of quality. When optimizing food recipes, it is usually advisable to use the desirability function using a two-way restriction: (3) (4) where Сi – is the mass fraction of the i-th nutrient (food substance) in the studied recipe; Li min, Li max – the limits of the standard value of the ith food substance. The parameter ni determines the characteristic of the curve, with ni →∞ the curve takes a rectangular shape. As a result of the research, a range of fish culinary products for a healthy nutrition was developed, including the "Fish Loaf", the recipe of which is presented in Table 1. Table 1. Formatting sections, subsections and subsubsections Name of the prescription component Content, % Cod fish fillet 30.0-40.0 Salmon fillet 15.0-25.0 Onion 5.0-8.0 Spinach 10.0-20.0 or eggplant or green peas or cauliflower Egg 3.5-4.5 Vegetable oil 4.0-6.0 Potato starch 2.0-3.0 Table salt 0.5-1.0 Spice mix 0.4-0.8 Water 12.0-30.0 Features: - functionality; -balance of amino acid, mineral and vitamin formulations; - high quality of organoleptic indicators. A culinary product has been developed for systematic use in order to increase the biological value and therapeutic value of diet food. It is part of dirts built on the principle of adapting the chemical composition and energy value of the diet to individual clinical and pathogenetic features of the disease. We compared indicators of the chemical composition of the developed culinary product with the daily requirement of an adult for food substances according to the balanced nutrition formula by A. A. Pokrovsky (Table 2). . , 1 1 , n j j n j jji i x xa C , 1 i k k i Y П p ,exp( , nc i i yp , (2 ) max min , max min i i ii i i LL LLC y 2 BIO Web of Conferences 30, 01009 (2021) https://doi.org/10.1051/bioconf/20213001009 ILS 2020

Table 2. Comparative indicators of the chemical composition of fish culinary products "fish loaf" with a daily requirement for them. Food substances Daily requirement Contents per 100 g of fish culinary product Contents per 100 g of fish culinary product, % of daily requirement 1 2 3 4 Protein, g 60-100 15.8000.600 20.0 Essential amino acids, g tryptophan 1 0.2340.039 23.0 leucine 4-6 1.6600.042 41.0 isoleucine 3-4 0,6440.056 21.0 valine 3-4 1.0610.090 30.0 threonine 2-3 0.9290.060 46.0 lysine 3-5 2.0390.080 70.0 methionine 2-4 0.3880.035 20.0 phenylalanine 2-4 0.7710.059 38.0 Nonessential amino acids, g histidine 1,5-2 1.1330.075 73.0 arginine 5-6 1.1040.088 22.0 cysteine 2-3 0.1900.022 10.0 tyrosine 3-4 0.7120.062 22.0 alanine 3 1.0130.098 30.0 serine 3 0.8960.078 30.0 glutamic acid 16 3.3530.020 21.0 asparagic acid 6 1.9850.130 33.0 proline 5 0.9070.070 18.0 Minerals, mg phosphorus 1200-1500 225.0000.001 16. 7 potassium 2500-5000 750.0000.001 30.0 magnesium 400-450 67.5000.001 16.9 Vitamins and vitamin-like substances, mg thiamine (vitamin B1) 1.1-2.0 0.2900.001 19.3 riboflavin (vitamin В2) 1.3-2.4 0.3100.001 23.8 pyridoxine (vitamin В6) 1.8-2.0 0.2800.001 15.6 cobalamin (vitamin В12) 0.003 0.00045+0.00001 15.0 References 1. O. Kutina, M. Mogil'nyj, T. Shlenskaya, and T. Sharova, Fish loaf with herbal components, Patent №2630314, Bul. 19 (2015) 2. M. Mogil'nyj (editor), Book of technical standards. Recipe book for the nutrition of workers at manufacturing enterprises and students in educational institutions of higher education (Deli plyus, Moscow, 2016) 3. The concept of long-term socio-economic development of the Russian Federation for the period up to 2020, approved by the order of the Government of the Russian Federation of November 17 (2008) 4. Fundamentals of the state policy of the Russian Federation in the field of healthy nutrition of the population for the period up to 2020 were approved by the Order of the Government of the Russian Federation of October 25 (2010) 5. Food Security Doctrine of the Russian Federation, approved by the Decree of the President of the Russian Federation of January 21, 20 (2020) 6. The development strategy of the food and processing industry of the Russian Federation for the period up to 2020, approved by the Order of the Government of the Russian Federation of April 17 (2012) 7. Food quality improvement strategy in the Russian Federation to 2030, approved by the order of the Government of the Russian Federation of June 29, (2016) 8. Action plan for the implementation of the food quality improvement strategy in the Russian Federation until 2030, approved by the decree of the Government of the Russian Federation of April 19, (2017) 9. GOST 7636-85. Fish, marine mammals, invertebrates and products of their processing. Methods of analysis, (Standartinform, Moscow, 2010) 3 BIO Web of Conferences 30, 01009 (2021) https://doi.org/10.1051/bioconf/20213001009 ILS 2020

10. GOST 31339-2006. Fish, non-fish objects and products of their processing. Acceptance rules and sampling methods, (Standartinform, Moscow, 2007) 11. GOST 25011-2017. Meat and meat products. Protein determination methods, Moscow: Standartinform, (Standartinform, Moscow, 2018) 12. GOST 32201-2013 (ISO13904:005). Feeds, compound feeds. Method for determination of tryptophan, (Standartinform, Moscow, 2014) 13. GOST 26449.1-85. Stationary distillation desalting units. Methods of saline water chemical analysis, (Izdatel'stvo standartov, Moscow, 1986) 14. GOST 26657-97. Fodders, mixed fodders, mixed fodder raw materials. Methods for determination of phosphorus content, (Interstate Council for Standardization, Metrology and Certification, Minsk, 1998) 15. A. Zaporozhskiy, V, Zaporozhskiy, Program for computer aided design, calculation and quality assessment of multicomponent food recipes (Generic-2.0), Certificate of program for computer, 2005611720 16. GOST R 55577-2013. Specialized and functional foodstuffs. Information about the distinctive signs and efficiency claims, (Standartinform, Moscow, 2014) 4 BIO Web of Conferences 30, 01009 (2021) https://doi.org/10.1051/bioconf/20213001009 ILS 2020

Study on the properties of modified starch and its feasibility in crystal dumpling skins Wang Jingwen1 , Oksana Melnyk1 , and Olha Ihnatieva1* 1Food Technology Department Sumy National Agrarian University, Sumy, Ukraine Abstract. Four kinds of modified starches that are expected to improve the quality of crystal dumpling skins were selected, namely Potato acetate starch, Potato hydroxypropyl starch, Octenyl succinate starch sodium, Acetate cassava starch. Through the determination of the physical indicators of the above four modified starches, and then explore the feasibility of applying them to crystal dumpling skins to improve their quality. Studies have found that the light transmittance of the four modified starches is higher than that of the original crystal dumpling mixed starch, which verifies the feasibility of applying these four modified starches to the crystal dumpling skin to improve its transparency and other characteristics. At the same time, it provides a certain reference value for related research on improving the characteristics of crystal dumpling skin. 1 Introduction At present, quick-frozen dumplings are developing rapidly in the quick-frozen food industry, accounting for 30% of the frozen food industry [1]. However, quick-frozen dumplings will have problems such as cracking of dumpling skins and muddy soup during processing, transportation or eating, especially for crystal dumplings made of whole starch, due to the poor water holding capacity of the epidermis, it is very easy to lose water during the production process and it is easy to freeze and crack during quick freezing at -50°C. Therefore, it is very important to find ways to improve its quality characteristics. Modified starch is a starch derivative obtained by processing physical and chemical methods on the basis of the original starch [2]. At present, a large number of studies have shown that the application of modified starch in dumplings can improve its transparency, reduce the rate of freeze cracking and cooking loss, etc. In view of the fact that there are more researches on the application of modified starch in quick-frozen dumplings made of flour [3-6], and there are relatively few researches on modified starch in quick-frozen crystal dumplings made of whole starch. The main research contents of this topic are: taking the commercially available crystal dumpling powder as the blank group and combining previous studies, four modified starches that are expected to improve the quality of crystal dumpling skins were selected: potato acetate starch, potato hydroxypropyl starch, octenyl succinate starch sodium, acetate cassava starch, by studying the physical properties of these starches, and then exploring the feasibility of applying it to crystal dumpling skins to improve its quality. 2 Experimental Wheat starch and corn starch Shandong Fanpu Food Co., Ltd. Potato acetate atarch Bolian Food Biotechnology Co., Ltd.; Potato hydroxypropyl starch Jinzun Food Ingredients Mall; Octenyl succinate starch sodium Shengda Food Business Department; Acetate cassava starch Henan Wanbang Industrial Co., Ltd: RVA-StarchMaster2 Rapid viscosity analyzer Shanghai Ruifen Company; L-550 Desktop low-speed largecapacity centrifuge Hunan Xiangyi Company; PH-070A Electric heating constant temperature blast drying oven Shanghai Yiheng Company; Electronic balance (0.0001g) Shanghai Qingke Tianmei Company; 722 Visible spectrophotometer Shanghai Youke Company; XW-80A vortex mixer Haimen Qilin Bell Company; HH-S2 digital display constant temperature water bath Jiangsu Jinyi Company. Determination of swelling degree and solubility of modified starch. Weigh 0.5g (dry basis, denoted as w0) of modified starch in a dry and clean centrifuge tube, weigh w1, add 20mL of distilled water, mix with a vortex mixer for 30s, respectively at 45, 55, 65, 75, 85, 95C keep in water bath for 30min, cool to room temperature, centrifuge at 5000r/min for 15min, pour off the supernatant, weigh w2, calculate the swelling power and solubility of modified starch according to formula (1) and (2) respectively [7]. Take 5 mL of the supernatant and place it in a drying dish at 105°C and dry it to a constant weight to obtain the quality w3 of the water-soluble starch. %100* 12 weightDry of starchraw ww Swelling Power %100* 4 lub 3 weightDry of starchraw w So ility Determination of transparency of modified starch paste. Add 1% (dry basis) modified starch suspension into *Corresponding author: [email protected] © The Authors, published by EDP Sciences. This is an open access article distributed under the terms of the Creative Commons Attribution License 4.0 (http://creativecommons.org/licenses/by/4.0/). BIO Web of Conferences 30, 01010 (2021) https://doi.org/10.1051/bioconf/20213001010 ILS 2020

a 20mL test tube, and then place it in a 95℃ water bath to heat for 30 min, mix with a vortex mixer every 5min for 30s, wait to cool to room temperature, and measure the starch paste at a wavelength of 650nm light transmittance, distilled water is used as a blank [8], the light transmittance reflects the transparency of starch paste. Determination of gelatinization characteristics of modified starch. The gelatinization characteristic value of starch is determined according to the method standard specified by the American Association of Cereal Chemists (AACC). Accurately weigh a certain amount of starch, the starch mass fraction is 6.0% (w/w, db), add it to an aluminum box containing 25g of distilled water, stir evenly with a rotating paddle, and place it in an RVA-rapid viscosity analyzer for measurement. Data processing and analysis. The statistical analysis of the data obtained in the experiment uses SPSS 24 software for statistical analysis of the data, and Origin Pro 9.1 software for drawing processing. All data are given in Mean ± SD, and the ANOVA method is used for the significance test. p<0.05 means statistically significant difference. 3 Results and discussion 3.1 Modified starch swelling power and solubility Swelling power reflects the water absorption capacity of starch during cooking, and reflects the characteristics of amylose [9]. Amylose inhibits the expansion of starch [10]; the influence of temperature on the swelling power of mixed starch and modified starch is shown in Figure 1. Within 45℃~95℃, the overall trend is that the swelling power of modified starch increases with increasing temperature. This is because as the temperature increases, more water molecules enter the amorphous area of the starch granule. When the temperature reaches its gelatinization temperature, the swollen amorphous area accelerates the destruction of the starch crystallization area, which in turn leads to an increase in starch swelling [11-13]. Solubility reflects the magnitude of the interaction force between starch and water, and has a greater impact on the processing characteristics of starch. The dissolution of starch is mainly due to the escape of amylose from the swollen granules [14]. The solubility of the original mixed starch and the modified starch is shown in Figure 2. The solubility of the modified starch increases with the increase of temperature, and the solubility increases rapidly from 65℃. This is because the starch is heated to absorb water and gelatinize rapidly, and high-energy water and heat destroys the internal hydrogen bonds of starch molecules, and part of the amylose gradually dissolves in water when heated, so the solubility of starch increases significantly [15-17]. Fig. 1 The effect of temperature on the swelling power of native starch and modified starch Fig. 2. The effect of temperature on the solubility of native starch and modified starch 3.2 Transparency of modified starch paste The transparency of modified starch paste is reflected by light transmittance, which reflects the mutual solubility of starch and water [18-19]. The light transmittance of the original mixed starch and modified starch is shown in above Figure 3. The light transmittance of the four modified starches is higher than that of the original mixed starch OS, and the light transmittance of MS2 is the highest, followed by the light transmittance of MS3 and MS1, MS4 has the lowest light transmittance but still higher than OS. This proves that the idea of applying it to crystal dumpling skins to improve its transparency is feasible. 2 BIO Web of Conferences 30, 01010 (2021) https://doi.org/10.1051/bioconf/20213001010 ILS 2020

Fig. 3. Light transmittance of different native starches and modified starches(OS-wheat starch: corn starch=3:1 mixed starch; MS1-potato acetate starch; MS2-potato hydroxypropyl starch; MS3- octenyl succinate starch sodium; MS4-cassava acetate starch) 3.3 Gelatinization properties of modified starch The gelatinization properties of starch are an important indicator reflecting the quality of starch, which affects the appearance, texture and mouthfeel of noodle products. The gelatinization characteristics of these starches were measured by RVA. RVA measures the process from the beginning of water absorption and swelling of starch granules to the destruction of the granular structure under the action of shear force and the leaching of starch molecules [20]. The gelatinization characteristic values of different raw starches are shown in Table 1. For the gelatinization temperature: the gelatinization temperature of the mixed powder with modified starch is higher than that of the blank group, and the gelatinization temperature except for the MS4-15% powder which are increase with the addition of modified starch; for peak viscosity and pow viscosity: the peak viscosity and low viscosity of the mixed powder with modified starch are lower than the blank group, and increase with the increase of the content of modified starch; Final viscosity: The final viscosity of the mixed powder with modified starch added is lower than that of the blank group. For attenuation value and retrogradation value: the attenuation value and retrogradation value of the mixed powder with modified starch added are lower than the blank group, and except for the fourth modified starch, the attenuation value and retrogradation value of other blended powders decreased with the increase of modified starch content. This may be due to the preferential water swelling of modified starch, which limits the water available for wheat starch, hinders the expansion of wheat starch, and leads to the decline of the gelatinization parameters (such as peak viscosity, attenuation value and retrogradation value, etc.) of the blended starch [21]. 4 Conclusion Based on previous studies, it is determined that the four modified starches selected are: Potato acetate starch, Sample Gelatinization temperature/cp Peak viscosity/cp Low viscosity/cp Final viscosity/cp Attenuation value/cp Regeneration value/cp 0 59.2±0.1 2248.3±0.6 1746.3±0.2 3179.0±0.9 562.0±0.1 1446.0±0.4 MS1-5% 77.7±0.3 1447.3±0.0 1054.0±0.7 2149.7±0.0 393.3±0.9 1095.7±0.4 MS1-10% 72.1±0.2 1478.3±0.5 1140.3±0.8 2141.3±0.3 338.0±0.5 1001.0±0.1 MS1-15% 71.2±0.7 1493.3±0.0 1157.3±0.1 2142.0±0.5 336.0±0.9 984.7±0.5 0 59.2±0.1 2248.3±0.6 1746.3±0.2 3179.0±0.9 562.0±0.1 1446.0±0.4 MS2-5% 86.0±0.7 1364.0±0.8 971.0±0.8 1996.0±0.7 393.0±0.7 1025.0±1.0 MS2-10% 79.8±0.1 1381.0±0.8 1033.7±0.4 1955.3±0.5 347.3±0.0 921.7±0.5 MS2-15% 73.5±0.3 1457.7±0.4 1125.3±0.1 1999.3±0.1 332.3±0.2 874.0±0.6 0 59.2±0.1 2248.3±0.6 1746.3±0.2 3179.0±0.9 562.0±0.1 1446.0±0.4 MS3-5% 85.6±0.5 1378.7±0.5 980.0±1.0 2031.7±0.3 398.7±0.5 1051.7±0.5 MS3-10% 75.8±0.2 1452.0±0.5 1068.7±0.9 2045.7±0.1 383.3±0.5 977.0±0.6 MS3-15% 74.6±0.0 1510.0±0.5 1165.3±0.5 2066.0±0.3 344.7±0.4 900.7±0.3 0 59.2±0.1 2248.3±0.6 1746.3±0.2 3179.0±0.9 562.0±0.1 1446.0±0.4 MS4-5% 86.9±0.5 1265.3±0.3 897.3±0.2 2000.0±0.8 368.0±0.1 1102.7±0.7 MS4-10% 87.0±0.7 1304.0±0.2 922.0±1.0 2077.3±0.6 382.0±0.0 1155.3±0.0 MS4-15% 87.4±0.8 1365.3±1.0 929.7±0.5 2118.0±1.0 435.7±0.5 1188.3±0.1 Table 1.. Effects of types and addition amount of modified starch on gelatinization characteristics of original mixed powder 3 BIO Web of Conferences 30, 01010 (2021) https://doi.org/10.1051/bioconf/20213001010 ILS 2020

Potato hydroxypropyl starch, Octenyl succinate starch sodium, Acetate cassava starch. The properties are compared and analyzed by measuring their physical indicators. The difference between them is found: Compared with the original mixed powder, the light transmittance of the four modified starches is higher than that of the original mixed starch. Among them, potato hydroxypropyl starch has the highest transparency; among the four modified starches, acetate cassava starch has the lowest light transmittance but is still higher than the transparency of the original mixed starch. The research results of gelatinization characteristics show that: modified starch has a great influence on the gelatinization characteristics of the original mixture: the gelatinization temperature is increased, and the remaining gelatinization parameters (peak viscosity, low viscosity, final viscosity, attenuation value, and retrogradation value) are reduced. This verifies the feasibility of the idea of applying these four modified starches to crystal dumpling skins, and at the same time provides a certain reference for related research on improving the characteristics of crystal dumpling skins. References 1. Z. Wenye. “Frozen convenience food processing technology and inspection, Beijing: Chemical Industry Press (2005) 2. Z. Yanping, Modified starch production and application [M]. Beijing: Chemical Industry Press (2007) 3. L. Wenjuan, H.. Shaokai, C. Yu et al. China Food Additives, 12, 60-64 (2015) 4. Y. Tuo, H. Juan, L. Yajie, etc. Gourmet Research, 36(02), 23-27 (2019) 5. J. Colive ,R.A. Carvalho. Industrial Crops & Products, 95 (2017) 6. Z. Sen, Z. Chunhua, H. Weixia, etc. Food Research and Development, 37(23), 21-25 (2016) 7. O.S. Lawal, K.O. Adebowale, B.M. Ogunsanwo, et al. International Journal of Biological Macromolecules, 35, 71-79 (2005) 8. F.O Onofre., Y.J Wang. International Journal of Pharmaceutics, 385(1-2), 104-112 (2010) 9. S.V Gomand., L. Lamberts, R.G.F Food Hydrocolloids, 24, 424-433 (2010) 10.R.F.Tester, W.R Morrison,. Cereal Chemistry, 67(6), 551-557 (1990) 11. Q.S. Lawal, J.Food Hydrocolloids, 23, 415-425 (2009) 12. S.G Choi, Starch/Stärke, 56, 181-189 (2004) 13. H.-J.Chung, , K-S Woo, S.-T Lim, Carbohydrate Polymers, 55, 9- 15 (2004) 14.J.S Lee., R.N. Kumar, H.D. Rozman,et al. Food Chemistry, 91, 203-211 (2005) 15. M. Xiangyan, Research on the physical and chemical properties of yellow rice starch. Chongqing: Southwest University (2008) 16.B. Berton, J. Scher, F. Villieras, et al. Power Technology, 128(2-3), 326-331 (2002) 17.R.S. Policegoudra, S.M Aradhya. Food Hydrocolloids, 22(4), 513-519 (2008) 18. D. Shuangkui, Z. Liqing, Y. Xiuzhu et al. Journal of the Chinese Cereals and Oils Association, 26(3), 34- 39 (2010) 19. D. Xianfeng, X. Shiying, W. Zhang. Transactions of the Chinese Society of Agricultural Engineering, 18(1), 129-132 (2002) 20. D. Cozzolino, S. Roumeliotis, J. Eglinton, Food Research International, 51(2), 444-449 (2013) 21. L. Ying, X. Zhiping. Agricultural Machinery, 5, 91-95 (2011) 4 BIO Web of Conferences 30, 01010 (2021) https://doi.org/10.1051/bioconf/20213001010 ILS 2020

Comparative analysis and antimicrobial action of some essential oils from plants Monica Mironescu1* , and Cecilia Georgescu 1 1Faculty of Agricultural Sciences Food Industry and Environmental Protection, Research Centre in Biotechnology and Food Engineering, University Lucian Blaga of Sibiu Sibiu, Romania Abstract. In this research, essential oils (EOs) were extracted through steaming from aerial parts of three plants: basil (Ocinum basilicum L.), peppermint (Mentha piperita L.) and oregano (Origanum vulgare L.) form Sibiu County, Romania. The GC-MS analysis indicated that eugenol and apiol were the major compounds in the basil EO. Timol, ɣ-terpinene, and p-cymene were the major compounds in the volatile oil extracted from oregano and menthol and methone the main components in the peppermint EO. In the first 48 h of analysis, basil EO inhibition action was the highest against Salmonella anatum (48%), followed by Aspergillus niger (26%), Bacillus cereus (21%) and Saccharomyces cerevisiae (15%); this action was maintained for 120 h for all samples. In the first 48 h, the peppermint EO showed the highest inhibition rate (50%) on B. cereus, followed by A. niger (45%), S. cerevisiae (20%) and no action against Salmonella; this action varied after 72h and 120 h by slightly increasing in case of B. cereus, strongly decreasing in case of A. niger and remaining constant in case of S. cerevisiae. Oregano EO showed a very small antimicrobial action, only on S. anatum (12%) and this action was maintained for 120 h. 1 Introduction Aromatic plants are rich sources of bioactive compounds or bio nutrients [1]. Their essential oils (EOs) are final products of plants metabolism, accumulated and stored in various plant organs, in cells with intracellular secretion or in secretory tissues. Essential oils are natural, volatile complex compounds characterized by the odor of their corresponding aromatic plants, which synthesize them as secondary metabolites [2, 3]. They have a lot of applications in food industry, cosmetics, perfumeries and pharmacy industry, due to their powerful antimicrobial and antioxidant activity [4]. The benefits resulting from the use of natural products rich in bioactive substances have promoted the growing interest of pharmaceutical, food and cosmetic industries as well as of individual consumers in the quality of herbal products [5, 6]. The EOs are complex mixtures that may contain over 300 different compounds [7]. Essential oils are natural mixtures of hydrocarbons (terpenes), oxygen-(alcohols, aldehydes, ketones, carboxylic acids, esthers, lactones) and sulphur-containing (sulphides, disulphides, trisulphides) organic substances. They are obtained mostly from plants by steam distillation or by extraction with carbon dioxide [8, 9]. Many articles present studies on volatile oils extracted from basil, peppermint or oregano [10, 11, 12]. Peppermint EO is one of the most usually used volatile oil in food and beverages, whereas basil and oregano oil are used in Mediterranean-origin food and in pharmaceutical and cosmetic market segments. Peppermint (Mentha piperita L.) essential oil is usually used topically as antimicrobial, antipruritic, antiseptic, rubefacient and astringent as well as for healing headaches, neuralgia, migraines and myalgia [13]. Basil (Ocimum basilicum L.) is an annual plant found in the wild tropical, subtropical and temperate regions of the world. It takes part from Labiatae (Lamiaceae) family. Basil is a common herb, grown also in many households with a broad range of therapeutic properties; it has reputed medicinal uses as antioxidant, antibacterial, antimicrobial, antifungal, antiviral, cytoprotective, anticonvulsant, hypoglycaemic, hypolipidemic, hepatoprotective, renoprotective, neuroprotective, spermicidal, dermatologic and insecticidal [14]. Oregano (Origanum vulgare L.) EO have been shown to possess antioxidant, antibacterial, antifungal, diaphoretic, carminative, antispasmodic and analgesic activities and, among these, the antimicrobial potential is of special interest [15, 16]. The aim of this study is to compare the composition and the antimicrobial activity of EOs extracted from aerial parts of the three plants considered (basil, peppermint and oregano) against microorganisms possible to poison food (pathogenic bacteria and molds) and microorganisms with positive role in food production (as the model yeast Saccharomyces cerevisiae). The antimicrobial activity is surveyed for a period of 4 days, in order to analyze the remanence of the antimicrobial action of the EOs.. 2 Experimental Plant materials Dried aerial parts of Ocimum basilicum L. (basil), Mentha piperita L. (peppermint), Origanum vulgare L. *Corresponding author : [email protected] © The Authors, published by EDP Sciences. This is an open access article distributed under the terms of the Creative Commons Attribution License 4.0 (http://creativecommons.org/licenses/by/4.0/). BIO Web of Conferences 30, 01011 (2021) https://doi.org/10.1051/bioconf/20213001011 ILS 2020

(oregano), were used for extraction the volatile oils. Arial parts from all this herbs were collected from Sibiu County, Romania in 2018 and they were dried in the shade. Essential oils extraction and characterization For obtaining and dosing the EOs, aerial parts were used. Extraction was realized through steaming for 5 h by using a neo-Clevenger equipment modified as by Moritz. The content was compared to the moisture less vegetal material [17]. The volatile oils were analyzed through by GC-MS. The analysis was performed with a Hewlett Packard 5890 III gas chromatograph equipped with a mass detector MS 5972. The chromatographic column used was a HP5-MS capillary column made of quartz, with a non-polar stationary phase consisting of 95 % methyl and 5 % phenyl polysillox. The constructive characteristics of the column are: length, 30 m, interior diameter, 0.25 mm and thickness of the stationary phase, 0.25 m. Helium was used as carrier gas (1:1 ml/min). The injection temperature was 60 C, incrementing with 3 C/min to 240 C. To identify the separated components, an integrated GC-MS software was used. Library Wiley 275, used to recognition of chemical compounds was carried out by comparing the acquired mass spectra with mass spectra of standards program [17]. Determination of the antimicrobial activity Antimicrobial action of the EOs was investigated on three types of microorganisms: pathogenic bacteria (Salmonella anatum ATCC 9270 and Bacillus cereus ATCC 10876), a mould contaminating food (Aspergillus niger ATCC 16404) and Saccharomyces cerevisiae ATCC 32701, a model non-pathogenic yeast. Before the analysis, the pure cultures of bacteria were maintained in Mueller-Hinton broth (MHB, Difco) for 48h at 45 C, the pure culture of yeast were maintained in nutrient broth (BD Difco Nutrient broth) for 120 h at 20 C (yeast); the liquid broth was used. The mould was cultivated first on malt agar (Difco) for 5 days at 20 C, then the spores were harvested in nutrient broth and the liquid was used for the analysis. Agar diffusion method was used to investigate the antimicrobial action of a quantity of 0.5 l of essential oil against microorganisms cultivated on nutrient agar (Difco Nntrient Agar); the inhibition percentage on the continuous film formed by the microorganism was determined by reporting the inhibition zone to the Petri dish area [18]. Five trials in parallel for each microorganism were made; the results were expressed as median value of all measurements. Because the EOs are volatile, the antimicrobial activity was analyzed after 48 h, 72 h and 120 h in order to investigate the remanence of the antimicrobial action in time. 3 Results and discussion EOs extracts from basil and peppermint have a white-yellowish color and the oregano oil has a yellow dark color. All EOs have characteristic flavors, slightly aromatic. The extraction yields are 0.66 % for Ocimum basilicum L. volatile oil, 0.72 % for Mentha piperita L. volatile oil, 0.16 % for Origanum vulgare volatile oil. The GC-MS analysis shows that all volatile oils analysed have a high number of components. The major compounds of the volatile oils are presented in Table 1. Twenty five components in the volatile oil extracted from Ocimum basilicum L. were separated and identified through GC-MS. Eugenol and apiol are the major compounds and mean components are alcohols. 19 components in the volatile oil extracted from Origanum vulgare L. were separated and identified through GCMS; timol, ɣ-terpinene, and p-cymene are the major compounds. 26 components in the volatile oil extract from Mentha piperita L were separated and identified through GC-MS; the main components of EO from peppermint are menthol and menthone. Table 1 presents the results obtained at the analysis of the antimicrobial activity of all three EOs on the chosen microorganisms. Basil essential oil is having antimicrobial action on all microorganisms. In the first 48 h of analysis, the action decreased in the order: S. anatum (48 %), A. niger (26), B. cereus (21 %) and S. cerevisiae (15 %); this action is maintained after 72 h or 120 h for all samples. In the first 48 h, the peppermint oil shows the highest inhibition (50 %) on B. cereus, followed by A. niger (45 %) and S. cerevisiae (20 %) and no inhibition on Salmonella; this action varies after 72h and 120 h by slightly increasing in case of B. cereus, strongly decreasing (to 20 %) in case of A. niger and remaining constant in case of S. cerevisiae. Oregano oil has the smallest antimicrobial action, only on S. anatum (12 %) and this action is maintained for 120 h. Scientific literature the differences in the extraction quantities and yields, due to variation in quantity, but also growing conditions (climate, soil constituents), part of the plant used for extraction, stage of ripening process. Also, the scientific information about EOs obtained from these plants are very different. For example, in case of EO extracted from American basil, the extraction yield varied between 0.07 % and 1.92 % and eugenol content was much smaller (5-29 %) [19], compared with our results. Regarding the peppermint EO, menthol and menthone were found by other authors, too, as the major components of the peppermint EO [20], whereas other researchers identified menthol, menthyl acetate and menthofurane as major compounds [11]. The extraction yield was found to be around 1 % by other authors [21], higher as the content obtained by us, but a real comparison is difficult to be made because of the high number of variables influencing the yield. Oregano essential oil yield (ranged from 0.114 % to 2.3 %, depending on the distillation time and carvacrol was the main component in other researches, too [22]. The highest action against S. anatum showed by the basil essential oil (Fig. 1a), combined with the very low inhibition action of the oregano EO is in agreement with other researches [23]. The EOs chemical component in basil with the inhibitory action is eugenol causing the increasing of permeability by decreasing the membrane integrity, with the Minimal Inhibitory Concentration MIC of 0.025 % against S. typhimurium [24]. 2 BIO Web of Conferences 30, 01011 (2021) https://doi.org/10.1051/bioconf/20213001011 ILS 2020