Sustainability Assessment of Pectin Extraction From Citrus Paradisi Peel and Application in Encapsulating Lactiplantibacillus Plantarum Cidca 83114

Sustainability Assessment of Pectin Extraction From Citrus Paradisi Peel and Application in Encapsulating Lactiplantibacillus Plantarum Cidca 83114 | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Sustainability Assessment of Pectin Extraction From Citrus Paradisi Peel and Application in Encapsulating Lactiplantibacillus Plantarum Cidca 83114 Natalia A. Di Clemente, Enzo La Cava, Sonia Sgroppo, Andrea Gomez-Zavaglia, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4637293/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Agro-industrial waste, including peels, pulps, pomace, and seeds, poses a significant global economic and environmental problem. Recovering these wastes to extract bioactive compounds ( e.g. , pecin, polyphenols, pigments, essential oils) offers a sustainable solution. Pectins have been increasingly used as delivery systems in the food industry. Green extractions have been developed to reduce the extraction time and environmental impact of conventional methods. However, little is known about their actual impact. Life Cycle Assessment (LCA) is a useful tool for assessing the environmental and energy impacts of a production cycle. This study aims to evaluate the environmental profile of pectin extraction from grapefruit peels using conventional heating (CHE) and thermosonication (TS) methods, and the application of the extracted pectin as a delivery system for encapsulating Lactiplantibacillus plantarum CIDCA 83114. The LCA was performed using Open LCA software version 2.0.1 modelled with ILCD 2011 method. The system boundaries were considered to be laboratory scale and the functional units were 1 kg of protected/dry encapsulated bacteria in pectin extracts from Citrus paradisi peel obtained by TS or CHE. The impact scores of the TS and CHE scenarios were similar in terms of millipoints (TS = 18.9 and CHE = 19.1 mPt). The main impact categories were climate change, human toxicity with carcinogenic effects and depletion of water resources contributing to deionized water and electricity consumption. The obtained results contribute to the decision-making process for the selection of a pectin extraction process on a laboratory scale, complemented by future economic impact studies. encapsulation Citrus paradisi pectins lactobacilli conventional heating thermosonication Life Cycle Assessment (LCA) Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 1. INTRODUCTION According to (FAO, 2019 ), about 14% of food is lost between harvest and distribution at a global level, and a further 17% is wasted during distribution and at the final consumer (UNEP, 2021). Approximately 40–50% of fruit and vegetable production is lost or wasted. These losses come from peel fractions, pulps, pomace, and seeds, representing about 16% of total food waste and contributing to around 6% of global greenhouse gas emissions. To promote sustainable food production, several agro-industrial wastes (peels, pulps, pomace, seeds) are being recovered to extract different bioactive compounds, including fiber (pectins, prebiotic oligosaccharides), polyphenols and pigments (Cassani & Gomez-Zavaglia, 2022 ). Bioactives from fruit and vegetable wastes and losses have been extensively researched, particularly regarding the implementation of high-yield and environmentally friendly extraction methods (Riyamol et al., 2023 ). The worldwide production of grapefruit [ Citrus paradisi (Macf.)] is estimated up slightly to 6.9 million tons, being China, the main productor forecast up slightly to 5.2 million tons (period 2023/24) (Foreign Agricultural Service/USDA, 2024, p. 6). Citrus peel is an agricultural waste from juice production, traditionally disposed of as solid waste or animal feed (Grassino et al., 2016 ), and accounts for almost 50% of the wet fruit mass after juicing (Sharma et al., 2017 ). One of the most important bioactives found in grapefruit peel is pectin, a polysaccharide composed of galacturonic acid units (Saini et al, 2022 ). Conventional methods for pectin extraction use an acidic or basic aqueous medium at high temperatures (Emaga et al., 2008 ; Geerkens et al, 2015 ). These processes are time consuming and energy intensive and result in serious environmental problems due to the production of acidic wastewater and corrosion of the equipment (Riyamol et al., 2023 ). Recently, pectin has been increasingly used in the food industry for developing various products as a biomaterial in the construction and application of micro/nano delivery systems for nutritional or pharmaceutical purposes (Liu et al, 2022 ). Ghibaudo et al. ( 2017 ) used pectate-iron beads for encapsulation of probiotic Lactobacillus plantarum CIDCA 83114, providing a source of iron and fibre. Similarly, pectate-calcium beads from citrus pectin were investigated as encapsulants of Lactobacillus plantarum CIDCA 83114 (La Cava et al., 2019 ). According to a recent report, the global pectin market reached US $ 1,170.8 million in 2023 and is forecast to grow to US $ 2,256 million by 2032 (IMARC, 2020 ). There is therefore a need for better extraction methods to maximize pectin yield and purity. Alternative green approaches, such as enzyme-assisted extraction (Lim et al., 2012 ), subcritical fluid extraction (Pattarapisitporn, et al., 2024 ), ultrasound-assisted extraction (Wang et al, 2014; 2015 ; La Cava et al, 2018 ; Polanco-Lugo et al.,2019), microwave-based extraction (Attard et al., 2014 ; Liu et al., 2017 ) and ohmic heating assisted extraction (Saberian et al., 2017 ; Sabanci et al., 2021 ) allow for a high yield of pectin extraction with the advantages of short extraction time, low solvent consumption and low environmental impact. In addition, the biodegradability and low toxicity of grapefruit pectin make it a promising biomaterial for micro/nano delivery applications, enhancing bioactivity and encapsulation ability (Liu et al, 2022 ). While numerous studies indicate that green extractions are effective in terms of extraction yield, pectin quality and reduced environmental impact, little research has been done on the actual value of their environmental impact. Life cycle assessment (LCA) is a valuable tool to evaluate and identify environmental impacts during product development or processing, considering resources and energy balances (Roy et al., 2008 ; Jacquemin et al., 2012 ; Ott et al., 2022 ). The goal of this study was to assess the environmental performance of different pectin extraction processes (conventional heating and thermosonication) and its application as delivery systems in the encapsulation of Lactiplantibacillus plantarum CIDCA 83114. 2. MATERIALS AND METHODS Different pink/red and white grapefruit ( Citrus paradisi Macf.) varieties, with a Brix/acid ratio of 5.5, uniform skin coloration and free of cuts, were provided by INTA Experimental Station in Bella Vista (Corrientes, Argentina 28° 30' 52.43'' N, 59° 1' 47.94'' S). Different citric combinations of "Parana" on different rootstocks (“Citrumelo Swingle,” “Duncan,” “Tangelo Orlando”, “Lima Rangpur”) were analyzed. The red/pink varieties investigated included Star Ruby” (pink), “Foster” (red), “Red Blush” (pink), and “Red Shambar” (pink). Sulfuric acid, hydrochloric acid, sodium hydroxide and calcium chloride (Cicarelli, Argentina) and MRS broth (Biokar Diagnostics, France) were used. 2.1 Description of the unit processes The stages of the process flow are described in Fig. 1 . Grapefruit peel preparation. Fruits were washed with tap water and sanitized with hypochlorite acid solution (HClO, 200 mg/L) for 5 min. Peel was separated and macerated in a water bath at 90°C for 5 min to inactivate enzymes and cut in 1 cm 2 -slices. Then, peels were dried in an oven under vacuum at 50°C overnight and milled for their homogenization. To extract pectin from the peels, deionized water adjusted with 2N H 2 SO 4 to reach pH 1.5 was used as the extraction solvent. 2.1.1 Pectin extraction. Two different methods were considered: conventional heating (CHE) and thermosonication (TS) according to La Cava et al. ( 2018 ). A Precytec AE-29 (CABA, Argentina) stirrer was used for CHE extraction at 90°C for 150 min. TS was conducted in a 6 L-ultrasonic tank (Cleanson model KT 1106, Argentina. 200 W, 40 KHz) with thermostatic control at 60°C for 30 min under periodic agitation (10 s on, 50 s off). 2.1.2 Purification. Filtered and extracted pectin were washed and precipitated with ethanol solution (96%) before final filtration in a nylon cloth. 2.1.2 Pectin hydrolysates preparation. HCl solution (0.2 N) was applied for 2 hours at 110°C for chemical hydrolysis, and the pH of the hydrolysates was adjusted to 6.5 with 1N NaOH, according to La Cava et al. ( 2019 ). Finally, hydrolysates were concentrated by rotary evaporation (Rotavapor® RE 120 – Buchi, Flawil, Switzerland) under reduced pressure at 50°C. 2.1.3 Fermentation. Lactiplantibacillus plantarum CIDCA 83114 was isolated from kefir grains (Garrote, Abraham, & De Antoni, 2001) and maintained frozen at − 80°C in 120 g/L nonfat milk solids (Difco, MI, USA). Microbial cells were activated for 24 hours in MRS broth (de Man, Rogosa, & Sharpe, 1960) at 37°C in aerobic conditions. The resulting culture was inoculated (1% v/v) in fresh MRS broth and incubated in the same conditions. Cultures in the stationary phase were used to inoculate pectin hydrolysates. 2.1.4 Encapsulation. Bacteria were encapsulated by ionotropic gelation, incorporating 4% w/v of fresh hydrolysates into the bacterial cultures and adjusting the pH to 6 with 1N NaOH, while shaking for 30 minutes. A 250 mmol/L CaCl 2 solution was dripped into the hydrolysates using a 0.3 mm needle (∼10 µL/drop) under continuous agitation for 30 min, according La Cava et al. ( 2019 ). The capsules were harvested by filtration through a stainless steel mesh of 0.10 mm, and rinsed three times with distilled water. 2.1.5 Dehydration and preservation. The capsules were frozen at − 80°C and freeze-dried for 48 hours on a freeze dryer (CHRIST Equipment, Germany) operating with the condenser at − 45°C at a chamber pressure of 0.04 mbar. The obtained samples were stored for 45 days at 4°C and the viability was monitored by plate counting on MRS-agar at days 0, 15, 30, and 45. 2.2. LCA methodology 2.2.1 Goal and scope . Identify the environmental profile of the encapsulating process of Lactiplantibacillus plantarum CIDCA 83114 in pectin extracts and compare two pectin extraction methods (conventional heating vs thermosonication). Functional unit: 1 kg of freeze-dried-encapsulated bacteria in pectin extracts obtained by conventional heating or thermosonication. 2.2.2 System boundaries. A gate-to-gate approach was considered in this laboratory study, assuming the impacts from end of life. The harvesting and transport of grapefruits and the treatment of waste were not considered. The system boundary was set to include only the process of pectin extraction from Citrus paradisi peel, followed by the encapsulation of L. plantarum CIDCA 83114 and the dehydration of the encapsulates. 2.2.3 Life Cycle Assessment software and impact categories. The present LCA was carried out using Open LCA software version 2.0.1 modelled with ILCD 2011 method, considering the midpoint indicators listed in Table 1 . For normalization and weighting, the EU27 2010 method was applied. 2.2.4 Life cycle inventory: Inventory data. Inventory data corresponding to production systems such as electricity, chemicals, tap water, deionized water, and waste treatment were taken from ELCD 3.2 database (Greendelta v2.18 correction20220908). Electrical consumption (MJ) was calculated based on the power ratings of the equipment (bath, grinder, oven, shaker, ultrasonic bath, vacuum pump, rotavapor, autoclave, refrigerant, freezer, freeze-dryer) and their duration of use (hour) (Gerbino et al., 2022). The leakage of refrigerant gas (refrigerant, freezer and freeze-dryer) was calculated according to Pénicaud et al (2018), using available data from providers. Inputs and outputs for each process, comparing the two pectin extraction methods [conventional heating (Scenario 1) or thermosonication (Scenario 2)] (Table 2 ). Table 1 ILCD 2011 midpoint indicators and their abbreviations and units. Impact categories Abbreviation Unit Acidification AP molc H + eq Climate change CCP kg CO 2 eq Freshwater ecotoxicity FETP CTUe Freshwater eutrophication FEP kg P eq Human toxicity, cancer effects HTPce CTUh Human toxicity, non-cancer effects HTPnce CTUh Ionizing radiation E (interim) IRP E CTUe Ionizing radiation HH IRP HH kBq U235 eq Land use LUP kg C deficit Marine eutrophication MEP kg N eq Mineral, fossil & ren resource depletion MFRP kg Sb eq Ozone depletion ODP kg CFC-11 eq Particulate matter PMP kg PM2.5 eq Photochemical ozone formation POP kg NMVOC eq Terrestrial eutrophication TEP molc N eq Water resource depletion WDP m 3 water eq Table 2 Inventory data. Inputs and outputs per functional unit for each process comparing two scenarios: conventional heating (Scenario 1) or thermosonication (Scenario 2). Process Inputs Scenario 1 Scenario 2 Grapefruit peel preparation Fruits (kg) 9000 9000 Electricity (MJ) 3263 3263 HClO (L) 4 4 Tap water (L) 20000 20000 Distillated water (L) 20000 20000 Pectins extraction Sulfuric acid (kg) 2.45 2.45 Deionized water (L) 24 29 Electricity (MJ) 2025 450 Purification Electricity (MJ) 223 223 Ethanol (kg) 4 4 Pectin Hydrolysates Deionized water (L) 8998 8998 Electricity (MJ) 13188 13188 Hydrogen chloride (Kg) 1.8 1.8 Fermentation Bacteria (kg) 25 25 Deionized water (L) 1000 1000 Electricity (MJ) 2772 2772 Encapsulating Deionized water (L) 2000 2000 CaC l2 (kg) 28 28 Electricity (MJ) 338 338 Dehydration and preservation Electricity (MJ) 1041 1041 Refrigerated gas (kg) 404 404 Outputs Grapefruit peel preparation Liquid waste (L) 40000 40000 Fruit waste (kg) 8000 8000 Pectins extraction Organic waste (kg) 547 547 Waste water (kg) - 5 Purification Organic waste (kg) 410 410 Chemical waste (kg) 40 40 Pectin Hydrolysates Chemical waste (kg) 9078 9078 Fermentation Waste water (kg) 1000 1000 Encapsulating Organic waste (kg) 999 999 Chemical waste (kg) 2000 2000 Dehydration and preservation Refrigerant leakage (kg) 6 6 Freeze-drying encapsulating bacteria (kg) 1 1 Sensitivity analysis was done for some of the input parameters (fruit, electricity, refrigerant), that did not cause significant changes to the results. 2.3. Antioxidant capacity As the different scenarios/extraction methods can affect the quality of pectin, antioxidant capacity was determined as a functional property using the DPPH • (2,2-Diphenyl-1-icrylhydrazyl) and ABTS •+ [2,2-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid)] assays [Magalhães et al. 2012]. The antioxidant capacity was expressed as EC50% (effective concentration), with a lower value indicating greater antioxidative activity. The EC50% values were calculated by non-linear regression of plots where the abscissa represented the concentration of the tested samples, and the ordinate, the average percentage of antioxidant activity. 3. RESULTS AND DISCUSSION Environmental profiles of the encapsulation process of Lactiplantibacillus plantarum CIDCA 83114 in pectin extracts are shown in Fig. 2 for two scenarios: CHE (A) and TS (B) extractions. The percentage contribution of each stage in the process to the total impact revealed that grapefruit peel preparation and pectin hydrolysates were the hotspots of both scenarios, in addition to pectin extraction when CHE is used (Fig. 2 a). Grapefruit peel preparation contributed mainly to HTPnce with 64.9% and 65.4% in Scenario 1 and Scenario 2, respectively, and 62% to ODP and WDP in both scenarios. Pectin hydrolysates process mainly contributed to MFRP 46.4% and 48.5% in Scenario 1and Scenario 2, respectively. These contributions are related to the water consumption, organic and chemical waste generated by these processes (Fig. 1 ). In Scenario 1, pectin extraction by CHE contributed particularly to MRFP (5.68%), CCP (3.10%) and HTPce (2.65%), influenced by the use of time and equipment. In Scenario 2, using TS, all the impact categories contributed less than 1.5%. Table 3 shows the raw values of the impact categories for Scenarios 1 and 2 and the percentage relation between them. Figure 3 represents the relative percentage normalized values of each impact category for both scenarios. According to the results presented in Table 3, Scenario 1 had the major environmental impact in all impact categories as positive values, except in IRP E. However, the total contribution, calculated in millipoints, showed similar values in both scenarios (CHE = 19.1 and TS = 18.9 mPt). Figure 3 presents the comparative profile of the environmental impact of the encapsulation process of Lactiplantibacillus plantarum CIDCA 83114 using two pectin extracts methods. The principal impact categories affected were acidification (AP), climate change (CCP), human toxicity cancer effects (HTPce), ionizing radiation HH (IRP HH), photochemical ozone formation (POP) and water resource depletion (WDP). The weighted results of the five principal impact categories were expressed in mPt for both scenarios (Fig. 4 ). The consumption of electricity during the production of deionized water and the fuel consumption for electricity production are associated with higher CO 2 emissions into the atmosphere, as well as increased emissions of nitrogen and sulfur oxides, leading to climate change and acidification impacts, respectively (Marzeddu et al., 2021 ). Consequently, volatile organic compounds (VOCs) were released, increasing the POP impact (Alisha, et al., 2019) which is correlated with human health impacts (Shajed & Jolliet, 2019). Beccali et al ( 2009 ) reported energy consumption as a hotspot in the eco-profiles of six citrus-based products in Italy, related to CO 2 emissions and water consumption. Deionized water and electricity consumption were mainly responsible for the environmental impact of Scenario 1 and Scenario 2, as shown in the Sankey diagram, where the thick lines represent the highest impact flux. (Fig. 1 S. 2S, 3S). Figure 5 shows the relative categories results of both scenarios. For each impact categories, the maximum result is set to 100%, corresponding to Scenario 1 (red) and the results of the Scenario 2 (blue) are displayed in relation to this result. As mentioned above, the environmental impact of Scenario 1 is higher than that of Scenario 2, but the differences do not exceed 6%. However, CHE (Scenario 1) showed a better yield (4.1 ± 1.7%) than TS (Scenario 2) (1.5 ± 0.4%), corresponding to the galacturonic acid concentration, which was higher for CHE (786.4 ± 58.2 g/kg) than for TS (605.9 ± 91.8 g/kg) (La Cava et al., 2018 ). In relation to antioxidant activity (Table 4 ), pectin extraction by CHE had the best performance. This suggests that the extraction method strongly determines the antioxidant capacity of the samples (Bozinou et al., 2019 ). In this regard, it has been reported that the acoustic cavitation generated during ultrasound treatment may lead to the production of free radicals, thus explaining the lower antioxidant capacity of TS extracts (Piyasena, Mohareb, & McKellar, 2003 ). Table 3. Impact assessment values for 1 kg of protected/dry-encapsulated bacteria in pectin extracts obtained by conventional heating (Scenario 1) or thermosonication (Scenario 2) and relative environmental difference between both scenarios. Impact categories Scenario 1 Scenario 2 Scenario 1/Scenario 2 (%) AP 1.2 1.1 0.323 CCP 449.6 439.0 2.365 FETP 15.9 15.6 2.057 FEP 5.3×10 -4 5.3×10 -4 0.101 HTPce 1.2×10 -6 1.2×10 -6 2.018 HTPnce 9.2×10 -6 9.1×10 -6 0.776 IRP E 3.2×10 -4 3.2×10 -4 -0.194 IRP HH 32.5 32.5 0.161 LUP 0.0 0.0 MEP 0.3 0.3 0.501 MFRP 5.2×10 -5 5.0×10 -5 4.390 ODP 2.7×10 -5 2.7×10 -5 0.119 PMP 0.1 0.1 0.191 POP 0.7 0.7 0.576 TEP 2.6 2.6 0.592 WDP 5.2 5.2 0.004 Differences in the various components of LCA studies (such as functional units, system boundaries, laboratory or industrial scale and impact assessment methods) will be considered for comparing the presented results with other studies. However, several studies show differences between traditional extraction methods and green ones, with the latter being more efficient and environmentally friendly (García-García et al., 2019; Pappas et al., 2023 ). In agreement with the results of this work, Wang et al. ( 2015 ) reported that ultrasound-assisted heating extraction was an efficient and economical method to obtain higher pectin yields at lower temperatures and shorter extraction times than CHE from grapefruit peel. Despite a similar environmental impact profile, the energy and time consumption in Scenario 1 were higher than that in Scenario 2, so this approach should be carefully managed when making decisions. Considering that the yield of pectin in Scenario 1 was more than double that of Scenario 2, a life cycle cost analysis should be carried out, also taking into account evaluations on an industrial scale (note that this work was carried out on a laboratory scale). Moreover, specific limitation in terms of collecting local and good quality inventory data is a major challenge. 4. CONCLUSION From the analysis presented above, it can be concluded that the environmental profiles of the encapsulation process of Lactiplantibacillus plantarum CIDCA 83114 in pectin extracts were similar when comparing conventional heating with thermosonication extraction method from grapefruit peel waste. The main resources contributing to the impacts are deionized water and electricity consumption. Therefore, it is important to consider using more energy efficient equipment to help achieve a sustainable approach to using energy. Furthermore, the present results, together with previous studies (La Cava et al., 2018 ; 2019 ), contribute to the development of an encapsulated probiotic in pectin as a potential alternative both to overcome the environmental problem of grapefruit peel and as a bioactive compound in the food and pharmaceutical industries. Declarations Acknowledgments This work was supported by the Argentinean Agency for Scientific and Technological Promotion (ANPCyT) (Project PICT/2020/0482 and PICT 2019-2428). E.L.L.C, A.G-Z. and E.G. are members of the research career of CONICET. N. D.C. is a postdoctoral fellow of ANPCyT. Author ' s Contributions N. D. C conducted the LCA analysis and the preparation of the original draft, as well as writing, reviewing, and editing the final manuscript. E. L: L. C contributed to the experimental work, data curation, reviewing and investigation. E. G. conducted the investigation, participated in LCA data curation, reviewing, and editing the final manuscript. A. G-Z and S. G. resources, reviewing, and editing the final manuscript and also supervised project administration and was responsible for funding acquisition. 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CyTA - Journal of Food , 17(1), 463–471. https://doi.org/10.1080/19476337.2019.1600036 Riyamol, N., Gada Chengaiyan, J., Rana, S. S., Ahmad, F., Haque, S., & Capanoglu, E. (2023). Recent Advances in the Extraction of Pectin from Various Sources and Industrial Applications. ACS Omega , 49 (8), 46309–46324. https://doi.org/10.1021/acsomega.3c04010 Roy, P., Nei, D., Orikasa, T., Xu, Q., Okadome, H., Nakamura, N., & Shiina, T. (2008). A review of life cycle assessment (LCA) on some food products. Journal of Food Engineering 90 (1), 1–10. https://doi.org/10.1016/j.jfoodeng.2008.06.016 Sabanci, S., Cevik, M., & Goksu, A. (2021). Investigation of time effect on pectin production from citrus wastes with ohmic heating assisted extraction process. Journal of Food Process Engineering , 44(6), 13689–13699. https://doi.org/10.1111/jfpe.13689 Saberian, H., Hamidi-Esfahani, Z., Gavlighi, H. A., & Barzegar, M. (2017). Optimization of pectin extraction from orange juice waste assisted by ohmic heating. Chemical Engineering and Processing-Process Intensification , 117, 154–161. https://doi.org/10.1016/j.cep.2017.03.025 Saini, R.K., Ranjit, A., Sharma, K., Prasad, P., Shang, X., Gowda, K. G. M., & Keum, Y. S. (2022). Bioactive Compounds of Citrus Fruits: A Review of Composition and Health Benefits of Carotenoids, Flavonoids, Limonoids, and Terpenes. Antioxidants (Basel). 11(2):239. doi: https://doi.org/10.3390%2Fantiox11020239 Shaked, S. & Jolliet, O. (2019). Global Life Cycle Impacts of Consumer Products, In Jerome Nriagu (Eds.), Encyclopedia of Environmental Health (2nd ed., pp. 331-342). Elsevier. ISBN 9780444639523. https://doi.org/10.1016/B978-0-12-409548-9.11700-2 Sharma, K., Mahato, N., Hwan Cho, M., & Rok Lee, Y. (2017). Converting citrus wastes into value-added products: Economic and environmently friendly approaches. Nutrition , 34, 29-46. https://doi.org/10.1016/j.nut.2016.09.006 United Nations Environment Programme (UNEP) (2021). Food waste index Report 2021 Nairobi. 99 p. ISBN 9789280738681. Wang, X., & Lu, X. (2014). Characterization of pectic polysaccharides extracted from apple pomace by hot-compressed water. Carbohydrate Polymers , 102, 174–184. https://doi.org/10.1016/j.carbpol.2013.11.012 Wang, W. J., Ma, X. B., Xu, Y. T., Cao, Y. Q., Jiang, Z. M., & Ding, T. (2015). Ultrasound-assisted heating extraction of pectin from grapefruit peel: Optimization and comparison with the conventional method. Food Chemistry , 178, 106–114. https://doi.org/10.1016/j.foodchem.2015.01.080 Wang, W. J., Ma, X., Jiang, P., Hu, L., Zhi, Z., Chen, J., & Liu, D. (2016). Characterization of pectin from grapefruit peel: A comparison of ultrasound-assisted and conventional heating extractions. Food Hydrocolloids , 61, 730–739. https://doi.org/10.1016/j.foodhyd.2016.06.019 Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-4637293","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":327841076,"identity":"385f9ecb-567a-400e-8e96-3bcfb14cfc41","order_by":0,"name":"Natalia A. Di Clemente","email":"data:image/png;base64,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","orcid":"","institution":"Centro de Investigación y Desarrollo en Criotecnología de Alimentos","correspondingAuthor":true,"prefix":"","firstName":"Natalia","middleName":"A. Di","lastName":"Clemente","suffix":""},{"id":327841077,"identity":"b7839574-8a2c-4224-ac40-48f7ef9723a5","order_by":1,"name":"Enzo La Cava","email":"","orcid":"","institution":"Universidad Nacional del Nordeste (UNNE) and Instituto de Química Básica y Aplicada del Nordeste Argentino (IQUIBA-NEA) UNNE-CONICET","correspondingAuthor":false,"prefix":"","firstName":"Enzo","middleName":"La","lastName":"Cava","suffix":""},{"id":327841078,"identity":"e73e040d-ccd5-4362-939b-5be426035d7d","order_by":2,"name":"Sonia Sgroppo","email":"","orcid":"","institution":"Universidad Nacional del Nordeste (UNNE) and Instituto de Química Básica y Aplicada del Nordeste Argentino (IQUIBA-NEA) UNNE-CONICET","correspondingAuthor":false,"prefix":"","firstName":"Sonia","middleName":"","lastName":"Sgroppo","suffix":""},{"id":327841079,"identity":"63c27246-d24e-4bdc-83eb-baa27cc4f294","order_by":3,"name":"Andrea Gomez-Zavaglia","email":"","orcid":"","institution":"Centro de Investigación y Desarrollo en Criotecnología de Alimentos","correspondingAuthor":false,"prefix":"","firstName":"Andrea","middleName":"","lastName":"Gomez-Zavaglia","suffix":""},{"id":327841080,"identity":"77a622a7-3f75-4afa-98c0-b95f7c2d2cba","order_by":4,"name":"Esteban Gerbino","email":"","orcid":"","institution":"Centro de Investigación y Desarrollo en Criotecnología de Alimentos","correspondingAuthor":false,"prefix":"","firstName":"Esteban","middleName":"","lastName":"Gerbino","suffix":""}],"badges":[],"createdAt":"2024-06-25 14:47:52","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4637293/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4637293/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":60671761,"identity":"286dbcdb-3790-4461-a788-c0ce97cb6ba8","added_by":"auto","created_at":"2024-07-19 10:23:50","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":344207,"visible":true,"origin":"","legend":"\u003cp\u003eSystem boundaries, inputs and outputs of the processes.\u003c/p\u003e","description":"","filename":"Figure1.png","url":"https://assets-eu.researchsquare.com/files/rs-4637293/v1/1910abee63f2c9807b05c139.png"},{"id":60672673,"identity":"517f4d7f-7948-43c9-b51f-74d7c89cf9fa","added_by":"auto","created_at":"2024-07-19 10:31:50","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":54909,"visible":true,"origin":"","legend":"\u003cp\u003eEnvironmental impact contributions of each stage of the process for Scenario 1 (a) and Scenario 2 (b).\u003c/p\u003e","description":"","filename":"Figure2.png","url":"https://assets-eu.researchsquare.com/files/rs-4637293/v1/81f5ebb40f559f1adf0b5a1b.png"},{"id":60671758,"identity":"293ad0ed-07f3-408f-a247-4a0dff46e02e","added_by":"auto","created_at":"2024-07-19 10:23:50","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":32090,"visible":true,"origin":"","legend":"\u003cp\u003eComparison of environmental impacts for Scenario 1 (CHE) and Scenario 2 (TS).\u003c/p\u003e","description":"","filename":"Figure3.png","url":"https://assets-eu.researchsquare.com/files/rs-4637293/v1/eb1a7e7aa719bcc610e26150.png"},{"id":60671759,"identity":"82736198-6112-427c-a55c-2a149e0232d6","added_by":"auto","created_at":"2024-07-19 10:23:50","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":15217,"visible":true,"origin":"","legend":"\u003cp\u003eComparison of the environmental performance of Scenarios 1 and 2 for principal impact categories.\u003c/p\u003e","description":"","filename":"Figure4.png","url":"https://assets-eu.researchsquare.com/files/rs-4637293/v1/a9b7019ee5e22b10d12af59c.png"},{"id":60671760,"identity":"70c9751d-b006-400c-943c-6ef0cc99a9fb","added_by":"auto","created_at":"2024-07-19 10:23:50","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":27010,"visible":true,"origin":"","legend":"\u003cp\u003eEnvironmental profile of Scenarios 1 and 2 for each midpoint.\u003c/p\u003e","description":"","filename":"Figure5.png","url":"https://assets-eu.researchsquare.com/files/rs-4637293/v1/2860b9453cbe9cc6f2a06f9f.png"},{"id":67224737,"identity":"070e0721-65f0-45b2-90e8-4441291fe056","added_by":"auto","created_at":"2024-10-22 15:02:21","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1084857,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4637293/v1/18852868-2fd5-4da1-8bfe-e8075bffb8db.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"\u003cp\u003eSustainability Assessment of Pectin Extraction From Citrus Paradisi Peel and Application in Encapsulating Lactiplantibacillus Plantarum Cidca 83114\u003c/p\u003e","fulltext":[{"header":"1. INTRODUCTION","content":"\u003cp\u003eAccording to (FAO, \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2019\u003c/span\u003e), about 14% of food is lost between harvest and distribution at a global level, and a further 17% is wasted during distribution and at the final consumer (UNEP, 2021). Approximately 40\u0026ndash;50% of fruit and vegetable production is lost or wasted. These losses come from peel fractions, pulps, pomace, and seeds, representing about 16% of total food waste and contributing to around 6% of global greenhouse gas emissions. To promote sustainable food production, several agro-industrial wastes (peels, pulps, pomace, seeds) are being recovered to extract different bioactive compounds, including fiber (pectins, prebiotic oligosaccharides), polyphenols and pigments (Cassani \u0026amp; Gomez-Zavaglia, \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Bioactives from fruit and vegetable wastes and losses have been extensively researched, particularly regarding the implementation of high-yield and environmentally friendly extraction methods (Riyamol et al., \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2023\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe worldwide production of grapefruit [\u003cem\u003eCitrus paradisi\u003c/em\u003e (Macf.)] is estimated up slightly to 6.9\u0026nbsp;million tons, being China, the main productor forecast up slightly to 5.2\u0026nbsp;million tons (period 2023/24) (Foreign Agricultural Service/USDA, 2024, p. 6). Citrus peel is an agricultural waste from juice production, traditionally disposed of as solid waste or animal feed (Grassino et al., \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2016\u003c/span\u003e), and accounts for almost 50% of the wet fruit mass after juicing (Sharma et al., \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). One of the most important bioactives found in grapefruit peel is pectin, a polysaccharide composed of galacturonic acid units (Saini et al, \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Conventional methods for pectin extraction use an acidic or basic aqueous medium at high temperatures (Emaga et al., \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2008\u003c/span\u003e; Geerkens et al, \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). These processes are time consuming and energy intensive and result in serious environmental problems due to the production of acidic wastewater and corrosion of the equipment (Riyamol et al., \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Recently, pectin has been increasingly used in the food industry for developing various products as a biomaterial in the construction and application of micro/nano delivery systems for nutritional or pharmaceutical purposes (Liu et al, \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Ghibaudo et al. (\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2017\u003c/span\u003e) used pectate-iron beads for encapsulation of probiotic \u003cem\u003eLactobacillus plantarum\u003c/em\u003e CIDCA 83114, providing a source of iron and fibre. Similarly, pectate-calcium beads from citrus pectin were investigated as encapsulants of \u003cem\u003eLactobacillus plantarum\u003c/em\u003e CIDCA 83114 (La Cava et al., \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). According to a recent report, the global pectin market reached US\u003cspan\u003e$\u003c/span\u003e 1,170.8\u0026nbsp;million in 2023 and is forecast to grow to US\u003cspan\u003e$\u003c/span\u003e 2,256\u0026nbsp;million by 2032 (IMARC, \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). There is therefore a need for better extraction methods to maximize pectin yield and purity. Alternative green approaches, such as enzyme-assisted extraction (Lim et al., \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2012\u003c/span\u003e), subcritical fluid extraction (Pattarapisitporn, et al., \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2024\u003c/span\u003e), ultrasound-assisted extraction (Wang et al, 2014; \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; La Cava et al, \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Polanco-Lugo et al.,2019), microwave-based extraction (Attard et al., \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Liu et al., \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2017\u003c/span\u003e) and ohmic heating assisted extraction (Saberian et al., \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Sabanci et al., \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2021\u003c/span\u003e) allow for a high yield of pectin extraction with the advantages of short extraction time, low solvent consumption and low environmental impact. In addition, the biodegradability and low toxicity of grapefruit pectin make it a promising biomaterial for micro/nano delivery applications, enhancing bioactivity and encapsulation ability (Liu et al, \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). While numerous studies indicate that green extractions are effective in terms of extraction yield, pectin quality and reduced environmental impact, little research has been done on the actual value of their environmental impact.\u003c/p\u003e \u003cp\u003eLife cycle assessment (LCA) is a valuable tool to evaluate and identify environmental impacts during product development or processing, considering resources and energy balances (Roy et al., \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2008\u003c/span\u003e; Jacquemin et al., \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Ott et al., \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). The goal of this study was to assess the environmental performance of different pectin extraction processes (conventional heating and thermosonication) and its application as delivery systems in the encapsulation of \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e CIDCA 83114.\u003c/p\u003e"},{"header":"2. MATERIALS AND METHODS","content":"\u003cp\u003eDifferent pink/red and white grapefruit (\u003cem\u003eCitrus paradisi\u003c/em\u003e Macf.) varieties, with a Brix/acid ratio of 5.5, uniform skin coloration and free of cuts, were provided by INTA Experimental Station in Bella Vista (Corrientes, Argentina 28\u0026deg; 30\u0026apos; 52.43\u0026apos;\u0026apos; N, 59\u0026deg; 1\u0026apos; 47.94\u0026apos;\u0026apos; S). Different citric combinations of \u0026quot;Parana\u0026quot; on different rootstocks (\u0026ldquo;Citrumelo Swingle,\u0026rdquo; \u0026ldquo;Duncan,\u0026rdquo; \u0026ldquo;Tangelo Orlando\u0026rdquo;, \u0026ldquo;Lima Rangpur\u0026rdquo;) were analyzed. The red/pink varieties investigated included Star Ruby\u0026rdquo; (pink), \u0026ldquo;Foster\u0026rdquo; (red), \u0026ldquo;Red Blush\u0026rdquo; (pink), and \u0026ldquo;Red Shambar\u0026rdquo; (pink).\u003c/p\u003e\n\u003cp\u003eSulfuric acid, hydrochloric acid, sodium hydroxide and calcium chloride (Cicarelli, Argentina) and MRS broth (Biokar Diagnostics, France) were used.\u003c/p\u003e\n\u003cdiv id=\"Sec3\"\u003e\n \u003ch2\u003e2.1 Description of the unit processes\u003c/h2\u003e\n \u003cp\u003eThe stages of the process flow are described in Fig. \u003cspan\u003e1\u003c/span\u003e. Grapefruit peel preparation. Fruits were washed with tap water and sanitized with hypochlorite acid solution (HClO, 200 mg/L) for 5 min. Peel was separated and macerated in a water bath at 90\u0026deg;C for 5 min to inactivate enzymes and cut in 1 cm\u003csup\u003e2\u003c/sup\u003e-slices. Then, peels were dried in an oven under vacuum at 50\u0026deg;C overnight and milled for their homogenization. To extract pectin from the peels, deionized water adjusted with 2N H\u003csub\u003e2\u003c/sub\u003eSO\u003csub\u003e4\u003c/sub\u003e to reach pH 1.5 was used as the extraction solvent.\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e2.1.1 Pectin extraction.\u003c/strong\u003e Two different methods were considered: conventional heating (CHE) and thermosonication (TS) according to La Cava et al. (\u003cspan\u003e2018\u003c/span\u003e). A Precytec AE-29 (CABA, Argentina) stirrer was used for CHE extraction at 90\u0026deg;C for 150 min. TS was conducted in a 6 L-ultrasonic tank (Cleanson model KT 1106, Argentina. 200 W, 40 KHz) with thermostatic control at 60\u0026deg;C for 30 min under periodic agitation (10 s on, 50 s off).\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e2.1.2 Purification.\u003c/strong\u003e Filtered and extracted pectin were washed and precipitated with ethanol solution (96%) before final filtration in a nylon cloth.\u003cspan\u003e\u003cstrong\u003e2.1.2 Pectin hydrolysates preparation.\u003c/strong\u003e HCl solution (0.2 N) was applied for 2 hours at 110\u0026deg;C for chemical hydrolysis, and the pH of the hydrolysates was adjusted to 6.5 with 1N NaOH, according to La Cava et al. (\u003cspan\u003e2019\u003c/span\u003e). Finally, hydrolysates were concentrated by rotary evaporation (Rotavapor\u0026reg; RE 120 \u0026ndash; Buchi, Flawil, Switzerland) under reduced pressure at 50\u0026deg;C.\u003cbr\u003e\u003c/span\u003e\u003cspan\u003e\u003cstrong\u003e2.1.3 Fermentation.\u003c/strong\u003e\u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e CIDCA 83114 was isolated from kefir grains (Garrote, Abraham, \u0026amp; De Antoni, 2001) and maintained frozen at \u0026minus;\u0026thinsp;80\u0026deg;C in 120 g/L nonfat milk solids (Difco, MI, USA). Microbial cells were activated for 24 hours in MRS broth (de Man, Rogosa, \u0026amp; Sharpe, 1960) at 37\u0026deg;C in aerobic conditions. The resulting culture was inoculated (1% v/v) in fresh MRS broth and incubated in the same conditions. Cultures in the stationary phase were used to inoculate pectin hydrolysates.\u003cbr\u003e\u003c/span\u003e\u003cspan\u003e\u003cstrong\u003e2.1.4 Encapsulation.\u003c/strong\u003e Bacteria were encapsulated by ionotropic gelation, incorporating 4% w/v of fresh hydrolysates into the bacterial cultures and adjusting the pH to 6 with 1N NaOH, while shaking for 30 minutes. A 250 mmol/L CaCl\u003csub\u003e2\u003c/sub\u003e solution was dripped into the hydrolysates using a 0.3 mm needle (\u0026sim;10 \u0026micro;L/drop) under continuous agitation for 30 min, according La Cava et al. (\u003cspan\u003e2019\u003c/span\u003e). The capsules were harvested by filtration through a stainless steel mesh of 0.10 mm, and rinsed three times with distilled water.\u003cbr\u003e\u003c/span\u003e\u003cspan\u003e\u003cstrong\u003e2.1.5 Dehydration and preservation.\u003c/strong\u003e The capsules were frozen at \u0026minus;\u0026thinsp;80\u0026deg;C and freeze-dried for 48 hours on a freeze dryer (CHRIST Equipment, Germany) operating with the condenser at \u0026minus;\u0026thinsp;45\u0026deg;C at a chamber pressure of 0.04 mbar. The obtained samples were stored for 45 days at 4\u0026deg;C and the viability was monitored by plate counting on MRS-agar at days 0, 15, 30, and 45.\u003cbr\u003e\u003c/span\u003e\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec4\"\u003e\n \u003ch2\u003e2.2. LCA methodology\u003c/h2\u003e\n \u003cp\u003e\u003cstrong\u003e2.2.1 Goal and scope\u003c/strong\u003e. Identify the environmental profile of the encapsulating process of \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e CIDCA 83114 in pectin extracts and compare two pectin extraction methods (conventional heating \u003cem\u003evs\u003c/em\u003e thermosonication).\u003c/p\u003e\n \u003cp\u003eFunctional unit: 1 kg of freeze-dried-encapsulated bacteria in pectin extracts obtained by conventional heating or thermosonication.\u003c/p\u003e\u003cstrong\u003e2.2.2 System boundaries.\u003c/strong\u003e A gate-to-gate approach was considered in this laboratory study, assuming the impacts from end of life. The harvesting and transport of grapefruits and the treatment of waste were not considered. The system boundary was set to include only the process of pectin extraction from \u003cem\u003eCitrus paradisi\u003c/em\u003e peel, followed by the encapsulation of \u003cem\u003eL. plantarum\u003c/em\u003e CIDCA 83114 and the dehydration of the encapsulates.\u003cp\u003e\u003cstrong\u003e2.2.3 Life Cycle Assessment software and impact categories.\u003c/strong\u003e The present LCA was carried out using Open LCA software version 2.0.1 modelled with ILCD 2011 method, considering the midpoint indicators listed in Table \u003cspan\u003e1\u003c/span\u003e. For normalization and weighting, the EU27 2010 method was applied.\u003cspan\u003e\u003cstrong\u003e2.2.4 Life cycle inventory: Inventory data.\u003c/strong\u003e Inventory data corresponding to production systems such as electricity, chemicals, tap water, deionized water, and waste treatment were taken from ELCD 3.2 database (Greendelta v2.18 correction20220908). Electrical consumption (MJ) was calculated based on the power ratings of the equipment (bath, grinder, oven, shaker, ultrasonic bath, vacuum pump, rotavapor, autoclave, refrigerant, freezer, freeze-dryer) and their duration of use (hour) (Gerbino et al., 2022). The leakage of refrigerant gas (refrigerant, freezer and freeze-dryer) was calculated according to P\u0026eacute;nicaud et al (2018), using available data from providers. Inputs and outputs for each process, comparing the two pectin extraction methods [conventional heating (Scenario 1) or thermosonication (Scenario 2)] (Table\u0026nbsp;\u003cspan\u003e2\u003c/span\u003e).\u003cbr\u003e\u003c/span\u003e\u003c/p\u003e\n \u003cdiv\u003e\u0026nbsp;\u003ctable id=\"Tab1\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv\u003eTable 1\u003c/div\u003e\n \u003cdiv\u003e\n \u003cp\u003eILCD 2011 midpoint indicators and their abbreviations and units.\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003eImpact categories\u003c/th\u003e\n \u003cth align=\"left\"\u003eAbbreviation\u003c/th\u003e\n \u003cth align=\"left\"\u003eUnit\u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003eAcidification\u003c/td\u003e\n \u003ctd align=\"left\"\u003eAP\u003c/td\u003e\n \u003ctd align=\"left\"\u003emolc H\u003csup\u003e+\u003c/sup\u003e eq\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003eClimate change\u003c/td\u003e\n \u003ctd align=\"left\"\u003eCCP\u003c/td\u003e\n \u003ctd align=\"left\"\u003ekg CO\u003csub\u003e2\u003c/sub\u003e eq\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003eFreshwater ecotoxicity\u003c/td\u003e\n \u003ctd align=\"left\"\u003eFETP\u003c/td\u003e\n \u003ctd align=\"left\"\u003eCTUe\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003eFreshwater eutrophication\u003c/td\u003e\n \u003ctd align=\"left\"\u003eFEP\u003c/td\u003e\n \u003ctd align=\"left\"\u003ekg P eq\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003eHuman toxicity, cancer effects\u003c/td\u003e\n \u003ctd align=\"left\"\u003eHTPce\u003c/td\u003e\n \u003ctd align=\"left\"\u003eCTUh\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003eHuman toxicity, non-cancer effects\u003c/td\u003e\n \u003ctd align=\"left\"\u003eHTPnce\u003c/td\u003e\n \u003ctd align=\"left\"\u003eCTUh\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003eIonizing radiation E (interim)\u003c/td\u003e\n \u003ctd align=\"left\"\u003eIRP E\u003c/td\u003e\n \u003ctd align=\"left\"\u003eCTUe\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003eIonizing radiation HH\u003c/td\u003e\n \u003ctd align=\"left\"\u003eIRP HH\u003c/td\u003e\n \u003ctd align=\"left\"\u003ekBq U235 eq\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003eLand use\u003c/td\u003e\n \u003ctd align=\"left\"\u003eLUP\u003c/td\u003e\n \u003ctd align=\"left\"\u003ekg C deficit\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003eMarine eutrophication\u003c/td\u003e\n \u003ctd align=\"left\"\u003eMEP\u003c/td\u003e\n \u003ctd align=\"left\"\u003ekg N eq\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003eMineral, fossil \u0026amp; ren resource depletion\u003c/td\u003e\n \u003ctd align=\"left\"\u003eMFRP\u003c/td\u003e\n \u003ctd align=\"left\"\u003ekg Sb eq\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003eOzone depletion\u003c/td\u003e\n \u003ctd align=\"left\"\u003eODP\u003c/td\u003e\n \u003ctd align=\"left\"\u003ekg CFC-11 eq\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003eParticulate matter\u003c/td\u003e\n \u003ctd align=\"left\"\u003ePMP\u003c/td\u003e\n \u003ctd align=\"left\"\u003ekg PM2.5 eq\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003ePhotochemical ozone formation\u003c/td\u003e\n \u003ctd align=\"left\"\u003ePOP\u003c/td\u003e\n \u003ctd align=\"left\"\u003ekg NMVOC eq\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003eTerrestrial eutrophication\u003c/td\u003e\n \u003ctd align=\"left\"\u003eTEP\u003c/td\u003e\n \u003ctd align=\"left\"\u003emolc N eq\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003eWater resource depletion\u003c/td\u003e\n \u003ctd align=\"left\"\u003eWDP\u003c/td\u003e\n \u003ctd align=\"left\"\u003em\u003csup\u003e3\u003c/sup\u003e water eq\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003c/div\u003e\u003cbr\u003e\n \u003cdiv\u003e\u0026nbsp;\u003ctable id=\"Tab2\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv\u003eTable 2\u003c/div\u003e\n \u003cdiv\u003e\n \u003cp\u003eInventory data. Inputs and outputs per functional unit for each process comparing two scenarios: conventional heating (Scenario 1) or thermosonication (Scenario 2).\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003eProcess\u003c/th\u003e\n \u003cth align=\"left\"\u003eInputs\u003c/th\u003e\n \u003cth align=\"left\"\u003eScenario 1\u003c/th\u003e\n \u003cth align=\"left\"\u003eScenario 2\u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"5\"\u003e\u003cem\u003eGrapefruit peel preparation\u003c/em\u003e\u003c/td\u003e\n \u003ctd align=\"left\"\u003eFruits (kg)\u003c/td\u003e\n \u003ctd align=\"left\"\u003e9000\u003c/td\u003e\n \u003ctd align=\"left\"\u003e9000\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003eElectricity (MJ)\u003c/td\u003e\n \u003ctd align=\"left\"\u003e3263\u003c/td\u003e\n \u003ctd align=\"left\"\u003e3263\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003eHClO (L)\u003c/td\u003e\n \u003ctd align=\"left\"\u003e4\u003c/td\u003e\n \u003ctd align=\"left\"\u003e4\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003eTap water (L)\u003c/td\u003e\n \u003ctd align=\"left\"\u003e20000\u003c/td\u003e\n \u003ctd align=\"left\"\u003e20000\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003eDistillated water (L)\u003c/td\u003e\n \u003ctd align=\"left\"\u003e20000\u003c/td\u003e\n \u003ctd align=\"left\"\u003e20000\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"3\"\u003e\u003cem\u003ePectins extraction\u003c/em\u003e\u003c/td\u003e\n \u003ctd align=\"left\"\u003eSulfuric acid (kg)\u003c/td\u003e\n \u003ctd align=\"left\"\u003e2.45\u003c/td\u003e\n \u003ctd align=\"left\"\u003e2.45\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003eDeionized water (L)\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u003cstrong\u003e24\u003c/strong\u003e\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u003cstrong\u003e29\u003c/strong\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003eElectricity (MJ)\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u003cstrong\u003e2025\u003c/strong\u003e\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u003cstrong\u003e450\u003c/strong\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\u003cem\u003ePurification\u003c/em\u003e\u003c/td\u003e\n \u003ctd align=\"left\"\u003eElectricity (MJ)\u003c/td\u003e\n \u003ctd align=\"left\"\u003e223\u003c/td\u003e\n \u003ctd align=\"left\"\u003e223\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003eEthanol (kg)\u003c/td\u003e\n \u003ctd align=\"left\"\u003e4\u003c/td\u003e\n \u003ctd align=\"left\"\u003e4\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"3\"\u003e\u003cem\u003ePectin Hydrolysates\u003c/em\u003e\u003c/td\u003e\n \u003ctd align=\"left\"\u003eDeionized water (L)\u003c/td\u003e\n \u003ctd align=\"left\"\u003e8998\u003c/td\u003e\n \u003ctd align=\"left\"\u003e8998\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003eElectricity (MJ)\u003c/td\u003e\n \u003ctd align=\"left\"\u003e13188\u003c/td\u003e\n \u003ctd align=\"left\"\u003e13188\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003eHydrogen chloride (Kg)\u003c/td\u003e\n \u003ctd align=\"left\"\u003e1.8\u003c/td\u003e\n \u003ctd align=\"left\"\u003e1.8\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"3\"\u003e\u003cem\u003eFermentation\u003c/em\u003e\u003c/td\u003e\n \u003ctd align=\"left\"\u003eBacteria (kg)\u003c/td\u003e\n \u003ctd align=\"left\"\u003e25\u003c/td\u003e\n \u003ctd align=\"left\"\u003e25\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003eDeionized water (L)\u003c/td\u003e\n \u003ctd align=\"left\"\u003e1000\u003c/td\u003e\n \u003ctd align=\"left\"\u003e1000\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003eElectricity (MJ)\u003c/td\u003e\n \u003ctd align=\"left\"\u003e2772\u003c/td\u003e\n \u003ctd align=\"left\"\u003e2772\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"3\"\u003e\u003cem\u003eEncapsulating\u003c/em\u003e\u003c/td\u003e\n \u003ctd align=\"left\"\u003eDeionized water (L)\u003c/td\u003e\n \u003ctd align=\"left\"\u003e2000\u003c/td\u003e\n \u003ctd align=\"left\"\u003e2000\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003eCaC\u003csub\u003el2\u003c/sub\u003e (kg)\u003c/td\u003e\n \u003ctd align=\"left\"\u003e28\u003c/td\u003e\n \u003ctd align=\"left\"\u003e28\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003eElectricity (MJ)\u003c/td\u003e\n \u003ctd align=\"left\"\u003e338\u003c/td\u003e\n \u003ctd align=\"left\"\u003e338\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\u003cem\u003eDehydration and preservation\u003c/em\u003e\u003c/td\u003e\n \u003ctd align=\"left\"\u003eElectricity (MJ)\u003c/td\u003e\n \u003ctd align=\"left\"\u003e1041\u003c/td\u003e\n \u003ctd align=\"left\"\u003e1041\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003eRefrigerated gas (kg)\u003c/td\u003e\n \u003ctd align=\"left\"\u003e404\u003c/td\u003e\n \u003ctd align=\"left\"\u003e404\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u003cstrong\u003eOutputs\u003c/strong\u003e\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\u003cem\u003eGrapefruit peel preparation\u003c/em\u003e\u003c/td\u003e\n \u003ctd align=\"left\"\u003eLiquid waste (L)\u003c/td\u003e\n \u003ctd align=\"left\"\u003e40000\u003c/td\u003e\n \u003ctd align=\"left\"\u003e40000\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003eFruit waste (kg)\u003c/td\u003e\n \u003ctd align=\"left\"\u003e8000\u003c/td\u003e\n \u003ctd align=\"left\"\u003e8000\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\u003cem\u003ePectins extraction\u003c/em\u003e\u003c/td\u003e\n \u003ctd align=\"left\"\u003eOrganic waste (kg)\u003c/td\u003e\n \u003ctd align=\"left\"\u003e547\u003c/td\u003e\n \u003ctd align=\"left\"\u003e547\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003eWaste water (kg)\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u003cstrong\u003e-\u003c/strong\u003e\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u003cstrong\u003e5\u003c/strong\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\u003cem\u003ePurification\u003c/em\u003e\u003c/td\u003e\n \u003ctd align=\"left\"\u003eOrganic waste (kg)\u003c/td\u003e\n \u003ctd align=\"left\"\u003e410\u003c/td\u003e\n \u003ctd align=\"left\"\u003e410\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003eChemical waste (kg)\u003c/td\u003e\n \u003ctd align=\"left\"\u003e40\u003c/td\u003e\n \u003ctd align=\"left\"\u003e40\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u003cem\u003ePectin Hydrolysates\u003c/em\u003e\u003c/td\u003e\n \u003ctd align=\"left\"\u003eChemical waste (kg)\u003c/td\u003e\n \u003ctd align=\"left\"\u003e9078\u003c/td\u003e\n \u003ctd align=\"left\"\u003e9078\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u003cem\u003eFermentation\u003c/em\u003e\u003c/td\u003e\n \u003ctd align=\"left\"\u003eWaste water (kg)\u003c/td\u003e\n \u003ctd align=\"left\"\u003e1000\u003c/td\u003e\n \u003ctd align=\"left\"\u003e1000\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\u003cem\u003eEncapsulating\u003c/em\u003e\u003c/td\u003e\n \u003ctd align=\"left\"\u003eOrganic waste (kg)\u003c/td\u003e\n \u003ctd align=\"left\"\u003e999\u003c/td\u003e\n \u003ctd align=\"left\"\u003e999\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003eChemical waste (kg)\u003c/td\u003e\n \u003ctd align=\"left\"\u003e2000\u003c/td\u003e\n \u003ctd align=\"left\"\u003e2000\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\u003cem\u003eDehydration and preservation\u003c/em\u003e\u003c/td\u003e\n \u003ctd align=\"left\"\u003eRefrigerant leakage (kg)\u003c/td\u003e\n \u003ctd align=\"left\"\u003e6\u003c/td\u003e\n \u003ctd align=\"left\"\u003e6\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003eFreeze-drying encapsulating bacteria (kg)\u003c/td\u003e\n \u003ctd align=\"left\"\u003e1\u003c/td\u003e\n \u003ctd align=\"left\"\u003e1\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003c/div\u003e\n \u003cp\u003eSensitivity analysis was done for some of the input parameters (fruit, electricity, refrigerant), that did not cause significant changes to the results.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec5\"\u003e\n \u003ch2\u003e2.3. Antioxidant capacity\u003c/h2\u003e\n \u003cp\u003eAs the different scenarios/extraction methods can affect the quality of pectin, antioxidant capacity was determined as a functional property using the DPPH\u003csup\u003e\u0026bull;\u003c/sup\u003e (2,2-Diphenyl-1-icrylhydrazyl) and ABTS\u003csup\u003e\u0026bull;+\u003c/sup\u003e [2,2-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid)] assays [Magalh\u0026atilde;es et al. 2012]. The antioxidant capacity was expressed as EC50% (effective concentration), with a lower value indicating greater antioxidative activity. The EC50% values were calculated by non-linear regression of plots where the abscissa represented the concentration of the tested samples, and the ordinate, the average percentage of antioxidant activity.\u003c/p\u003e\n\u003c/div\u003e"},{"header":"3. RESULTS AND DISCUSSION","content":"\u003cp\u003eEnvironmental profiles of the encapsulation process of \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e CIDCA 83114 in pectin extracts are shown in Fig.\u0026nbsp;\u003cspan\u003e2\u003c/span\u003e for two scenarios: CHE (A) and TS (B) extractions. The percentage contribution of each stage in the process to the total impact revealed that grapefruit peel preparation and pectin hydrolysates were the hotspots of both scenarios, in addition to pectin extraction when CHE is used (Fig.\u0026nbsp;\u003cspan\u003e2\u003c/span\u003ea). Grapefruit peel preparation contributed mainly to HTPnce with 64.9% and 65.4% in Scenario 1 and Scenario 2, respectively, and 62% to ODP and WDP in both scenarios. Pectin hydrolysates process mainly contributed to MFRP 46.4% and 48.5% in Scenario 1and Scenario 2, respectively. These contributions are related to the water consumption, organic and chemical waste generated by these processes (Fig.\u0026nbsp;\u003cspan\u003e1\u003c/span\u003e). In Scenario 1, pectin extraction by CHE contributed particularly to MRFP (5.68%), CCP (3.10%) and HTPce (2.65%), influenced by the use of time and equipment. In Scenario 2, using TS, all the impact categories contributed less than 1.5%.\u003c/p\u003e\n\u003cp\u003eTable\u0026nbsp;3 shows the raw values of the impact categories for Scenarios 1 and 2 and the percentage relation between them. Figure\u0026nbsp;\u003cspan\u003e3\u003c/span\u003e represents the relative percentage normalized values of each impact category for both scenarios. According to the results presented in Table\u0026nbsp;3, Scenario 1 had the major environmental impact in all impact categories as positive values, except in IRP E. However, the total contribution, calculated in millipoints, showed similar values in both scenarios (CHE\u0026thinsp;=\u0026thinsp;19.1 and TS\u0026thinsp;=\u0026thinsp;18.9 mPt).\u003c/p\u003e\n\u003cp\u003eFigure\u0026nbsp;\u003cspan\u003e3\u003c/span\u003e presents the comparative profile of the environmental impact of the encapsulation process of \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e CIDCA 83114 using two pectin extracts methods. The principal impact categories affected were acidification (AP), climate change (CCP), human toxicity cancer effects (HTPce), ionizing radiation HH (IRP HH), photochemical ozone formation (POP) and water resource depletion (WDP). The weighted results of the five principal impact categories were expressed in mPt for both scenarios (Fig.\u0026nbsp;\u003cspan\u003e4\u003c/span\u003e). The consumption of electricity during the production of deionized water and the fuel consumption for electricity production are associated with higher CO\u003csub\u003e2\u003c/sub\u003e emissions into the atmosphere, as well as increased emissions of nitrogen and sulfur oxides, leading to climate change and acidification impacts, respectively (Marzeddu et al., \u003cspan\u003e2021\u003c/span\u003e). Consequently, volatile organic compounds (VOCs) were released, increasing the POP impact (Alisha, et al., 2019) which is correlated with human health impacts (Shajed \u0026amp; Jolliet, 2019). Beccali et al (\u003cspan\u003e2009\u003c/span\u003e) reported energy consumption as a hotspot in the eco-profiles of six citrus-based products in Italy, related to CO\u003csub\u003e2\u003c/sub\u003e emissions and water consumption. Deionized water and electricity consumption were mainly responsible for the environmental impact of Scenario 1 and Scenario 2, as shown in the Sankey diagram, where the thick lines represent the highest impact flux. (Fig.\u0026nbsp;\u003cspan\u003e1\u003c/span\u003eS. 2S, 3S).\u003c/p\u003e\n\u003cp\u003eFigure\u0026nbsp;\u003cspan\u003e5\u003c/span\u003e shows the relative categories results of both scenarios. For each impact categories, the maximum result is set to 100%, corresponding to Scenario 1 (red) and the results of the Scenario 2 (blue) are displayed in relation to this result. As mentioned above, the environmental impact of Scenario 1 is higher than that of Scenario 2, but the differences do not exceed 6%. However, CHE (Scenario 1) showed a better yield (4.1\u0026thinsp;\u0026plusmn;\u0026thinsp;1.7%) than TS (Scenario 2) (1.5\u0026thinsp;\u0026plusmn;\u0026thinsp;0.4%), corresponding to the galacturonic acid concentration, which was higher for CHE (786.4\u0026thinsp;\u0026plusmn;\u0026thinsp;58.2 g/kg) than for TS (605.9\u0026thinsp;\u0026plusmn;\u0026thinsp;91.8 g/kg) (La Cava et al., \u003cspan\u003e2018\u003c/span\u003e). In relation to antioxidant activity (Table\u0026nbsp;\u003cspan\u003e4\u003c/span\u003e), pectin extraction by CHE had the best performance. This suggests that the extraction method strongly determines the antioxidant capacity of the samples (Bozinou et al., \u003cspan\u003e2019\u003c/span\u003e). In this regard, it has been reported that the acoustic cavitation generated during ultrasound treatment may lead to the production of free radicals, thus explaining the lower antioxidant capacity of TS extracts (Piyasena, Mohareb, \u0026amp; McKellar, \u003cspan\u003e2003\u003c/span\u003e).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 3.\u003c/strong\u003e Impact assessment values for 1 kg of protected/dry-encapsulated bacteria in pectin\u003c/p\u003e\n\u003cp\u003eextracts obtained by conventional heating (Scenario 1) or thermosonication (Scenario 2)\u003c/p\u003e\n\u003cp\u003eand relative environmental difference between both scenarios.\u003c/p\u003e\n\u003cdiv\u003e\n \u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\" width=\"499\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd width=\"26.452905811623246%\"\u003e\n \u003cp\u003e\u003cstrong\u003eImpact categories\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20.841683366733466%\"\u003e\n \u003cp\u003e\u003cstrong\u003eScenario 1\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.43486973947896%\"\u003e\n \u003cp\u003e\u003cstrong\u003eScenario 2\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"35.27054108216433%\"\u003e\n \u003cp\u003e\u003cstrong\u003eScenario 1/Scenario 2 (%)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"26.452905811623246%\"\u003e\n \u003cp\u003eAP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20.841683366733466%\" valign=\"bottom\"\u003e\n \u003cp\u003e1.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.43486973947896%\" valign=\"bottom\"\u003e\n \u003cp\u003e1.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"35.27054108216433%\" valign=\"top\"\u003e\n \u003cp\u003e0.323\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"26.452905811623246%\"\u003e\n \u003cp\u003eCCP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20.841683366733466%\" valign=\"bottom\"\u003e\n \u003cp\u003e449.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.43486973947896%\" valign=\"bottom\"\u003e\n \u003cp\u003e439.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"35.27054108216433%\" valign=\"top\"\u003e\n \u003cp\u003e2.365\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"26.452905811623246%\"\u003e\n \u003cp\u003eFETP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20.841683366733466%\" valign=\"bottom\"\u003e\n \u003cp\u003e15.9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.43486973947896%\" valign=\"bottom\"\u003e\n \u003cp\u003e15.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"35.27054108216433%\" valign=\"top\"\u003e\n \u003cp\u003e2.057\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"26.452905811623246%\"\u003e\n \u003cp\u003eFEP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20.841683366733466%\" valign=\"bottom\"\u003e\n \u003cp\u003e5.3\u0026times;10\u003csup\u003e-4\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.43486973947896%\" valign=\"bottom\"\u003e\n \u003cp\u003e5.3\u0026times;10\u003csup\u003e-4\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"35.27054108216433%\" valign=\"top\"\u003e\n \u003cp\u003e0.101\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"26.452905811623246%\"\u003e\n \u003cp\u003eHTPce\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20.841683366733466%\" valign=\"bottom\"\u003e\n \u003cp\u003e1.2\u0026times;10\u003csup\u003e-6\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.43486973947896%\" valign=\"bottom\"\u003e\n \u003cp\u003e1.2\u0026times;10\u003csup\u003e-6\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"35.27054108216433%\" valign=\"top\"\u003e\n \u003cp\u003e2.018\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"26.452905811623246%\"\u003e\n \u003cp\u003eHTPnce\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20.841683366733466%\" valign=\"bottom\"\u003e\n \u003cp\u003e9.2\u0026times;10\u003csup\u003e-6\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.43486973947896%\" valign=\"bottom\"\u003e\n \u003cp\u003e9.1\u0026times;10\u003csup\u003e-6\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"35.27054108216433%\" valign=\"top\"\u003e\n \u003cp\u003e0.776\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"26.452905811623246%\"\u003e\n \u003cp\u003eIRP E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20.841683366733466%\" valign=\"bottom\"\u003e\n \u003cp\u003e3.2\u0026times;10\u003csup\u003e-4\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.43486973947896%\" valign=\"bottom\"\u003e\n \u003cp\u003e3.2\u0026times;10\u003csup\u003e-4\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"35.27054108216433%\" valign=\"top\"\u003e\n \u003cp\u003e-0.194\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"26.452905811623246%\"\u003e\n \u003cp\u003eIRP HH\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20.841683366733466%\" valign=\"bottom\"\u003e\n \u003cp\u003e32.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.43486973947896%\" valign=\"bottom\"\u003e\n \u003cp\u003e32.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"35.27054108216433%\" valign=\"top\"\u003e\n \u003cp\u003e0.161\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"26.452905811623246%\"\u003e\n \u003cp\u003eLUP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20.841683366733466%\" valign=\"bottom\"\u003e\n \u003cp\u003e0.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.43486973947896%\" valign=\"bottom\"\u003e\n \u003cp\u003e0.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"35.27054108216433%\" valign=\"top\"\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"26.452905811623246%\"\u003e\n \u003cp\u003eMEP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20.841683366733466%\" valign=\"bottom\"\u003e\n \u003cp\u003e0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.43486973947896%\" valign=\"bottom\"\u003e\n \u003cp\u003e0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"35.27054108216433%\" valign=\"top\"\u003e\n \u003cp\u003e0.501\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"26.452905811623246%\"\u003e\n \u003cp\u003eMFRP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20.841683366733466%\" valign=\"bottom\"\u003e\n \u003cp\u003e5.2\u0026times;10\u003csup\u003e-5\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.43486973947896%\" valign=\"bottom\"\u003e\n \u003cp\u003e5.0\u0026times;10\u003csup\u003e-5\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"35.27054108216433%\" valign=\"top\"\u003e\n \u003cp\u003e4.390\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"26.452905811623246%\"\u003e\n \u003cp\u003eODP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20.841683366733466%\" valign=\"bottom\"\u003e\n \u003cp\u003e2.7\u0026times;10\u003csup\u003e-5\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.43486973947896%\" valign=\"bottom\"\u003e\n \u003cp\u003e2.7\u0026times;10\u003csup\u003e-5\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"35.27054108216433%\" valign=\"top\"\u003e\n \u003cp\u003e0.119\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"26.452905811623246%\"\u003e\n \u003cp\u003ePMP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20.841683366733466%\" valign=\"bottom\"\u003e\n \u003cp\u003e0.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.43486973947896%\" valign=\"bottom\"\u003e\n \u003cp\u003e0.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"35.27054108216433%\" valign=\"top\"\u003e\n \u003cp\u003e0.191\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"26.452905811623246%\"\u003e\n \u003cp\u003ePOP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20.841683366733466%\" valign=\"bottom\"\u003e\n \u003cp\u003e0.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.43486973947896%\" valign=\"bottom\"\u003e\n \u003cp\u003e0.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"35.27054108216433%\" valign=\"top\"\u003e\n \u003cp\u003e0.576\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"26.452905811623246%\"\u003e\n \u003cp\u003eTEP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20.841683366733466%\" valign=\"bottom\"\u003e\n \u003cp\u003e2.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.43486973947896%\" valign=\"bottom\"\u003e\n \u003cp\u003e2.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"35.27054108216433%\" valign=\"top\"\u003e\n \u003cp\u003e0.592\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"26.452905811623246%\"\u003e\n \u003cp\u003eWDP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20.841683366733466%\" valign=\"bottom\"\u003e\n \u003cp\u003e5.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"17.43486973947896%\" valign=\"bottom\"\u003e\n \u003cp\u003e5.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"35.27054108216433%\" valign=\"top\"\u003e\n \u003cp\u003e0.004\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\u003cimg src=\"https://myfiles.space/user_files/122228_c8a1650c59388082/122228_custom_files/img1721384296.png\"\u003e\u003cbr\u003e\u003c/p\u003e\n\u003cdiv\u003e \u0026nbsp; \u003c/div\u003e\n\u003cdiv\u003e \u0026nbsp; \u003c/div\u003e\n\u003cp\u003eDifferences in the various components of LCA studies (such as functional units, system boundaries, laboratory or industrial scale and impact assessment methods) will be considered for comparing the presented results with other studies. However, several studies show differences between traditional extraction methods and green ones, with the latter being more efficient and environmentally friendly (Garc\u0026iacute;a-Garc\u0026iacute;a et al., 2019; Pappas et al., \u003cspan\u003e2023\u003c/span\u003e). In agreement with the results of this work, Wang et al. (\u003cspan\u003e2015\u003c/span\u003e) reported that ultrasound-assisted heating extraction was an efficient and economical method to obtain higher pectin yields at lower temperatures and shorter extraction times than CHE from grapefruit peel.\u003c/p\u003e\n\u003cp\u003eDespite a similar environmental impact profile, the energy and time consumption in Scenario 1 were higher than that in Scenario 2, so this approach should be carefully managed when making decisions. Considering that the yield of pectin in Scenario 1 was more than double that of Scenario 2, a life cycle cost analysis should be carried out, also taking into account evaluations on an industrial scale (note that this work was carried out on a laboratory scale). Moreover, specific limitation in terms of collecting local and good quality inventory data is a major challenge.\u003c/p\u003e"},{"header":"4. CONCLUSION","content":"\u003cp\u003eFrom the analysis presented above, it can be concluded that the environmental profiles of the encapsulation process of \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e CIDCA 83114 in pectin extracts were similar when comparing conventional heating with thermosonication extraction method from grapefruit peel waste. The main resources contributing to the impacts are deionized water and electricity consumption. Therefore, it is important to consider using more energy efficient equipment to help achieve a sustainable approach to using energy. Furthermore, the present results, together with previous studies (La Cava et al., \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2019\u003c/span\u003e), contribute to the development of an encapsulated probiotic in pectin as a potential alternative both to overcome the environmental problem of grapefruit peel and as a bioactive compound in the food and pharmaceutical industries.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgments\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis work was supported by the Argentinean Agency for Scientific and Technological Promotion (ANPCyT) (Project PICT/2020/0482 and PICT 2019-2428). E.L.L.C, A.G-Z. and E.G. are members of the research career of CONICET. N. D.C. is a postdoctoral fellow of ANPCyT.\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003cstrong\u003eAuthor\u003c/strong\u003e\u003cstrong\u003e\u0026apos;\u003c/strong\u003e\u003cstrong\u003es Contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eN. D. C conducted the LCA analysis and the preparation of the original draft, as well as writing, reviewing, and editing the final manuscript. E. L: L. C contributed to the experimental work, data curation, reviewing and investigation. E. G. conducted the investigation, participated in LCA data curation, reviewing, and editing the final manuscript. A. G-Z and S. G. resources, reviewing, and editing the final manuscript and also supervised project administration and was responsible for funding acquisition.\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003cstrong\u003eCompeting interests:\u0026nbsp;\u003c/strong\u003eThe authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eAlishah, A., Motevali, A., Tabatabaeekoloor, R., \u0026amp; Hashemi, S. J. (2019). Multiyear life energy and life cycle assessment of orange production in Iran. \u003cem\u003eEnvironmental Science and Pollution Research\u003c/em\u003e, 26(31), 32432\u0026ndash;32445. https://doi.org/10.1007/s11356-019-06344-y \u003c/li\u003e\n\u003cli\u003eAttard, T. M., Watterson, B., Budarin, V. L., Clark, J. H., \u0026amp; Hunt, A. J. (2014). 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Characterization of pectin from grapefruit peel: A comparison of ultrasound-assisted and conventional heating extractions. \u003cem\u003eFood Hydrocolloids\u003c/em\u003e, 61, 730\u0026ndash;739. https://doi.org/10.1016/j.foodhyd.2016.06.019 \u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"encapsulation, Citrus paradisi pectins, lactobacilli, conventional heating, thermosonication, Life Cycle Assessment (LCA)","lastPublishedDoi":"10.21203/rs.3.rs-4637293/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4637293/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eAgro-industrial waste, including peels, pulps, pomace, and seeds, poses a significant global economic and environmental problem. Recovering these wastes to extract bioactive compounds (\u003cem\u003ee.g.\u003c/em\u003e, pecin, polyphenols, pigments, essential oils) offers a sustainable solution. Pectins have been increasingly used as delivery systems in the food industry. Green extractions have been developed to reduce the extraction time and environmental impact of conventional methods. However, little is known about their actual impact. Life Cycle Assessment (LCA) is a useful tool for assessing the environmental and energy impacts of a production cycle. This study aims to evaluate the environmental profile of pectin extraction from grapefruit peels using conventional heating (CHE) and thermosonication (TS) methods, and the application of the extracted pectin as a delivery system for encapsulating \u003cem\u003eLactiplantibacillus plantarum\u003c/em\u003e CIDCA 83114. The LCA was performed using Open LCA software version 2.0.1 modelled with ILCD 2011 method. The system boundaries were considered to be laboratory scale and the functional units were 1 kg of protected/dry encapsulated bacteria in pectin extracts from \u003cem\u003eCitrus paradisi\u003c/em\u003e peel obtained by TS or CHE. The impact scores of the TS and CHE scenarios were similar in terms of millipoints (TS\u0026thinsp;=\u0026thinsp;18.9 and CHE\u0026thinsp;=\u0026thinsp;19.1 mPt). The main impact categories were climate change, human toxicity with carcinogenic effects and depletion of water resources contributing to deionized water and electricity consumption. The obtained results contribute to the decision-making process for the selection of a pectin extraction process on a laboratory scale, complemented by future economic impact studies.\u003c/p\u003e","manuscriptTitle":"Sustainability Assessment of Pectin Extraction From Citrus Paradisi Peel and Application in Encapsulating Lactiplantibacillus Plantarum Cidca 83114","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-07-19 10:23:45","doi":"10.21203/rs.3.rs-4637293/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"3fc721d7-a0a0-46a3-8415-0bcbd3c40123","owner":[],"postedDate":"July 19th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-10-22T14:54:11+00:00","versionOfRecord":[],"versionCreatedAt":"2024-07-19 10:23:45","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-4637293","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4637293","identity":"rs-4637293","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}