Sequestering Banana- A horticulture plant and its leaf wax for its sustainable development goals

Sequestering Banana- A horticulture plant and its leaf wax for its sustainable development goals | Authorea try { document.documentElement.classList.add('js'); } catch (e) { } var _gaq = _gaq || []; _gaq.push(['_setAccount', 'G-8VDV14Y67G']); _gaq.push(['_trackPageview']); (function() { var ga = document.createElement('script'); ga.type = 'text/javascript'; ga.async = true; ga.src = ('https:' == document.location.protocol ? 'https://ssl' : 'http://www') + '.google-analytics.com/ga.js'; var s = document.getElementsByTagName('script')[0]; s.parentNode.insertBefore(ga, s); })(); Skip to main content Preprints Collections Wiley Open Research IET Open Research Ecological Society of Japan All Collections About About Authorea FAQs Contact Us Quick Search anywhere Search for preprint articles, keywords, etc. Search Search ADVANCED SEARCH SCROLL This is a preprint and has not been peer reviewed. Data may be preliminary. 26 February 2025 V1 Latest version Share on Sequestering Banana- A horticulture plant and its leaf wax for its sustainable development goals Author : Shweta S 0000-0001-6639-2201 [email protected] Authors Info & Affiliations https://doi.org/10.22541/au.174054962.29869777/v1 622 views 203 downloads Contents Abstract Supplementary Material Information & Authors Metrics & Citations View Options References Figures Tables Media Share Abstract The potential use of the Banana (Musa paradisiaca L.) leaf which is commonly used in southern parts of India. The leaves are used in various functions, festivals for eating, packing. Most of them goes wasted to the land and gets polluted. The leaves have a wax coating on the surface of the leaf which is natural wax and are esters of fatty acids. This natural wax can be used for various commercial purposes as a coating, cosmetics, food coating, confections, medicines. In the present study the banana leaf wax was extracted and quantified for various studies anticancer, antidiabetic, XRD, IR, UV VIS, TGA, Homo Lumo analysis. The banana leaves were cut into small pieces and with solvent hexane, wax was extracted by reflux method. The precipitated wax was collected and analysed for anticancer, antidiabetic, IR, XRD, UV VIS, TGA, Homo Lumo spectral analysis for characterization. The wax was sent for analysis of anticancer and antidiabetic assay. The results obtained revealed that the anticancer activity exhibited against MDAMB-231 cell-line and results compared with the standard cisplatin drug. The IC50 of cell inhibition of banana wax was found to be 168.07 µg/ml and diabetic alpha amylase activity was measured with the standard acarbose which resulted a positive response with the plant extract 186.02 µg/ml. The presence of Arachidonic acid, Beharic acid, EPA, Lignoceric acid, linoleic acid, linolenic acid, Myristic acid, Oleic acid, Palmitic acid and Stearic acid revealed a significant use in the various useful products. Sequestering Banana- A horticulture plant and its leaf wax for its sustainable development goals Shweta J Sabannavar 1 1 Assistant Professor, Botany Department, K. L. E’s P. C Jabin Science College, Hubballi- 580031, Karnataka, India, [email protected] , +919731102426, ORCID: https://orcid.org/0000-0001-6639-2201 Abstract The potential use of the Banana ( Musa paradisiaca L.) leaf which is commonly used in southern parts of India. The leaves are used in various functions, festivals for eating, packing. Most of them goes wasted to the land and gets polluted. The leaves have a wax coating on the surface of the leaf which is natural wax and are esters of fatty acids. This natural wax can be used for various commercial purposes as a coating, cosmetics, food coating, confections, medicines. In the present study the banana leaf wax was extracted and quantified for various studies anticancer, antidiabetic, XRD, IR, UV VIS, TGA, Homo Lumo analysis. The banana leaves were cut into small pieces and with solvent hexane, wax was extracted by reflux method. The precipitated wax was collected and analysed for anticancer, antidiabetic, IR, XRD, UV VIS, TGA, Homo Lumo spectral analysis for characterization. The wax was sent for analysis of anticancer and antidiabetic assay. The results obtained revealed that the anticancer activity exhibited against MDAMB-231 cell-line and results compared with the standard cisplatin drug. The IC 50 of cell inhibition of banana wax was found to be 168.07 µg/ml and diabetic alpha amylase activity was measured with the standard acarbose which resulted a positive response with the plant extract 186.02 µg/ml. The presence of Arachidonic acid, Beharic acid, EPA, Lignoceric acid, linoleic acid, linolenic acid, Myristic acid, Oleic acid, Palmitic acid and Stearic acid revealed a significant use in the various useful products. Research Implications: These studies can reveal that the banana wax has a significant potential to be explored in various fields. The plant-based waxes are also readily decomposed than the polymer material. The market of wax industry is also expanding and growing at an encouraging rate. It is also noted that natural waxes are increasing in demand, especially plant-based waxes. Utilization of banana wax provides a great benefit by creating a high value product from the unutilized biomass resources of banana cultivation in the tropical developing countries. The present study helps to bring an insight to the new ecofriendly use of banana wax. The wax showed response to the anticancer activity and antidiabetic activity. The presence of various fatty acids leads to conclude that the banana wax can be used in various fields of industries for its sustainable approaches for ecofriendly environment. Keywords: Banana, Wax, commercial purpose, diabetic alpha amylase, anticancer activity, sustainable goals. 1. INTRODUCTION Banana ( Musa paradisiaca L.) belongs to the family Musaceae and is one among the herbaceous plants with thick rhizome, pseudo stem fleshy, succulent formed by the imbricate leaf sheaths and grown in tropical regions. The leaves are large, oblong, petioles long, bright and glossy green leaves (Varsha, 2019). In India the production of bananas is about 30.4 million tons, once the bananas are harvested some leaves are used for various purposes like cooking, wrapping and food serving in tropical and subtropical areas but not all the leaves so many of them gets wasted. The leaves can be used for extraction of wax (Anuj et al ., 2016). The use of banana leaves is dominant over other parts because of its varied health benefits. Many health experts says that the banana leaves have antibacterial properties which kills the germs present in the food, when it is served on the banana leaf. This plant is widely exploited, not only for its fruits, but also for peels, leaves, pseudo–stem, stalk and inflorescence. These are used in various food and non–food applications such as thickening agent, colorant and flavor, an alternative source for macro and micronutrients, nutraceuticals, livestock feed, fibres, and a source of natural bioactive compounds and bio–fertilizers (Padam et al ., 2014). The wax extracted from the leaves can serve as the alternative source for creams, ointments, cosmetics. There are many other different waxes which are used such as Carnauba wax which is extracted from palm plant, bees wax from honey bee, soya wax from soya bee etc. The diabetes is another ailment which is increasing day by day in all the human beings. Currently, the use of modern medicines for glycemic control has caused numerous adverse side effects, resulting in an increasing demand for safe and cost-effective measures (Widyawati 2019, Peesa, 2013). In the same way the cancer is also another chronic disease. The majority of the anticancer medications utilized in chemotherapy are cytotoxic to normal cells (Zandi et al ., 2010). The natural plant-based drugs have been considered has safe and cost effective but for the same, plants have been exploited for the preparation of the drugs. The natural resources will be destroyed in order to extract drugs from plants. An attempt has been made to extract the wax by another natural source as it is more important and needed to save our natural sources for our future generations. 2. THEORETICAL FRAMEWORK The plant leaf waxes contain hydroxy-β-diketones, oxo-βdiketones, alkenes, branched alkanes, acids, esters, acetates and benzoates of aliphatic alcohols, methyl, phenylethyl and triterpenoid esters. This leaf surface wax serves as a mechanical barrier which protects the plant tissues against UV radiation, bacterial or fungal attacks and also reduces water loss in summer time (Yanagida, 2005). The plant-based waxes are also readily decomposed than the polymer material. The market of wax industry is also expanding and growing at an encouraging rate. It is also noted that natural waxes are increasing in demand, especially plant-based waxes (Pashova, 2001). Utilization of banana wax provides a great benefit by creating a high value product from the unutilized biomass resources of banana cultivation in the tropical developing countries. The objectives of this study were to investigate and characterize the chemical composition of banana leaf wax. In this study an attempt has been made to extract wax, characterize the compounds from plant wax. 3. METHODOLOGY The banana wax was extracted by reflux method (. The leaves were cut into small pieces and placed in a round bottom flask containing hexane until all the leaves are immersed in the solvent. The banana leaves and solvent mixture were set on a water bath and maintained at a constant temperature at 60 \RL–֯ C for 10 minutes. A glass condenser was directly connected to the top of the flask. After refluxing, the solvent mixture was collected into the conical flask through the filter. The flask was allowed to cool down at room temperature for 2 hours and placed in a freezer for 6 hours. The precipitated wax was collected and analysed for anticancer, antidiabetic, IR, XRD, UV VIS, TGA, Homo Lumo spectral analysis for characterization. The wax was sent for analysis of anticancer and antidiabetic assay. 3.1 Anticancer assay 3.1.1. MTT assay This assay was carried out in Cytoxon Biosolutions Pvt. Ltd Hubballi, India (Alley et al ., 1986, Ali et al ., 2022). 3.1.2. Cell Culture The cells of the MCF-7(Epithelial cells of breast cancer) lines were culture in DMEM medium (Cytoxon Biosolutions Pvt. Ltd., Hubballi, India) supplemented with 1 % of penicillin and 10 % feta were purchased from NCCS (National Centre For Cell Sciences) Pune. Cells were nourished with DMEM-HG media and kept for incubation at 37 0 C, 5% CO 2 until they reach confluence. Upon the cells reaching 80- 90% confluency, cells were passaged. After passaging cells were used for experiment. MDAMB-231 cells were seeded in 96 well plate at 2.2x104 cells/well. Cells were allowed to attach and grow for 24 hours. Number of wells in 96 well plate was considered depending on experimental setup and concentrations of plant extract considered. Five different concentrations of plant extracts were considered to test the cyto-compatibility. Standard drug (Cisplatin) and untreated cultures were considered as control groups. Experiments were performed in duplicates. The seeded cells observed under microscope and confirmed the attachment of cells and formed a monolayer. After 24 hours of drug treatment cells were taken for performing MMT assay and cytotoxic effects of drug on MDAMB-231 cells was done. Briefly, cells were treated with pre-defined concentration of plant extract and controls for 24 hours. Later, 0.5mg /ml MTT was added to each well and incubated for 3 hours. Post MTT incubation, culture plate was observed under microscope to check the needle like formazon crystals formation. Formazon crystals were dissolved by adding DMSO to cells and supernatant media was read for optical density at 625nm using spectrophotometer. The percentage growth inhibition was calculated using the following formula and concentration of test drug needed to inhibit cell growth by 50% (IC50) values is generated from the dose-response curves for each cell line. 3.2. Alpha-amylase inhibitory assay The Alpha- amylase inhibitory assay of banana extract was evaluated according to a method by Shettar et al ., 2017. In an appendorf tube, 1ml of PBS solution was mixed with 0.5 ml of different concentrations (50,100, 150, 200 and 250 µg/ml) of samples with the standard solution and 200µl of 0.5mg/ml α-amylase was added followed by 200µl of 5mg/ml starch solution and incubated for 10 minutes at room temperature. Control was takes as starch with amylase and without amylase. Then the reaction mixture was stopped by adding 400µl of DNS solution heating the mixture in boiling water bath for 5 min and cooled. The absorbance was measured at 540 nm (Labman UV Visible Spectrophotometer). The percent of enzyme inhibition was calculated using the following formula: % of α-amylase inhibition = [(A c − A s )/A c ] × 100 where, A c and A s are the absorbance of control and sample, respectively. Acarbose was used as standard. 4. RESULTS AND DISCUSSION The anticancer activity revealed that the concentration of the plant extract showed the inhibition of the cancer cells. The MTT assay results revealed morphological features with abnormalities such as apoptotic bodies, cell shrinkage, membrane blabbing, and cell turgidity. All these features of the cells undergoing apoptosis. MTT and microscopic study results show that the test samples inducing apoptosis shown in Table I and Figure 1. The test sample has shown that it might be inducing apoptosis in lower concentrations of plant wax with the MDAMB-231 cell line (Ali et al ., 2022). The banana leaf wax was subjected to α- amylase inhibitory assay along with standard acarbose. The wax showed higher activity. The α-amylase inhibitory activities of differed concentrations are recorded in Table 2, Figure 2 (Lakshmi et al ., 2014). Alpha-amylase is type of the intestinal enzyme which play important role in carbohydrate digestion and glucose absorption. Suppression of the activity of digestive enzymes such as α-amylase, would delay the digestion of starch and oligosaccharides, which in turn decreases the absorption of glucose and consequently reduce the blood glucose. Aqueous extract of banana wax exhibited highest percentage of inhibition (Figure 2). This significant anti-diabetic activity was comparable to the standard drug inhibition. This technique is one of the anti-diabetic therapeutic approaches to reduce the post prandial glucose level in blood by inhibiting activity of alpha-amylase enzyme and it can be used as a strategy in management of blood glucose. The IR Spectroscopy is used to identify the functional groups present in a molecule in Figure 3. Therefore, the characteristic peaks for common functional groups from the spectrum are as described in the Table 3. The absorption peaks revealed the presence of alcohol, carboxylic acid, carbonyl, alkene groups. The results are in findings with the Kanna and Stephen, 2021. The UV/Visible analysis was carried between 300 – 600 nm. The λ max for the wax was found at 411 nm and 469 nm as may be seen in the Figure 4. The UV-Visible spectrum of banana leaf wax peel shows three absorption bands at 411 nm, 469 nm, and 533 nm. These findings are in accordance with (Kaana and Stephen, 2021). These bands can be attributed to the presence of carotenoids in the wax peel. Carotenoids are natural pigments that are found in many fruits and vegetables, including bananas. They are responsible for the yellow, orange, and red colors of these fruits and vegetables. Carotenoids offer health advantages as they function as antioxidants shielding cells, from the effects of free radicals. Free radicals are molecules that can cause cell damage and contribute to the development of ailments like cancer and heart disease. The carotenoids play a role in maintaining vision and supporting immune function. To identify carotenoids in banana leaf wax peel researchers have utilized the UV spectrum analysis method (Smith and Brown, 2009). The specific absorption bands at wavelengths can further aid in identifying the types of carotenoids present. The characteristic absorption band at 411 nm signifies beta carotene while an absorption band at 469 nm indicates lutein. These results indicate the presence of unsaturated compounds. Thermogravimetric analysis is an important technique, which gives idea on the rate of change of mass as a function of time or temperature in the various atmospheres such as nitrogen, oxygen, carbon-di-oxide, hydrogen, etc. The results indicate that the banana wax sample undergoes a significant weight loss of 44.63 % starting at approximately 446.38 °C (Figure 5). This weight loss seems to occur relatively rapidly in a single step. The remaining 55.37% of the sample mass remains stable even at higher temperatures. This information helps the use of banana wax in various fields. The identification of crystallinity of polymers, recognition of crystalline phases (polymorphism), and orientation of polymers can be identified by XRD analysis. It is represented in Figure 6 and the reports are in accordance with Wada, 2001. The SEM analysis of the banana leaf wax indicates the presence of amorphous nature which are in accordance with the Rahmatia, 2022, (Figure 7). The Homo Lumo analysis revealed the Arachidonic acid, Beharic acid, EPA, Lignoceric acid, linoleic acid, linolenic acid, Myristic acid, Oleic acid, Palmitic acid and Stearic acid (Table 4, Figure 8). These acids indicates that the banana wax can be used in the production of various useful products. 5. CONCLUSION The present study helps to bring an insight to the new ecofriendly use of banana wax. The wax showed response to the anticancer activity and antidiabetic activity. The presence of various fatty acids leads to conclude that the banana wax can be used in various fields of industries. 6. ACKNOWLEDGEMENT This work was funded by the Karnataka Science and Technology Academy, Bengaluru, Karnataka, India and the author is very much thankful. 7. CONFLICT OF INTEREST The author declare no competing interests. 8. REFERENCES Ali Mohamed Alshabi., Saad Ahmed Alkahtani., Ibrahim Ahmed Shaikh., Mohamed A.A., Orabi., Basel A. (2022). Journal of King Saud Uni Sci. 34:102238-102245. Alley, M. 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Supplementary Material File (figures.docx) Download 1.83 MB File (tables.docx) Download 33.94 KB Information & Authors Information Version history V1 Version 1 26 February 2025 Copyright This work is licensed under a Non Exclusive No Reuse License. Keywords anticancer commercial purpose diabetic alpha amylase homo lumo wax Authors Affiliations Shweta S 0000-0001-6639-2201 [email protected] P C Jabin Science College View all articles by this author Metrics & Citations Metrics Article Usage 622 views 203 downloads .FvxKWukQNSOunydq8rnd { width: 100px; } Citations Download citation Shweta S. Sequestering Banana- A horticulture plant and its leaf wax for its sustainable development goals. Authorea . 26 February 2025. DOI: https://doi.org/10.22541/au.174054962.29869777/v1 If you have the appropriate software installed, you can download article citation data to the citation manager of your choice. Simply select your manager software from the list below and click Download. 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