Comparison of the effect of hydroethanolic and aqueous solvents on functional potential and evaluation of active compounds in pumpkin extract (Cucurbita moschata) by GC/MS method
Subject Areas : Food Science and Technology
Z. Latifi
1
,
S. Abediankenari
2
,
Aliakbar Mashayekh
3
1 - Ph.D. Student of Food Science and Technology, Islamic Azad University, Nour Branch, Mazandaran, Iran
2 - Professor of immunology (phD), Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran
3 - Assistant Professor of Physical Education and Sport Sciences, Faculty of Humanities and Social Sciences, Islamic Azad University, Nour, Iran
Keywords: Hydroethanolic extract, Aqueous extract, Bioactive compounds, Pumpkin,
Abstract :
Pumpkin (Cucurbita moschata) is one of the vegetables that has high nutritional value and bioactive compounds. The aim of this study was to compare the effect of hydroethanolic and aqueous solvents on the functional potential and to investigate the bioactive compounds of Pumpkin extract. For functional evaluation of extracts, the amount of ascorbic acid, total phenol and flavonoids, antioxidant capacity were performed by DPPH and FRAP methods and also the chemical compounds in the extracts were identified by GC/MS. The results showed highest levels of ascorbic acid, total phenol and flavonoids in the hydroethanolic extract were 98.66±13.29 mg/100g, 1.663±0.004 mg GA/100g and 0.381±0.01 mg QU/100g, respectively. Hydroethanolic extract showed the highest ability to inhibit DPPH radicals compared to aqueous extract in all prepared concentrations; so that hydroethanolic extract with a concentration of 800 μg/ml with 61.866% inhibition had the highest inhibitory properties. Detected chemical compounds in hydroethanolic and aqueous extracts using GC/MS were identified 16 and 7 chemical compounds, respectively, when the highest compound content in hydroethanolic extract equal to 34.86767% related to D-erythro-pentose, 2-deoxy and the aqueous extract was related to 5,2-dichlorophenol (74,2053%). The results of this study suggest the use of hydroethanolic solvent for higher extraction of bioactive compounds from pumpkin.
References:
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Akpuaka, A., Ekwenchi, M., Dashak, D., and Dildar, A. (2012). Gas Chromatography-Mass Spectrometry (GC/MS) analysis of phthalate isolates in n-hexane extract of Azadirachta indica A. Juss (Neem) leaves. Journal of American Science, 8(12), 146-155.
Al-Qaisy, M. R., and Rathi, M. H. 2019. Total Phenolic Content and Antioxidant Efficacy of Three parts of the pumpkin Cucurbita moschata and the effect of the drying method on them. World Journal of Pharmacy and Pharmaceutical Sciences. 8(4): 1679-1689.
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Albishi, T., John, J. A., Al-Khalifa, A. S., and Shahidi, F. (2013). Phenolic content and antioxidant activities of selected potato varieties and their processing by-products. Journal of Functional Foods, 5(2), 590-600.
Alothman, M., Bhat, R., and Karim, A. (2009). Antioxidant capacity and phenolic content of selected tropical fruits from Malaysia, extracted with different solvents. Food chemistry, 115(3), 785-788.
Ani, V., Varadaraj, M., and Naidu, K. A. (2006). Antioxidant and antibacterial activities of polyphenolic compounds from bitter cumin (Cuminum nigrum). European Food Research and Technology, 224(1), 109-115.
Badr, S. E., Shaaban, M., Elkholy, Y. M., Helal, M. H., Hamza, A. S., Masoud, M. S., and El Safty, M. M. (2011). Chemical composition and biological activity of ripe pumpkin fruits (Cucurbita pepo) cultivated in Egyptian habitats. Natural product research, 25(16), 1524-1539.
Bhandary, S. K., Kumari, S., Bhat, V. S., Sharmila, K., and Bekal, M. P. (2012). Preliminary phytochemical screening of various extracts of Punica granatum peel, whole fruit and seeds. J Health Sci, 2(4), 35-38.
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Chauhan, H., and Singh, M. 2019. Phytochemical characterization and antibacterial potential of Indian and Chinese cabbage genotypes against human pathogens in Uttarakhand, India. International Journal of Recent Scientific Research, 10(12): 36462-36466.
Chigayo, K., Mojapelo, P. E. L., Mnyakeni-Moleele, S., and Misihairabgwi, J. M. (2016). Phytochemical and antioxidant properties of different solvent extracts of Kirkia wilmsii tubers. Asian Pacific Journal of Tropical Biomedicine, 6(12), 1037-1043.
Dai, J., and Mumper, R. J. (2010). Plant phenolics: extraction, analysis and their antioxidant and anticancer properties. Molecules, 15(10), 7313-7352.
Dhiman, A. K., Sharma, K., and Attri, S. (2009). Functional constitutents and processing of pumpkin: A review. Journal of Food Science and Technology, 46(5), 411.
Elfalleh, W., Kirkan, B., and Sarikurkcu, C. (2019). Antioxidant potential and phenolic composition of extracts from Stachys tmolea: An endemic plant from Turkey. Industrial Crops and Products, 127, 212-216.
Ellong, E. N., Billard, C., Adenet, S., and Rochefort, K. (2015). Polyphenols, carotenoids, vitamin C content in tropical fruits and vegetables and impact of processing methods. Food and Nutrition Sciences, 6(03), 299.
Enneb, S., Drine, S., Bagues, M., Triki, T., Boussora, F., Guasmi, F., Ferchichi, A. (2020). Phytochemical profiles and nutritional composition of squash (Cucurbita moschata) from Tunisia. South African Journal of Botany, 130, 165-171.
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Jayasundara, C., Deraniyagala, S. A., Hettiarachchi, C., and Thiripuranathar, G. (2018). Antioxidant and antibacterial activities of leaves, skin, flesh and seeds of Sri Lankan variety of cucurbita moschata. International Journal of Ayurveda and Pharma Research.
Jung, C.-H., Seog, H.-M., Choi, I.-W., Park, M.-W., and Cho, H.-Y. (2006). Antioxidant properties of various solvent extracts from wild ginseng leaves. LWT-Food Science and Technology, 39(3), 266-274.
Kartal, N., Sokmen, M., Tepe, B., Daferera, D., Polissiou, M., and Sokmen, A. (2007). Investigation of the antioxidant properties of Ferula orientalis L. using a suitable extraction procedure. Food chemistry, 100(2), 584-589.
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Kim, M. Y., Kim, E. J., Kim, Y.-N., Choi, C., and Lee, B.-H. (2012). Comparison of the chemical compositions and nutritive values of various pumpkin (Cucurbitaceae) species and parts. Nutrition research and practice, 6(1), 21-27.
Kim, S.-J., Cho, A. R., and Han, J. (2013). Antioxidant and antimicrobial activities of leafy green vegetable extracts and their applications to meat product preservation. Food control, 29(1), 112-120.
Krishnaiah, D., Sarbatly, R., and Nithyanandam, R. (2011). A review of the antioxidant potential of medicinal plant species. Food and bioproducts processing, 89(3), 217-233.
Kulczyński, B., and Gramza-Michałowska, A. (2016). Goji berry (Lycium barbarum): composition and health effects–a review. Polish Journal of Food and Nutrition Sciences, 66(2), 67-76.
Kulczyński, B., and Gramza-Michałowska, A. (2019a). The profile of carotenoids and other bioactive molecules in various pumpkin fruits (Cucurbita maxima Duchesne) cultivars. Molecules, 24(18), 3212.
Kulczyński, B., and Gramza-Michałowska, A. (2019b). The profile of secondary metabolites and other bioactive compounds in Cucurbita pepo L. and Cucurbita moschata pumpkin cultivars. Molecules, 24(16), 2945.
Kulczyński, B., Gramza-Michałowska, A., and Królczyk, J. B. (2020). Optimization of Extraction Conditions for the Antioxidant Potential of Different Pumpkin Varieties (Cucurbita maxima). Sustainability, 12(4), 1305.
Mala, K. S., and Kurian, A. E. (2016). Nutritional composition and antioxidant activity of pumpkin wastes. International Journal of Pharmaceutical, Chemical & Biological Sciences, 6(3).
Montesano, D., Blasi, F., Simonetti, M. S., Santini, A., and Cossignani, L. (2018). Chemical and nutritional characterization of seed oil from Cucurbita maxima(var. Berrettina) pumpkin. Foods, 7(3), 30.
Okwu, D. E., and Ohenhen, O. (2010). Isolation and characterization of Steroidal Glycosides from the leaves of Stachytarpheta Jamaicensis Linn Vahl. Der Chemica Sinica, 1(2), 6-14.
Ordonez, A., Gomez, J., and Vattuone, M. (2006). Antioxidant activities of Sechium edule (Jacq.) Swartz extracts. Food chemistry, 97(3), 452-458.
Pandey, S., Singh, J., Upadhyay, A., Ram, D., and Rai, M. (2003). Ascorbate and carotenoid content in an Indian collection of pumpkin (Cucurbita moschata ex Poir.). Cucurbit Genetics Cooperative Report, 26, 51-53.
Ramroudi, M., Kazemitabar, S. K., Esmaeilzadeh Kenari, R., and Najafi Zarini, H. (2020). Evaluation of Antioxidant Activity of Hydroalcoholic Extracts from Pumpkin (Cucurbita moschata) Seed, Flesh and Skin. Iranian Journal of Biosystems Engineering, 51(3), 663-672.
Rane, Z., Anish-Kumar, P., and Bhaskar, A. (2012). Phytochemical evaluation by GC-MS and in vitro antioxidant activity of Punica granatum fruit rind extract. Journal of Chemical and Pharmaceutical Research, 4(6), 2869-2873.
Rathi, M., and Abdulhay, H. (2018). Total phenolic contents and antioxidant activity of extracts of Tea (Black, Green and White). Asian Jr. of Microbiol. Biotech. Env. Sc, 20(2018), 45-49.
Rezig, L., Chouaibi, M., Meddeb, W., Msaada, K., and Hamdi, S. (2019). Chemical composition and bioactive compounds of Cucurbitaceae seeds: Potential sources for new trends of plant oils. Process Safety and Environmental Protection, 127, 73-81.
Sauer, M., Porro, D., Mattanovich, D., and Branduardi, P. (2008). Microbial production of organic acids: expanding the markets. Trends in biotechnology, 26(2), 100-108.
Singh, J., Singh, V., Shukla, S., and Rai, A. (2016). Phenolic content and antioxidant capacity of selected cucurbit fruits extracted with different solvents. J Nutr Food Sci, 6(6), 1-8.
Singh, S., Singh, R., Thakur, P., and Kumar, R. (2018). Phytochemicals, functionality and breeding for enrichment of cole vegetables (Brassica oleracea). Phytochemicals in vegetables: a valuable source of bioactive compounds. Bentham Science Publishers, UAE, 256-295.
Stevenson, D. G., Eller, F. J., Wang, L., Jane, J.-L., Wang, T., and Inglett, G. E. (2007). Oil and tocopherol content and composition of pumpkin seed oil in 12 cultivars. Journal of agricultural and food chemistry, 55(10), 4005-4013.
Vale, A., Santos, J., Melia, N., Peixoto, V., Brito, N., and Oliveira, M. B. P. (2015). Phytochemical composition and antimicrobial properties of four varieties of Brassica oleracea sprouts. Food Control, 55, 248-256.
Wang, X., Wang, C., Zha, X., Mei, Y., Xia, J., and Jiao, Z. (2017). Supercritical carbon dioxide extraction of β-carotene and α-tocopherol from pumpkin: a Box–Behnken design for extraction variables. Analytical Methods, 9(2), 294-303.
Wu, H., Zhu, J., Diao, W., and Wang, C. (2014). Ultrasound-assisted enzymatic extraction and antioxidant activity of polysaccharides from pumpkin (Cucurbita moschata). Carbohydrate polymers, 113, 314-324.
Xie, X., Deng, T., Duan, J., Ding, S., Yuan, J., and Chen, M. (2019). Comparing the effects of diethylhexyl phthalate and dibutyl phthalate exposure on hypertension in mice. Ecotoxicology and environmental safety, 174, 75-82.
Yakoub, A. R. B., Abdehedi, O., Jridi, M., Elfalleh, W., Nasri, M., and Ferchichi, A. (2018). Flavonoids, phenols, antioxidant, and antimicrobial activities in various extracts from Tossa jute leave (Corchorus olitorus). Industrial Crops and Products, 118, 206-213.
Zaccari, F., and Galietta, G. (2015). α-Carotene and β-carotene content in raw and cooked pulp of three mature stage winter squash “type butternut”. Foods, 4(3), 477-486.
Zinash, A., and Woldetsadik, K. (2013). Effect of accessions on the chemical quality of fresh pumpkin. African Journal of Biotechnology, 12(51), 7092-7098.
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Ahmad, G., and Khan, A. A. (2019). Pumpkin: horticultural importance and its roles in various forms; a review. Int J Hortic Agric, 4(1), 1-6.
Akpuaka, A., Ekwenchi, M., Dashak, D., and Dildar, A. (2012). Gas Chromatography-Mass Spectrometry (GC/MS) analysis of phthalate isolates in n-hexane extract of Azadirachta indica A. Juss (Neem) leaves. Journal of American Science, 8(12), 146-155.
Al-Qaisy, M. R., and Rathi, M. H. 2019. Total Phenolic Content and Antioxidant Efficacy of Three parts of the pumpkin Cucurbita moschata and the effect of the drying method on them. World Journal of Pharmacy and Pharmaceutical Sciences. 8(4): 1679-1689.
Al-Rikabi, A. K. J. (2007). Extraction of phenolic compounds from wheat bran and its assessment as antioxidants. Journal of Basrah Researches (Sciences), 33(2B).
Albishi, T., John, J. A., Al-Khalifa, A. S., and Shahidi, F. (2013). Phenolic content and antioxidant activities of selected potato varieties and their processing by-products. Journal of Functional Foods, 5(2), 590-600.
Alothman, M., Bhat, R., and Karim, A. (2009). Antioxidant capacity and phenolic content of selected tropical fruits from Malaysia, extracted with different solvents. Food chemistry, 115(3), 785-788.
Ani, V., Varadaraj, M., and Naidu, K. A. (2006). Antioxidant and antibacterial activities of polyphenolic compounds from bitter cumin (Cuminum nigrum). European Food Research and Technology, 224(1), 109-115.
Badr, S. E., Shaaban, M., Elkholy, Y. M., Helal, M. H., Hamza, A. S., Masoud, M. S., and El Safty, M. M. (2011). Chemical composition and biological activity of ripe pumpkin fruits (Cucurbita pepo) cultivated in Egyptian habitats. Natural product research, 25(16), 1524-1539.
Bhandary, S. K., Kumari, S., Bhat, V. S., Sharmila, K., and Bekal, M. P. (2012). Preliminary phytochemical screening of various extracts of Punica granatum peel, whole fruit and seeds. J Health Sci, 2(4), 35-38.
Biesalski, H.-K., Dragsted, L. O., Elmadfa, I., Grossklaus, R., Müller, M., Schrenk, D., Weber, P. (2009). Bioactive compounds: Definition and assessment of activity. Nutrition, 25(11-12), 1202-1205.
Chauhan, H., and Singh, M. 2019. Phytochemical characterization and antibacterial potential of Indian and Chinese cabbage genotypes against human pathogens in Uttarakhand, India. International Journal of Recent Scientific Research, 10(12): 36462-36466.
Chigayo, K., Mojapelo, P. E. L., Mnyakeni-Moleele, S., and Misihairabgwi, J. M. (2016). Phytochemical and antioxidant properties of different solvent extracts of Kirkia wilmsii tubers. Asian Pacific Journal of Tropical Biomedicine, 6(12), 1037-1043.
Dai, J., and Mumper, R. J. (2010). Plant phenolics: extraction, analysis and their antioxidant and anticancer properties. Molecules, 15(10), 7313-7352.
Dhiman, A. K., Sharma, K., and Attri, S. (2009). Functional constitutents and processing of pumpkin: A review. Journal of Food Science and Technology, 46(5), 411.
Elfalleh, W., Kirkan, B., and Sarikurkcu, C. (2019). Antioxidant potential and phenolic composition of extracts from Stachys tmolea: An endemic plant from Turkey. Industrial Crops and Products, 127, 212-216.
Ellong, E. N., Billard, C., Adenet, S., and Rochefort, K. (2015). Polyphenols, carotenoids, vitamin C content in tropical fruits and vegetables and impact of processing methods. Food and Nutrition Sciences, 6(03), 299.
Enneb, S., Drine, S., Bagues, M., Triki, T., Boussora, F., Guasmi, F., Ferchichi, A. (2020). Phytochemical profiles and nutritional composition of squash (Cucurbita moschata) from Tunisia. South African Journal of Botany, 130, 165-171.
Hayouni, E. A., Abedrabba, M., Bouix, M., and Hamdi, M. (2007). The effects of solvents and extraction method on the phenolic contents and biological activities in vitro of Tunisian Quercus coccifera L. and Juniperus phoenicea L. fruit extracts. Food chemistry, 105(3), 1126-1134.
Indrianingsih, A., Rosyida, V., Apriyana, W., Hayati, S. N., Nisa, K., Darsih, C., Indirayati, N. (2019). Comparisons of antioxidant activities of two varieties of pumpkin (Cucurbita moschata and Cucurbita maxima) extracts. Paper presented at the IOP Conference Series: Earth and Environmental Science.
Jacobo-Valenzuela N, de Jesus Zazueta-Morales J, Gallegos-Infante JA, Aguilar-Gutierrez F, Camacho-Hernandez IL, Rocha-Guzman NE, et al. (2011). Chemical and physicochemical characterization of winter squash (Cucurbita moschata). Notulae Botanicae Horti Agrobotanici Cluj-Napoca, 39(1), 34-40.
Jayasundara, C., Deraniyagala, S. A., Hettiarachchi, C., and Thiripuranathar, G. (2018). Antioxidant and antibacterial activities of leaves, skin, flesh and seeds of Sri Lankan variety of cucurbita moschata. International Journal of Ayurveda and Pharma Research.
Jung, C.-H., Seog, H.-M., Choi, I.-W., Park, M.-W., and Cho, H.-Y. (2006). Antioxidant properties of various solvent extracts from wild ginseng leaves. LWT-Food Science and Technology, 39(3), 266-274.
Kartal, N., Sokmen, M., Tepe, B., Daferera, D., Polissiou, M., and Sokmen, A. (2007). Investigation of the antioxidant properties of Ferula orientalis L. using a suitable extraction procedure. Food chemistry, 100(2), 584-589.
Kaur, G. (2018). Development of Functionalprobiotic Beverage from Pumpkin. Lovely Professional University,
Kim, M. Y., Kim, E. J., Kim, Y.-N., Choi, C., and Lee, B.-H. (2012). Comparison of the chemical compositions and nutritive values of various pumpkin (Cucurbitaceae) species and parts. Nutrition research and practice, 6(1), 21-27.
Kim, S.-J., Cho, A. R., and Han, J. (2013). Antioxidant and antimicrobial activities of leafy green vegetable extracts and their applications to meat product preservation. Food control, 29(1), 112-120.
Krishnaiah, D., Sarbatly, R., and Nithyanandam, R. (2011). A review of the antioxidant potential of medicinal plant species. Food and bioproducts processing, 89(3), 217-233.
Kulczyński, B., and Gramza-Michałowska, A. (2016). Goji berry (Lycium barbarum): composition and health effects–a review. Polish Journal of Food and Nutrition Sciences, 66(2), 67-76.
Kulczyński, B., and Gramza-Michałowska, A. (2019a). The profile of carotenoids and other bioactive molecules in various pumpkin fruits (Cucurbita maxima Duchesne) cultivars. Molecules, 24(18), 3212.
Kulczyński, B., and Gramza-Michałowska, A. (2019b). The profile of secondary metabolites and other bioactive compounds in Cucurbita pepo L. and Cucurbita moschata pumpkin cultivars. Molecules, 24(16), 2945.
Kulczyński, B., Gramza-Michałowska, A., and Królczyk, J. B. (2020). Optimization of Extraction Conditions for the Antioxidant Potential of Different Pumpkin Varieties (Cucurbita maxima). Sustainability, 12(4), 1305.
Mala, K. S., and Kurian, A. E. (2016). Nutritional composition and antioxidant activity of pumpkin wastes. International Journal of Pharmaceutical, Chemical & Biological Sciences, 6(3).
Montesano, D., Blasi, F., Simonetti, M. S., Santini, A., and Cossignani, L. (2018). Chemical and nutritional characterization of seed oil from Cucurbita maxima(var. Berrettina) pumpkin. Foods, 7(3), 30.
Okwu, D. E., and Ohenhen, O. (2010). Isolation and characterization of Steroidal Glycosides from the leaves of Stachytarpheta Jamaicensis Linn Vahl. Der Chemica Sinica, 1(2), 6-14.
Ordonez, A., Gomez, J., and Vattuone, M. (2006). Antioxidant activities of Sechium edule (Jacq.) Swartz extracts. Food chemistry, 97(3), 452-458.
Pandey, S., Singh, J., Upadhyay, A., Ram, D., and Rai, M. (2003). Ascorbate and carotenoid content in an Indian collection of pumpkin (Cucurbita moschata ex Poir.). Cucurbit Genetics Cooperative Report, 26, 51-53.
Ramroudi, M., Kazemitabar, S. K., Esmaeilzadeh Kenari, R., and Najafi Zarini, H. (2020). Evaluation of Antioxidant Activity of Hydroalcoholic Extracts from Pumpkin (Cucurbita moschata) Seed, Flesh and Skin. Iranian Journal of Biosystems Engineering, 51(3), 663-672.
Rane, Z., Anish-Kumar, P., and Bhaskar, A. (2012). Phytochemical evaluation by GC-MS and in vitro antioxidant activity of Punica granatum fruit rind extract. Journal of Chemical and Pharmaceutical Research, 4(6), 2869-2873.
Rathi, M., and Abdulhay, H. (2018). Total phenolic contents and antioxidant activity of extracts of Tea (Black, Green and White). Asian Jr. of Microbiol. Biotech. Env. Sc, 20(2018), 45-49.
Rezig, L., Chouaibi, M., Meddeb, W., Msaada, K., and Hamdi, S. (2019). Chemical composition and bioactive compounds of Cucurbitaceae seeds: Potential sources for new trends of plant oils. Process Safety and Environmental Protection, 127, 73-81.
Sauer, M., Porro, D., Mattanovich, D., and Branduardi, P. (2008). Microbial production of organic acids: expanding the markets. Trends in biotechnology, 26(2), 100-108.
Singh, J., Singh, V., Shukla, S., and Rai, A. (2016). Phenolic content and antioxidant capacity of selected cucurbit fruits extracted with different solvents. J Nutr Food Sci, 6(6), 1-8.
Singh, S., Singh, R., Thakur, P., and Kumar, R. (2018). Phytochemicals, functionality and breeding for enrichment of cole vegetables (Brassica oleracea). Phytochemicals in vegetables: a valuable source of bioactive compounds. Bentham Science Publishers, UAE, 256-295.
Stevenson, D. G., Eller, F. J., Wang, L., Jane, J.-L., Wang, T., and Inglett, G. E. (2007). Oil and tocopherol content and composition of pumpkin seed oil in 12 cultivars. Journal of agricultural and food chemistry, 55(10), 4005-4013.
Vale, A., Santos, J., Melia, N., Peixoto, V., Brito, N., and Oliveira, M. B. P. (2015). Phytochemical composition and antimicrobial properties of four varieties of Brassica oleracea sprouts. Food Control, 55, 248-256.
Wang, X., Wang, C., Zha, X., Mei, Y., Xia, J., and Jiao, Z. (2017). Supercritical carbon dioxide extraction of β-carotene and α-tocopherol from pumpkin: a Box–Behnken design for extraction variables. Analytical Methods, 9(2), 294-303.
Wu, H., Zhu, J., Diao, W., and Wang, C. (2014). Ultrasound-assisted enzymatic extraction and antioxidant activity of polysaccharides from pumpkin (Cucurbita moschata). Carbohydrate polymers, 113, 314-324.
Xie, X., Deng, T., Duan, J., Ding, S., Yuan, J., and Chen, M. (2019). Comparing the effects of diethylhexyl phthalate and dibutyl phthalate exposure on hypertension in mice. Ecotoxicology and environmental safety, 174, 75-82.
Yakoub, A. R. B., Abdehedi, O., Jridi, M., Elfalleh, W., Nasri, M., and Ferchichi, A. (2018). Flavonoids, phenols, antioxidant, and antimicrobial activities in various extracts from Tossa jute leave (Corchorus olitorus). Industrial Crops and Products, 118, 206-213.
Zaccari, F., and Galietta, G. (2015). α-Carotene and β-carotene content in raw and cooked pulp of three mature stage winter squash “type butternut”. Foods, 4(3), 477-486.
Zinash, A., and Woldetsadik, K. (2013). Effect of accessions on the chemical quality of fresh pumpkin. African Journal of Biotechnology, 12(51), 7092-7098.
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