Foods and Raw Materials Foods and Raw Materials Foods and Raw Materials 2308-4057 2310-9599 83156 10.21603/2308-4057-2025-1-630 Research Article Research Article Research Article Pomegranate leaves, buds, and flowers: phytochemical, antioxidant, and comparative solvent analyzes Pomegranate leaves, buds, and flowers: phytochemical, antioxidant, and comparative solvent analyzes https://orcid.org/0000-0003-3682-3044 Tekin Zehra Tekin Zehra ztekin@adiyaman.edu.tr https://orcid.org/0000-0001-7784-4138 Kucukbay F.Zehra Kucukbay F.Zehra zehra.kucukbay@inonu.edu.tr Adıyaman University Adıyaman Турция Adıyaman University Adıyaman Turkey İnönü University Malatya Турция İnönü University Malatya Turkey 07 11 2024 07 11 2024 13 1 155 171 29 09 2023 05 12 2023 https://jfrm.ru/en/issues/22431/22627/

Punica granatum L. possesses significant nutritional and medicinal potential. Its pharmacological activities have been investigated, but no comparative evaluation has been reported regarding the effect of different extraction solvents on the phytochemical content and antioxidant activity of its leaf, bud, and flower extracts. This research involved seven various solvents, namely methanol, ethanol, water, acidified methanol, acidified ethanol, acidified water, and hexane. A set of experiments made it possible to define the effect of each of these solvents on the contents of phenolics, flavonoids, flavanols, flavonols, anthocyanins, and tannins, as well as on the antioxidant activity of pomegranate leaf, bud and flower tissues. The research objective was to identify the optimal solvent for the most effective extraction of the abovementioned functional compounds. The antioxidant activity tests involved DPPH free radical scavenging, metal chelating, iron (III) reducing power, and CUPRAC assays. The aqueous extract of P. granatum leaves demonstrated the highest total phenolic content (192.57 mg GAE/g extract) while the greatest flavonoid content belonged to the acidified methanol extract of P. granatum buds (73.93 mg RE/g extract). The HPLC analysis detected such significant phenolic compounds as punicalagin in buds and flowers, as well as gallic acid in leaves. All the extracts showed good antioxidant activity; however, the bud extracts had a better antioxidant profile than the extracts from leaves and flowers. The pomegranate leaf, bud, and flower extracts demonstrated excellent phytochemical and antioxidant properties, which makes it possible to recommend these plant tissues as raw materials to be used in pharmaceutical, food, nutraceutical, and cosmetic industries.

Punica granatum L. possesses significant nutritional and medicinal potential. Its pharmacological activities have been investigated, but no comparative evaluation has been reported regarding the effect of different extraction solvents on the phytochemical content and antioxidant activity of its leaf, bud, and flower extracts. This research involved seven various solvents, namely methanol, ethanol, water, acidified methanol, acidified ethanol, acidified water, and hexane. A set of experiments made it possible to define the effect of each of these solvents on the contents of phenolics, flavonoids, flavanols, flavonols, anthocyanins, and tannins, as well as on the antioxidant activity of pomegranate leaf, bud and flower tissues. The research objective was to identify the optimal solvent for the most effective extraction of the abovementioned functional compounds. The antioxidant activity tests involved DPPH free radical scavenging, metal chelating, iron (III) reducing power, and CUPRAC assays. The aqueous extract of P. granatum leaves demonstrated the highest total phenolic content (192.57 mg GAE/g extract) while the greatest flavonoid content belonged to the acidified methanol extract of P. granatum buds (73.93 mg RE/g extract). The HPLC analysis detected such significant phenolic compounds as punicalagin in buds and flowers, as well as gallic acid in leaves. All the extracts showed good antioxidant activity; however, the bud extracts had a better antioxidant profile than the extracts from leaves and flowers. The pomegranate leaf, bud, and flower extracts demonstrated excellent phytochemical and antioxidant properties, which makes it possible to recommend these plant tissues as raw materials to be used in pharmaceutical, food, nutraceutical, and cosmetic industries.

 Punica granatum L. pomegranate buds pomegranate leaves pomegranate flowers antioxidant activity phytochemical analysis pharmacological potential Punica granatum L. pomegranate buds pomegranate leaves pomegranate flowers antioxidant activity phytochemical analysis pharmacological potential The research was supported by the İnönü University Research Fund (TDK-2020-2100). The research was supported by the İnönü University Research Fund (TDK-2020-2100).

Das K, Roychoudhury A. Reactive oxygen species (ROS) and response of antioxidants as ROS-scavengers during environmental stress in plants. Frontiers in Environmental Science. 2014;2:53. https://doi.org/10.3389/fenvs.2014.00053 Das K, Roychoudhury A. Reactive oxygen species (ROS) and response of antioxidants as ROS-scavengers during environmental stress in plants. Frontiers in Environmental Science. 2014;2:53. https://doi.org/10.3389/fenvs.2014.00053 Demirci-Çekiç S, Özkan G, Avan AN, Uzunboy S, Çapanoğlu E, Apak R. Biomarkers of oxidative stress and antioxidant defense. Journal of Pharmaceutical and Biomedical Analysis. 2022;209:114477. https://doi.org/10.1016/j.jpba.2021.114477 Demirci-Çekiç S, Özkan G, Avan AN, Uzunboy S, Çapanoğlu E, Apak R. Biomarkers of oxidative stress and antioxidant defense. Journal of Pharmaceutical and Biomedical Analysis. 2022;209:114477. https://doi.org/10.1016/j.jpba.2021.114477 Juan CA, Pérez de la Lastra JM, Plou FJ, Pérez-Lebeña E. The chemistry of reactive oxygen species (ROS) revisited: Outlining their role in biological macromolecules (DNA, lipids and proteins) and induced pathologies. International Journal of Molecular Sciences 2021;22(9):4642. https://doi.org/10.3390/ijms22094642 Juan CA, Pérez de la Lastra JM, Plou FJ, Pérez-Lebeña E. The chemistry of reactive oxygen species (ROS) revisited: Outlining their role in biological macromolecules (DNA, lipids and proteins) and induced pathologies. International Journal of Molecular Sciences 2021;22(9):4642. https://doi.org/10.3390/ijms22094642 Redza-Dutordoir M, Averill-Bates DA. Activation of apoptosis signalling pathways by reactive oxygen species. Biochimica et Biophysica Acta (BBA) – Molecular Cell Research. 2016;1863(12):2977–2992. https://doi.org/10.1016/j.bbamcr.2016.09.012 Redza-Dutordoir M, Averill-Bates DA. Activation of apoptosis signalling pathways by reactive oxygen species. Biochimica et Biophysica Acta (BBA) – Molecular Cell Research. 2016;1863(12):2977–2992. https://doi.org/10.1016/j.bbamcr.2016.09.012 Birben E, Sahiner UM, Sackesen C, Erzurum S, Kalayci O. Oxidative stress and antioxidant defense. World Allergy Organization Journal. 2012;5(1):9–19. https://doi.org/10.1097/WOX.0b013e3182439613 Birben E, Sahiner UM, Sackesen C, Erzurum S, Kalayci O. Oxidative stress and antioxidant defense. World Allergy Organization Journal. 2012;5(1):9–19. https://doi.org/10.1097/WOX.0b013e3182439613 Faskhutdinova ER, Sukhikh AS, Le VM, Minina VI, Khelef MEA, Loseva AI. Effects of bioactive substances isolated from Siberian medicinal plants on the lifespan of Caenorhabditis elegans. Foods and Raw Materials. 2022;10(2):340–352. https://doi.org/10.21603/2308-4057-2022-2-544 Faskhutdinova ER, Sukhikh AS, Le VM, Minina VI, Khelef MEA, Loseva AI. Effects of bioactive substances isolated from Siberian medicinal plants on the lifespan of Caenorhabditis elegans. Foods and Raw Materials. 2022;10(2):340–352. https://doi.org/10.21603/2308-4057-2022-2-544 Pizzino G, Irrera N, Cucinotta M, Pallio G, Mannino F. Arcoraci V, et al. Oxidative stress: Harms and benefits for human health. Oxidative Medicine and Cellular Longevity. 2017;2017:8416763. https://doi.org/10.1155/2017/8416763 Pizzino G, Irrera N, Cucinotta M, Pallio G, Mannino F. Arcoraci V, et al. Oxidative stress: Harms and benefits for human health. Oxidative Medicine and Cellular Longevity. 2017;2017:8416763. https://doi.org/10.1155/2017/8416763 Sharifi-Rad M, Anil Kumar NV, Zucca P, Varoni EM, Dini L, Panzarini E, et al. Lifestyle, Oxidative stress, and antioxidants: Back and forth in the pathophysiology of chronic diseases. Frontiers in Physiology. 2020;11:694. https://doi.org/10.3389/fphys.2020.00694 Sharifi-Rad M, Anil Kumar NV, Zucca P, Varoni EM, Dini L, Panzarini E, et al. Lifestyle, Oxidative stress, and antioxidants: Back and forth in the pathophysiology of chronic diseases. Frontiers in Physiology. 2020;11:694. https://doi.org/10.3389/fphys.2020.00694 Xu X, Liu A, Hu S, Ares I, Martínez-Larrañaga M-R, Wang X, et al. Synthetic phenolic antioxidants: Metabolism, hazards and mechanism of action. Food Chemistry. 2021;353:129488. https://doi.org/10.1016/j.foodchem.2021.129488 Xu X, Liu A, Hu S, Ares I, Martínez-Larrañaga M-R, Wang X, et al. Synthetic phenolic antioxidants: Metabolism, hazards and mechanism of action. Food Chemistry. 2021;353:129488. https://doi.org/10.1016/j.foodchem.2021.129488 Wang W, Xiong P, Zhang H, Zhu Q, Liao C, Jiang G. Analysis, occurrence, toxicity and environmental health risks of synthetic phenolic antioxidants: A review. Environmental Research. 2021;201:111531. https://doi.org/10.1016/j.envres.2021.111531 Wang W, Xiong P, Zhang H, Zhu Q, Liao C, Jiang G. Analysis, occurrence, toxicity and environmental health risks of synthetic phenolic antioxidants: A review. Environmental Research. 2021;201:111531. https://doi.org/10.1016/j.envres.2021.111531 Lourenço SC, Moldão-Martins M, Alves VD. Antioxidants of natural plant origins: From sources to food industry applications. Molecules. 2019;24(22):4132. https://doi.org/10.3390/molecules24224132 Lourenço SC, Moldão-Martins M, Alves VD. Antioxidants of natural plant origins: From sources to food industry applications. Molecules. 2019;24(22):4132. https://doi.org/10.3390/molecules24224132 Kandylis P, Kokkinomagoulos E. Food applications and potential health benefits of pomegranate and its derivatives. Foods. 2020;9(2):122. https://doi.org/10.3390/foods9020122 Kandylis P, Kokkinomagoulos E. Food applications and potential health benefits of pomegranate and its derivatives. Foods. 2020;9(2):122. https://doi.org/10.3390/foods9020122 Dandachi F, Hamadeh B, Youssef H, Chahine H, Chalak L. Diversity assessment of the Lebanese germplasm of pomegranate (Punica granatum L.) by morphological and chemical traits. Annals of Agricultural Sciences. 2017;62(1):89–98. https://doi.org/10.1016/j.aoas.2017.05.004 Dandachi F, Hamadeh B, Youssef H, Chahine H, Chalak L. Diversity assessment of the Lebanese germplasm of pomegranate (Punica granatum L.) by morphological and chemical traits. Annals of Agricultural Sciences. 2017;62(1):89–98. https://doi.org/10.1016/j.aoas.2017.05.004 Karimi M, Sadeghi R, Kokini J. Pomegranate as a promising opportunity in medicine and nanotechnology. Trends in Food Science and Technology. 2017;69:59–73. https://doi.org/10.1016/j.tifs.2017.08.019 Karimi M, Sadeghi R, Kokini J. Pomegranate as a promising opportunity in medicine and nanotechnology. Trends in Food Science and Technology. 2017;69:59–73. https://doi.org/10.1016/j.tifs.2017.08.019 Middha SK, Usha P, Pande V. HPLC evaluation of phenolic profile, nutritive content, and antioxidant capacity of extracts obtained from Punica granatum fruit peel. Advences in Pharmacological and Pharmaceutical Sciences. 2013;2013:296236. https://doi.org/10.1155/2013/296236 Middha SK, Usha P, Pande V. HPLC evaluation of phenolic profile, nutritive content, and antioxidant capacity of extracts obtained from Punica granatum fruit peel. Advences in Pharmacological and Pharmaceutical Sciences. 2013;2013:296236. https://doi.org/10.1155/2013/296236 Derakhshan Z, Ferrante M, Tadi M, Ansari F, Heydari A. Hosseini MS, et al. Antioxidant activity and total phenolic content of ethanolic extract of pomegranate peel, juice and seeds. Food and Chemical Toxicology. 2018;114:108–111. https://doi.org/10.1016/j.fct.2018.02.023 Derakhshan Z, Ferrante M, Tadi M, Ansari F, Heydari A. Hosseini MS, et al. Antioxidant activity and total phenolic content of ethanolic extract of pomegranate peel, juice and seeds. Food and Chemical Toxicology. 2018;114:108–111. https://doi.org/10.1016/j.fct.2018.02.023 Ricci D, Giamperi L, Bucchini A, Fraternale D. Antioxidant activity of Punica granatum fruits. Fitoterapia. 2006;77(4):310–312. https://doi.org/10.1016/j.fitote.2006.01.008 Ricci D, Giamperi L, Bucchini A, Fraternale D. Antioxidant activity of Punica granatum fruits. Fitoterapia. 2006;77(4):310–312. https://doi.org/10.1016/j.fitote.2006.01.008 Küçükbay FZ, Tekin Z. Evaluation of phytochemical contents and antioxidant activity of pomegranate flower. Journal of the Turkish Chemical Society Section A: Chemistry. 2020;7(1):37–42. https://doi.org/10.18596/jotcsa.628615 Küçükbay FZ, Tekin Z. Evaluation of phytochemical contents and antioxidant activity of pomegranate flower. Journal of the Turkish Chemical Society Section A: Chemistry. 2020;7(1):37–42. https://doi.org/10.18596/jotcsa.628615 Hafizov SG, Musina ON, Hafizov GK. Extracting hydrophilic components from pomegranate peel and pulp. Food Processing: Techniques and Technology. 2023;53(1):168–182. (In Russ.). https://doi.org/10.21603/2074-9414-2023-1-2425 Hafizov SG, Musina ON, Hafizov GK. Extracting hydrophilic components from pomegranate peel and pulp. Food Processing: Techniques and Technology. 2023;53(1):168–182. (In Russ.). https://doi.org/10.21603/2074-9414-2023-1-2425 Cheurfa M, Achouche M, Azouzi A, Abdalbasit MA. Antioxidant and anti-diabetic activity of pomegranate (Punica granatum L.) leaves extracts. Foods and Raw Materials. 2020;8(2):329–336. https://doi.org/10.21603/2308-4057-2020-2-329-336 Cheurfa M, Achouche M, Azouzi A, Abdalbasit MA. Antioxidant and anti-diabetic activity of pomegranate (Punica granatum L.) leaves extracts. Foods and Raw Materials. 2020;8(2):329–336. https://doi.org/10.21603/2308-4057-2020-2-329-336 Ge S, Duo L, Wang J, Zhula G, Yang J, Li Z, et al. A unique understanding of traditionally medicine of pomegranate, Punica granatum L. and its current research status. Journal of Ethnopharmacology. 2021;271:113877. https://doi.org/10.1016/j.jep.2021.113877 Ge S, Duo L, Wang J, Zhula G, Yang J, Li Z, et al. A unique understanding of traditionally medicine of pomegranate, Punica granatum L. and its current research status. Journal of Ethnopharmacology. 2021;271:113877. https://doi.org/10.1016/j.jep.2021.113877 Balamurugan C, Karuppasamy R, Sivaraj C, Saraswathi K, Arumugam P. Cytotoxic activity of leaves extract of pomegranate (Punica granatum L.). Journal of Pharmacognosy and Phytochemistry. 2021;10(1):1982–1985. https://doi.org/10.22271/phyto.2021.v10.i1ab.13640 Balamurugan C, Karuppasamy R, Sivaraj C, Saraswathi K, Arumugam P. Cytotoxic activity of leaves extract of pomegranate (Punica granatum L.). Journal of Pharmacognosy and Phytochemistry. 2021;10(1):1982–1985. https://doi.org/10.22271/phyto.2021.v10.i1ab.13640 Balamurugan C, Karuppasamy R, Sivaraj C, Saraswathi K, Arumugam P. Punica granatum L. (Pomegranate) leaves extract: The study of antioxidant and antibacterial activity. Journal of Pharmacognosy and Phytochemistry. 2020;9(6):397–402. https://doi.org/10.22271/phyto.2020.v9.i6f.12915 Balamurugan C, Karuppasamy R, Sivaraj C, Saraswathi K, Arumugam P. Punica granatum L. (Pomegranate) leaves extract: The study of antioxidant and antibacterial activity. Journal of Pharmacognosy and Phytochemistry. 2020;9(6):397–402. https://doi.org/10.22271/phyto.2020.v9.i6f.12915 Bekir J, Mars M, Souchard JP, Bouajila J. Assessment of antioxidant, anti-inflammatory, anti-cholinesterase and cytotoxic activities of pomegranate (Punica granatum) leaves. Food and Chemical Toxicology. 2013;55:470–475. https://doi.org/10.1016/j.fct.2013.01.036 Bekir J, Mars M, Souchard JP, Bouajila J. Assessment of antioxidant, anti-inflammatory, anti-cholinesterase and cytotoxic activities of pomegranate (Punica granatum) leaves. Food and Chemical Toxicology. 2013;55:470–475. https://doi.org/10.1016/j.fct.2013.01.036 Dassprakash MV, Arun R, Abraham SK, Premkumar K. In vitro and in vivo evaluation of antioxidant and antigenotoxic potential of Punica granatum leaf extract. Pharmaceutical Biology. 2012;50(12):1523–1530. https://doi.org/10.3109/13880209.2012.689771 Dassprakash MV, Arun R, Abraham SK, Premkumar K. In vitro and in vivo evaluation of antioxidant and antigenotoxic potential of Punica granatum leaf extract. Pharmaceutical Biology. 2012;50(12):1523–1530. https://doi.org/10.3109/13880209.2012.689771 Huang THW, Peng G, Kota BP, Li GQ, Yamahara J, Roufogalis BD, et al. Anti-diabetic action of Punica granatum flower extract: Activation of PPAR-γ and identification of an active component. Toxicology and Applied Pharmacology. 2005;207(2):160–169. https://doi.org/10.1016/j.taap.2004.12.009 Huang THW, Peng G, Kota BP, Li GQ, Yamahara J, Roufogalis BD, et al. Anti-diabetic action of Punica granatum flower extract: Activation of PPAR-γ and identification of an active component. Toxicology and Applied Pharmacology. 2005;207(2):160–169. https://doi.org/10.1016/j.taap.2004.12.009 Xu J, Zhao Y, Aisa HA. Anti-inflammatory effect of pomegranate flower in lipopolysaccharide (LPS) - stimulated RAW264.7 macrophages. Pharmaceutical Biology. 2017;55(1):2095–2101. https://doi.org/10.1080/13880209.2017.1357737 Xu J, Zhao Y, Aisa HA. Anti-inflammatory effect of pomegranate flower in lipopolysaccharide (LPS) - stimulated RAW264.7 macrophages. Pharmaceutical Biology. 2017;55(1):2095–2101. https://doi.org/10.1080/13880209.2017.1357737 Sarker M, Das SC, Saha SK, Al Mahmud Z, Bachar SC. Analgesic and anti-inflammatory activities of flower extracts of Punica granatum Linn. (Punicaceae). Journal of Applied Pharmaceutical Science. 2012;02(04):133–136. https://doi.org/10.7324/JAPS.2012.2408 Sarker M, Das SC, Saha SK, Al Mahmud Z, Bachar SC. Analgesic and anti-inflammatory activities of flower extracts of Punica granatum Linn. (Punicaceae). Journal of Applied Pharmaceutical Science. 2012;02(04):133–136. https://doi.org/10.7324/JAPS.2012.2408 Abdolahi N, Soltani A, Mirzaali A, Soltani S, Balakheyli H, Aghaei M. Antibacterial and antioxidant activities and phytochemical properties of Punica granatum flowers in Iran. Iranian Journal of Science and Technology, Transactions A: Science. 2018;42:1105–1110. https://doi.org/10.1007/s40995-017-0413-8 Abdolahi N, Soltani A, Mirzaali A, Soltani S, Balakheyli H, Aghaei M. Antibacterial and antioxidant activities and phytochemical properties of Punica granatum flowers in Iran. Iranian Journal of Science and Technology, Transactions A: Science. 2018;42:1105–1110. https://doi.org/10.1007/s40995-017-0413-8 Celik I, Temur A, Isik I. Hepatoprotective role and antioxidant capacity of pomegranate (Punica granatum) flowers infusion against trichloroacetic acid-exposed in rats. Food and Chemical Toxicology. 2009;47(1):145–149. https://doi.org/10.1016/j.fct.2008.10.020 Celik I, Temur A, Isik I. Hepatoprotective role and antioxidant capacity of pomegranate (Punica granatum) flowers infusion against trichloroacetic acid-exposed in rats. Food and Chemical Toxicology. 2009;47(1):145–149. https://doi.org/10.1016/j.fct.2008.10.020 Ahangarpour A, Heidari R, Abdolahzadeh M, Oroojan AA. Antispasmodic effects of aqueous and hydroalcoholic Punica granatum flower extracts on the uterus of non-pregnant rats. Journal of Reproduction and Infertility. 2012;13(2):138–142. Ahangarpour A, Heidari R, Abdolahzadeh M, Oroojan AA. Antispasmodic effects of aqueous and hydroalcoholic Punica granatum flower extracts on the uterus of non-pregnant rats. Journal of Reproduction and Infertility. 2012;13(2):138–142. Barwal SB, Nirmal Sunil A, Dhasade VV, Patil MJ, Pal SC, Mandal Subhash C. Antihistaminic effect of various extracts of Punica granatum Linn. flowers buds. Journal of Young Pharmacists. 2009;1(4):332–335. https://doi.org/10.4103/0975-1483.59321 Barwal SB, Nirmal Sunil A, Dhasade VV, Patil MJ, Pal SC, Mandal Subhash C. Antihistaminic effect of various extracts of Punica granatum Linn. flowers buds. Journal of Young Pharmacists. 2009;1(4):332–335. https://doi.org/10.4103/0975-1483.59321 Orgil O, Schwartz E, Baruch L, Matityahu I, Mahajna J, Amir R. The antioxidative and anti-proliferative potential of non-edible organs of the pomegranate fruit and tree. LWT – Food Science and Technology 2014;58(2):571–577. https://doi.org/10.1016/j.lwt.2014.03.030 Orgil O, Schwartz E, Baruch L, Matityahu I, Mahajna J, Amir R. The antioxidative and anti-proliferative potential of non-edible organs of the pomegranate fruit and tree. LWT – Food Science and Technology 2014;58(2):571–577. https://doi.org/10.1016/j.lwt.2014.03.030 Montefusco A, Durante M, Migoni D, de Caroli M, Ilahy R, Pék Z, et al. Analysis of the phytochemical composition of pomegranate fruit juices, peels and kernels: A comparative study on four cultivars grown in southern Italy. Plants. 2021;10(11):2521. https://doi.org/10.3390/plants10112521 Montefusco A, Durante M, Migoni D, de Caroli M, Ilahy R, Pék Z, et al. Analysis of the phytochemical composition of pomegranate fruit juices, peels and kernels: A comparative study on four cultivars grown in southern Italy. Plants. 2021;10(11):2521. https://doi.org/10.3390/plants10112521 Boussaa F, Zaouay F, Burlo-Carbonell F, Noguera-Artiaga L, Carbonell-Barrachina A, Melgarejo P, et al. Growing location affects physical properties, bioactive compounds, and antioxidant activity of pomegranate fruit (Punica granatum L. var. Gabsi). International Journal of Fruit Science. 2020;20:508–523. https://doi.org/10.1080/15538362.2020.1741058 Boussaa F, Zaouay F, Burlo-Carbonell F, Noguera-Artiaga L, Carbonell-Barrachina A, Melgarejo P, et al. Growing location affects physical properties, bioactive compounds, and antioxidant activity of pomegranate fruit (Punica granatum L. var. Gabsi). International Journal of Fruit Science. 2020;20:508–523. https://doi.org/10.1080/15538362.2020.1741058 Melgarejo-Sánchez P, Núñez-Gómez D, Martínez-Nicolás JJ, Hernández F, Legua P, Melgarejo P. Pomegranate variety and pomegranate plant part, revelance from bioactive point of view: A review. Bioresources and Bioprocessing. 2021;8:2. https://doi.org/10.1186/s40643-020-00351-5 Melgarejo-Sánchez P, Núñez-Gómez D, Martínez-Nicolás JJ, Hernández F, Legua P, Melgarejo P. Pomegranate variety and pomegranate plant part, revelance from bioactive point of view: A review. Bioresources and Bioprocessing. 2021;8:2. https://doi.org/10.1186/s40643-020-00351-5 Dai J, Mumper RJ. Plant phenolics: Extraction, analysis and their antioxidant and anticancer properties. Molecules. 2010;15(10):7313–7352. https://doi.org/10.3390/molecules15107313 Dai J, Mumper RJ. Plant phenolics: Extraction, analysis and their antioxidant and anticancer properties. Molecules. 2010;15(10):7313–7352. https://doi.org/10.3390/molecules15107313 Wakeel A, Jan SA, Ullah I, Shinwari ZK, Xu M. Solvent polarity mediates phytochemical yield and antioxidant capacity of Isatis tinctoria. PeerJ. 2019;7:e7857. https://doi.org/10.7717/peerj.7857 Wakeel A, Jan SA, Ullah I, Shinwari ZK, Xu M. Solvent polarity mediates phytochemical yield and antioxidant capacity of Isatis tinctoria. PeerJ. 2019;7:e7857. https://doi.org/10.7717/peerj.7857 Elfalleh W, Hannachi H, Tlili N, Yahia Y, Nasri N, Ferchichi A. Total phenolic contents and antioxidant activities of pomegranate peel, seed, leaf and flower. Journal of Medicinal Plants Research. 2012;6:4724–4730. https://doi.org/10.5897/JMPR11.995 Elfalleh W, Hannachi H, Tlili N, Yahia Y, Nasri N, Ferchichi A. Total phenolic contents and antioxidant activities of pomegranate peel, seed, leaf and flower. Journal of Medicinal Plants Research. 2012;6:4724–4730. https://doi.org/10.5897/JMPR11.995 Singleton VL, Orthofer R, Lamuela-Raventos RM. Analysis of total phenols and other oxidation substrates and antioxidants by means of Folin-Ciocalteu reagent. Methods in Enzymology. 1999;299:152–178. https://doi.org/10.1016/S0076-6879(99)99017-1 Singleton VL, Orthofer R, Lamuela-Raventos RM. Analysis of total phenols and other oxidation substrates and antioxidants by means of Folin-Ciocalteu reagent. Methods in Enzymology. 1999;299:152–178. https://doi.org/10.1016/S0076-6879(99)99017-1 Zhishen J, Mengcheng T, Jianming W. The determination of flavonoid contents in mulberry and their scavenging effects on superoxide radicals. Food Chemistry. 1999;64(4):555–559. https://doi.org/10.1016/S0308-8146(98)00102-2 Zhishen J, Mengcheng T, Jianming W. The determination of flavonoid contents in mulberry and their scavenging effects on superoxide radicals. Food Chemistry. 1999;64(4):555–559. https://doi.org/10.1016/S0308-8146(98)00102-2 Kumaran A, Karunakaran RJ. In vitro antioxidant activities of methanol extracts of five Phyllanthus species from India. LWT – Food Science and Technology. 2007;40(2):344–352. https://doi.org/10.1016/j.lwt.2005.09.011 Kumaran A, Karunakaran RJ. In vitro antioxidant activities of methanol extracts of five Phyllanthus species from India. LWT – Food Science and Technology. 2007;40(2):344–352. https://doi.org/10.1016/j.lwt.2005.09.011 McMurrough I, McDowell J. Chromatographic separation and automated analysis of flavanols. Analytical Biochemistry. 1978;91(1):92–100. https://doi.org/10.1016/0003-2697(78)90819-9 McMurrough I, McDowell J. Chromatographic separation and automated analysis of flavanols. Analytical Biochemistry. 1978;91(1):92–100. https://doi.org/10.1016/0003-2697(78)90819-9 Lee J, Durst RW, Wrolstad RE. Determination of total monomeric anthocyanin pigment content of fruit juices, beverages, natural colorants, and wines by the pH differential method: Collaborative study. Journal of AOAC International. 2005;88(5):1269–1278. https://doi.org/10.1093/jaoac/88.5.1269 Lee J, Durst RW, Wrolstad RE. Determination of total monomeric anthocyanin pigment content of fruit juices, beverages, natural colorants, and wines by the pH differential method: Collaborative study. Journal of AOAC International. 2005;88(5):1269–1278. https://doi.org/10.1093/jaoac/88.5.1269 Broadhurst RB, Jones WT. Analysis of condensed tannins using acidified vanilin. Journal of the Science of Food and Agriculture. 1978;29(9):788–794. https://doi.org/10.1002/jsfa.2740290908 Broadhurst RB, Jones WT. Analysis of condensed tannins using acidified vanilin. Journal of the Science of Food and Agriculture. 1978;29(9):788–794. https://doi.org/10.1002/jsfa.2740290908 Gupta C, Verma R. Visual estimation and spectrophotometric determination of tannin content and antioxidant activity of three common vegetable. International Journal Pharmaceutical Sciences and Research. 2011;2(1):175–182. https://doi.org/10.13040/IJPSR.0975-8232.2(1).175-82 Gupta C, Verma R. Visual estimation and spectrophotometric determination of tannin content and antioxidant activity of three common vegetable. International Journal Pharmaceutical Sciences and Research. 2011;2(1):175–182. https://doi.org/10.13040/IJPSR.0975-8232.2(1).175-82 Blois MS. Antioxidant determinations by the use of a stable free radical. Nature. 1958;181(4617):1199–1200. https://doi.org/10.1038/1811199a0 Blois MS. Antioxidant determinations by the use of a stable free radical. Nature. 1958;181(4617):1199–1200. https://doi.org/10.1038/1811199a0 Carter P. Spectrophotometric determination of serum iron at the submicrogram level with a new reagent (ferrozine). Analytical Biocehmistry. 1971;40(2):450–458. https://doi.org/10.1016/0003-2697(71)90405-2 Carter P. Spectrophotometric determination of serum iron at the submicrogram level with a new reagent (ferrozine). Analytical Biocehmistry. 1971;40(2):450–458. https://doi.org/10.1016/0003-2697(71)90405-2 Oyaizu M. Studies on product of browning reaction. Antioxidative activities of products of browning reaction prepared from glucosamine. The Japanese Society of Nutrition and Dietetics. 1986;44(6):307–315. https://doi.org/10.5264/eiyogakuzashi.44.307 Oyaizu M. Studies on product of browning reaction. Antioxidative activities of products of browning reaction prepared from glucosamine. The Japanese Society of Nutrition and Dietetics. 1986;44(6):307–315. https://doi.org/10.5264/eiyogakuzashi.44.307 Apak R, Güçlü K, Ozyürek M, Karademir SE. Novel total antioxidant capacity index for polyphenols and vitamins C and E, using their cupric ion reducing capability in the presence of neocuproine: CUPRAC method. Journal of Agricultural and Food Chemistry. 2004;52(26):7970–7981. https://doi.org/10.1021/jf048741x Apak R, Güçlü K, Ozyürek M, Karademir SE. Novel total antioxidant capacity index for polyphenols and vitamins C and E, using their cupric ion reducing capability in the presence of neocuproine: CUPRAC method. Journal of Agricultural and Food Chemistry. 2004;52(26):7970–7981. https://doi.org/10.1021/jf048741x Trabelsi A, El Kaibi MA, Abbassi A, Horchani A, Chekir-Ghedira L, Ghedira K. Phytochemical study and antibacterial and antibiotic modulation activity of Punica granatum (Pomegranate) leaves. Scientifica. 2020;2020:8271203. https://doi.org/10.1155/2020/8271203 Trabelsi A, El Kaibi MA, Abbassi A, Horchani A, Chekir-Ghedira L, Ghedira K. Phytochemical study and antibacterial and antibiotic modulation activity of Punica granatum (Pomegranate) leaves. Scientifica. 2020;2020:8271203. https://doi.org/10.1155/2020/8271203 Kaur G, Jabbar Z, Athar M, Alam MS. Punica granatum (pomegranate) flower extract possesses potent antioxidant activity and abrogates Fe-NTA induced hepatotoxicity in mice. Food and Chemical Toxicology. 2006;44(7):984–993. https://doi.org/10.1016/j.fct.2005.12.001 Kaur G, Jabbar Z, Athar M, Alam MS. Punica granatum (pomegranate) flower extract possesses potent antioxidant activity and abrogates Fe-NTA induced hepatotoxicity in mice. Food and Chemical Toxicology. 2006;44(7):984–993. https://doi.org/10.1016/j.fct.2005.12.001 Abubakar AR, Haque M. Preparation of medicinal plants. Basic extraction and fractionation procedures for experimental purposes. Journal of Pharmacy and BioAllied Sciences. 2020;12(1):1–10. https://doi.org/10.4103/jpbs.JPBS_175_19 Abubakar AR, Haque M. Preparation of medicinal plants. Basic extraction and fractionation procedures for experimental purposes. Journal of Pharmacy and BioAllied Sciences. 2020;12(1):1–10. https://doi.org/10.4103/jpbs.JPBS_175_19 Kaneria MJ, Bapodara MB, Chanda SV. Effect of extraction techniques and solvents on antioxidant activity of pomegranate (Punica granatum L.) leaf and stem. Food Analytical Methods. 2012;5:396–404. https://doi.org/10.1007/s12161-011-9257-6 Kaneria MJ, Bapodara MB, Chanda SV. Effect of extraction techniques and solvents on antioxidant activity of pomegranate (Punica granatum L.) leaf and stem. Food Analytical Methods. 2012;5:396–404. https://doi.org/10.1007/s12161-011-9257-6 Zheng J, Yu X, Maninder M, Xu B. Total phenolic and antioxidants profiles of commonly consumed edible flowers in China. International Journal of Food Properties. 2018;21(1):1524–1540. https://doi.org/10.1080/10942912.2018.1494195 Zheng J, Yu X, Maninder M, Xu B. Total phenolic and antioxidants profiles of commonly consumed edible flowers in China. International Journal of Food Properties. 2018;21(1):1524–1540. https://doi.org/10.1080/10942912.2018.1494195 Attanayake R, Rajapaksha R, Weerakkody P, Bandaranayake PCG. The effect of maturity status on biochemical composition, antioxidant acitivity, and antocyanin biosynthesis gene expression in a pomegranate (Punica granatum L.) cultivar with red flowers, yellow peel, and pinkish arils. Journal of Plant Growth Regulation. 2019;38:992–1006. https://doi.org/10.1007/s00344-018-09909-2 Attanayake R, Rajapaksha R, Weerakkody P, Bandaranayake PCG. The effect of maturity status on biochemical composition, antioxidant acitivity, and antocyanin biosynthesis gene expression in a pomegranate (Punica granatum L.) cultivar with red flowers, yellow peel, and pinkish arils. Journal of Plant Growth Regulation. 2019;38:992–1006. https://doi.org/10.1007/s00344-018-09909-2 Mekni M, Azez R, Tekaya M, Mechri B, Hammami M. Phenolic, non-phenolic compounds and antioxidant activity of pomegranate flower, leaf and bark extracts of four Tunisian cultivars. Journal of Medicinal Plants Research. 2013;7(17):1100–1107. Mekni M, Azez R, Tekaya M, Mechri B, Hammami M. Phenolic, non-phenolic compounds and antioxidant activity of pomegranate flower, leaf and bark extracts of four Tunisian cultivars. Journal of Medicinal Plants Research. 2013;7(17):1100–1107. Kopjar M, Orsolic M, Pilizota V. Anthocyanins, phenols, and antioxidant activity of sour cherry puree extracts and their stability during storage. International Journal of Food Properties. 2014;17(6):1393–1405. https://doi.org/10.1080/10942912.2012.714027 Kopjar M, Orsolic M, Pilizota V. Anthocyanins, phenols, and antioxidant activity of sour cherry puree extracts and their stability during storage. International Journal of Food Properties. 2014;17(6):1393–1405. https://doi.org/10.1080/10942912.2012.714027 Lee C-J, Chen L-G, Liang W-L, Wang C-C. Anti-inflammatory effects of Punica granatum Linne in vitro and in vivo. Food Chemistry. 2010;118(2):315–322. https://doi.org/10.1016/j.foodchem.2009.04.123 Lee C-J, Chen L-G, Liang W-L, Wang C-C. Anti-inflammatory effects of Punica granatum Linne in vitro and in vivo. Food Chemistry. 2010;118(2):315–322. https://doi.org/10.1016/j.foodchem.2009.04.123 Castañeda-Ovando A, Pacheco-Hernández ML, Páez-Hernández ME, Rodríguez JA, Galán-Vidal CA. Chemical studies of anthocyanins: A review. Food Chemistry. 2009;113(4):859–871. https://doi.org/10.1016/j.foodchem.2008.09.001 Castañeda-Ovando A, Pacheco-Hernández ML, Páez-Hernández ME, Rodríguez JA, Galán-Vidal CA. Chemical studies of anthocyanins: A review. Food Chemistry. 2009;113(4):859–871. https://doi.org/10.1016/j.foodchem.2008.09.001 Iloki-Assanga SB, Lewis-Luján LM, Lara-Espinoza CL, Gil-Salido AA, Fernandez-Angulo D, Rubio-Pino JL, et al. Solvent effect on phytochemical constituent profiles and antioxidant activities, using four different extraction formulations for analysis of Bucida buceras L. and Phoradendron californicum. BMC Research Notes. 2015;8:396. https://doi.org/10.1186/s13104-015-1388-1 Iloki-Assanga SB, Lewis-Luján LM, Lara-Espinoza CL, Gil-Salido AA, Fernandez-Angulo D, Rubio-Pino JL, et al. Solvent effect on phytochemical constituent profiles and antioxidant activities, using four different extraction formulations for analysis of Bucida buceras L. and Phoradendron californicum. BMC Research Notes. 2015;8:396. https://doi.org/10.1186/s13104-015-1388-1 Medini F, Fellah H, Ksouri R, Abdelly C. Total phenolic, flavonoid and tannin contents and antioxidant and antimicrobial activities of organic extracts of shoots of the plant Limonium delicatulum. Journal of Taibah University for Science. 2014;8(3):216–224. https://doi.org/10.1016/j.jtusci.2014.01.003 Medini F, Fellah H, Ksouri R, Abdelly C. Total phenolic, flavonoid and tannin contents and antioxidant and antimicrobial activities of organic extracts of shoots of the plant Limonium delicatulum. Journal of Taibah University for Science. 2014;8(3):216–224. https://doi.org/10.1016/j.jtusci.2014.01.003 Zheng J, Meenu M, Xu B. A systematic investigation on free phenolics acids and flavonoids profiles of commonly consumed edible flowers in China. Journal of Pharmaceutical and Biomedical Analysis. 2019;172:268–277. https://doi.org/10.1016/j.jpba.2019.05.007 Zheng J, Meenu M, Xu B. A systematic investigation on free phenolics acids and flavonoids profiles of commonly consumed edible flowers in China. Journal of Pharmaceutical and Biomedical Analysis. 2019;172:268–277. https://doi.org/10.1016/j.jpba.2019.05.007 Feng L, Yin Y, Fang Y, Yang X. Quantitative determination of punicalagin and related substances in different parts of pomegranate. Food Analytical Methods. 2017;10:3600–3606. https://doi.org/10.1007/s12161-017-0916-0 Feng L, Yin Y, Fang Y, Yang X. Quantitative determination of punicalagin and related substances in different parts of pomegranate. Food Analytical Methods. 2017;10:3600–3606. https://doi.org/10.1007/s12161-017-0916-0 Ghafoor K, Al Juhaimi F, Özcan MM, Uslu N, Babiker EE, Mohamed Ahmed IA. Bioactive properties and phenolic compounds in bud, sprout, and fruit of Capparis spp. plants. Journal of Food Processing and Preservation. 2020;44(3):e14357. https://doi.org/10.1111/jfpp.14357 Ghafoor K, Al Juhaimi F, Özcan MM, Uslu N, Babiker EE, Mohamed Ahmed IA. Bioactive properties and phenolic compounds in bud, sprout, and fruit of Capparis spp. plants. Journal of Food Processing and Preservation. 2020;44(3):e14357. https://doi.org/10.1111/jfpp.14357 Donno D, Turrini F, Boggia R, Guido M, Gamba G, Mellano MG, et al. Vitis vinifera L. pruning waste for bud-preparations as source of phenolic compounds-traditional and innovative extraction techniques to produce new natural products. Plants. 2021;10(11):2233. https://doi.org/10.3390/plants10112233 Donno D, Turrini F, Boggia R, Guido M, Gamba G, Mellano MG, et al. Vitis vinifera L. pruning waste for bud-preparations as source of phenolic compounds-traditional and innovative extraction techniques to produce new natural products. Plants. 2021;10(11):2233. https://doi.org/10.3390/plants10112233 Gang Y, Eom T-Y, Marasinghe SD, Lee Y, Jo E, Oh C. Optimising the DPPH assay for cell-free marine microorganism supernatants. Marine Drugs. 2021;19(5):256. https://doi.org/10.3390/md19050256 Gang Y, Eom T-Y, Marasinghe SD, Lee Y, Jo E, Oh C. Optimising the DPPH assay for cell-free marine microorganism supernatants. Marine Drugs. 2021;19(5):256. https://doi.org/10.3390/md19050256 Baliyan S, Mukherjee R, Priyadarshini A, Vibhuti A, Gupta A, Pandey RP, et al. Determination of antioxidants by DPPH radical scavenging activity and quantitative phytochemical analysis of Ficus religiosa. Molecules. 2022;27(4):1326. https://doi.org/10.3390/molecules27041326 Baliyan S, Mukherjee R, Priyadarshini A, Vibhuti A, Gupta A, Pandey RP, et al. Determination of antioxidants by DPPH radical scavenging activity and quantitative phytochemical analysis of Ficus religiosa. Molecules. 2022;27(4):1326. https://doi.org/10.3390/molecules27041326 Yu M, Gouvinhas I, Rocha J, Barros AIRNA. Phytochemical and antioxidant analysis of medicinal and food plants towards bioactive food and pharmaceutical resources. Scientific Reports. 2021;11:10041. https://doi.org/10.1038/s41598-021-89437-4 Yu M, Gouvinhas I, Rocha J, Barros AIRNA. Phytochemical and antioxidant analysis of medicinal and food plants towards bioactive food and pharmaceutical resources. Scientific Reports. 2021;11:10041. https://doi.org/10.1038/s41598-021-89437-4 Gulcin İ, Alwasel SH. Metal ions, metal chelators and metal chelating assay as antioxidant method. Processes. 2022;10(1):132. https://doi.org/10.3390/pr10010132 Gulcin İ, Alwasel SH. Metal ions, metal chelators and metal chelating assay as antioxidant method. Processes. 2022;10(1):132. https://doi.org/10.3390/pr10010132 Gulcin İ, Buyukokuroglu ME, Kufrevioglu OI. Metal chelating and hydrogen peroxide scavenging effects of melatonin. Journal of Pineal Research. 2003;34(4):278–281. https://doi.org/10.1034/j.1600-079x.2003.00042.x Gulcin İ, Buyukokuroglu ME, Kufrevioglu OI. Metal chelating and hydrogen peroxide scavenging effects of melatonin. Journal of Pineal Research. 2003;34(4):278–281. https://doi.org/10.1034/j.1600-079x.2003.00042.x Rummun N, Somanah J, Ramsaha S, Bahorun T, Neergheen-Bhujun VS. Bioactivity of nonedible parts of Punica granatum L.: A potential source of functional ingredients. International Journal of Functional Ingredients. 2013;2013:602312. https://doi.org/10.1155/2013/602312 Rummun N, Somanah J, Ramsaha S, Bahorun T, Neergheen-Bhujun VS. Bioactivity of nonedible parts of Punica granatum L.: A potential source of functional ingredients. International Journal of Functional Ingredients. 2013;2013:602312. https://doi.org/10.1155/2013/602312 Zhang L, Yang X, Zhang Y, Wang L, Zhang R. In vitro antioxidant properties of different parts of pomegranate flower. Food and Bioproducts Processing. 2011;89(3):234–240. https://doi.org/10.1016/j.fbp.2010.04.007 Zhang L, Yang X, Zhang Y, Wang L, Zhang R. In vitro antioxidant properties of different parts of pomegranate flower. Food and Bioproducts Processing. 2011;89(3):234–240. https://doi.org/10.1016/j.fbp.2010.04.007 Ghazzawi HA, Al-Sayyed HF, Al-Kurd RA, Mwalla MM, Arafat TA, AbdelQader SM. Effect of different extraction solvents on the antioxidant content and capacity of nine seasonal fruits. Clinical Nutrition Open Science. 2021;38:33–42. https://doi.org/10.1016/j.nutos.2021.06.003 Ghazzawi HA, Al-Sayyed HF, Al-Kurd RA, Mwalla MM, Arafat TA, AbdelQader SM. Effect of different extraction solvents on the antioxidant content and capacity of nine seasonal fruits. Clinical Nutrition Open Science. 2021;38:33–42. https://doi.org/10.1016/j.nutos.2021.06.003 Sanda MA, Zengin G, Aktumsek A, Cakmak YS. Evaluation of antioxidant potential of two Daphne species (D. gnidioides and D. pontica) from Turkey. Emirates Journal of Food and Agriculture. 2015;27(6):488–494. https://doi.org/10.9755/ejfa.2015.04.030 Sanda MA, Zengin G, Aktumsek A, Cakmak YS. Evaluation of antioxidant potential of two Daphne species (D. gnidioides and D. pontica) from Turkey. Emirates Journal of Food and Agriculture. 2015;27(6):488–494. https://doi.org/10.9755/ejfa.2015.04.030 Alsataf S, Başyiğit B, Karaaslan M. Mutivariate analyses of the antioxidant, antidiabetic, antimicrobial activity of pomegranate tissues with respect to pomegranate juice. Waste and Biomass Valorization. 2021;12:5909–5921. https://doi.org/10.1007/s12649-021-01427-9 Alsataf S, Başyiğit B, Karaaslan M. Mutivariate analyses of the antioxidant, antidiabetic, antimicrobial activity of pomegranate tissues with respect to pomegranate juice. Waste and Biomass Valorization. 2021;12:5909–5921. https://doi.org/10.1007/s12649-021-01427-9 Uysal S, Zengin G, Aktumsek A, Karatas S. Chemical and biological approaches of nine fruit tree leaves collected from the Mediterranean region of Turkey. Journal of Functional Foods. 2016;22:518–532. https://doi.org/10.1016/j.jff.2016.02.006 Uysal S, Zengin G, Aktumsek A, Karatas S. Chemical and biological approaches of nine fruit tree leaves collected from the Mediterranean region of Turkey. Journal of Functional Foods. 2016;22:518–532. https://doi.org/10.1016/j.jff.2016.02.006