Effect of different solvents on the extraction of phytochemical compounds of Neem (Azadirachta indica A. Juss)
Subject Areas : BiochemistryMaryam Rahimi 1 , Javad Ali Soufiyan 2 , Zeynab Mohkami 3 , Ali Ali Ghorbani Ranjbari 4 , Fatemeh Bidarnamani 5
1 - University of zabol,agriculture faculty
2 - Department of Horticulture, Faculty of Agriculture, University of Zabol, Zabol, Iran
3 - Faculty member, Agricultural Research Institute, University of Zabol, Zabol, Iran
4 - Faculty of Veterinary , University of Tehran, Tehran, Iran
5 - Department of Agriculture and Plant Breeding, Institute of Agricultural Research, University of Zabol, Zabol, Iran
Keywords: Medicinal Plants, Extraction methods, antioxidant activity, Total phenols, total flavonoeids,
Abstract :
Neem (Azadirachta indica) that belongs to the Meliaceae family has many medicinal properties such as treatment of skin diseases, hypoglycemia, hypertension, cancer, and AIDS. In this study, to investigate the effect of different solvents on the quantity and quality of phytochemical properties of neem collected from Bahoklat area in Chabahar city, a factorial experiment was conducted in a completely randomized design in 2021. Ethanol, methanolic, n-hexane and acetone extracts of neem leaves and fruits were prepared by cold maceration method. Total phenol content, total flavonoid and antioxidant activity were measured by folin-cicalto reagent, aluminum chloride colorimetric and diphenylpicrylhydrazyl free radical scaving (DPPH) methods, respectively. The results showed that the effect of organ type, solvent type and their interaction on all measured traits was significant. Based on the results, the highest amount of total phenol (48.22 mg Gallic acid / g dry weight) was reported in acetone leaf extract, and the lowest amount (2.76 mg Gallic acid / g dry weight) was reported in hexane extract of fruit. In addition, the highest amount of total flavonoids was observed in acetoin fruit and leaf extracts (4.71 and 4.61 mg Quercetin / g dry weight, respectively) and the lowest amount (3.22 mg quercetin / g dry weight) was obtained in the hexane extract of fruit. The highest and the lowest antioxidant activity was detected in ethanolic leaf extract (94.77%) and acetone extract of fruit (12.02%), respectively. Therefore, neem leaf is suggested as a useful source of phenolic compounds and natural antioxidants.
Alkadi, H. 2020. A review on free radicals and antioxidants. Infectious Disorders-Drug Targets (Formerly Current Drug Targets-Infectious Disorders), 20(1): 16-26.
2.Barros, L., Baptista, P. and Ferreira, I.C.F.R. 2007. Effect of fruiting body maturity stage on antioxidant activity measured by several biochemical assays Lactarius piperatus. Food and Chemical Toxicology, 45(9): 1731-1737.
3.Chung, I.M., Thiruvengadam, M., Rekha, K. and Rajakumar, G. 2016. Elicitation enhanced the production of phenolic compounds and biological activities in hairy root cultures of bitter melon (Momordica charantia L.). Brazilian Archives of Biology and Technology, 59(3): 1-10.
4.Deng, Y., Shi, D., Yin, Z., Guo, J., Jia, R., Xu, J., Song, X., Lv, C., Fan, Q., Lian, X., Shi, F., Ye, G. and Zhan, W. 2012. Acaricidal activity of Petroleum ether extract of neem (Azadirachta indica) oil and its four fractions separated by column chromatography against Sarcoptes scabiei var.cniculi larvae in vitro. Experimental Parasitology, 130: 475-457.
5.Elumalai, P., Gunadharini, D.N., Senthilkumar, K., Banudevi, S., Arunkumar, R., Benson, C.S., Sharmila, G. and Arunakaran, J. 2011. Ethanolic neem (Azadirachta indica A. Juss) leaf extract induces apoptosis and inhibits the IGF signaling pathway in breast cancer cell lines. Biomedicine & Preventive Nutrition, 2(1): 59-68.
6.Farjaminezhad, R. and Garoosi, Gh. 2020. Establishment of green analytical method for ultrasound-assisted extraction of azadirachtin, mevalonic acid and squalene from cell suspension culture of Azadirachta indica using response surface methodology. Industrial Crops and Products, 144: 111946.
7.Firoozkoohi, F., Esmailzadeh Bahabadi, S., Mohkami, Z. and Yousefzaei, F. 2018. The effect of different solvents on the amount of phenol and total flavonoids and antioxidant activity of different organs of Momordica charantia L. in Sistan region. Journal of Ecophytochemistry of Medicinal Plants, 20(4): 85-74.
8.Frankel, E.N. and Meyer, A.S. 2000. The problems of using one‐dimensional methods to evaluate multifunctional food and biological antioxidants. Journal of the Science of Food and Agriculture, 80(13): 1925-1941.
9.Ghasemzadeh, A., Jaafar, H.Z. and Rahmat, A. 2011. Effects of solvent type on phenolics and flavonoids content and antioxidant activities in two varieties of young ginger (Zingiber officinale Roscoe) extracts. Journal of Medicinal Plants Research, 5(7): 1147-1154.
10.Ghorbani Ghozhdi, H., Mohkami, Z. and Estaji, A. 2021. Medicinal plants in Iranian traditional medicine (Matching the name of native name with the scientific name). Marz danesh publisher. 629 p. (In Persian).
11.Hossain, M.A., Al-Toubi, W.A.S., Weli, A.M., Al-Rahimi, Q. and Al-Sabani, J.N. 2013. Identification and characterization of chemical compounds in different crude extracts from leaves of Omani neem. Journal of Taibah University for Science, 7(4): 181-188.
12.Islas, J.F., Acosta, E., G-Buentello, Z., Delgado-Gallegos, J.L., Moreno-Treviño, M.J., Escalante, B.E. and Moreno-Cuevas, J. 2020. An overview of Neem (Azadirachta indica) and its potential impact on health. Journal of Functional Foods, 74: 104171.
13.Jonathan, S.G., Olawuyi, O.J., Aina, D.A., Odeniyi, S.O., Adediji, I.O. and Ikhedia, A. 2012. Comparative studies on antifungal, anti-oxidant and phytochemical potential of Momordica charantia and Moringa oleifera. New York Science Journal, 5(12): 17-28.
14.Khanavi, M., Hasanloo, T. and Haji Mahdipour, H. 2019. Quantitative determination of Azadirachtin in Melia indica and M. azedarach. 2019. Medicinal plants journal, 69: 185-193. (In Persian)
15.Li, F.J., Liu, X.Y., Xu, G.J. and Guo, H.Y. 2015. Effects of extraction solvents on antioxidant activity of Bitter Gourd (Momordica charantia L.) fruits in vitro. Innovations in Food Research, 1: 1-3.
16.López, C.J., Caleja, C., Prieto, M.A., Filomena Barreiro M., Barros, L. and C.F.R. Ferreira, I. 2018. Optimization and comparison of heat and ultrasound assisted extraction techniques to obtain anthocyanin compounds from Arbutus unedo L. Fruits, Food Chemistry, 264:81-91.
17.Omidbeigi, R. 2015. Production and processing of medicinal plants, Vol. 4. Quds Razavi Province Publications. 428 p. (In Persian)
18.Parsons, B.J. 2017. Antioxidants in food, the significance of characterization identification, chemical and biological assays in determining the role of antioxidants in food. Foods, 6(8): 68-72.
19.Patel, S., Patel, T., Parmar, K., Patel, B. and Patel, P. 2011. Evaluation of antioxidant activity, phenol and flavonoid contents of Momordica charantia Linn fruit. Advance Research in Pharmaceuticals and Biologicals, 1: 120- 129.
20.Pham, H.N.T., Nguyen, V.T., Vuong, Q.V., Bowyer, M.C. and Scarlett, C.J. 2015. Effect of extraction solvents and drying methods on the physicochemical and antioxidant properties of Helicteres hirsuta Lour. leaves. Technologies, 3(4): 285-301.
21.Sabura, A., Ahmadi, A., Zinali, A. and Parsa, M. 2014. Comparison of phenolic, flavonoid content and antioxidant activity of shoots of two populations of Scutellaria pinnaifida in northern Iran. Journal of Rafsanjan University of Medical Sciences, 13: 266-249. (In persian)
22.Sadeghi, Z., Valizadeh, J. and Aziziyan Shermeh, A. 2016. Investigation of phenol content, total flavonoids and antioxidant activity of Pistacia atlantica gum from Saravan region (Sistan and Baluchestan province). Journal of Hypophytochemistry of Medicinal Plants, 10(2): 18-27. (In persian)
23.Sahrawat, A., Sharma, J., Siddarth, N., Tiwari, S., Datt Joshi, M., Pundhir, A., Kumar, R., Radhika, A., Shivani, B., Nilesh, Sh. and Akash, G. 2018. Phytochemical analysis and antibacterial properties of Azadirachta indica (Neem) leaves extract against E. coli. Journal of Pharmacognosy and Phytochemistry, 7(4): 1368-1371.
24.Sanchez-Moreno, C., Larrauri, J.A. and Saura- Calixto, F. 1999. Free radical scavenging capacity and inhibition of lipid oxidation of wines, grape juices and related polyphenolic constituents. Food Research International, 32: 407-412.
25.Subapriya, R. and Nagini, S. 2005. Medicinal properties of neem leave a review. Curr Med Chem Anticancer Agents, 5(2):149-156.
26.Sultana, B., Anwar, F. and Przybylski, R. 2007. Antioxidant activity of phenolic components present in barks of Azadirachta indica, Terminalia arjuna, Acacia nilotica, and Eugenia jambolana Lam. trees. Food Chemistry, 104(3): 1106-1114.
27.Wen, L.J. and Liu, W.F. 2007. Study on extracting and antioxidant activity of flavonoids from Momordica charantia L. Food Science, 9: 042.
28.Zhu, Zh., Guan, Q., Koubaa, M.J., Barba, F., Roohinejad, Sh., Cravotto, G., Yang, X., Sh., Li, and He, J. 2017. HPLC-DAD-ESI-MS2 analytical profile of extracts obtained from purple sweet potato after green ultrasound-assisted extraction, Food Chemistry, 215: 391-400.
_||_Alkadi, H. 2020. A review on free radicals and antioxidants. Infectious Disorders-Drug Targets (Formerly Current Drug Targets-Infectious Disorders), 20(1): 16-26.
2.Barros, L., Baptista, P. and Ferreira, I.C.F.R. 2007. Effect of fruiting body maturity stage on antioxidant activity measured by several biochemical assays Lactarius piperatus. Food and Chemical Toxicology, 45(9): 1731-1737.
3.Chung, I.M., Thiruvengadam, M., Rekha, K. and Rajakumar, G. 2016. Elicitation enhanced the production of phenolic compounds and biological activities in hairy root cultures of bitter melon (Momordica charantia L.). Brazilian Archives of Biology and Technology, 59(3): 1-10.
4.Deng, Y., Shi, D., Yin, Z., Guo, J., Jia, R., Xu, J., Song, X., Lv, C., Fan, Q., Lian, X., Shi, F., Ye, G. and Zhan, W. 2012. Acaricidal activity of Petroleum ether extract of neem (Azadirachta indica) oil and its four fractions separated by column chromatography against Sarcoptes scabiei var.cniculi larvae in vitro. Experimental Parasitology, 130: 475-457.
5.Elumalai, P., Gunadharini, D.N., Senthilkumar, K., Banudevi, S., Arunkumar, R., Benson, C.S., Sharmila, G. and Arunakaran, J. 2011. Ethanolic neem (Azadirachta indica A. Juss) leaf extract induces apoptosis and inhibits the IGF signaling pathway in breast cancer cell lines. Biomedicine & Preventive Nutrition, 2(1): 59-68.
6.Farjaminezhad, R. and Garoosi, Gh. 2020. Establishment of green analytical method for ultrasound-assisted extraction of azadirachtin, mevalonic acid and squalene from cell suspension culture of Azadirachta indica using response surface methodology. Industrial Crops and Products, 144: 111946.
7.Firoozkoohi, F., Esmailzadeh Bahabadi, S., Mohkami, Z. and Yousefzaei, F. 2018. The effect of different solvents on the amount of phenol and total flavonoids and antioxidant activity of different organs of Momordica charantia L. in Sistan region. Journal of Ecophytochemistry of Medicinal Plants, 20(4): 85-74.
8.Frankel, E.N. and Meyer, A.S. 2000. The problems of using one‐dimensional methods to evaluate multifunctional food and biological antioxidants. Journal of the Science of Food and Agriculture, 80(13): 1925-1941.
9.Ghasemzadeh, A., Jaafar, H.Z. and Rahmat, A. 2011. Effects of solvent type on phenolics and flavonoids content and antioxidant activities in two varieties of young ginger (Zingiber officinale Roscoe) extracts. Journal of Medicinal Plants Research, 5(7): 1147-1154.
10.Ghorbani Ghozhdi, H., Mohkami, Z. and Estaji, A. 2021. Medicinal plants in Iranian traditional medicine (Matching the name of native name with the scientific name). Marz danesh publisher. 629 p. (In Persian).
11.Hossain, M.A., Al-Toubi, W.A.S., Weli, A.M., Al-Rahimi, Q. and Al-Sabani, J.N. 2013. Identification and characterization of chemical compounds in different crude extracts from leaves of Omani neem. Journal of Taibah University for Science, 7(4): 181-188.
12.Islas, J.F., Acosta, E., G-Buentello, Z., Delgado-Gallegos, J.L., Moreno-Treviño, M.J., Escalante, B.E. and Moreno-Cuevas, J. 2020. An overview of Neem (Azadirachta indica) and its potential impact on health. Journal of Functional Foods, 74: 104171.
13.Jonathan, S.G., Olawuyi, O.J., Aina, D.A., Odeniyi, S.O., Adediji, I.O. and Ikhedia, A. 2012. Comparative studies on antifungal, anti-oxidant and phytochemical potential of Momordica charantia and Moringa oleifera. New York Science Journal, 5(12): 17-28.
14.Khanavi, M., Hasanloo, T. and Haji Mahdipour, H. 2019. Quantitative determination of Azadirachtin in Melia indica and M. azedarach. 2019. Medicinal plants journal, 69: 185-193. (In Persian)
15.Li, F.J., Liu, X.Y., Xu, G.J. and Guo, H.Y. 2015. Effects of extraction solvents on antioxidant activity of Bitter Gourd (Momordica charantia L.) fruits in vitro. Innovations in Food Research, 1: 1-3.
16.López, C.J., Caleja, C., Prieto, M.A., Filomena Barreiro M., Barros, L. and C.F.R. Ferreira, I. 2018. Optimization and comparison of heat and ultrasound assisted extraction techniques to obtain anthocyanin compounds from Arbutus unedo L. Fruits, Food Chemistry, 264:81-91.
17.Omidbeigi, R. 2015. Production and processing of medicinal plants, Vol. 4. Quds Razavi Province Publications. 428 p. (In Persian)
18.Parsons, B.J. 2017. Antioxidants in food, the significance of characterization identification, chemical and biological assays in determining the role of antioxidants in food. Foods, 6(8): 68-72.
19.Patel, S., Patel, T., Parmar, K., Patel, B. and Patel, P. 2011. Evaluation of antioxidant activity, phenol and flavonoid contents of Momordica charantia Linn fruit. Advance Research in Pharmaceuticals and Biologicals, 1: 120- 129.
20.Pham, H.N.T., Nguyen, V.T., Vuong, Q.V., Bowyer, M.C. and Scarlett, C.J. 2015. Effect of extraction solvents and drying methods on the physicochemical and antioxidant properties of Helicteres hirsuta Lour. leaves. Technologies, 3(4): 285-301.
21.Sabura, A., Ahmadi, A., Zinali, A. and Parsa, M. 2014. Comparison of phenolic, flavonoid content and antioxidant activity of shoots of two populations of Scutellaria pinnaifida in northern Iran. Journal of Rafsanjan University of Medical Sciences, 13: 266-249. (In persian)
22.Sadeghi, Z., Valizadeh, J. and Aziziyan Shermeh, A. 2016. Investigation of phenol content, total flavonoids and antioxidant activity of Pistacia atlantica gum from Saravan region (Sistan and Baluchestan province). Journal of Hypophytochemistry of Medicinal Plants, 10(2): 18-27. (In persian)
23.Sahrawat, A., Sharma, J., Siddarth, N., Tiwari, S., Datt Joshi, M., Pundhir, A., Kumar, R., Radhika, A., Shivani, B., Nilesh, Sh. and Akash, G. 2018. Phytochemical analysis and antibacterial properties of Azadirachta indica (Neem) leaves extract against E. coli. Journal of Pharmacognosy and Phytochemistry, 7(4): 1368-1371.
24.Sanchez-Moreno, C., Larrauri, J.A. and Saura- Calixto, F. 1999. Free radical scavenging capacity and inhibition of lipid oxidation of wines, grape juices and related polyphenolic constituents. Food Research International, 32: 407-412.
25.Subapriya, R. and Nagini, S. 2005. Medicinal properties of neem leave a review. Curr Med Chem Anticancer Agents, 5(2):149-156.
26.Sultana, B., Anwar, F. and Przybylski, R. 2007. Antioxidant activity of phenolic components present in barks of Azadirachta indica, Terminalia arjuna, Acacia nilotica, and Eugenia jambolana Lam. trees. Food Chemistry, 104(3): 1106-1114.
27.Wen, L.J. and Liu, W.F. 2007. Study on extracting and antioxidant activity of flavonoids from Momordica charantia L. Food Science, 9: 042.
28.Zhu, Zh., Guan, Q., Koubaa, M.J., Barba, F., Roohinejad, Sh., Cravotto, G., Yang, X., Sh., Li, and He, J. 2017. HPLC-DAD-ESI-MS2 analytical profile of extracts obtained from purple sweet potato after green ultrasound-assisted extraction, Food Chemistry, 215: 391-400.
