ارزیابی فعالیت آنتی اکسیدانتی و کاربرد تکنیک HPLC جهت آنالیز ترکیبات فنلی عصاره برگ درخت بادام زاگرسی (Amygdalus haussknechtii L.)
محورهای موضوعی : ژنتیکعلی اصغر حاتم نیا 1 , ربیع شریفی 2
1 - عضو هیات علمی گروه زیست‌شناسی، دانشکده علوم، دانشگاه ایلام
2 - گروه زیست‌شناسی، دانشکده علوم پایه، دانشگاه ایلام، ایلام، ایران
کلید واژه: ترکیبات فنلی, کروماتوگرافی مایع با کارایی بالا, برگ, فعالیت آنتی‌اکسیدانتی, بادام زاگرسی,
چکیده مقاله :
این تحقیق با هدف بررسی و مقایسه میزان ترکیبات آنتی اکسیدانتی و فعالیت آنتی اکسیدانتی سه ژنوتیپ بادام زاگرسی در رویشگاه های طبیعی استان ایلام انجام گرفت. محتوی فنل کل با استفاده از روش فولین- سیکالتئو و فعالیت آنتی اکسیدانتی با استفاده از دو سنجش ظرفیت جمع آوری رادیکال DPPH و ظرفیت جمع‌آوری رادیکال نیتریت ارزیابی شد. جهت تعیین کمی ترکیبات فنلی از دستگاه کروماتوگرافی مایع با کارایی بالا استفاده گردید. نتایج نشان داد که فعالیت پاداکسایشی عصاره برگ ژنوتیپ H3 به طور معنی داری بیشتر از ژنوتیپ های H1 و H2 بوده که این فعالیت بالا را می توان به میزان بالای محتوی فنلی و فلاوونوئیدی نسبت داد، به طوری که همبستگی بالایی بین محتوی فنل کل با ظرفیت جمع آوری رادیکال DPPH (902/0 =R) و ظرفیت جمع آوری رادیکال نیتریت (806/0 =R) مشاهده شد. آنالیز ترکیبات فنلی توسط HPLC وجود ترکیبات کاتچین، سیناپیک اسید، کوئرستین و آپیژنین در ژنوتیپ H3 را تایید نمود. همچنین ترکیب آپژنین در هر سه ژنوتیپ مورد مطالعه مشاهده شد. به طور کلی نتایج پیشنهاد می دهند که عصاره برگ ژنوتیپ H3 با محتوی بالایی از ترکیبات فنلی و فلاوونوئیدی و فعالیت پاداکسایشی بالا می تواند به‌عنوان منبعی از ترکیبات پاداکسایشی در صنعت داروسازی مورد استفاده قرار گیرد.
This study was conducted to investigate antioxidant compounds contents and antioxidant activity of three Amygdalus haussknechtii genotypes in the natural habitat of Ilam Province. Total phenolic content was measured using Folin–Ciocalteu method and antioxidant activity was estimated by two methods, namely DPPH (2,2-diphenyl- 1- picrylhydrazyl) and nitric oxide radical scavenging. High- performance chromatography was used for quantitative estimation of the phenolic compounds. Results showed that antioxidant activity of leaf extract of H3 genotype was significantly higher than that of H1 and H2 genotypes. The high antioxidant activity can be attributed to higher total phenolic and flavonoid contents, so that high correlation coefficient was observed between total phenolic content and DPPH and nitric oxide radical scavenging, R = 0.902 and R= 0.806, respectively. Analyzing the phenolic compounds by HPLC method indicated that catechin, sinapic acid, quercetin, and apigenin were found in H3 genotype. Also, apigenin compound was observe in all three investigated genotypes. Generally, the results suggested that leaf extract of H3 genotype with high phenolic and flavonoid content and antioxidant activity can be presented as a source of natural antioxidant and used in pharmaceutical industries.
Ang, L.Z.P., Hashim, R., Sulaiman, S.F., Coulibaly, A.Y., Sulaiman, O., Kawamura, F. and Salleh, K.M. (2015). In vitro antioxidant and antidiabetic activites of Gluta torquata. Industrial Crops and Products, 76: 755-760.
Barreira, J.C., Ferreira, I.C., Oliveira, M.B.P. and Pereira, J.A. (2008). Antioxidant activity and bioactive compounds of ten Portuguese regional and commercial almond cultivars. Food and Chemical Toxicology, 46(6): 2230-2235.
Browicz, K. and Zohary, D. (1996). The genus Amygdalus L. (Rosaceae): species relationships, distribution and evolution under domestication. Genetic Resources and Crop Evolution. 43, 229–247.
Carvalho, M., Ferreira, P.J., Mendes, V.S., Silva, R., Pereira, J.A., Jerónimo, C. and Silva, B.M. (2010). Human cancer cell antiproliferative and antioxidant activities of Juglans regia L. Food and chemical toxicology, 48(1): 441-447.
Del Rio, D., Rodriguez-Mateos, A., Spencer, J.P., Tognolini, M., Borges, G. and Crozier, A. (2013). Dietary (poly) phenolics in human health: structures, bioavailability, and evidence of protective effects against chronic diseases. Antioxidants & redox signaling, 18(14): 1818-1892.
Garrat, D.C. (1964). The quantitative analysis of drugs (vol 3). pp. 456–458. Japan: Chapman and Hall.
Hatamnia, A.A., Abbaspour, N. and Darvishzadeh, R. (2014). Antioxidant activity and phenolic profile of different parts of Bene (Pistacia atlantica subsp. kurdica) fruits. Food chemistry, 145: 306-311.
Hatamnia, A.A., malekzadeh, P., Nourollahi, Kh. and Valadbeigi, V. (2015). A study on phenolic compounds content and antioxidant activity of kolkhoung (Pistacia khinjuk Stocks) leaf in natural habitat of Ilam province. Journal of Iranian Plant Ecophysiological Research, 40: 31-40.
Hertog, M.G., Hollman, P.C. and Venema, D.P. (1992). Optimization of a quantitative HPLC determination of potentially anticarcinogenic flavonoids in vegetables and fruits. Journal of Agricultural and Food Chemistry, 40(9): 1591-1598.
Heim, K.E., Tagliaferro, A.R. and Bobilya, D.J. (2002). Flavonoid antioxidants: chemistry, metabolism and structure-activity relationships. The Journal of nutritional biochemistry, 13(10): 572-584.
Hu, F.B. and Willett, W.C. (2002). Optimal diets for prevention of coronary heart disease. Jama, 288(20): 2569-2578.
Huang, D., Ou, B. and Prior, R.L. (2005). The chemistry behind antioxidant capacity assays. Journal of agricultural and food chemistry, 53(6): 1841-1856.
Kaur, C. and Kapoor, H.C. (2001). Antioxidants in fruits and vegetables–the millennium’s health. International journal of food science & technology, 36(7): 703-725.
Lenucci, M.S., Cadinu, D., Taurino, M., Piro, G. and Dalessandro, G. (2006). Antioxidant composition in cherry and high-pigment tomato cultivars. Journal of agricultural and food chemistry, 54(7): 2606-2613.
Marcocci, L., Packer, L., Droy-Lefai, M.T., Sekaki, A. and Gardes-Albert, M. (1994). Antioxidant action of Ginkgo biloba extracts EGb 761. Methods in Enzymology, 234: 462–475.
Milbury, P.E., Chen, C.Y., Dolnikowski, G.G. and Blumberg, J.B. (2006). Determination of flavonoids and phenolics and their distribution in almonds. Journal of Agricultural and Food Chemistry, 54(14): 5027-5033.
Moure, A., Pazos, M., Medina, I., Domínguez, H. and Parajó, J.C. (2007). Antioxidant activity of extracts produced by solvent extraction of almond shells acid hydrolysates. Food Chemistry, 101(1): 193-201.
Rababah, T.M., Hettiarachchy, N.S. and Horax, R. (2004). Total phenolics and antioxidant activities of fenugreek, green tea, black tea, grape seed, ginger, rosemary, gotu kola, and ginkgo extracts, vitamin E, and tert-butylhydroquinone. Journal of Agricultural and Food Chemistry, 52(16): 5183-5186.
Rahemi, A., Fattahi Moghaddam, M.R., Ebadi, A., Taghavi, T. and Hassani, D. (2011). Fruit Characteristics of some Wild Almonds in Iran. Seed and Plant Improvement Journal, 27(4): 459-481.
Rajaei, A., Barzegar, M., Mobarez, A.M., Sahari, M.A. and Esfahani, Z.H. (2010). Antioxidant, anti-microbial and antimutagenicity activities of pistachio (Pistachia vera) green hull extract. Food and Chemical Toxicology, 48(1): 107-112.
Ryan, D., Robards, K., Prenzler, P. and Antolovich, M. (1999). Applications of mass spectrometry to plant phenols. TrAC Trends in Analytical Chemistry, 18(5): 362-372.
Sánchez-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(6): 407-412.
Sfahlan, A.J., Mahmoodzadeh, A., Hasanzadeh, A., Heidari, R. and Jamei, R. (2009). Antioxidants and antiradicals in almond hull and shell (Amygdalus communis L.) as a function of genotype. Food Chemistry, 115(2): 529-533.
Tlili, N., Kirkan, B. and Sarikurkcu, C. (2019). LC–ESI–MS/MS characterization, antioxidant power and inhibitory effects on α-amylase and tyrosinase of bioactive compounds from hulls of Amygdalus communis: The influence of the extracting solvents. Industrial crops and products, 128: 147-152.
Tomaino, A., Martorana, M., Arcoraci, T., Monteleone, D., Giovinazzo, C. and Saija, A. (2010). Antioxidant activity and phenolic profile of pistachio (Pistacia vera L., variety Bronte) seeds and skins. Biochimie, 92(9): 1115-1122.
Tsantili, E., Shin, Y., Nock, J.F. and Watkins, C.B. (2010). Antioxidant concentrations during chilling injury development in peaches. Postharvest Biology and Technology, 57(1): 27-34.
Wijeratne, S.S., Abou-Zaid, M.M. and Shahidi, F. (2006). Antioxidant polyphenols in almond and its coproducts. Journal of Agricultural and Food Chemistry, 54(2): 312-318.
Wu, H.C., Chen, H.M. and Shiau, C.Y. (2003). Free amino acids and peptides as related to antioxidant properties in protein hydrolysates of mackerel (Scomber austriasicus). Food Research International, 36(9-10): 949-957.
_||_Ang, L.Z.P., Hashim, R., Sulaiman, S.F., Coulibaly, A.Y., Sulaiman, O., Kawamura, F. and Salleh, K.M. (2015). In vitro antioxidant and antidiabetic activites of Gluta torquata. Industrial Crops and Products, 76: 755-760.
Barreira, J.C., Ferreira, I.C., Oliveira, M.B.P. and Pereira, J.A. (2008). Antioxidant activity and bioactive compounds of ten Portuguese regional and commercial almond cultivars. Food and Chemical Toxicology, 46(6): 2230-2235.
Browicz, K. and Zohary, D. (1996). The genus Amygdalus L. (Rosaceae): species relationships, distribution and evolution under domestication. Genetic Resources and Crop Evolution. 43, 229–247.
Carvalho, M., Ferreira, P.J., Mendes, V.S., Silva, R., Pereira, J.A., Jerónimo, C. and Silva, B.M. (2010). Human cancer cell antiproliferative and antioxidant activities of Juglans regia L. Food and chemical toxicology, 48(1): 441-447.
Del Rio, D., Rodriguez-Mateos, A., Spencer, J.P., Tognolini, M., Borges, G. and Crozier, A. (2013). Dietary (poly) phenolics in human health: structures, bioavailability, and evidence of protective effects against chronic diseases. Antioxidants & redox signaling, 18(14): 1818-1892.
Garrat, D.C. (1964). The quantitative analysis of drugs (vol 3). pp. 456–458. Japan: Chapman and Hall.
Hatamnia, A.A., Abbaspour, N. and Darvishzadeh, R. (2014). Antioxidant activity and phenolic profile of different parts of Bene (Pistacia atlantica subsp. kurdica) fruits. Food chemistry, 145: 306-311.
Hatamnia, A.A., malekzadeh, P., Nourollahi, Kh. and Valadbeigi, V. (2015). A study on phenolic compounds content and antioxidant activity of kolkhoung (Pistacia khinjuk Stocks) leaf in natural habitat of Ilam province. Journal of Iranian Plant Ecophysiological Research, 40: 31-40.
Hertog, M.G., Hollman, P.C. and Venema, D.P. (1992). Optimization of a quantitative HPLC determination of potentially anticarcinogenic flavonoids in vegetables and fruits. Journal of Agricultural and Food Chemistry, 40(9): 1591-1598.
Heim, K.E., Tagliaferro, A.R. and Bobilya, D.J. (2002). Flavonoid antioxidants: chemistry, metabolism and structure-activity relationships. The Journal of nutritional biochemistry, 13(10): 572-584.
Hu, F.B. and Willett, W.C. (2002). Optimal diets for prevention of coronary heart disease. Jama, 288(20): 2569-2578.
Huang, D., Ou, B. and Prior, R.L. (2005). The chemistry behind antioxidant capacity assays. Journal of agricultural and food chemistry, 53(6): 1841-1856.
Kaur, C. and Kapoor, H.C. (2001). Antioxidants in fruits and vegetables–the millennium’s health. International journal of food science & technology, 36(7): 703-725.
Lenucci, M.S., Cadinu, D., Taurino, M., Piro, G. and Dalessandro, G. (2006). Antioxidant composition in cherry and high-pigment tomato cultivars. Journal of agricultural and food chemistry, 54(7): 2606-2613.
Marcocci, L., Packer, L., Droy-Lefai, M.T., Sekaki, A. and Gardes-Albert, M. (1994). Antioxidant action of Ginkgo biloba extracts EGb 761. Methods in Enzymology, 234: 462–475.
Milbury, P.E., Chen, C.Y., Dolnikowski, G.G. and Blumberg, J.B. (2006). Determination of flavonoids and phenolics and their distribution in almonds. Journal of Agricultural and Food Chemistry, 54(14): 5027-5033.
Moure, A., Pazos, M., Medina, I., Domínguez, H. and Parajó, J.C. (2007). Antioxidant activity of extracts produced by solvent extraction of almond shells acid hydrolysates. Food Chemistry, 101(1): 193-201.
Rababah, T.M., Hettiarachchy, N.S. and Horax, R. (2004). Total phenolics and antioxidant activities of fenugreek, green tea, black tea, grape seed, ginger, rosemary, gotu kola, and ginkgo extracts, vitamin E, and tert-butylhydroquinone. Journal of Agricultural and Food Chemistry, 52(16): 5183-5186.
Rahemi, A., Fattahi Moghaddam, M.R., Ebadi, A., Taghavi, T. and Hassani, D. (2011). Fruit Characteristics of some Wild Almonds in Iran. Seed and Plant Improvement Journal, 27(4): 459-481.
Rajaei, A., Barzegar, M., Mobarez, A.M., Sahari, M.A. and Esfahani, Z.H. (2010). Antioxidant, anti-microbial and antimutagenicity activities of pistachio (Pistachia vera) green hull extract. Food and Chemical Toxicology, 48(1): 107-112.
Ryan, D., Robards, K., Prenzler, P. and Antolovich, M. (1999). Applications of mass spectrometry to plant phenols. TrAC Trends in Analytical Chemistry, 18(5): 362-372.
Sánchez-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(6): 407-412.
Sfahlan, A.J., Mahmoodzadeh, A., Hasanzadeh, A., Heidari, R. and Jamei, R. (2009). Antioxidants and antiradicals in almond hull and shell (Amygdalus communis L.) as a function of genotype. Food Chemistry, 115(2): 529-533.
Tlili, N., Kirkan, B. and Sarikurkcu, C. (2019). LC–ESI–MS/MS characterization, antioxidant power and inhibitory effects on α-amylase and tyrosinase of bioactive compounds from hulls of Amygdalus communis: The influence of the extracting solvents. Industrial crops and products, 128: 147-152.
Tomaino, A., Martorana, M., Arcoraci, T., Monteleone, D., Giovinazzo, C. and Saija, A. (2010). Antioxidant activity and phenolic profile of pistachio (Pistacia vera L., variety Bronte) seeds and skins. Biochimie, 92(9): 1115-1122.
Tsantili, E., Shin, Y., Nock, J.F. and Watkins, C.B. (2010). Antioxidant concentrations during chilling injury development in peaches. Postharvest Biology and Technology, 57(1): 27-34.
Wijeratne, S.S., Abou-Zaid, M.M. and Shahidi, F. (2006). Antioxidant polyphenols in almond and its coproducts. Journal of Agricultural and Food Chemistry, 54(2): 312-318.
Wu, H.C., Chen, H.M. and Shiau, C.Y. (2003). Free amino acids and peptides as related to antioxidant properties in protein hydrolysates of mackerel (Scomber austriasicus). Food Research International, 36(9-10): 949-957.