Phytochemical study and comparison of natural compounds in essential oil and extract of Epilobium hirsutum by chromatographic and spectroscopic methods
Subject Areas : PhytochemistryFatemeh Sadat Tafrishi 1 , Mahboubeh Taherkhani 2
1 - 1MSc student, Department of Phytochemistry and Essential Oils Technology, Faculty of Pharmaceutical Chemistry, Pharmaceutical Sciences Branch, Islamic Azad University (IAUPS), Tehran, Iran
2 - Department of Chemistry,College of Science, Takestan Branch, Islamic Azad University, Takestan, Iran
Keywords: Mazandaran, Phytochemistry, flavonoid, Essential oil, Epilobium hirsutum, Epigallocatechin gallate,
Abstract :
Epilobium belonging to the Onagraceae family has many medicinal properties such as anti-inflammatory, anti-oxidant, antimicrobial, anti-tumor, analgesic and anti-androgenic in traditional medicine. In this study, the aerial parts of Epilobium hirsutum L.were collected at the flowering stage from the heights of central Alborz, Mazandaran province, between Siah Bisheh and Zangoleh Bridge on July 19, 2017 and phytochemically studied for effective volatile compounds in essential oils and natural products in the extract. The essential oil was extracted by hydrodistillation using celvenger apparatus and was analyzed qualitatively and quantitatively by GC-MS method. Based on the results, the main compounds in the essential oil were related to alpha-pinene with 21.65%, delta-cadinen (15.29%) and 1 -8-cineole (12.24%). Hydrocarbon terpenes had the highest concentration in the essential oil. The natural compound in E. hirsutum extract was purified by column chromatography (CC) and thin layer chromatography (TLC). As a result, a natural compound called epigallocatechin gallate was extracted and identified by 13C-NMR, 1H-NMR and DEPT spectrophotometric techniques and its molecular structure was determined. In addition, the plant extract was investigated for the presence of phytochemical content and the presence of flavonoids (Shinoda method), alkaloids (Mayer test), tannins (Ferric chloride method), saponins (Froth method) and glycosides (Keller-Kiliani test). The presence of flavonoids (red color), Alkaloids (yellow precipitate) saponins (foam layer), glycosides (bluish green layer) and tannins (dark green for Catechol tannin) in the extract was aproved.
Auwal, M.S., Saka, S., Mairiga, I.A., Sanda, K.A., Shuaibu, A. and Ibrahim, A. 2014. Preliminary phytochemical and elemental analysis of aqueous and fractionated pod extracts of Acacia nilotica (Thorn mimosa), Veterinary Research Forum, 5(2): 95-100.
2.Azizian, D. 2005. Flora of Iran. Research Institute of forests and rangelands. Tehran. Iran.
3.Bartfay, W.J., Bartfay, E. and Johnson, J.G. 2012. Gram-negative and gram-positive antibacterial properties of the whole plant extract of Willow herb (Epilobium angustifolium). Biological Research for Nursing, 14(1): 85-89.
4.Battinelli, L., Tita, B., Evandri, M.G. and Mazzanti, G. 2001. Antimicrobial activity of Epilobium spp. extracts. Il Farmaco, 56(5-7): 345-348.
5.Borchardt, J.R., Wyse, D.L., Sheaffer, C.C., Kauppi, K.L., Fulcher, R.G., Ehlke, N.J., Biesboer, D.D. and Bey, R.F. 2008. Antimicrobial activity of native and naturalized plants of Minnesota and Wisconsin. Journal of Medicinal Plants Research, 2(5): 98-110.
6.Cando, D., Morcuende, D., Utrera, M. and Estévez, M. 2014. Phenolic-rich extracts from Willowherb (Epilobium hirsutum L.) inhibit lipid oxidation but accelerate protein carbonylation and discoloration of beef patties. European Food Research and Technology, 238(5): 741-751.
7.Dzhafar, S.S., Dalar, A., Mukemre, M., Ekin, S., Yıldız, D. and Yunusoglu, O. 2020. Phytochemical profile and in vitro and in vivo anticonvulsant and antioxidant activities of Epilobium hirsutum, International Journal of Secondary Metabolite, 7(2): 63-76.
8.Eghmazi, E., Akhgar, M.R. and Kariminik, A. 2015. Chemical constituents and antibacterial activity of the essential oil from Epilobium hirsutum, Journal of Biodiversity and Environmental Sciences, 7(1): 338-344.
9.Granica, S., Piwowarski, J.P., Czerwińska, M.E. and Kiss, A.K. 2014. Phytochemistry, pharmacology and traditional uses of different Epilobium species (Onagraceae): a review, Journal of Ethnopharmacology, 156: 316-346.
10.Hafez Ghoran, S., Mighani, H. and Ebrahimi, P. 2014. InVitro anti-bacterial activity of chloroform, ethyl acetate and hydroalcoholic extracts of Scilla persica Hausskn. Journal of Gorgan University of Medical Sciences, 16(1): 106-113.
11.Hevesi, B.T., Houghton, P.J., Habtemariam, S. and Kéry, Á. 2009. Antioxidant and anti-inflammatory effect of Epilobium parviflorum Schreb. Phytotherapy Research, 23(5): 719-724.
12.Jürgenson, S., Matto, V. and Raal, A. 2012. Vegetational variation of phenolic compounds in Epilobium angustifolium. Natural Product Research, 26(20): 1951-1953.
13.Kiss, A.K., Bazylko, A., Filipek, A., Granica, S., Jaszewska, E., Kiarszys, U., Kośmider, A. and Piwowarski, J. 2011. Oenothein Bʼs contribution to the anti-inflammatory and antioxidant activity of Epilobium sp. Phytomedicine, 18(7): 557-560.
14.Kosalec, I., Zovko, M., Sankovic, K., Kremer, D. and Pepeljnjak, S. 2008. Antioxidant and antimicrobial activity of willow herb (Epilobium angustifolium L.), Planta Medica, 74 - PA43.
16.Krishnaiah, D., Sarbatly, R. and Bono, A. 2007. Phytochemical antioxidants for health and medicine– A move towards nature. Biotechnology and Molecular Biology Reviews, 1(4): 97-104.
15.Kunduhoglu, B., Pilatin, S. and Caliskan, F. 2011. Antimicrobial screening of some medicinal plants collected from Eskisehir, Turkey. Fresenius Environmental Bulletin, 20(4): 945-952.
17.Mohammadi Bazargani, M. 24-26 Feb 2015, Investigation of antibacterial, antioxidant activity and production potential of extracts obtained from different solvents in Epilobium hirsutum, International conference on sustainable development, strategies and challenges With a focus on Agriculture, Natural Resources, Environment and Tourism, Tabriz, Iran, Article COI: ICSDA01_0504.
18.Mohammadi Bazargani, M., Falahati-Anbaran, M. and Rohloff, J. 2021. Comparative analyses of phytochemical variation within and between congeneric species of willow herb, Epilobium hirsutum and E. parviflorum: Contribution of Environmental Factors. Frontiers in Plant Science, 11: 595190.
19.Nawwar, MAM., Marzouk, M.S., Nigge W. and Linscheid, M. 1998. High‐performance liquid chromatographic / electrospray ionization mass spectrometric screening for polyphenolic compounds of Epilobium hirsutum- the structure of the unique ellagitannin epilobamide‐a . Journal of mass spectrometry, 32(6): 645-654.
20.Nurdiani, R., Firdaus, M. and Prihanto, A.A. 2012. Phytochemical screening and antibacterial activity of methanol extract of mangrove plant (Rhyzophora mucronata) from Porong River Estuary. Journal Basic Science and Technology, 1(2): 27-29.
21.Pakravan, S., Hajimoradloo, A. and Ghorbani, R. 2012. Effect of dietary willow herb, Epilobium hirsutum extract on growth performance, body composition, haematological parameters and aeromonas hydrophila challenge on common carp, Cyprinus carpio. Aquaculture Research, 43(6): 861-869.
22.Pirvu, L., Albu,B., Niță, S., Sha’at, F., Pavaloiu, R., Bazdoaca, C., Staras, A., Tihăuan, B., Kamerzan, C.M. and Perisoara, A. 2019. Epilobium hirsutum L. and Malva neglecta L. propylene glycol extracts for cosmetic use; chemical composition, microbiological stability and diffusion aspects. Scholars Academic Journal of Pharmacy, 8(8): 424-432.
23.Pirvu, L., Nicorescu, V., Hlevca, C., Udeanu, D.I. and Nicorescu, I. 2015. Antimicrobial and synergistic activity of some whole and selective Epilobium hirsutum L. (great willowherb) extracts tested on standard and wild Staphylococcus aureus strains. Farmacia, 63: 690-695.
24.Padmaja, M., Sravanthi, M. and Hemalatha, K.P.J. 2011. Evaluation of antioxidant activity of two indian medicinal plants. Journal of Phytology, 3(3):86-91.
25.Pourmorad, F., Ebrahimzadeh, M.A., Mahmoudi, M. and Yasini, S. 2007. Antinociceptive activity of methanolic extract of Epilobium hirsutum. Pakistan Journal of Biological Sciences, 10(16): 2764-2767.
26.Safavi, F., Ebrahimi, P. and Mighani, H. 2013. In vitro antibacterial activity of root and aerial parts of Scrophularia Striata Bioss. on Escherichia Coli, Staphylococcus Aureus and Bacillus Cereus. Armaghane-danesh, Yasuj University of Medical Sciences Journal, 1(8):603-614.
27.Steenkamp, V., Gouws, M.C., Gulumian, M., Elgorashi, E.E. and Van Standen, J. 2006. Studies on antibacterial, anti-inflammatory and antioxidant activity of herbal remedies used in the treatment of benign prostatic hyperplasia and prostatitis. Journal of Ethnopharmacology, 103(1): 71-75.
28.Tita, B., Abdel-Haq, H., Vitalone, A., Mazzanti, G. and Saso, L. 2001. Analgesic properties of Epilobium angustifolium, evaluated by the hot plate test and the writhing test. Il Farmaco, 56(5-7): 341-343.
29.Tiwari, P., Kumar, B., Kaur, M., Kaur, G. and Kaur, H. 2011. Phytochemical screening and extraction: A Review. Internationale Pharmaceutica Sciencia, 1(1): 98-106.
30.Tóth, B.H., Blazics, B. and Kéry, Á. 2009. Polyphenol composition and antioxidant capacity of Epilobium species. Journal of Pharmaceutical and Biomedical Analysis, 49(1): 26-31.
31.Tóth, B.H., Balázs, A., Vukics, V., Szőke, É. and Kéry, Á. 2006. Identification of Epilobium species and willow-herbs (Onagraceae) by HPLC analysis of flavonoids as chemotaxonomic markers. Chromatographia, 63(13), pp.S119-S123.
33.Vitalone, A. and Allkanjari, O. 2018. Epilobium spp: Pharmacology and phytochemistry. phytotherapy research, 32(7): 1229-1240.
34.Vitalone, A., Bordi, F., Baldazzi, C., Mazzanti, G., Saso, L. and Tita, B. 2001. Anti-proliferative effect on a prostatic epithelial cell line (PZ-HPV-7) by Epilobium angustifolium L. Il Farmaco, 56(5-7): 483-489.
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Auwal, M.S., Saka, S., Mairiga, I.A., Sanda, K.A., Shuaibu, A. and Ibrahim, A. 2014. Preliminary phytochemical and elemental analysis of aqueous and fractionated pod extracts of Acacia nilotica (Thorn mimosa), Veterinary Research Forum, 5(2): 95-100.
2.Azizian, D. 2005. Flora of Iran. Research Institute of forests and rangelands. Tehran. Iran.
3.Bartfay, W.J., Bartfay, E. and Johnson, J.G. 2012. Gram-negative and gram-positive antibacterial properties of the whole plant extract of Willow herb (Epilobium angustifolium). Biological Research for Nursing, 14(1): 85-89.
4.Battinelli, L., Tita, B., Evandri, M.G. and Mazzanti, G. 2001. Antimicrobial activity of Epilobium spp. extracts. Il Farmaco, 56(5-7): 345-348.
5.Borchardt, J.R., Wyse, D.L., Sheaffer, C.C., Kauppi, K.L., Fulcher, R.G., Ehlke, N.J., Biesboer, D.D. and Bey, R.F. 2008. Antimicrobial activity of native and naturalized plants of Minnesota and Wisconsin. Journal of Medicinal Plants Research, 2(5): 98-110.
6.Cando, D., Morcuende, D., Utrera, M. and Estévez, M. 2014. Phenolic-rich extracts from Willowherb (Epilobium hirsutum L.) inhibit lipid oxidation but accelerate protein carbonylation and discoloration of beef patties. European Food Research and Technology, 238(5): 741-751.
7.Dzhafar, S.S., Dalar, A., Mukemre, M., Ekin, S., Yıldız, D. and Yunusoglu, O. 2020. Phytochemical profile and in vitro and in vivo anticonvulsant and antioxidant activities of Epilobium hirsutum, International Journal of Secondary Metabolite, 7(2): 63-76.
8.Eghmazi, E., Akhgar, M.R. and Kariminik, A. 2015. Chemical constituents and antibacterial activity of the essential oil from Epilobium hirsutum, Journal of Biodiversity and Environmental Sciences, 7(1): 338-344.
9.Granica, S., Piwowarski, J.P., Czerwińska, M.E. and Kiss, A.K. 2014. Phytochemistry, pharmacology and traditional uses of different Epilobium species (Onagraceae): a review, Journal of Ethnopharmacology, 156: 316-346.
10.Hafez Ghoran, S., Mighani, H. and Ebrahimi, P. 2014. InVitro anti-bacterial activity of chloroform, ethyl acetate and hydroalcoholic extracts of Scilla persica Hausskn. Journal of Gorgan University of Medical Sciences, 16(1): 106-113.
11.Hevesi, B.T., Houghton, P.J., Habtemariam, S. and Kéry, Á. 2009. Antioxidant and anti-inflammatory effect of Epilobium parviflorum Schreb. Phytotherapy Research, 23(5): 719-724.
12.Jürgenson, S., Matto, V. and Raal, A. 2012. Vegetational variation of phenolic compounds in Epilobium angustifolium. Natural Product Research, 26(20): 1951-1953.
13.Kiss, A.K., Bazylko, A., Filipek, A., Granica, S., Jaszewska, E., Kiarszys, U., Kośmider, A. and Piwowarski, J. 2011. Oenothein Bʼs contribution to the anti-inflammatory and antioxidant activity of Epilobium sp. Phytomedicine, 18(7): 557-560.
14.Kosalec, I., Zovko, M., Sankovic, K., Kremer, D. and Pepeljnjak, S. 2008. Antioxidant and antimicrobial activity of willow herb (Epilobium angustifolium L.), Planta Medica, 74 - PA43.
16.Krishnaiah, D., Sarbatly, R. and Bono, A. 2007. Phytochemical antioxidants for health and medicine– A move towards nature. Biotechnology and Molecular Biology Reviews, 1(4): 97-104.
15.Kunduhoglu, B., Pilatin, S. and Caliskan, F. 2011. Antimicrobial screening of some medicinal plants collected from Eskisehir, Turkey. Fresenius Environmental Bulletin, 20(4): 945-952.
17.Mohammadi Bazargani, M. 24-26 Feb 2015, Investigation of antibacterial, antioxidant activity and production potential of extracts obtained from different solvents in Epilobium hirsutum, International conference on sustainable development, strategies and challenges With a focus on Agriculture, Natural Resources, Environment and Tourism, Tabriz, Iran, Article COI: ICSDA01_0504.
18.Mohammadi Bazargani, M., Falahati-Anbaran, M. and Rohloff, J. 2021. Comparative analyses of phytochemical variation within and between congeneric species of willow herb, Epilobium hirsutum and E. parviflorum: Contribution of Environmental Factors. Frontiers in Plant Science, 11: 595190.
19.Nawwar, MAM., Marzouk, M.S., Nigge W. and Linscheid, M. 1998. High‐performance liquid chromatographic / electrospray ionization mass spectrometric screening for polyphenolic compounds of Epilobium hirsutum- the structure of the unique ellagitannin epilobamide‐a . Journal of mass spectrometry, 32(6): 645-654.
20.Nurdiani, R., Firdaus, M. and Prihanto, A.A. 2012. Phytochemical screening and antibacterial activity of methanol extract of mangrove plant (Rhyzophora mucronata) from Porong River Estuary. Journal Basic Science and Technology, 1(2): 27-29.
21.Pakravan, S., Hajimoradloo, A. and Ghorbani, R. 2012. Effect of dietary willow herb, Epilobium hirsutum extract on growth performance, body composition, haematological parameters and aeromonas hydrophila challenge on common carp, Cyprinus carpio. Aquaculture Research, 43(6): 861-869.
22.Pirvu, L., Albu,B., Niță, S., Sha’at, F., Pavaloiu, R., Bazdoaca, C., Staras, A., Tihăuan, B., Kamerzan, C.M. and Perisoara, A. 2019. Epilobium hirsutum L. and Malva neglecta L. propylene glycol extracts for cosmetic use; chemical composition, microbiological stability and diffusion aspects. Scholars Academic Journal of Pharmacy, 8(8): 424-432.
23.Pirvu, L., Nicorescu, V., Hlevca, C., Udeanu, D.I. and Nicorescu, I. 2015. Antimicrobial and synergistic activity of some whole and selective Epilobium hirsutum L. (great willowherb) extracts tested on standard and wild Staphylococcus aureus strains. Farmacia, 63: 690-695.
24.Padmaja, M., Sravanthi, M. and Hemalatha, K.P.J. 2011. Evaluation of antioxidant activity of two indian medicinal plants. Journal of Phytology, 3(3):86-91.
25.Pourmorad, F., Ebrahimzadeh, M.A., Mahmoudi, M. and Yasini, S. 2007. Antinociceptive activity of methanolic extract of Epilobium hirsutum. Pakistan Journal of Biological Sciences, 10(16): 2764-2767.
26.Safavi, F., Ebrahimi, P. and Mighani, H. 2013. In vitro antibacterial activity of root and aerial parts of Scrophularia Striata Bioss. on Escherichia Coli, Staphylococcus Aureus and Bacillus Cereus. Armaghane-danesh, Yasuj University of Medical Sciences Journal, 1(8):603-614.
27.Steenkamp, V., Gouws, M.C., Gulumian, M., Elgorashi, E.E. and Van Standen, J. 2006. Studies on antibacterial, anti-inflammatory and antioxidant activity of herbal remedies used in the treatment of benign prostatic hyperplasia and prostatitis. Journal of Ethnopharmacology, 103(1): 71-75.
28.Tita, B., Abdel-Haq, H., Vitalone, A., Mazzanti, G. and Saso, L. 2001. Analgesic properties of Epilobium angustifolium, evaluated by the hot plate test and the writhing test. Il Farmaco, 56(5-7): 341-343.
29.Tiwari, P., Kumar, B., Kaur, M., Kaur, G. and Kaur, H. 2011. Phytochemical screening and extraction: A Review. Internationale Pharmaceutica Sciencia, 1(1): 98-106.
30.Tóth, B.H., Blazics, B. and Kéry, Á. 2009. Polyphenol composition and antioxidant capacity of Epilobium species. Journal of Pharmaceutical and Biomedical Analysis, 49(1): 26-31.
31.Tóth, B.H., Balázs, A., Vukics, V., Szőke, É. and Kéry, Á. 2006. Identification of Epilobium species and willow-herbs (Onagraceae) by HPLC analysis of flavonoids as chemotaxonomic markers. Chromatographia, 63(13), pp.S119-S123.
33.Vitalone, A. and Allkanjari, O. 2018. Epilobium spp: Pharmacology and phytochemistry. phytotherapy research, 32(7): 1229-1240.
34.Vitalone, A., Bordi, F., Baldazzi, C., Mazzanti, G., Saso, L. and Tita, B. 2001. Anti-proliferative effect on a prostatic epithelial cell line (PZ-HPV-7) by Epilobium angustifolium L. Il Farmaco, 56(5-7): 483-489.
