Polyphenolic Profile and Antioxidant Activities of Nigella Sativa Seed Extracts In Vitro and In Vivo
Nigella sativa L. is an aromatic plant belonging to the
family Ranunculaceae. It has been used traditionally, especially in the
middle East and India, for the treatment of asthma, cough, bronchitis,
headache, rheumatism, fever, influenza and eczema. Several
biological activities have been reported in Nigella sativa seeds,
including antioxidant. In this context we tried to estimate the
antioxidant activity of various extracts prepared from Nigella sativa
seeds, methanolic extract (ME), chloroformic extract (CE), hexanic
extract (HE : fixed oil), ethyl acetate extract (EAE) water extract
(WE). The Folin-Ciocalteu assay showed that CE and EAE contained
high level of phenolic compounds 81.31 and 72.43μg GAE/mg of
extract respectively. Similarly, the CE and EAE exhibited the highest
DPPH radical scavenging activity, with IC50 values of 106.56μg/ml
and 121.62μg/ml respectively. In addition, CE and HE showed the
most scavenging activity against superoxide radical generated in the
PMS-NADH-NBT system with respective IC50 values of 361.86
μg/ml and 371.80 μg/ml, which is comparable to the activity of the
standard antioxidant BHT (344.59 μg/ml). Ferrous ion chelating
capacity assay showed that WE, EAE and ME are the most active
with 40.57, 39.70 and 22.02 mg EDTA-E/g of extract. The inhibition
of linoleic acid/ß-carotene coupled oxidation was estimated by ßcarotene
bleaching assay, this showed a highest relative antioxidant
activity with CE and EAE (69.82% of inhibition). The antioxidant
activities of the methanolic extract and the fixed oil are confirmed by
an in vivo assay in mice, the daily oral administration of methanolic
extract (500 and 800 mg/kg/day) and fixed oil (2 and 4 ml/kg/day)
during 21 days, resulted in a significant enhancement of the blood
total antioxidant capacity (measured by KRL test) and the plasmatic
antioxidant capacity towards DPPH radical.
[1] G. B. N. Ames, M. K. Shigenaga and T. M. Hagen, "Oxidants,
antioxidants, and the degenerative diseases of aging," Proceedings of
the National Academy of Sciences of the United States of America, vol.
90, pp. 7915-793, 1993.
[2] J. S. Bland, "Oxidants and antioxidants in clinical medicine: Past,
present and future potential," J Nutr Environ Med,vol. 5, pp. 255-280,
1995.
[3] A. Halliwell and J. M .C. Gutteridge, "The antioxidant of human
extracellular fluids,"Arch. Biochem. Biophys, vol. 280, pp. 1-8, 1990.
[4] R. Collin, "Oxidative DNA damage, antioxidants, and cancer," Bio-
Essays, vol. 21, pp. 238-246, 1999.
[5] R. A. Floyd, "Antioxidants, oxidative stress, and degenerative
neurological disorders," Proceedings of the Society for Experimental
Biology and Medicine, vol. 222, pp. 236-245, 1999.
[6] C. A. Rice-Evans, N. J. Miller, and G. Paganga, "Structure-antioxidant
activity relationships of flavonoids and phenolic acids," Free Rad Biol
Med, vol. 20, pp. 933-956, 1996.
[7] R. Amarowicz, M. Naczk and F. Shahidi, "Antioxidant activity of
various fractions of non-tannin phenolics of canola hulls," J. Agric.
Food Chem, vol. 48, pp. 2755-2759, 2000.
[8] O. I. Aruoma, B. Halliwell, R. Aeschbach and J. Loligers, "Antioxidant
and pro-oxidant properties of active rosemary constituents: Carnosol
and carnosic acid," Xenobiotica, vol. 22, no. 2, pp. 257-268, 1992.
[9] M. Burits and F. Bucar, Antioxidant activity of Nigella sativa essential
oil," Phytother. Res, vol. 14, pp. 323-328, 2000.
[10] H.B. Li, K.W. Cheng, C.C. Wong, K.W. Fan, F, Chen and Y. Jiang,
"Evaluation of antioxidant capacity and total phenolic content of
different fractions of selected microalgae," Food Chem,vol. 102, pp.
771-776, 2007.
[11] T. Bahorun, B. Gressier,F. Trotin, C.Brunete,T. Dine, J. Vasseur, J.C.
Gazin, M. Pinkas, M. Luycky and M.Gazin, "Oxygen species
scavenging activity of phenolic extracts from hawthorn fresh plant
organs and pharmaceutical preparations," Drug Res, vol.46, pp. 1086-
1089,1996.
[12] A. Mansouri, G. Embarek,E. Kokkalou and P. Kefalas, "Phenolic profile
and antioxidant activity of the Algerian ripe date palm fruit (Phoenix
dactylifera)," Food Chem, vol. 89, pp. 411-420, 2005.
[13] Ani, M.C. Varadaraj and K. Akhilender Naidu, "Antioxidant and
antibacterial activities of polyphenolic compounds from bitter cumin
(Cuminum nigrum L.)," Eur. Food Res. Technol, vol. 224, pp. 109-115,
2006
[14] K. Le, F,Chiu F and K. Ng, "Identification and quantification of
antioxidants in Fructus lycii," Food Chem.vol.105, pp. 353-363, 2007.
[15] N. Kartal, M. Sokmen, B. Tepe, D. Daferera,M. Polissiou and A.
Sokmen, "Investigation of the antioxidant properties of Ferula orientalis
L using a suitable extraction procedure," Food Chem, vol. 100, pp. 584-
589, 2007.
[16] A. Girard, S. Madani, F Boukortt, M. Cherkaoui-Malki, J. Belleville and
J. Prost, "Fructose-aenriched diet modifies antioxidant status and lipid
metabolism in spontaneously hypertensive rats," Nutrition, vol. 22, pp.
758-766, 2006.
[17] C. Manna, S. D-angelo, V. Migliardi, E. Loffredi, O.Mazzoni, P.
Morrica, P. Galletti and V. Zappia, "Protective effect of the phenolic
fraction from virgin olive oils against oxidative stress in human cells," J.
Agric. Food Chem, vol.50, pp. 6521-6526, 2002.
[18] P. Hasani, N. Yasa, S. Vosough-Ghanbari, A. Mohammadirad, G.
Dehghan and M. Abdollahi, " In vivo antioxidant potential of Teucrium
polium, as compared to ╬▒-tocopherol," Acta Pharm, vol.57, pp. 123-
127, 2007.
[19] S. Garg, G. P. Talwar, and S. N. Upadhyay, "Immunocontraceptive
activity guide fractionation and characterization of active constituents of
neem (Azadirachta indica) seed extracts," J Ethnopharmacol, vol. 60,
pp. 235-246, 1998.
[20] E. Yesilada, K. Tsuchiya, Y. Takaishi and K. Kawazoe, "Isolation and
characterization of free radical scavenging flavonoid glycosides from
the flowers of Spartium junceum by activity-guided fractionation," J
Ethnopharmacol , vol.73, pp. 471-478, 2000.
[21] K. Suresh Kumar, K. Ganesan and P.V. Subba Rao, "Antioxidant
potential of solvent extracts of Kappaphycus alvarezii (Doty) Doty - An
edible seaweed," Food Chem, vol. 107, pp. 289-295, 2008.
[22] W. Brand-Williams, M. E. Cuvelier and C. Berset, "Use of a free radical
method to evaluate antioxidant activity," LWT, vol. 28, pp. 25-30,
1995.
[23] S.V. Javanovic, S. Steenken, M. Tosic and B. Marjanovic, M.J. Simic,
"Flavonoids as antioxidants," J. Am. Chem. Soc, vol. 116, pp. 4846-
4851, 1994.
[24] F. Benkaci-Ali, A. Baaliouamer and B.Y. Meklati, "Kinetic study of
microwave extraction of essential oil of Nigella sativa L. Seeds,"
Chromatographia, vol. 64, pp. 227-231, 2006.
[25] O.A. Badary, R.A. Taha, A.M. Gamal El-Din and M.H. Abdel-Wahab,
"Thymoquinone is a potent superoxide anion scavenger," Drug Chem.
Toxicol, vol. 26, pp. 87-98, 2003.
[26] H. Zhao,W. Fan, J. Dong, J.Lu, J. Chen, L.Shan, Y. Lin and W Kong,
"Evaluation of antioxidant activities and total phenolic contents of
typical malting barley varieties," Food Chem, vol. 107, pp. 296-304,
2008.
[27] S. Bourgou, R. Ksouri, A. Bellila, I. Skandrani, H. Falleh and B.
Marzouk, "Phenolic composition and biological activities of Tunisian
Nigella sativa L. shoots and roots," C R Biol , vol. 331, pp. 48-55,
2008.
[28] C.J. Morris, J.R. Earl, C.W. Trenam and D.R Blake,"Reactive oxygen
species and iron-a dangerous partnership in inflammation," Int. J.
Biochem. Cell Biol, vol. 27, pp. 109-122, 1995.
[29] J.E. Brown, H. Khodr, R.C. Hider and C. Rice-Evans, "Structural
dependence of flavonoïds interactions with Cu2+ ions: implication for
their antioxidant properties," Biochem. J, vol. 330, pp. 1173-1178,
1998.
[30] S.P.Wong, L.P. Leong and J.H. William Koh, "Antioxidant activities of
aqueous extracts of selected plants," Food Chem, vol. 99, pp. 775-783,
2006.
[31] S.A.B.E. Van Acker, D.J. Van Den Berg, M.N.L. Tromp, D.H.
Griffioen, W.P.V. Bennenkom, W.J.F. Van Der Vijgh and A. Bast,
"Structural aspects of antioxidant activity of flavonoids," Free Radical
Biol. Med , vol. 20, pp. 331-342, 1996.
[32] C.M. Liyana-Pathirana and F. Shahidi, "Antioxydant propreties of
commercial soft and hard winter wheats (Triticum aestivium L.) and
their milling fractions, J Sci Food Agric, vol. 86, pp. 477-485, 2006.
[33] E.N. Frankel and A.S. Meyer, "The problems of using one-dimensional
methods to evaluate multifunctional food and biological antioxidants," J
Sci Food Agric, vol. 80, pp.1925-1940, 2000.
[34] Y.F. Chu, J. Sun, X. Wu and R.H. Liu, "Antioxidant and
antiproliferative activities of common vegetables," J. Agric. Food
Chem, vol. 50, pp. 6910-6916, 2002.
[35] P. Stocker, J. F. Lesgards, N. Vidal, F. Chalier and M. Prost, "ESR
study of a biological assay on whole blood: antioxidant efficiency of
various vitamins," Biochim. Biophys. Acta , vol. 1621, pp. 1-8, 2003.
[36] G.Kökdil, L. Tamer, B. Ercan, M. ├çelik and U. Atik, "Effects of Nigella
orientalis and N. segetalis fixed oils on blood biochemistry in rats,"
Phytother. Res, vol. 20: 71-75, 2006.
[37] G.Kökdil, L.Tamer, B. Ercan, A. Ilcim, N.Aras and U. Atik, "Effects of
Nigella unguicularis fixed oil on blood biochemistry and
oxidant/antioxidant balance in rats," J Ethnopharmacol , vol. 99, pp.
131-135, 2005.
[38] S.M. Suboh, Y.Y. Bilto and T.A. Aburjai, "Protective effects of selected
medicinal plants against protein degradation, lipid peroxidation and
deformability loss of oxidatively stressed human erythrocytes,"
Phytother. Res, vol.18, pp. 280-284, 2004.
[39] A. Janaszewska and G. Bartosz, "Assay of total antioxidant capacity:
comparison of four methods as applied to human blood plasma," Scand
J Clin Lab Investig, vol. 62, pp. 231-236, 2002.
[40] D. Huang, B. Ou and R.L. Prior, "The Chemistry behind Antioxidant
Capacity Assays," J. Agric. Food Chem, vol. 53, pp.1841-1856, 2005
[1] G. B. N. Ames, M. K. Shigenaga and T. M. Hagen, "Oxidants,
antioxidants, and the degenerative diseases of aging," Proceedings of
the National Academy of Sciences of the United States of America, vol.
90, pp. 7915-793, 1993.
[2] J. S. Bland, "Oxidants and antioxidants in clinical medicine: Past,
present and future potential," J Nutr Environ Med,vol. 5, pp. 255-280,
1995.
[3] A. Halliwell and J. M .C. Gutteridge, "The antioxidant of human
extracellular fluids,"Arch. Biochem. Biophys, vol. 280, pp. 1-8, 1990.
[4] R. Collin, "Oxidative DNA damage, antioxidants, and cancer," Bio-
Essays, vol. 21, pp. 238-246, 1999.
[5] R. A. Floyd, "Antioxidants, oxidative stress, and degenerative
neurological disorders," Proceedings of the Society for Experimental
Biology and Medicine, vol. 222, pp. 236-245, 1999.
[6] C. A. Rice-Evans, N. J. Miller, and G. Paganga, "Structure-antioxidant
activity relationships of flavonoids and phenolic acids," Free Rad Biol
Med, vol. 20, pp. 933-956, 1996.
[7] R. Amarowicz, M. Naczk and F. Shahidi, "Antioxidant activity of
various fractions of non-tannin phenolics of canola hulls," J. Agric.
Food Chem, vol. 48, pp. 2755-2759, 2000.
[8] O. I. Aruoma, B. Halliwell, R. Aeschbach and J. Loligers, "Antioxidant
and pro-oxidant properties of active rosemary constituents: Carnosol
and carnosic acid," Xenobiotica, vol. 22, no. 2, pp. 257-268, 1992.
[9] M. Burits and F. Bucar, Antioxidant activity of Nigella sativa essential
oil," Phytother. Res, vol. 14, pp. 323-328, 2000.
[10] H.B. Li, K.W. Cheng, C.C. Wong, K.W. Fan, F, Chen and Y. Jiang,
"Evaluation of antioxidant capacity and total phenolic content of
different fractions of selected microalgae," Food Chem,vol. 102, pp.
771-776, 2007.
[11] T. Bahorun, B. Gressier,F. Trotin, C.Brunete,T. Dine, J. Vasseur, J.C.
Gazin, M. Pinkas, M. Luycky and M.Gazin, "Oxygen species
scavenging activity of phenolic extracts from hawthorn fresh plant
organs and pharmaceutical preparations," Drug Res, vol.46, pp. 1086-
1089,1996.
[12] A. Mansouri, G. Embarek,E. Kokkalou and P. Kefalas, "Phenolic profile
and antioxidant activity of the Algerian ripe date palm fruit (Phoenix
dactylifera)," Food Chem, vol. 89, pp. 411-420, 2005.
[13] Ani, M.C. Varadaraj and K. Akhilender Naidu, "Antioxidant and
antibacterial activities of polyphenolic compounds from bitter cumin
(Cuminum nigrum L.)," Eur. Food Res. Technol, vol. 224, pp. 109-115,
2006
[14] K. Le, F,Chiu F and K. Ng, "Identification and quantification of
antioxidants in Fructus lycii," Food Chem.vol.105, pp. 353-363, 2007.
[15] N. Kartal, M. Sokmen, B. Tepe, D. Daferera,M. Polissiou and A.
Sokmen, "Investigation of the antioxidant properties of Ferula orientalis
L using a suitable extraction procedure," Food Chem, vol. 100, pp. 584-
589, 2007.
[16] A. Girard, S. Madani, F Boukortt, M. Cherkaoui-Malki, J. Belleville and
J. Prost, "Fructose-aenriched diet modifies antioxidant status and lipid
metabolism in spontaneously hypertensive rats," Nutrition, vol. 22, pp.
758-766, 2006.
[17] C. Manna, S. D-angelo, V. Migliardi, E. Loffredi, O.Mazzoni, P.
Morrica, P. Galletti and V. Zappia, "Protective effect of the phenolic
fraction from virgin olive oils against oxidative stress in human cells," J.
Agric. Food Chem, vol.50, pp. 6521-6526, 2002.
[18] P. Hasani, N. Yasa, S. Vosough-Ghanbari, A. Mohammadirad, G.
Dehghan and M. Abdollahi, " In vivo antioxidant potential of Teucrium
polium, as compared to ╬▒-tocopherol," Acta Pharm, vol.57, pp. 123-
127, 2007.
[19] S. Garg, G. P. Talwar, and S. N. Upadhyay, "Immunocontraceptive
activity guide fractionation and characterization of active constituents of
neem (Azadirachta indica) seed extracts," J Ethnopharmacol, vol. 60,
pp. 235-246, 1998.
[20] E. Yesilada, K. Tsuchiya, Y. Takaishi and K. Kawazoe, "Isolation and
characterization of free radical scavenging flavonoid glycosides from
the flowers of Spartium junceum by activity-guided fractionation," J
Ethnopharmacol , vol.73, pp. 471-478, 2000.
[21] K. Suresh Kumar, K. Ganesan and P.V. Subba Rao, "Antioxidant
potential of solvent extracts of Kappaphycus alvarezii (Doty) Doty - An
edible seaweed," Food Chem, vol. 107, pp. 289-295, 2008.
[22] W. Brand-Williams, M. E. Cuvelier and C. Berset, "Use of a free radical
method to evaluate antioxidant activity," LWT, vol. 28, pp. 25-30,
1995.
[23] S.V. Javanovic, S. Steenken, M. Tosic and B. Marjanovic, M.J. Simic,
"Flavonoids as antioxidants," J. Am. Chem. Soc, vol. 116, pp. 4846-
4851, 1994.
[24] F. Benkaci-Ali, A. Baaliouamer and B.Y. Meklati, "Kinetic study of
microwave extraction of essential oil of Nigella sativa L. Seeds,"
Chromatographia, vol. 64, pp. 227-231, 2006.
[25] O.A. Badary, R.A. Taha, A.M. Gamal El-Din and M.H. Abdel-Wahab,
"Thymoquinone is a potent superoxide anion scavenger," Drug Chem.
Toxicol, vol. 26, pp. 87-98, 2003.
[26] H. Zhao,W. Fan, J. Dong, J.Lu, J. Chen, L.Shan, Y. Lin and W Kong,
"Evaluation of antioxidant activities and total phenolic contents of
typical malting barley varieties," Food Chem, vol. 107, pp. 296-304,
2008.
[27] S. Bourgou, R. Ksouri, A. Bellila, I. Skandrani, H. Falleh and B.
Marzouk, "Phenolic composition and biological activities of Tunisian
Nigella sativa L. shoots and roots," C R Biol , vol. 331, pp. 48-55,
2008.
[28] C.J. Morris, J.R. Earl, C.W. Trenam and D.R Blake,"Reactive oxygen
species and iron-a dangerous partnership in inflammation," Int. J.
Biochem. Cell Biol, vol. 27, pp. 109-122, 1995.
[29] J.E. Brown, H. Khodr, R.C. Hider and C. Rice-Evans, "Structural
dependence of flavonoïds interactions with Cu2+ ions: implication for
their antioxidant properties," Biochem. J, vol. 330, pp. 1173-1178,
1998.
[30] S.P.Wong, L.P. Leong and J.H. William Koh, "Antioxidant activities of
aqueous extracts of selected plants," Food Chem, vol. 99, pp. 775-783,
2006.
[31] S.A.B.E. Van Acker, D.J. Van Den Berg, M.N.L. Tromp, D.H.
Griffioen, W.P.V. Bennenkom, W.J.F. Van Der Vijgh and A. Bast,
"Structural aspects of antioxidant activity of flavonoids," Free Radical
Biol. Med , vol. 20, pp. 331-342, 1996.
[32] C.M. Liyana-Pathirana and F. Shahidi, "Antioxydant propreties of
commercial soft and hard winter wheats (Triticum aestivium L.) and
their milling fractions, J Sci Food Agric, vol. 86, pp. 477-485, 2006.
[33] E.N. Frankel and A.S. Meyer, "The problems of using one-dimensional
methods to evaluate multifunctional food and biological antioxidants," J
Sci Food Agric, vol. 80, pp.1925-1940, 2000.
[34] Y.F. Chu, J. Sun, X. Wu and R.H. Liu, "Antioxidant and
antiproliferative activities of common vegetables," J. Agric. Food
Chem, vol. 50, pp. 6910-6916, 2002.
[35] P. Stocker, J. F. Lesgards, N. Vidal, F. Chalier and M. Prost, "ESR
study of a biological assay on whole blood: antioxidant efficiency of
various vitamins," Biochim. Biophys. Acta , vol. 1621, pp. 1-8, 2003.
[36] G.Kökdil, L. Tamer, B. Ercan, M. ├çelik and U. Atik, "Effects of Nigella
orientalis and N. segetalis fixed oils on blood biochemistry in rats,"
Phytother. Res, vol. 20: 71-75, 2006.
[37] G.Kökdil, L.Tamer, B. Ercan, A. Ilcim, N.Aras and U. Atik, "Effects of
Nigella unguicularis fixed oil on blood biochemistry and
oxidant/antioxidant balance in rats," J Ethnopharmacol , vol. 99, pp.
131-135, 2005.
[38] S.M. Suboh, Y.Y. Bilto and T.A. Aburjai, "Protective effects of selected
medicinal plants against protein degradation, lipid peroxidation and
deformability loss of oxidatively stressed human erythrocytes,"
Phytother. Res, vol.18, pp. 280-284, 2004.
[39] A. Janaszewska and G. Bartosz, "Assay of total antioxidant capacity:
comparison of four methods as applied to human blood plasma," Scand
J Clin Lab Investig, vol. 62, pp. 231-236, 2002.
[40] D. Huang, B. Ou and R.L. Prior, "The Chemistry behind Antioxidant
Capacity Assays," J. Agric. Food Chem, vol. 53, pp.1841-1856, 2005
@article{"International Journal of Biological, Life and Agricultural Sciences:50319", author = "Asma Meziti and Hicham Meziti and Kaouthar Boudiaf and Benboubetra Mustapha and Hemama Bouriche.", title = "Polyphenolic Profile and Antioxidant Activities of Nigella Sativa Seed Extracts In Vitro and In Vivo", abstract = "Nigella sativa L. is an aromatic plant belonging to the
family Ranunculaceae. It has been used traditionally, especially in the
middle East and India, for the treatment of asthma, cough, bronchitis,
headache, rheumatism, fever, influenza and eczema. Several
biological activities have been reported in Nigella sativa seeds,
including antioxidant. In this context we tried to estimate the
antioxidant activity of various extracts prepared from Nigella sativa
seeds, methanolic extract (ME), chloroformic extract (CE), hexanic
extract (HE : fixed oil), ethyl acetate extract (EAE) water extract
(WE). The Folin-Ciocalteu assay showed that CE and EAE contained
high level of phenolic compounds 81.31 and 72.43μg GAE/mg of
extract respectively. Similarly, the CE and EAE exhibited the highest
DPPH radical scavenging activity, with IC50 values of 106.56μg/ml
and 121.62μg/ml respectively. In addition, CE and HE showed the
most scavenging activity against superoxide radical generated in the
PMS-NADH-NBT system with respective IC50 values of 361.86
μg/ml and 371.80 μg/ml, which is comparable to the activity of the
standard antioxidant BHT (344.59 μg/ml). Ferrous ion chelating
capacity assay showed that WE, EAE and ME are the most active
with 40.57, 39.70 and 22.02 mg EDTA-E/g of extract. The inhibition
of linoleic acid/ß-carotene coupled oxidation was estimated by ßcarotene
bleaching assay, this showed a highest relative antioxidant
activity with CE and EAE (69.82% of inhibition). The antioxidant
activities of the methanolic extract and the fixed oil are confirmed by
an in vivo assay in mice, the daily oral administration of methanolic
extract (500 and 800 mg/kg/day) and fixed oil (2 and 4 ml/kg/day)
during 21 days, resulted in a significant enhancement of the blood
total antioxidant capacity (measured by KRL test) and the plasmatic
antioxidant capacity towards DPPH radical.", keywords = "Antioxidant Capacity, Chelating, Phenolic
Compounds, Nigella Sativa, Scavenger", volume = "6", number = "4", pages = "117-9", }
