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Antioxidant activity of banana flavonoids
Fitoterapia 79 (2008) 279 – 282 www.elsevier.com/locate/fitote
Antioxidant activity of banana flavonoids S. Vijayakumar a,⁎, G. Presannakumar b , N.R. Vijayalakshmi b a b
Department of Zoology, NSS College Manjeri, Malappuram Kerala Pin, 676122, India Department of Biochemistry, University of Kerala, Thiruvananthapuram, 695581 India Received 15 March 2007; accepted 21 January 2008 Available online 9 February 2008
Abstract The antioxidant activity of flavonoids from banana (Musa paradisiaca) was studied in rats fed normal as well as high fat diets. Concentrations of peroxidation products namely malondialdehyde, hydroperoxides and conjugated diens were significantly decreased whereas the activities of catalase and superoxide dismutase were enhanced significantly. Concentrations of glutathione were also elevated in the treated animals. © 2008 Elsevier B.V. All rights reserved. Keywords: Musa paradisiaca; Catalase; Superoxide dismutase; Malondialdehyde
1. Introduction Flavonoids constitute one of the largest groups of naturally occuring phenols. They are ubiquitous in all parts of green plants and as such are likely to be encountered in any work involving plant extracts. It is estimated that 2% of all carbon photosynthesised by plants is converted into flavonoids. Consumption of flavonoids may be benificial because they interact with various biological systems and show anti-inflammatory, hypolipidemic, hypoglycemic and antioxidant activities [1–7]. Research works have revealed that lipid peroxidation is involved in various physiological and pathological abnormalities such as inflammation, heart diseases and ageing. There is an increasing evidence that oxidised lower density lipoprotein(LDL and VLDL) may be involved in the pathogenesis of atherosclerosis[8]. Flavonoids have been shown to inhibit LDL oxidation [9,10]. The antioxidant properties of flavonoids from Solanum melongena, Punica granatum, Solenostemon rotundifolius, have been reported[11,12]. Though banana is regularly consumed by many people, the effect of flavonoids present in them have not been studied so far. Therefore it is significant to study the antioxidant activities of flavonoids present in the banana.
⁎ Corresponding author. E-mail address: [email protected] (S. Vijayakumar). 0367-326X/$ - see front matter © 2008 Elsevier B.V. All rights reserved. doi:10.1016/j.fitote.2008.01.007
280
S. Vijayakumar et al. / Fitoterapia 79 (2008) 279–282
Table 1 Concentrations of lipid peroxides in rats treated with M. paradisica flavonoids Groups Doses Malondialdehyde
I II III IV
Hydroperoxides
mg/ Liver 100 g BW/ day
Heart
Kidney
– 1.0 – 1.0
0.47 ± 0.011 0.37 ± 0.007⁎ 0.62 ± 0.015 0.41 ± 0.008⁎
1.22 ± 0.030 9.84 ± 0.24 42.39 ± 1.05 1.96 ± 0.049 0.71 ± 0.014⁎ 5.96 ± 0.11⁎ 29.4 ± 0.58⁎ 1.35 ± 0.027⁎ 0.99 ± 0.02 19.22 ± 0.48 50.59 ± 1.26 2.0 ± 0.05 0.71 ± 0.014⁎ 10.6 ± 0.2 42.65 ± 0.85⁎ 1.6 ± 0.032⁎
0.74 ± 0.018 0.51 ± 0.010⁎ 0.72 ± 0.018 0.46 ± 0.009⁎
Liver
Heart
Conjugated dienes Kidney
Liver
Heart
Kidney
74.82 ± 1.87 13.22 ± 0.33 12.96 ± 0.32 64.71 ± 1.29⁎ 7.64 ± 0.15⁎ 8.43 ± 0.16 74.84 ± 1.87 11.76 ± 0.29 14.0 ± 0.35 60.61 ± 1.21⁎ 8.42 ± 0.16⁎ 3.04 ± 0.28
Values expressed as mM/100 g wet tissues. Average of the values of six rats in each group ± SE. Group I was compared with groups II and III was compared with group IV. ⁎P b 0.01.
Experiments were conducted in normal as well as high fat diet fed rats to study the antioxidant effect and understand the mechanisms in both groups. 2. Experimental 2.1. Plant Musa paradisiaca (Musaceae) was purchased from the local market. 2.2. Extracts preparation Flavonoids were extracted from raw banana according to Markham [13]. Raw bananas were extracted with MeOH: H2O (9:1) and 50% aq. MeOH. Combined extracts, evaporated to remove MeOH, were cleared off low polarity contaminants by repeated extractions with hexane or CHCl3. The aqueous layer containing the bulk of flavonoids was then concentrated and used in the experiment. Estimation of flavonoids was performed according to Eskin et al. [14] using catechin as standard. 2.3. Animals Male Sprague–Dawley rats weighing 100–125 g were used. They were housed in standard environmental conditions and fed with rodent diet and water ad libitum. 2.4. Study on antiperoxidative activities Animals were divided into 4 groups, consisting of 6 rats in each group. The rats were fed with a normal laboratory diet (group I), a normal laboratory diet with flavonoids at a dose of 10 mg BW (group II), a high fat diet (group III) and Table 2 Activities of catalase and superoxide dismutase in rats treated with M. paradisiaca flavonoids Groups
I II III IV
Dose
Catalase a
Superoxide dismutase b
mg/100 g BW/day
Liver
Heart
Kidney
Liver
Heart
Kidney
– 1.0 – 1.0
62.24 ± 1.24 90.97 ± 2.7⁎ 68.01 ± 1.36 85.81 ± 2.14⁎
12.12 ± 0.24 17.15 ± 0.42⁎ 12.03 ± 0.24 16.65 ± 0.41⁎
25.16 ± 0.5 34.01 ± 0.85⁎ 22.0 ± 0.44 31.14 ± 0.77⁎
12.14 ± 0.24 19.07 ± 0.47⁎ 12.96 ± 0.3 19.45 ± 0.51⁎
13.65 ± 0.27 20.11 ± 0.5⁎ 13.25 ± 0.35 6.48 ± 0.49⁎
11.65 ± 0.23 18.47 ± 0.46⁎ 10.20 ± 0.20 14.30 ± 030⁎
Values expressed as mM/100 g wet tissues. Average of the values of six rats in each group ± SE. Group I was compared with groups II and III was compared with group IV. ⁎Pb0.01. a Values × 10− 3 units/mg protein. b Units/mg protein.
S. Vijayakumar et al. / Fitoterapia 79 (2008) 279–282
281
Table 3 Concentration of glutathione in tissues of rats treated with M. paradisiaca flavonoids Groups
Dose
Liver
Heart
Kidney
375.19 ± 7.5 419.6 ± 10.49⁎ 310.90 ± 6.21 380.38 ± 9.5⁎
487.1 ± 9.74 546.74 ± 13.66⁎ 300.8 ± 6.01 342.16 ± 8.21⁎
320.12 ± 7.68 373.2 ± 10.44⁎ 400.6 ± 8.01 455.31 ± 12.74⁎
mg/100 g BW/day – 1.0 – 1.0
I II III IV
Values expressed as mM/100 g wet tissues. Average of the values of six rats in each group ± SE. Group I was compared with groups II and group III was compared with group IV. ⁎P b 0.01.
high fat diet with flavonoids at a dose of 10 mg (group IV). Animals were treated orally by gastric intubation for 90 days. At the end of the experimental periods, the liver, kidney and heart were removed for the estimations of malondialdehyde (MDA), hydroperoxides, conjugated diens and glutathione according to John AB Steven [15]. The activities of superoxide dismutase (SOD) and catalase were determined by the method of Kakkar et al. [16]. Protein estimation was done by the method of Lowry et al. [17]. 2.5. Statistical analysis Statistical significance was calculated using Student's t-test [18]. 3. Results and discussion Food intake and weight gain were almost similar in both control as well as experimental groups. Concentrations of malondialdehyde, hydroperoxides and conjugated diens were decreased significantly (Table 1) whereas the activities of superoxide dismutase (SOD) and catalase were enhanced significantly in the groups (II and IV) treated with flavonoids (Table 2). No significant change in the concentration of conjugated diens was noticed in the kidney of rats fed high fat diet. Concentration of glutathione showed significant increase in the tissues of rats fed normal as well as high fat diets (Table 3). The higher activity of SOD indicating its free radical scavenging activity [19]. The highly stimulated activities of SOD and catalase in rats fed flavonoids probably account for the decreased levels of peroxidation products such as malondialdehyde, hydroperoxides and conjugated diens. The antiperoxidative activity of flavonoids was reported to be directly proportional to the number of free phenolic hydroxy groups [20].The present study emphasis that the flavonoids from banana act as an effective antioxidant. Therefore, the supplementation of natural antioxidants through a balanced diet containing banana could be effective in protecting the body against various oxidative stresses. References [1] [2] [3] [4] [5] [6] [7] [8] [9] [10] [11] [12] [13] [14]
Mookerjee BK, Lee TP, Lipper HA, Middleton E. J Immunopharmacol 1986;8:371. De Whalley CV, Rankin SM, Hoult Jessup W, Leake DS. Biochem Pharmacol 1990;39:1743. Abad MJ, Bermejo P, Villas A, Valverde S. J Natl Prod 1993;56:1164. Borissova P, Valcheva S, Belcheva A. Acta Physiol Pharmacol Bulg 1994;20:25. Sailaja A, Scalese M, Lanza M, Marzullo D, Bonina F, Castelli F. Free Radical Biol Med 1995;19:481. Vinodhkumar R, Augusyt KT. Indian J Biochem Biophys 1989;26:400. Charles STH, Simon LH. Diabetes 1984;33:291. Steinberg D, Parthasarathy S, Carew TE, Khoo JC, Witzum JL. New Engl J Med 1980;320:915. Mangiapane H, Thomson J, Salter A, Brown S, Bell GD, White DA. Biochem Pharmacol 1992;43:45. Rankin S, De Whalley CV, Hoult RS, Jessup W, Wilkins GM, Collard J, et al. Biochem Pharmacol 1993;45:67. Sudheesh S, Presanna Kumar G, Vijayakumar S, Vijaylakshmi NR. Plant Foods Hum Nutr 1997;51:321. Sudheesh S, Sandhya C, Koshy AS, Vijaylakshmi NR. Phytother Res 1999;13:393. Markham KR. Techniques of flavonoid identification. London: Academic Press; 1982. Eskin NAM, Hoehn E, Frenket C. J Agric Food Chem 1978;26:973.
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S. Vijayakumar et al. / Fitoterapia 79 (2008) 279–282
[15] Steven John AB. Microsomal lipid peroxidation. In: Colowick SP, Kaplan NO, editors. Methods in enzymology. London: Academic Press; 1978. [16] Kakkar P, Das B, Viswanathan PN. Indian J Biochem Biophys 1984;21:310. [17] Lowry OH, Rosebrough NJ, Farr AL, Randail RJ. Biol Chem 1951;193:265. [18] Bennet CA, Franklin NL. Statistical analysis in chemistry and chemical industry. New York: Wiley and Sons; 1967. [19] Sudheesh S, Vijayalakshmi NR. Fitoterapia 2005;76:181. [20] Joyeux M, Lobstein N, Anton R, Mortier F. Planta Med 1995;61:126.
Antioxidant activity of banana flavonoids S. Vijayakumar a,⁎, G. Presannakumar b , N.R. Vijayalakshmi b a b
Department of Zoology, NSS College Manjeri, Malappuram Kerala Pin, 676122, India Department of Biochemistry, University of Kerala, Thiruvananthapuram, 695581 India Received 15 March 2007; accepted 21 January 2008 Available online 9 February 2008
Abstract The antioxidant activity of flavonoids from banana (Musa paradisiaca) was studied in rats fed normal as well as high fat diets. Concentrations of peroxidation products namely malondialdehyde, hydroperoxides and conjugated diens were significantly decreased whereas the activities of catalase and superoxide dismutase were enhanced significantly. Concentrations of glutathione were also elevated in the treated animals. © 2008 Elsevier B.V. All rights reserved. Keywords: Musa paradisiaca; Catalase; Superoxide dismutase; Malondialdehyde
1. Introduction Flavonoids constitute one of the largest groups of naturally occuring phenols. They are ubiquitous in all parts of green plants and as such are likely to be encountered in any work involving plant extracts. It is estimated that 2% of all carbon photosynthesised by plants is converted into flavonoids. Consumption of flavonoids may be benificial because they interact with various biological systems and show anti-inflammatory, hypolipidemic, hypoglycemic and antioxidant activities [1–7]. Research works have revealed that lipid peroxidation is involved in various physiological and pathological abnormalities such as inflammation, heart diseases and ageing. There is an increasing evidence that oxidised lower density lipoprotein(LDL and VLDL) may be involved in the pathogenesis of atherosclerosis[8]. Flavonoids have been shown to inhibit LDL oxidation [9,10]. The antioxidant properties of flavonoids from Solanum melongena, Punica granatum, Solenostemon rotundifolius, have been reported[11,12]. Though banana is regularly consumed by many people, the effect of flavonoids present in them have not been studied so far. Therefore it is significant to study the antioxidant activities of flavonoids present in the banana.
⁎ Corresponding author. E-mail address: [email protected] (S. Vijayakumar). 0367-326X/$ - see front matter © 2008 Elsevier B.V. All rights reserved. doi:10.1016/j.fitote.2008.01.007
280
S. Vijayakumar et al. / Fitoterapia 79 (2008) 279–282
Table 1 Concentrations of lipid peroxides in rats treated with M. paradisica flavonoids Groups Doses Malondialdehyde
I II III IV
Hydroperoxides
mg/ Liver 100 g BW/ day
Heart
Kidney
– 1.0 – 1.0
0.47 ± 0.011 0.37 ± 0.007⁎ 0.62 ± 0.015 0.41 ± 0.008⁎
1.22 ± 0.030 9.84 ± 0.24 42.39 ± 1.05 1.96 ± 0.049 0.71 ± 0.014⁎ 5.96 ± 0.11⁎ 29.4 ± 0.58⁎ 1.35 ± 0.027⁎ 0.99 ± 0.02 19.22 ± 0.48 50.59 ± 1.26 2.0 ± 0.05 0.71 ± 0.014⁎ 10.6 ± 0.2 42.65 ± 0.85⁎ 1.6 ± 0.032⁎
0.74 ± 0.018 0.51 ± 0.010⁎ 0.72 ± 0.018 0.46 ± 0.009⁎
Liver
Heart
Conjugated dienes Kidney
Liver
Heart
Kidney
74.82 ± 1.87 13.22 ± 0.33 12.96 ± 0.32 64.71 ± 1.29⁎ 7.64 ± 0.15⁎ 8.43 ± 0.16 74.84 ± 1.87 11.76 ± 0.29 14.0 ± 0.35 60.61 ± 1.21⁎ 8.42 ± 0.16⁎ 3.04 ± 0.28
Values expressed as mM/100 g wet tissues. Average of the values of six rats in each group ± SE. Group I was compared with groups II and III was compared with group IV. ⁎P b 0.01.
Experiments were conducted in normal as well as high fat diet fed rats to study the antioxidant effect and understand the mechanisms in both groups. 2. Experimental 2.1. Plant Musa paradisiaca (Musaceae) was purchased from the local market. 2.2. Extracts preparation Flavonoids were extracted from raw banana according to Markham [13]. Raw bananas were extracted with MeOH: H2O (9:1) and 50% aq. MeOH. Combined extracts, evaporated to remove MeOH, were cleared off low polarity contaminants by repeated extractions with hexane or CHCl3. The aqueous layer containing the bulk of flavonoids was then concentrated and used in the experiment. Estimation of flavonoids was performed according to Eskin et al. [14] using catechin as standard. 2.3. Animals Male Sprague–Dawley rats weighing 100–125 g were used. They were housed in standard environmental conditions and fed with rodent diet and water ad libitum. 2.4. Study on antiperoxidative activities Animals were divided into 4 groups, consisting of 6 rats in each group. The rats were fed with a normal laboratory diet (group I), a normal laboratory diet with flavonoids at a dose of 10 mg BW (group II), a high fat diet (group III) and Table 2 Activities of catalase and superoxide dismutase in rats treated with M. paradisiaca flavonoids Groups
I II III IV
Dose
Catalase a
Superoxide dismutase b
mg/100 g BW/day
Liver
Heart
Kidney
Liver
Heart
Kidney
– 1.0 – 1.0
62.24 ± 1.24 90.97 ± 2.7⁎ 68.01 ± 1.36 85.81 ± 2.14⁎
12.12 ± 0.24 17.15 ± 0.42⁎ 12.03 ± 0.24 16.65 ± 0.41⁎
25.16 ± 0.5 34.01 ± 0.85⁎ 22.0 ± 0.44 31.14 ± 0.77⁎
12.14 ± 0.24 19.07 ± 0.47⁎ 12.96 ± 0.3 19.45 ± 0.51⁎
13.65 ± 0.27 20.11 ± 0.5⁎ 13.25 ± 0.35 6.48 ± 0.49⁎
11.65 ± 0.23 18.47 ± 0.46⁎ 10.20 ± 0.20 14.30 ± 030⁎
Values expressed as mM/100 g wet tissues. Average of the values of six rats in each group ± SE. Group I was compared with groups II and III was compared with group IV. ⁎Pb0.01. a Values × 10− 3 units/mg protein. b Units/mg protein.
S. Vijayakumar et al. / Fitoterapia 79 (2008) 279–282
281
Table 3 Concentration of glutathione in tissues of rats treated with M. paradisiaca flavonoids Groups
Dose
Liver
Heart
Kidney
375.19 ± 7.5 419.6 ± 10.49⁎ 310.90 ± 6.21 380.38 ± 9.5⁎
487.1 ± 9.74 546.74 ± 13.66⁎ 300.8 ± 6.01 342.16 ± 8.21⁎
320.12 ± 7.68 373.2 ± 10.44⁎ 400.6 ± 8.01 455.31 ± 12.74⁎
mg/100 g BW/day – 1.0 – 1.0
I II III IV
Values expressed as mM/100 g wet tissues. Average of the values of six rats in each group ± SE. Group I was compared with groups II and group III was compared with group IV. ⁎P b 0.01.
high fat diet with flavonoids at a dose of 10 mg (group IV). Animals were treated orally by gastric intubation for 90 days. At the end of the experimental periods, the liver, kidney and heart were removed for the estimations of malondialdehyde (MDA), hydroperoxides, conjugated diens and glutathione according to John AB Steven [15]. The activities of superoxide dismutase (SOD) and catalase were determined by the method of Kakkar et al. [16]. Protein estimation was done by the method of Lowry et al. [17]. 2.5. Statistical analysis Statistical significance was calculated using Student's t-test [18]. 3. Results and discussion Food intake and weight gain were almost similar in both control as well as experimental groups. Concentrations of malondialdehyde, hydroperoxides and conjugated diens were decreased significantly (Table 1) whereas the activities of superoxide dismutase (SOD) and catalase were enhanced significantly in the groups (II and IV) treated with flavonoids (Table 2). No significant change in the concentration of conjugated diens was noticed in the kidney of rats fed high fat diet. Concentration of glutathione showed significant increase in the tissues of rats fed normal as well as high fat diets (Table 3). The higher activity of SOD indicating its free radical scavenging activity [19]. The highly stimulated activities of SOD and catalase in rats fed flavonoids probably account for the decreased levels of peroxidation products such as malondialdehyde, hydroperoxides and conjugated diens. The antiperoxidative activity of flavonoids was reported to be directly proportional to the number of free phenolic hydroxy groups [20].The present study emphasis that the flavonoids from banana act as an effective antioxidant. Therefore, the supplementation of natural antioxidants through a balanced diet containing banana could be effective in protecting the body against various oxidative stresses. References [1] [2] [3] [4] [5] [6] [7] [8] [9] [10] [11] [12] [13] [14]
Mookerjee BK, Lee TP, Lipper HA, Middleton E. J Immunopharmacol 1986;8:371. De Whalley CV, Rankin SM, Hoult Jessup W, Leake DS. Biochem Pharmacol 1990;39:1743. Abad MJ, Bermejo P, Villas A, Valverde S. J Natl Prod 1993;56:1164. Borissova P, Valcheva S, Belcheva A. Acta Physiol Pharmacol Bulg 1994;20:25. Sailaja A, Scalese M, Lanza M, Marzullo D, Bonina F, Castelli F. Free Radical Biol Med 1995;19:481. Vinodhkumar R, Augusyt KT. Indian J Biochem Biophys 1989;26:400. Charles STH, Simon LH. Diabetes 1984;33:291. Steinberg D, Parthasarathy S, Carew TE, Khoo JC, Witzum JL. New Engl J Med 1980;320:915. Mangiapane H, Thomson J, Salter A, Brown S, Bell GD, White DA. Biochem Pharmacol 1992;43:45. Rankin S, De Whalley CV, Hoult RS, Jessup W, Wilkins GM, Collard J, et al. Biochem Pharmacol 1993;45:67. Sudheesh S, Presanna Kumar G, Vijayakumar S, Vijaylakshmi NR. Plant Foods Hum Nutr 1997;51:321. Sudheesh S, Sandhya C, Koshy AS, Vijaylakshmi NR. Phytother Res 1999;13:393. Markham KR. Techniques of flavonoid identification. London: Academic Press; 1982. Eskin NAM, Hoehn E, Frenket C. J Agric Food Chem 1978;26:973.
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S. Vijayakumar et al. / Fitoterapia 79 (2008) 279–282
[15] Steven John AB. Microsomal lipid peroxidation. In: Colowick SP, Kaplan NO, editors. Methods in enzymology. London: Academic Press; 1978. [16] Kakkar P, Das B, Viswanathan PN. Indian J Biochem Biophys 1984;21:310. [17] Lowry OH, Rosebrough NJ, Farr AL, Randail RJ. Biol Chem 1951;193:265. [18] Bennet CA, Franklin NL. Statistical analysis in chemistry and chemical industry. New York: Wiley and Sons; 1967. [19] Sudheesh S, Vijayalakshmi NR. Fitoterapia 2005;76:181. [20] Joyeux M, Lobstein N, Anton R, Mortier F. Planta Med 1995;61:126.