Back
 FNS  Vol.4 No.2 , February 2013
Preliminary Evaluation for Comparative Antioxidant Activity in the Water and Ethanol Extracts of Dried Citrus Fruit (Citrus unshiu) Peel Using Chemical and Biochemical in Vitro Assays
Abstract: Comparative antioxidant activities of the water and ethanol extracts obtained from dried citrus fruit (Citrus unshiu) peel were determined using chemical and biochemical in vitro assays. Chemical assays were used for evaluation of 1,1-diphenyl-2-picryl hydrazyl (DPPH) radical scavenging, hydrogen peroxide scavenging, and reducing power of both extracts and their total phenolic, flavonoid and tannin contents and antioxidant activities were investigated. Biochemical assays were performed to evaluate the inhibition activities of AAPH-induced rat RBC hemolysis and Fe2+-induced lipid peroxidation using rat brain tissue cells. Linear correlation between the antioxidant activities of both extracts were determined by chemical assays, and total phenolic, flavonoid and tannin contents was observed in concentration-dependent mode. Both extracts showed >95% DPPH radical scavenging and >85% hydrogen peroxide scavenging, and higher reducing capacity at the same level of 3200 ppm. In the inhibition activity of AAPH-induced hemolysis, water extracts showed a strong activity in concentration-dependent mode up to 1600 ppm with no statistical difference found between 1600 and 3200 ppm (P > 0.05). In the inhibition activity of Fe2+-induced lipid peroxidation, ethanol extracts showed the higher inhibition percentage of lipid peroxidation than those of water extracts at the same concentration with no significant difference (P > 0.05) found in the range of 800 to 3200 ppm. The extracts of dried Citrus unshiu peel may be considered as potential antioxidant ingredients of functional food depending on the conditions at which reactive oxygen species are implicated.
Cite this paper: J. Kim, "Preliminary Evaluation for Comparative Antioxidant Activity in the Water and Ethanol Extracts of Dried Citrus Fruit (Citrus unshiu) Peel Using Chemical and Biochemical in Vitro Assays," Food and Nutrition Sciences, Vol. 4 No. 2, 2013, pp. 177-188. doi: 10.4236/fns.2013.42025.
References

[1]   M. Lohrasbi, M. Pourbafrani, C. Niklasson and M. J. Taherzadeh, “Process Design and Economic Analysis of a Citrus Waste Biorefinery with Biofuels and Limonene as Products,” Bioresource Technology, Vol. 101, No. 19. 2010, pp. 7382-7388. doi:10.1016/j.biortech.2010.04.078

[2]   G. H. Xu, J. C. Chen, D. H. Liu, Y. H. Zhang, P. Jiang and X. Q. Ye, “Minerals, Phenolic Compounds, and Antioxidant Capacity of Citrus Peel Extract by Hot Water,” Journal of Food Science, Vol. 73, No. 1, 2008, pp. C11-C18. doi:10.1111/j.1750-3841.2007.00546.x

[3]   M. B. Kim, J. S. Park and S. B. Lim, “Antioxidant Activity and Cell Toxicity of Pressurized Liquid Extracts from 20 Selected Plant Species in Jeju, Korea,” Food Chemistry, Vol. 122, No. 3, 2010, pp. 546-552. doi:10.1016/j.foodchem.2010.03.007

[4]   U. Imeh and S. Khokhar, “Distribution of Conjugated and Free Phenols in Fruits: Antioxidant Activity and Cultivar Variations,” Journal of Agricultural and Food Chemistry, Vol. 50, No. 22, 2002, pp. 6301-6306. doi:10.1021/jf020342j

[5]   J. A. Ross and C. M. Kasum, “Dietary Flavonoids: Bioavailability Metabolic Effects, and Safety,” Annual Review of Nutrition, Vol. 22, 2002, pp. 19-34. doi:10.1146/annurev.nutr.22.111401.144957

[6]   J. Silalahi, “Anticancer and Health Protective Properties of Citrus Fruit Components,” Asia pacific Journal of Clinical Nutrition, Vol. 11, No. 1, 2002, pp. 79-84. doi:10.1046/j.1440-6047.2002.00271.x

[7]   B. Halliwell, “Free Radicals, Antioxidants, and Human Disease: Curiosity, Cause, or Consequence?” Lancet, Vol. 344, No. 8924, 1994, pp. 721-724. doi:10.1016/S0140-6736(94)92211-X

[8]   J. J. Pincemail, “Free Radicals and Antioxidants in Human Diseases,” In: A. E. Favier, J. Cadet, B. Kalyanaraman, M. Fontecave, J. L. Pierre, Eds., Analysis of Free Radicals in Biological Systems, Birkhauser Verlag, Berlin, 1995, pp. 83-98.

[9]   D. Dreher and A. F. Junod, “Role of Oxygen Free Radicals in Cancer Development,” European Journal of Cancer, Vol. 32A, No. 1, 1996, pp. 30-38. doi:10.1016/0959-8049(95)00531-5

[10]   S. Maxwell, “Anti-Oxidant Therapy: Does It Have a Role in the Treatment of Human Disease?” Expert Opinion on Investgational Drugs, Vol. 6, No. 3, 1997, pp. 211-236. doi:10.1517/13543784.6.3.211

[11]   V. Lopez, S. Akerreta, E. Casanova, J. M. Garcia-Mina, R. Y. Cavero and M. I. Calvo, “In Vitro Antioxidant and Anti-Rhizopus Activities of Lamiaceae Herbal Extracts,” Plant Foods for Human Nutrtion, Vol. 62, No. 4, 2007, pp. 151-155. doi:10.1007/s11130-007-0056-6

[12]   G. Satpathy, Y. K. Tyagi and R. K. Gupta, “Preliminary Evaluation of Nutraceutical and Therapeutic Potential of Raw Spondias Pinnata K., an Exotic Fruit of India,” Food Research International, Vol. 44, No. 7, 2011, pp. 2076-2087. doi:10.1016/j.foodres.2011.01.040

[13]   M. G. L. Hertog, E. J. M. Feskeens, C. H. Holmann, K. B. Katan and D. Kromhout, “Dietary Antioxidant Flavonoids and Risk of Coronary Heart Disease: The Zutphen Elderly Study,” Lancet, Vol. 342, No. 8878, 1993, pp. 1007-1011. doi:10.1016/0140-6736(93)92876-U

[14]   V. Elangovan, N. Sekar and S. Govindasamy, “Chemoprotective Potential of Dietary Bioflavonoids against 20-Methylcholanthrene-Induced Tumorigenesis,” Cancer Letter, Vol. 87, No. 1, 1994, pp. 107-113. doi:10.1016/0304-3835(94)90416-2

[15]   F. R. Marin, M. J. Frutos, J. A. Perez-Alvarez, F. Martinez-Sanchez and J. A. Del Rio, “Flavonoids as Nutraceuticals: Structure Related Antioxidant Properties and Their Role on Ascorbic Acid Preservation,” In: Atta-Ur-Rahman, Ed., Studies in Natural Products Chemistry, Elsevier Science, Amsterdam, 2002, pp. 324-389.

[16]   D. D. Majo, M. Giammanco, M. L. Guardia, E. Tripoli, S. Giammanco and E. Finotti, “Flavanones in Citrus Fruit: Structure-Antioxidant Activity Relationships,” Food Research International, Vol. 38, No. 10, 2005, pp. 1161-1166. doi:10.1016/j.foodres.2005.05.001

[17]   M. A. Anagnostopoulou, P. Kefalas, V. P. Papageorgiou, A. N. Assimopoulou and D. Boskou, “Radical Scavenging Activity of Various Extracts and Fractions of Sweet Orange Peel (Citrus sinensis),” Food Chemistry, Vol. 94, No. 1, 2006, pp. 19-25. doi:10.1016/j.foodchem.2004.09.047

[18]   A. Bocco, M. E. Cuvelier, H. Richard and C. Berset, “Antioxidant Activity and Phenolic Composition of Citrus Peel and Seed Extracts,” Journal of Agricultural and Food Chemistry, Vol. 46, No. 6, 1998, pp. 2123-2129. doi:10.1021/jf9709562

[19]   S. Gorinstein, O. Martín-Belloso, Y. S. Park, R. Haruenkit, A. Lojek, M. Cíz, A. Gaspi, I. Libman and S. Trakhtenberg, “Comparison of Some Biochemical Characteristics of Different Citrus Fruits,” Food Chemistry, Vol. 74, No. 3, 2001, pp. 309-315. doi:10.1016/S0308-8146(01)00157-1

[20]   J. A. Manthey and K. Grohmann, “Phenols in Citrus Peel Byproducts Concentrations of Hydroxycinnamates and Polymethoxylated Flavones in Citrus Peel Molasses,” Journal of Agricultural and Food Chemistry, Vol. 49, No. 7, 2001, pp. 3268-3273. doi:10.1021/jf010011r

[21]   D. K. Asami, Y. J. Hong, D. M. Barrett and A. E. Mitchell, “Comparison of the Total Phenolic and Ascorbic Acid Content of Freeze-Dried and Air-Dried Marionberry, Strawberry, and Corn Grown Using Conventional, Organic, and Sustainable Agricultural Practice,” Journal of Agricultural and Food Chemistry, Vol. 51, No. 5, 2003, pp. 1237-1241. doi:10.1021/jf020635c

[22]   M. Y. Choi, C. Chai, J. H. Park, J. Lim, J. Lee and S. M. Kwon, “Effects of Storage Period and Heat Treatment on Phenolic Compound Composition in Dried Citrus Peels (Chenpi) and Discrimination of Chenpi with Different Storage Periods Through Targeted Metabolomic Study Using HPLC-DAD Analysis,” Journal of Paarmaceutical and Biomedical Analysis, Vol. 54, No. 4, 2011, pp. 638-645. doi:10.1016/j.jpba.2010.09.036

[23]   M. Ou, “Regular Chinese Medicine Handbook,” Warmth Publishing Ltd., Taiwan, 1999.

[24]   H. J. Oh, H. M. Ahn, S. S. Kim, P. Y. Yun, K. L. Jeon, Y. H. Ko and K. Z. Riu, “Composition and Antimicrobial Activities of Essential Oils in the Peel of Citrus Fruits. Journal of Applied Biological Chemistry, Vol. 50, No. 3, 2007, pp. 148-154.

[25]   B. B. Li, B. Smith and M. Hossain, “Extraction of Phenolics from Citrus Peels I. Solvent Extraction Method,” Separation and Purification Technology, Vol. 48, No. 2, 2006, pp. 182-188. doi:10.1016/j.seppur.2005.07.005

[26]   K. Ghasemi, Y. Ghasemi and M. A. Ebrahimzadeh, “Antioxidant Activity, Phenol and Flavonoid Contents of 13 Citrus Species Peels and Tissues,” Pakistan Journal of Pharmaceutical Sciences, Vol. 22, No. 3, 2009, pp. 277-281.

[27]   J. C. Tang, J. N. Zhang, Y. T. Wu and Z. X. Li, “Effect of the Water Extract and Ethanol Extract from Traditional Chinese Medicines Angelica sinensis (Oliv.) Diels, Ligusticum chuanxiong Hort. and Rheum palmatum L. on Rat Liver Cytochrome P450 Activity,” Phytotherapy Research, Vol. 20, No. 12, 2006, pp. 1046-1051. doi:10.1002/ptr.1974

[28]   J.-S. Kim, “Radical Scavenging Capacity and Antioxidant Activity of the E Vitamer Fraction in Rice Bran,” Journal of Food Science, Vol. 70, No. 3, 2005, pp. C208-213. doi:10.1111/j.1365-2621.2005.tb07127.x

[29]   T. Hatano, H. Kagawa, T. Yasuhara and T. Okuda, “Two New Flavonoids and Other Constituents in Licorice Root: Their Relative Astringency and Radical Scavenging Effects,” Chemical and Pharmaceutical Bulletin, Vol. 36, No. 6, 1988, pp. 2090-2097. doi:10.1248/cpb.36.2090

[30]   D. Atmani, N. Chaher, M. Berboucha, K. Ayouni, H. Lounis, H. Boudaoud, N. Debbache and D. Atmani, “Antioxidant Capacity and Phenol Content of Selected Algerian Medicinal Plants,” Food Chemistry, Vol. 112, No. 2, 2009, pp. 303-309. doi:10.1016/j.foodchem.2008.05.077

[31]   M. Oyaizu, “Studies on Product of Browning Reaction Prepared from Glucose Amine,” Japanese Journal of Nutrition, Vol. 44, No. 6, 1986, pp. 307-315. doi:10.5264/eiyogakuzashi.44.307

[32]   S. N. Lim, P. C. K. Cheung, V. E. C. Ooi and P. O. Ang, “Evaluation of Antioxidative Activity of Extracts from a Brown Seaweed Sargassum siliquastrum,” Journal of Agricultural and Food Chemistry, Vol. 50, No. 13, 2002, pp. 3862-3866. doi:10.1021/jf020096b

[33]   R. Julkunen-Tiitto, “Phenolic Contents in the Leaves of Northern Willows: Methods for the Analysis of Certain Phenolics,” Journal of Agricultural and Food Chemistry, Vol. 33, No. 2, 1985, pp. 213-217. doi:10.1021/jf00062a013

[34]   Z. Marksimovic, D. Malencic and N. Kovacevic, “Polyphenol Contents and Antioxidant Activity of Maydis stigma Extracts,” Bioresource Technology, Vol. 96, No. 8, 2005, pp. 873-877. doi:10.1016/j.biortech.2004.09.006

[35]   T. B. Ng, F. Liu and Z. T. Wang, “Antioxidant Activity of Natural Products from Plants,” Life Sciences, Vol. 66, No. 8, 2000, pp. 709-723. doi:10.1016/S0024-3205(99)00642-6

[36]   F. Liu, V. E. C. Ooi and S. T. Chang, “Free Radical Scavenging Activities of Mushroom Polysaccharide Extracts,” Life Sciences, Vol. 60, No. 10, 1997, pp. 763-771. doi:10.1016/S0024-3205(97)00004-0

[37]   C. Sánchez-Moreno, J. A. Larrauri and F. Saura-Calixto, “Free Radical Scavenging Capacity and Inhibition of Lipid Oxidation of Wines, Grape Juices and Related Polyphenolic Constituents,” Food Research International, Vol. 32, No. 6, 1999, pp. 407-412. doi:10.1016/S0963-9969(99)00097-6

[38]   R. P. Singh, K. N. C. Murthy and G. K. Jayaprakasha, “Studies on the Antioxidant Activity of Pomegranate (Punica granatum) Peel and Seed Extracts Using in Vitro Models,” Journal of Agricultural and Food Chemistry, Vol. 50, No. 1, 2002, pp. 81-86. doi:10.1021/jf010865b

[39]   E. R. Sherwin, “Oxidation and Antioxidants in Fat and Oil Processing,” Journal of the American Oil Chemists’ Society, Vol. 55, No.11, 1978, pp. 809-814.

[40]   E. Tripoli, M. L. Guardia, S. Giammanco, D. D. Majo and M. Giammanco, “Citrus Flavonoids: Molecular Structure, Biological Activity and Nutritional Properties: A Review,” Food Chemistry, Vol. 104, No. 2, 2007, pp. 466-479. doi:10.1016/j.foodchem.2006.11.054

[41]   M. E. Cuverlier, H. Richard and C. Berset, “Comparison of the Antioxidative Activity of Some Acid Phenols: Structure Activity Relationship,” Bioscience Biotechnology and Biochemistry, Vol. 56, No. 2, 1992, pp. 324-325. doi:10.1271/bbb.56.324

[42]   V. Bondet, W. Brand-Williams and C. Berset, “Kinetics and Mechanisms of Antioxidant Activity Using the DPPH? Free Radical Method,” Lebensmittel Wissenschaft und Technologie, Vol. 30, No. 6, 1997, pp. 609-615.

[43]   H. Y. Lin and C. C. Chou, “Antioxidative Activities of Water-Soluble Disaccharide Chitosan Derivatives,” Food Research International, Vol. 37, No. 9, 2004, pp. 883889. doi:10.1016/j.foodres.2004.04.007

[44]   B. Halliwell, “Reactive Oxygen Species in Living Systems; Source, Biochemistry, and Role in Human Disease,” The American Journal of medicine, Vol. 91, No. 3, 1991, pp. 14-22.

[45]   S. M. Nabavi, M. A. Ebrahimzadeh, S. F. Navabi and M. Jafari, “Free Radical Scavenging Activity and Antioxidant Capacity of Eryngium caucasicum Trautv and Froripia subpinnata,” Pharmacologyonline, Vol. 3, 2008, pp. 19-25.

[46]   M. A. Ebrahimzadeh, S. F. Navabi and S. M. Navabi, “Antioxidant Activities of Methanol Extract of Sambucus ebulus L. Flower,” Pakistan Journal of Biological Sciences, Vol. 12, No. 5, 2009, pp. 447-450. doi:10.3923/pjbs.2009.447.450

[47]   M. L. Calabrò, V. Galtieri, P. Cutroneo, S. Tommasini, P. Ficarra and R. Ficarra, “Study of the Extraction Procedure by Experimental Design and Validation of a LC Method for Determination of Flavonoids in Citrus bergamia Juice,” Journal Pharmaceutical and Biomedical Analysis, Vol. 35, No. 2, 2004, pp. 349-363. doi:10.1016/S0731-7085(03)00585-5

[48]   Z. Yi, Y. Yu, Y. Liang and B. Zeng, “In Vitro Antioxidant and Antimicrobial Activities of the Extract of Percarpium citri Reticulatae of a New Citrus Cultivar and Its Main Flavonoids,” Food Science and Technology, Vol. 41, No. 4, 2008, pp. 597-603. doi:10.1016/j.lwt.2007.04.008

[49]   C. L. Hsu, W. L. Chen, Y. M. Weng and C. Y. Tseng, “Chemical Composition, Physical Properties, and Antioxidant Activities of Yam Flours as Affected by Different Drying Methods,” Food Chemistry, Vol. 83, No. 1, 2003, pp. 85-92. doi:10.1016/S0308-8146(03)00053-0

[50]   J. S. Rathee, S. A. Hassarajani and S. Chattopadhyay, “Antioxidant Activity of Nyctanthes arbor-tristis Leaf Extract,” Food Chemistry, Vol. 103, No. 4, 2007, pp. 1350- 1357. doi:10.1016/j.foodchem.2006.10.048

[51]   M. A. Ebrahimzadeh, S. M. Navabi, A. F. Navabi, F. Bahramian and A. R. Bekhradnia, “Antioxidant and Free Radical Scavenging Activity of H. officinalis L. var. angustifolius, V. odorata, B. hyrcana and C. speciosum,” Pakistan Journal of Pharmaceutical Sciences, Vol. 23, No. 1, 2010, pp. 29-34.

[52]   H. Liu, N. Qiu, H. Ding and R. Yao, “Polyphenols Contents and Antioxidant Capacity of 68 Chinese Herbals Suitable for Medical or Food Uses,” Food Research International, Vol. 41, No. 4, 2008, pp. 363-370. doi:10.1016/j.foodres.2007.12.012

[53]   Y. Q. Ma, X. Q. Ye, Z. X. Fang, J. C. Chen, G. H. Xu and D. H. Liu, “Phenolic Compounds and Antioxidant Activity of Extracts from Ultrasonic Treatment of Satsuma Mandarin (Citrus unshiu Marc.) Peels,” Journal of Agricultural and Food Chemistry, Vol. 56, No. 14, 2008, pp. 5682-5690. doi:10.1021/jf072474o

[54]   V. W. Bowry and R. Stocker, “Tocopherol-Mediated Peroxidation. The Peroxidation Effect of Vitamin E on the Radical-Initiated Oxidation of Human Low-Density Lipoprotein,” Journal of the American Chemistry Society, Vol. 115, No. 14, 1993, pp. 6029-6044. doi:10.1021/ja00067a019

[55]   Z. Q. Liu, L. P. Ma, B. Zhou, L. Yang and Z. L. Liu, “Antioxidative Effects of Green Tea Polyphenols on Free Radical Initiated and Photosensitized Peroxidation of Human Low Density Lipoprotein,” Chemistry and Physics of Lipids, Vol. 106, No. 1, 2000, pp. 53-63. doi:10.1016/S0009-3084(00)00133-X

[56]   F. Dai, Q. Miao, B. Zhou, L. Yang and Z. L. Liu, “Protective Effects of Flavonols and Their Glycosides against Free Radical-Induced Oxidative Hemolysis of Red Blood Cells,” Life Sciences, Vol. 78, No. 21, 2006, pp. 2488- 2493. doi:10.1016/j.lfs.2005.10.009

[57]   C. A. Rice-Evans, N. J. Miller and G. Paganga, “Antioxidant Properties of Phenolic Compounds,” Trends in Plant Science, Vol. 2, No. 4, 1997, pp. 152-158. doi:10.1016/S1360-1385(97)01018-2

[58]   A. Djeridane, M. Yousfi, B. Nadjemi, D. Boutassouna, P. Stocker and N. Vidal, “Antioxidant Activity of Some Algerian Medicinal Plants Extracts Containing Phenolic Compounds,” Food Chemistry, Vol. 97, No. 4, 2006, pp. 654-660. doi:10.1016/j.foodchem.2005.04.028

[59]   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,” Journal of Agricultural Food Chemistry, Vol. 50, No. 22, 2002, pp. 6521-6526. doi:10.1021/jf020565+

[60]   B. A. Igor, I. D. Anatolli, V. B. Aleksander, A. K. Valadimer and I. P. Alla, “Chelating and Free Radical Scavenging Mechanisms of Inhibiting Action of Rutin and Quercetin in Lipid Peroxidation,” Biochemical Pharmacology, Vol. 38, No. 11, 1989, pp. 1763-1769. doi:10.1016/0006-2952(89)90410-3

[61]   N. P. Seeram and M. G. Nair, “Inhibition of Lipid Peroxidation and Structure-Activity-Related Studies of the Dietary Constitutes Anthocyanins, Anthocyanidins, and Catechins,” Journal of Agricultural and Food Chemistry, Vol. 50, No. 19, 2002, pp. 5308-5312. doi:10.1021/jf025671q

 
 
Top