• Rohan Sharadanand Phatak Krishna Institute of Medical Sciences Deemed University
  • Asha Krishnaji
  • Anup Subhash



Background: India is endowed with enormous varieties of spices grown in the majority of the country and one of the largest exporters of spices in the
world. Many household spices are being used integrally in the Indian foods. Spices are normally added in the food to impart flavor. They are naturally
occurring antioxidants, which have the potential capacity to counteract aging process in the body, to stabilize the cell membrane by scavenging free
radicals in small doses.
Objectives: The current study was designed to determine antioxidant potential of some selected spices and their mixture based on the established
scientific evidences and oxygen radical absorbance capacity values using different in vitro models and correlating.
Methods: The powder of assigned spices and their mixture were alcoholically extracted by a simple maceration method. They were evaluated for
their total phenolic and flavonoid contents. Antioxidative abilities of the extracts of spices individually and their mixtures extracts were analyzed by
phosphomolybdenum assay, cupric ions reducing antioxidant capacity (CUPRAC) and ferric ions reducing ability power methods. The free radical
scavenging activities such as hydrogen peroxide, ABTS, anti-peroxidation like TBARS (thiobarbituric acid reactive substance), crocin bleaching and
metal chelation capacity were assayed through in vitro models.
Results and Discussion: Antioxidant and antiradical effects of the selected spices extracts and spices mixture extract (SME) was ascertained through
different in vitro models. Results of individual tests demonstrated good correlation among themselves. CUPRAC has strong correlation with all other
assays except to diphenyl-1-picrylhydrazyl (DPPH) and thiobarbituric acid reactive substances (TBARS). DPPH and TBARS were correlated with each
other however they demonstrated no relationship with other assays.
Conclusion: Extracts of selected Indian spices extracts and SME have shown dissimilarity to each other. Study for the 1st time establishes aggregate
index for evaluation of Indian spices. On basis of aggregate index, nutmeg has highest value among the selected spices and no significant synergistic
effect has been found in SME. CUPRAC as antioxidant assay and hydroxyl free radical scavenging assay by using Smirnoff reagent are most appropriate
assays to ascertain antioxidative and free scavenger properties of natural products.

Keywords: Antioxidants, Antiradical, Oxygen Radical Absorbance Capacity, Spices, Spices Mixture Extract.


Halliwell B, Gutreridge JM. Free Radicals in Biology and Medicine. 2nd ed. Oxford, UK. Clarendon Press; 1989.

Halliwell B, Aruoma OI. DNA damage by oxygen-derived species. Its mechanism and measurement in mammalian systems. FEBS Lett 1991;281(1-2):9-19.

Ames BN, Shigenaga MK, Hagen TM. Oxidants, antioxidants, and the degenerative diseases of aging. Proc Natl Acad Sci U S A 1993;90(17):7915-22.

Sultana S, Ripa FA, Hamid K. Comparative antioxidant activity study of some commonly used spices in Bangladesh. Pak J Biol Sci 2010;13(7):340-3.

Rahman K. Studies on free radicals, antioxidants, and co-factors. Clin Interv Aging 2007;2(2):219-36.

Barlow SM. Toxicological aspects of antioxidants used as food additives. In: Hudson BJ, editor. Food Antioxidants. Amsterdam, The Netherlands: Elsevier; 1990. p. 253-307.

Tirzitis G, Bartosz G. Determination of antiradical and antioxidant activity: Basic principles and new insights. Acta Biochim Pol 2010;57(2):139-42.

Haytowitz DB, Bhagwat S. USDA Database for the Oxygen Radical Absorbance Capacity (ORAC) of Selected Foods. Release 2. 2010. p. 1-46.

Singleton VL, Rossi JA. Colorimetry of total phenolics withphosphomolybdic phosphotungstic acid reagents. Am J Enol Vitic 1965;16:144-58.

Zhishen J, Mengcheng T, Jianming W. The determination of flavonoid contents in mulberry and their scavenging effect on superoxide radicals. Food Chem 1999;64:555-9.

Prieto P, Pineda M, Aguilar M. Spectrophotometric quantitation of antioxidant capacity through the formation of a phosphomolybdenum complex: Specific application to the determination of vitamin E. Anal Biochem 1999;269(2):337-41.

Apak R, Güçlü K, Özyürek, M, Karademir SE. A novel total antioxidant capacity index for dietary polyphenols, vitamins c and e, using their cupric ion reducing capability in the presence of neocuproine: Cuprac method. J Agric Food Chem 2004;52(26):7970-81.

Oyaizu M. Studies on products of browning reaction: Antioxidant activities of products of browning reaction prepared from glucosamine. Jpn J Nutr 1986;44:307-15.

Ruch RJ, Cheng SJ, Klaunig JE. Prevention of cytotoxicity and inhibition of intercellular communication by antioxidant catechins isolated from Chinese green tea. Carcinogenesis 1989;10(6):1003-8.

Panda BN, Raj AB, Shrivastava NR, Prathani AR. The evaluation of nitric oxide scavenging activity of Acalypha Indica Linn Root. Asian J Res Chem 2009;2(2):148-50.

Smirnoff N, Cumbes QJ. Hydroxyl radical scavenging activity of compatible solutes. Phytochemistry 1989;28:1057-60.

Duan XW, Jiang YM, Su XG, Zhang ZQ, Shi J. Antioxidant properties of anthocyanins extracted from litchi (Litchi chinenesis Sonn.) fruit pericarp tissues in relation to their role in the pericarp browning. Food Chem 2007;101:1365-71.

Anandjiwala S, Bagul MS, Parabia M, Rajani M. Evaluation of free radical scavenging activity of an ayurvedic formulation, panchvalkala. Indian J Pharm Sci 2008;70(1):31-5.

Re R, Pellegrini N, Proteggente A, Pannala A, Yang M, Rice-Evans C. Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radic Biol Med 1999;26(9-10):1231-7.

Banerjee A, Dasgupta N, De B. In vitro study of antioxidant activity of Syzygium cumini fruit. Food Chem 2005;90:727-33.

Peng S. Methods and applications of anticancer bioassays. In: Peng S, Zhao M. editors. Pharmaceutical Bioassays: Methods and Applications. Hoboken, NJ, USA: John Wiley & Sons, Inc.; 2009.

Ichikawa H, Konishi T. In vitro antioxidant potentials of traditional Chinese medicine, Shengmai San and their relation to in vivo protective effect on cerebral oxidative damage in rats. Biol Pharm Bull 2002;25(7):898-903.

Lussignoli S, Fraccaroli M, Andrioli G, Brocco G, Bellavite P. A microplate-based colorimetric assay of the total peroxyl radical trapping capability of human plasma. Anal Biochem 1999;269(1):38‑44.

Dinis TC, Maderia VM, Almeida LM. Action of phenolic derivatives (acetaminophen, salicylate, and 5-aminosalicylate) as inhibitors of membrane lipid peroxidation and as peroxyl radical scavengers. Arch Biochem Biophys 1994;315(1):161-9.

Gupta SK, Prakash J, Srivastava S. Validation of traditional claim of Tulsi, Ocimum sanctum Linn. as a medicinal plant. Indian J Exp Biol 2002;40(7):765-73.

Prakash P, Gupta N. Therapeutic uses of Ocimum sanctum Linn (Tulsi) with a note on eugenol and its pharmacological actions: A short review. Indian J Physiol Pharmacol 2005;49(2):125-31.

Gu¨lcin I, Elmastas M, Aboul-Enein HY. Antioxidant activity of clove oil – A powerful antioxidant source. Arab J Chem 2012;5(4):489-99.

Cortés-Rojas DF, de Souza CR, Oliveira WP. Clove (Syzygium aromaticum): A precious spice. Asian Pac J Trop Biomed 2014;4(2):90‑6.

Subash Babu P, Prabuseenivasan S, Ignacimuthu S. Cinnamaldehyde – A potential antidiabetic agent. Phytomedicine 2007;14(1):15-22.

Prabuseenivasan S, Jayakumar M, Ignacimuthu S. In vitro antibacterial activity of some plant essential oils. BMC Complement Altern Med 2006;6:39.

Roussel AM, Hininger I, Benaraba R, Ziegenfuss TN, Anderson RA. Antioxidant effects of a cinnamon extract in people with impaired fasting glucose that are overweight or obese. J Am Coll Nutr 2009;28(1):16-21.

Menon VP, Sudheer AR. Antioxidant and anti-inflammatory properties of curcumin. Adv Exp Med Biol 2007;595:105-25.

Ramadas D, Srinivas A. Antioxidant effects of 28KDA antioxidant protein from turmeric (Curcuma longa l). Asian J Pharm Clin Res 2011;4 Suppl 1:75-9.

Surh YJ, Lee E, Lee JM. Chemoprotective properties of some pungent ingredients present in red pepper and ginger. Mutat Res 1998;402(1‑2):259-67.

Manju V, Nalini N. Effect of ginger on lipid peroxidation and antioxidant status in 1, 2-dimethyl hydrazine induced experimental colon carcinogenesis. J Biochem Technol 2010;2(2):161-7.

Wilson T, Singh AP, Freeman MM, Singh V, Olson RM, Vorsa N, et al. Characterization of curry leaf polyphenolics and their antioxidant activity. FASEB J 2009;23 (Meeting Abstract Supplement):718.4.

Handral HK, Pandith A, Shruthi SD. A review on Murraya koenigii: Multipotential medicinal plant. Asian J Pharm Clin Res 2012;5(4):5-14.

Kiasalari Z, Khalili M, Roghani M, Sadeghian A. Antiepileptic and antioxidant effect of Brassica nigra on pentylenetetrazol-induced kindling in mice. Iran J Pharm Res 2012;11(4):1209-17.

Dua A, Gupta SK, Mittal A, Mahajan R. A study of antioxidant properties and antioxidant compounds of cumin (Cuminum cyminum). Int J Pharm Biol Arch 2012;3(5):1110-6.

Allahghadri T, Rasooli I, Owlia P, Nadooshan MJ, Ghazanfari T, Taghizadeh M, et al. Antimicrobial property, antioxidant capacity, and cytotoxicity of essential oil from cumin produced in Iran. J Food Sci 2010;75(2):H54-61.

Jeena K, Lijua VB, Umadevia NP, Kuttan R. Antioxidant, anti‑inflammatory and antinociceptive properties of black pepper essential oil (Piper nigrum Linn). J Essent Oil Bearing Plants 2014;17(1):1-12.

Srinivasan K. Black pepper and its pungent principle-piperine: A review of diverse physiological effects. Crit Rev Food Sci Nutr 2007;47(8):735-48.

Gupta AD, Bansa VK, Vikash B, Maithil N. Chemistry, antioxidant and antimicrobial potential of nutmeg (Myristica fragrans Houtt). J Genet Eng Biotechnol 2013;11(1):25-31.

Prior R. Antioxidant food databases? Valuable or not? Brunswick Laboratories website. Response to the USDA ORAC Statement.pdf. Accessed December 1, 2012.

Chapple IL, Matthews JB. The role of reactive oxygen and antioxidant species in periodontal tissue destruction. Periodontol 2000 2007;43:160‑232.

Katsube T, Tabata H, Ohta Y, Yamasaki Y, Anuurad E, Shiwaku K, et al. Screening for antioxidant activity in edible plant products: Comparison of low-density lipoprotein oxidation assay, DPPH radical scavenging assay, and Folin-Ciocalteu assay. J Agric Food Chem 2004;52(8):2391‑6.

He Z, Lan M, Lu D, Zhao H, Yuan H. Antioxidant activity of 50 traditional chinese medicinal materials varies with total phenolics. Chin Med 2013;4(4):148-56.

Wojdyłoa A, Oszmian´ski J, Czemerys R. Antioxidant activity and phenolic compounds in 32 selected herbs. Food Chem 2007;105:940-9.

Abdul MR, Marka V, Prameela Y. A study on antioxidant activity of some commonly used spices in India. Int J Life Sci Biotechnol Pharma Res 2013;2(4):145-50.

Deepa G, Ayesha S, Nishtha K, Thankamani M. Comparative evaluation of various total antioxidant capacity applied to phytochemical compounds of Indian culinary spices. Int Food Res J 2013;20(4):1711‑6.

Chan EW, Lim YY, Lim TY. Total phenolic content and antioxidant activity of leaves and rhizomes of some ginger species in Peninsular Malaysia. Gard Bull Singapore 2007;59(1&2):47-56.

Tupe RS, Kemse, NG, Khaire AA. Evaluation of antioxidant potentials and total phenolic contents of selected Indian herbs powder extracts. Int Food Res J 2013;20(3):1053-63.

Muselík J, García-Alonso M, Martín-López MP, ŽemliÄka M, Rivas-Gonzalo JC. Measurement of antioxidant activity of wine catechins, procyanidins, anthocyanins and pyranoanthocyanins. Int J Mol Sci 2007;8:797-809.

Apak R, Güçlü K, Demirata B, Ozyürek M, Celik SE, Bektasoglu B, et al. Comparative evaluation of various total antioxidant capacity assays applied to phenolic compounds with the CUPRAC assay. Molecules 2007;12(7):1496-547.

Floegel A, Kim D, Chung S, Koo SI, Chu OK. Comparison of ABTS/DPPH assays to measure antioxidant capacity in popular antioxidant-rich US foods. J Food Comp Anal 2011;24:1043-8.

Sultana B, Anwar F, Przybylski R. Antioxidant activity of phenolic components present in barks of Azadirachta indica, Terminalia arjuna, Acacia nilotica, and Eugenia jambolana Lam. Trees. Food Chem 2007;104:1106-14.



How to Cite

Phatak, R. S., A. Krishnaji, and A. Subhash. “SCREENING OF SOME INDIAN HOUSEHOLD SPICES FOR COMPARATIVE STUDIES OF ANTIOXIDANT AND ANTIRADICAL ACTIVITIES BY USING IN-VITRO MODELS”. Asian Journal of Pharmaceutical and Clinical Research, vol. 8, no. 2, Mar. 2015, pp. 431-8,



Original Article(s)