Evaluation of anti-oxidant status in-vitro and in-vivo in hydro-alcoholic extract of Eugenia caryophyllus


  • Roma Ghai KIET Kiet School of Pharmacy Ghaziabad,Uttar Pradesh, India
  • Kandasamy Nagarajan KIET School of Pharmacy
  • Jitendra Singh Jamia hamdard
  • Shiwam Swarup KIET school of Pharmacy
  • Minu Keshari Department fo life schience,shridhar university




DPPH Assay, Eugenia caryophyllus, Hyperlipidemia, Lipid Peroxidation, Triton Induced.


Free radicals mediated oxidative stress is the major risk factor for many chronic diseases like atherosclerosis, diabetes mellitus, arthritis, cancer, ageing and neurodegenerative diseases. Therapy with anti-oxidants is gradually gaining lot of importance for treatment of such diseases. Hydro-alcoholic extract of Eugenia caryophyllus was studied for its in-vivo antioxidant activity using two different animal models viz. Triton induced hyperlipidemia and High fat diet induced hyperlipidemia. Total phenolic content and total flavonoid content, DPPH assay was also carried out for in vitro anti-oxidant efficacy. Total protein, lipid peroxidation (MDA), reduced glutathione, superoxide dismutase and catalase were evaluated in the liver tissue in Triton induced hyperlipidemia and diet induced hyperlipidemia models. The study findings indicated significant in-vivo and in-vitro antioxidant property that may be related to the amount of polyphenols and flavonoids present in the extract. These results clearly indicate that Eugenia caryophyllus is effective against free radical mediated oxidative stress.


[1] Abou Ghalia AH & Fouad IM (2000) Glutathione and its metabolizing enzymes in patients with different benign and malignant diseases. Clinical Biochemistry 33, 657-662 http://dx.doi.org/10.1016/S0009-9120(00)00181-8.

[2] Cai, L. & Wu, C. D. (1996) Compounds from Syzygium aromaticum possessing growth inhibitory activity against oral pathogens. Journal of Natural Products 59, 987-90. http://dx.doi.org/10.1021/np960451q.

[3] Chaieb K, Hajlaoui H, Zmantar T, Kahla-Nakbi AB, Rouabhia M, Mahdouani K & Bakhrouf A (2007) The Chemical Composition and Biological Activity of Clove Essential Oil, Eugenia caryophyllata (Syzigium aromaticum L Myrtaceae) :A Short Review. Phytotherapy Research 21, 501-06 http://dx.doi.org/10.1002/ptr.2124.

[4] Claiborne A. (1985) Catalase activity. In Handbook of Methods for Oxygen Free Radical Research (Greenwald RA, ed) Boca Raton, FL: CRC Press, pp 283 –284.

[5] Ellman, G. L. (1959) Tissue sulfhydryl groups. Archive of Biochemistry and Biophysics 82, 70-77. http://dx.doi.org/10.1016/0003-9861(59)90090-6.

[6] Kalonia, H. & Kumar, A. (2006) Protective Effect of Br-16a (Poly Herbal Preparation) on the behavioral and biochemical alterations in sleep disturbed mice (grid suspended over water method). Annals of Neurosciences 13, 94-98. http://dx.doi.org/10.5214/ans.0972.7531.2006.130401.

[7] Kataki MS, Ahmed MZ, Awasthi D, Tomar B., Mehra P, Yadav RS & Rajak P (2012) In vitro antioxidant profile of Wedelia calandulaceae leaves. Pharmacologia 3, 75-83 http://dx.doi.org/10.5567/pharmacologia.2012.75.83.

[8] Kumar G, Srivastava, a, Sharma, SK & Gupta Y K (2013) the hypolipidemic activity of Ayurvedic medicine, Arogyavardhini vati in Triton WR-1339-induced hyperlipidemic rats: A comparison with fenofibrate. Journal of Ayurveda and Integrative Medicine, 4, 165-70 http://dx.doi.org/10.4103/0975-9476.118707.

[9] Lowry OH, Rosebrough N J, Farr AL, & Randall RJ (1951) Protein Measurement with the Folin Phenol Reagent. The Journal of Biological Chemistry, 193, 265-275.

[10] Marklund S & Marklund G (1974) Involvement of the superoxide anion radical in the autoxidation of pyrogallol and a convenient assay for superoxide dismutase. European Journal of Biochemistry, 47, 469-474. http://dx.doi.org/10.1111/j.1432-1033.1974.tb03714.x.

[11] Mittal M, Siddiqui MR, Tran K, Reddy SP & Malik AB (2014) Reactive Oxygen Species in Inflammation and Tissue Injury. Antioxidants and Redox Signaling 20, 1126-1167 http://dx.doi.org/10.1089/ars.2012.5149.

[12] Nourooz-Zadeh J, Tajaddini-Sarmadi J, McCarthy S, Betteridge DJ & Wolf SP (1995). Elevated levels of authentic plasma hydroperoxides in NIDDM. Diabetes, 44, 1054-1058. http://dx.doi.org/10.2337/diab.44.9.1054.

[13] Ohkawa H, Ohishi N &Yagi K (1979). Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Analytical Biochemistry, 95, 351-58. http://dx.doi.org/10.1016/0003-2697(79)90738-3.

[14] Olayinka AA & Anthony IO (2009) Phyochemical screening and polyphenolic antioxidant activity of aqueous crude leaf extract of Helichrysum pedulanculatum. International Journal of Molecular Science, 10, 46-58.

[15] Poljsak B (2011). Strategies for reducing or preventing the generation of oxidative Stress. Oxidative Medicine and Cellular Longevity, Article ID 194586, http://dx.doi.org/10.1155/2011/194586.

[16] Poljsak B, Å uput D. & Milisav I (2013). Achieving the Balance between ROS and Antioxidants: When to Use the Synthetic Antioxidants. Oxidative Medicine and Cellular Longevity, Article ID 956792 http://dx.doi.org/10.1155/2013/956792.

[17] Pourgholami MH, Kamalinejad M, Javadi M, Majzoob S. & Sayyah M (1999) Evaluation of the anticonvulsant activity of the essential oil of Eugenia caryophyllata in male mice. Journal of Ethnopharmacology, 64, 167-71. http://dx.doi.org/10.1016/S0378-8741(98)00121-4.

[18] Pourmorad F, Hosseinimetr SJ & Shahadimajd N (2006) Antioxidant activity, phenol and flavanoid contents of some selected Iranian medicinal plants. African Journal of Biotechnology, 6, 1142-1145.

[19] Singh AK, Dhamanigi SS. & Asad M (2009) Anti-stress activity of hydroalcoholic extract of Eugenia caryophyllus buds (Clove). Indian Journal of Pharmacology 41, 28-31. http://dx.doi.org/10.4103/0253-7613.48889.

[20] Srivastava KC (1993) Antiplatelet principles from a food spice clove (Syzygium aromaticum L.) Prostaglandins Leukotrienes and Essential Fatty Acids 48, 363-372. http://dx.doi.org/10.1016/0952-3278(93)90116-E.

[21] Thampi BS, Manoj G, Leelamma S & Menon VG (1991) Dietary fibre and lipid peroxidation: effects of dietary fibre on levels of lipids and lipid peroxides in high fat diet. Indian Journal of Experimental Biology 29, 563-67.

[22] Zorov DB, Juhaszova M, & Sollott SJ (2014) Mitochondrial reactive Oxygen Species (ROS) and ROS-Induced ROS Release. Physiology Reviews 94, 909-950 http://dx.doi.org/10.1152/physrev.00026.2013.