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THE ANTI-OXIDANT ACTIVITY OF TURMERIC (CURCUMA LONGA) 1
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2 ABSTRACT TTurmeric antioxidant protein had been isolated from the aq. extract of turmeric. TThe antioxidant principle was found to be a heat stable protein. TThe antioxidant protein showed a concentration – dependent inhibitory activity on the promoter induced lipid peroxidation. TThe protection of Ca 2+ - ATPase activity was found to be associated with the prevention of loss of –SH groups.
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3 KEYWORDS TTURMERIC ANTI-OXIDANT PROTEIN LLIPID PEROXIDATION TTHIOLS
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4 INTRODUCTION SSeveral processes are involved in the adaptation of organisms to environment. MMainly used for defence against free radicals. PPeroxidation of unsaturated lipids has been used in wide range of diseases. TTurmeric (curcuma longa ),one of the major species used as potent anti-oxidant. TThe aqueous and alcoholic extract of turmeric have been shown to be anti-oxidant.
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5 ANTIOXIDANTS LLevamisole BButylated hydroxy toulene SSantoquin BButylated hydroxy anisole
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6 MATERIALS AND METHODS FFresh cod liver oil supplied as seven seas by Universal Generics Pvt. Ltd. and male Wistar rat brain 10% homogenate in Tris HCl buffer (0.01M) were used as substrates. PPreparation of turmeric extract- Powdered drug(1.5gm.) dissolved in 75 ml of boiling distilled water, centrifuged and supernatnt was selected. IIsolation of turmeric antioxidant protein (TAP)- aqueous extract was concentrated,insoluble material was centrifuged. One fifth extract containing 15 mg proteins was loaded on a SEPHADEX-G-200 column. TThe protein was simultaneously assayed. FFurther assay of liquid peroxidation using brain tissue and cod liver oil. AAssay Ca 2+ ATP ase activity. DDetermination of total sulphahydryl content.
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7 SEPRATION OF TAP ON SEPHADEX-G-200 COLUMN ONE ANTIOXIDENT UNIT = 50% INHIBITION OF LIPID PEROXIDATION IN PRESENCE OF TAP S:NO. PARTIC -ULARS VOL. (ml) ACTIVIT Y (antioxid ant activity in units/ml) TOTA L UNIT S PROTIE N (mg/ml) SPECIFI C ACTIVIT Y % YIEL D 1.Aqueou s extract 7512.129091.48.66100.0 0 2.Concen tr-ate 1078.807888.759.086.68 3.Sephad ex-G- 200 column effluent 66.639.80.4216.4721.99
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8 RESULTS TThe antioxidant principal was found to be heat stable and no loss of activity during extraction. IInhibition of lipid peroxidation by TAP- TAP had concentration dependent inhibitory effect. 50% inhibitory activity at protein conc. of 50µg/ml, 50% inhibitory effect on ascorbate Fe 2+ /TBH- induced systems in rat brain at conc. 100 µg/ml. IInfluence of TAP Ca2+ - ATPase activity in rat brain homogenate- The activity was found to decreased by 40% with increase in TBARS release. EEffect of TAP on sulphahydryl content depletion- In presence of TAP 38% of depletion was seen after 60 min. as compared to 45% in absence of TAP.
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9 EFFECT OF TAP ON NON- ENZYMATIC LIPID PEROXIDATION OF COD LIVER OIL values are mean±S.D. of 6 determanation expressed as nmoles TBARS released/50 mg oil in 30 min. S.N O. PARTICULARSWITHOUT TAP WITH TAP % INHIBITI ON 1.Oil10.63±1.7 6 6.38±0.9240.00 2.Oil+1 mM Fe 2+ 21.25±2.4 5 15.75±1.3 7 25.88 3.Oil+1 mM Ascorbate- Fe 2+ 15.00±2.0 4 10.88±1.0 2 27.50 4.Oil+1 mM TBH34.95±3.4 2 19.47±2.1 3 44.29
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10 DISCUSSION AND CONCLUSION TThe antioxidant principal is protein in nature by its maximal absorbance at 280nm. and loss of its antioxidant property on trypsin treatment. TTAP prevents Ca 2+ - ATPase from inactivation in the presence of promoters of lipid peroxidtion or thiol reagents. TTAP prevents cellular –SH depletion during peroxidation. CConsumption of high concentration of turmeric reported to be non-toxic as compared to the other dietary antioxidants like levamisole, butylated hydroxy anisole and santoquin which are known to enhance cellular and humoral immunity with age. PPresence of a heat stable antiioxidant protein in the aqueous extract of turmeric is highly significant in relevance to its dietary consumption.
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