The objective of the present study was to evaluate the safety of long term consumption of ethanolic fraction of Neurocalyx calycinus leaves (NCEF) in rodents. The NCEF was subjected to detect the presence of various phytoconstituents. In acute oral toxicity study, graded doses of NCEF was administered in mice and were observed up to 14 days. In sub-chronic oral toxicity study, NCEF was administered to Wistar rats at doses of 50, 500 and 1000 mg/kg b.w. per day for 90 days and after that, observed up to 28 days. NCEF showed the presence of alkaloids, steroids, phenolics and glycosides. In acute toxicity study, there was no mortality and no behavioural signs of toxicity at the highest dose level (6400 mg/kg b.w.). In sub-chronic oral toxicity study, there were no significant difference observed in the consumption of food and water, body weight and relative organ weights. Haematological, serum biochemical, hepatic oxidative stress marker analysis and urine analysis revealed the non-adverse effects of prolonged oral consumption of NCEF. The histopathologic examination did not show any differences in vital organs. Based on our findings, NCEF, at dosage levels up to 1000 mg/kg b.w., is non-toxic and safe for long term oral consumption.
Tetracera akara (Burm. f.) Merr. (Dilleniaceae) is a woody climber, locally known as 'Pattuvalli', distributed in the Western Ghats region of Kerala and Tamil Nadu. The roots of T. akara is reported to be used by the Kani tribe of Kerala to cure various liver disorders [1] . The results from previous studies of comparative in vitro antioxidant studies revealed that T. akara ethanol fraction is rich in bioactive phytoconstituents like phenols and flavonoids with good antioxidant potential [2] and is safe for oral administration in experimental animals [3] . Hepatoprotective effect of ethanol fraction of roots of T. akara against paracetamol-induced acute liver damage has been reported earlier, which revealed that the ethanol fraction of T. akara showed antihepatotoxic effect against drug over dose in Wistar rats [4] . However, the protective effect of T. akara against chemicalinduced liver injury has not been evaluated yet which is necessary to scientifically validate the hepatoprotective effect of T. akara against various models of hepatic injury.The use of many halogenated alkanes such as carbon tetrachloride (CCl 4 ), chloroform (CHCl 3 ) or iodoform (CHI 3 ) was banned or severely restricted because of their distinct toxicity. Yet CCl 4 continues to be used
Neurocalyx calycinus (R. Br. ex Benn.) Rob. is an endemic species of the family Rubiaceae, found in Western Ghats of India and Sri Lanka. The leaf paste of N. calycinus has been used by Cholanaickan tribe as medicine to heal wounds, burns, inflammation and pain. Many of the human diseases are manifested due to the accumulation of free radicals. Plant derived antioxidants have the power to scavenge these free radicals without any internal complications. Aqueous, n-hexane, chloroform, ethyl acetate and ethanol extracts of the leaves of N. calycinus were selected for qualitative phytochemical screening. DPPH, nitric oxide, superoxide and hydroxyl radical scavenging assays were carried out. Total phenolic, flavonoid and antioxidant content of all the extract was measured. Phytochemical screening showed that alkaloids, steroids, phenolic compounds, glycosides and saponins were found in the ethanolic extract. Ethanol extract showed better antioxidant activity than the other solvent extracts. The reducing power of all the extracts was increased dose dependently. Highest amount of phenolic (56.27 mg of gallic acid equivalent /gm of dried extract), flavonoid (36.92 mg of quercetin equivalent/gm of dried extract) and antioxidant (85.68 mg of ascorbic acid equivalent/gm of dried extract) content was found in the ethanolic extract. A positive correlation (R2) was found between the total phenolic content and free radical scavenging activity of the leaf extract of N. calycinus. The results confirmed that the phenolic compounds in the ethanolic extract of the leaves of N. calycinus are potent source of natural antioxidants.
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