Alok Kumar Hazra

Antiradical and antidiabetic properties of standardized extract of Sunderban mangrove Rhizophora mucronata.

Background: Mangroves have the ability to grow where no other vascular plants survive. Rhizophora mucronata is a true mangrove and traditionally used to treat diabetes and its allied complications. Objectives: In the present study, we standardized the 80% methanolic standardized extract of R. mucronata leaves (RH) and found out its antiradical and antidiabetic activities. Materials and Methods: The methanolic extract of R. mucronata leaves (RH) was standardized and quantified for phenolics, flavonoinds, gallic acid, quercetin, and coumarin. The reducing abilities and antiradical activities of RH were performed in vitro methods like, 1,1-diphenyl-2-picrylhydrazyl, nitric oxides, superoxides, hydroxyl, and ABTS (2,2’-azino-bis-3-ethyl benzthiazoline-6-sulphonic acid). Thereafter, RH was evaluated for it antidiabetic potentialities on streptozotocin (STZ)-induced type-2 diabetes. STZ (90 mg/kg, intraperitoneal) was administered to 2 days old pups to induce diabetes. RH was fed at doses of 50 and 100 mg/kg and glibenclamide (positive control) at 5 mg/kg, when the rats were 6 weeks old and continued for 10 weeks. Fasting glucose was monitored before and after the treatment. Further, lipid peroxides and reduced glutathione level were estimated on rat liver. Results: The results obtained from this study revealed RH possesses flavonoinds and also gallic acid, quercetin, and coumarin. Further, it has antiradical activities. It has also reduced blood glucose level in type-2 diabetic rats and reduced the formation of lipid peroxidation in liver. RH enhanced the level of glutathione in liver tissue. Conclusion: RH exhibits source of natural antioxidants and great potentialities as an antidiabetic agent by improving the hyperglycemia through its antiradical action.

Antioxidant and hepatoprotective properties of Indian Sunderban mangrove Bruguiera gymnorrhiza L. leave

Background: Bruguiera gymnorrhiza L. (family Rhizophoraceae) is a true mangrove habitat in Indian Sunderban and traditionally uses for liver disorders. Objectives: The aim was to evaluate antioxidant and hepatoprotective actions of leave extract of B. gymnorrhiza L. Materials and Methods: Hydro - methanolic extract of mangrove leaves (BR) was standardized using spectrophotometric and high-performance thin layer chromatography methods. Radical scavenging activities were assessed in different in vitro methods, like 1,1-diphenyl-2-picrylhydrazyl, 2,2′-azino-bis-3-ethyl benzthiazoline-6-sulphonic acid + , superoxides, nitric oxides and hydroxyl radicals. Hepatoprotective efficacy of BR (125 mg/kg and 250 mg/kg, p.o) was measured in D-galactosamine (GalN) induced (200 mg/kg, i.p) hepatitis in Wistar rats. Silymarin (25 mg/kg, p.o) was used as known hepatoprotective agent. Results: Polyphenols such as gallic acid, quercetin, and coumarin obtained from BR exhibited powerful antioxidant properties. Moreover, it produced dose-dependent protection against GalN induced hepatitis in rats. It significantly reduced GalN induced elevation of enzymes (alanine transaminase, aspartate aminotransferase, and alkaline phosphatase) in serum and resist oxidative stress marked by lipid peroxides, glutathione, and catalase in hepatic parenchyma. Conclusions: Polyphenols rich B. gymnorrhiza L. leaves ameliorate hepatic tissue injury through its antioxidant effects.

HPLC ANALYSIS OF PHENOLIC ACIDS AND ANTIOXIDANT ACTIVITY OF SOME CLASSICAL AYURVEDIC GUGGULU FORMULATIONS

Classical guggulu formulations have been used as mainstay therapeutics in chronic inflammatory diseases in Ayurvedic system of medicine since centuries. The present study included six guggulu formulations such as Kaishora guggulu, Kanchanara guggulu, Yogaraj guggulu, Punrnava guggulu, Amritadi guggulu and Trayodasanga guggulu.Phenolic compounds play a vital role as evident from phenolic and flavonoid content in guggulu drugs as well as antioxidant activity assayed by DPPH radical scavenging activity and FRAP antioxidant power. Yograj guggulu was found to have strong reducing ability (1156.76 μM Fe(II)/mg) among six drugs chosen in this investigation and Amritadi guggulu has revealed powerful scavenger of DPPH radicals (IC50:1.05 mg/ml). Phenolic acid composition for each guggulu drug was analyzed and twelve phenolic acid compounds were identified and estimated by HPLC. Gallic acid (8.29-49.28 mg/g) was found in high concentration and ellagic acid (0.69-6.02 mg/g) was found to the next in all formulations. Other phenolic acid components estimated were protocatechuic acid, p-hydroxybenzoic acid, gentisic acid, chlorogenic acid, vanillic acid, caffeic acid, syringic acid, p-coumaric acid, ferulic acid, sinapic acid in all guggulu formulations. A synergistic activity towards therapeutic efficacy was proposed on the basis of phenolic acid composition in each guggulu formulation.

Detection of arecoline by simple high-performance thin-layer chromatographic method in Indian nontobacco pan masala.

Chewing the habit of blended pan masala containing areca nut with or without tobacco is a common practice in the Indian subcontinent. Arecoline, a pyridine alkaloid presence in areca nut alarmed for oral carcinogenesis and strictly prohibited in the western world. However, in India using blended pan masala is very popular among young and old individuals. In this context, we aimed to detect arecoline in Indian blended nontobacco pan masala sold in Kolkata using a simple densitometric high-performance thin-layer chromatographic (HPTLC) method and for alarming their use in common people. Eleven popularly Indian blended nontobacco pan masala were collected from the territory of Kolkata and isolated arecoline, following solvent extraction method derived for pyridine alkaloid. The quantitative analysis of arecoline was measured using automated software-based HPTLC instruments and validated the method according to International Conference on Harmonization guidelines. Arecoline was detected in all 11 blended nontobacco pan masala samples in a range of minimum 130 to maximum 415 μg/g dry samples. Arecoline is hazardous carcinogenic compound, so the use of Indian blended nontobacco pan masala should be restricted. Further, the method was found suitable for routine quantitative analysis of arecoline in areca nut containing substances.

Acute and sub-chronic oral toxicity study of black tea in rodents

Objectives: Systematic oral toxicity study for black tea (Camellia sinensis), the most commonly consumed variety of tea, is lacking. The present study was undertaken to assess the iron load on black tea (Camellia sinensis) and its safety aspects in animals. Materials and Methods: The analysis of iron was done in six tea samples as per American Public Health Association method using flame atomic absorption spectrophotometer. Maximum physical iron-loaded tea sample was identified on black tea sample 2 (BTS-2), and this was further studied for acute and 90-day sub-chronic toxicity following Organisation for Economic Co-operation and Development guidelines. Results: Black tea sample 2 did not show any signs of toxicity or mortality at up to 2 g/kg per oral dose in Swiss albino mice. 90-day toxicity studies in Wistar rats did not reveal any evidence of toxicity at up to 250 mg/kg/day (2.5% infusion of BTS-2) oral dose as exhibited by regular observations, body weight, food consumption, hematology, serum chemistry, organ weights, and histopathology. Further, serum iron, total iron binding capacity, unsaturated iron binding capacity, and ferritin were not altered after 90 days of treatment. Masson trichrome staining and Perls’ staining did not reveal any abnormalities in hepatic tissue following 90-day treatment of high iron-loaded BTS-2. Conclusions: This safety study provides evidence that BTSs, in spite of relatively high iron content, show no significant iron-related toxicity on acute or sub-chronic oral administration in animals.

A rapid HPTLC method to estimate piperine in Ayurvedic formulations containing plant ingredients of Piperaceae family

Background Trikatu, Sitopaladi, Hingavastaka, Avipattikara, Sringyadi and Talisadya are very popular Ayurvedic (churna) medicines practiced in India; however, unfortunately, they possess several quality control issues. Objective The aim of this study was to find out a simple, accurate and sensitive HPTLC method for the detection and quantification of marker molecule, piperine (alkaloid) on these Ayurvedic formulations for standardization. Materials and methods Methanolic extraction (reflux) was performed from the above six churnas as well as three single ingredients Piper longum (pipul), Piper nigrum (marich) and Piper chaba (chai). HPTLC was done using piperine as a standard. The mobile phase was a mixture of toluene-ethyl acetate (7:3, v/v) and detection at 342λ. Results The Rf was detected at 0.39. Piperine was quantified in all samples. P. nigrum showed higher piperine than P. longum and P. chaba. The maximum piperine was noted in Hingavastaka churna and followed by Sringyadi churna, Sitopaladi churna, Talisadya churna, Trikatu churna and Avipattikara churna. Conclusion This method can be successfully employed for standardization and quantitative analysis of piperine in Ayurvedic formulations (churnas) and also be helpful to clinicians and pharmacists to draw significant role of piperine present in all these samples.

Risk of Betel Nut Chewing: Chemical and Toxicological Studies

Last few decades epidemiological reports warn that the development of oral submucous fibrosis − a precancerous state, is universal among betel nut or areca nut users. Betel nut contains arecoline and other microelements which are released into the buccal cavity during the process of areca nut chewing. The present effort was undertaken to find out the correlations between chemical ingredients, like arecoline, magnesium, iron and copper present in betel nut with toxicological incidences in experimental animals. Arecoline was estimated by HPLC (Perkin Elmer, USA) and other micronutrients like magnesium, iron and copper was assessed by AAS (Perkin Elmer, USA). Repeated dose 90-day sub-acute oral toxicity of betel nut water extract (BNE) at the dose of 0.5X10-2 ml/g/day and 1.0X10-2 ml/g/day was performed on Wistar rats as per the Organization for Economic Co-operation and Development (OECD) guidelines No. 408. The result indicates that betel nut contains Group 1 human carcinogen arecoline with significant high level of magnesium, iron and copper. Moreover, consumption of betel nut induces liver damage in rats by indicating significant enhancement of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP). It may conclude that chemical ingredients presences in betel nut are harmful for health and it should be restricted.

Antioxidant properties of polyphenolic rich HPLC standardized extract of Beta vulgaris L. roots

OBJECTIVES: The present study was planned to evaluate the antioxidant effects of standardized Beta vulgaris L. or beetroot. METHODS: The polyphenols present in lyophilized water extract of beetroot (BR) were quantified and standardized by HPLC. Deactivation of free radicals by BR was studied by in vitro biochemical assays including 1,1-diphenyl-2-picrylhydrazyl (DPPH), hydrogen peroxide and superoxide radical scavenging activities. Nitrite content of BR was estimated. Moreover, inhibition of protein denaturation properties of BR was carried out. RESULTS : HPLC chromatogram revealed presence of nine bioactive polyphenols in BR. Further, strong dose-dependent antioxidant properties were noted. IC50 (Inhibitory Concentration) of BR for DPPH, hydrogen peroxide and superoxide radical were 104.44μg/ml, 4.06μg/ml and 71.43μg/ml respectively. BR also showed enriched in nitrite content 3.359 mM/mg and it inhibited protein denaturation (IC50 594μg/ml). CONCLUSION: Beetroot was found to possess strong antioxidant property that may be due to presence of polyphenols.