Viscum capitellatum is a globose annual evergreen shrub with haustoria in the host. The diameter of globe was found to be 62-79-100 cm and height was 15-30 cm. Pair of scales is produced by reduction of leaves. Scales are transformed into oval shaped stems. Stems are dark green in colour and are woody. Fresh material has faint sweet taste and is odourless while dried material has a characteristic smell.
Transverse sections of leaf and transverse and longitudinal sections of stems were taken and lastly stained with various staining reagents as per standard procedures.
The cork is not well developed and epidermal cells are arranged in single row with thick walled semi rectangular beaded walls. Chloroplast was found in epidermis; epidermal cells are covered with yellowish cuticle having papilla like projection Numerous parasitic stomata were found on the both sides of leaves. Chloroplasts were found in guard cells of stomata. The average number of stomata per sq.mm is 49. The average stomatal index was found to be 15.9. Calcium oxalate crystals were observed in globose masses with cluster shape in outer layer of cortex. Collateral vascular bundles were observed.
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Total ash, acid insoluble ash and water soluble ash of V. capitellatum Smith. were obtained by reported methods. Ash value gives the about of presence of inorganic matter present in the plant material. Total ash, acid insoluble ash and water-soluble ash was found to be 7.00%, 1.50% and 2.90% respectively. Extractive values give an account of nature of chemical constituent present in crude drug. It may predict that higher amount of carbohydrates or proteins and phenolic compounds present in V. capitellatum from the Water and methanol soluble components, which were found in higher amount. Loss on drying gives the idea about amount of water present in crude drug. Higher water content may prone to have chemical or microbial decomposition of crude drug. Moisture content was found to be 9.97% in the dried plant material. Ash was further utilized for elemental analysis present in crude drug and showed the presence of Magnesium, Calcium, Chlorides, Sodium and Nitrate ions. Rarely starch grains are observed in cortical region. The starch grains are simple, ovoid to spherical in shape and 7.5um in diameter with centric hilum. Crude fiber content was determined according to the method reported in AOAC. 20.46% of fiber content was found in the plant material. Crude fibers are helpful in digestion and lowering cholesterol in the body.
The florescence analysis revealed that the plant Viscum capitellatum showed brown, faint brown, orange, light green, faint yellow and yellowish green florescence when exposed to day light and UV light.
Five prominent peaks were observed in HPLC profile while four major peaks were observed as flavonoids in HPTLC fingerprints. Two single compounds were isolated with the help of preparative chromatography from petroleum ether extract and methanol extract (ethyl acetate fraction) respectively. The compounds are given for spectral analysis (UV, IR, NMR and Mass) and identified as β- sitosterol and Quercetin according to their spectral characteristics respectively.
The carbohydrate estimation was done according to the established method. In hot acidic media glucose is dehydrated to hydroxy methyl furfural. This forms a green coloured product with phenol and has absorption maximum at 490 nm. The total carbohydrate content of Viscum capitellatum was found to be 0.87 %w/w. Total phenolic content of methanolic extract was estimated using phenol reagent. In this method, the blue colour formed due to the polyphenol present in the extract was measured at 760 nm using UV spectrophotometer. The phenolic content was estimated (47.837 µg) from standard calibration curve of gallic acid. Total flavonoid content of methanol extract of was determined using reported method. The absorbance of reaction mixture was measured at 415 nm. Total flavonoid content was estimated (29.24 µg) using standard calibration curve of quercetin. Total proanthocyanidin content of methanol extract was determined using reported method. Red coloured solutions so obtained were read at 500 nm. Total proanthocyanidin content was estimated (15.59 µg) using standard calibration curve of epicatechin.
Acute toxicity study was followed by using OECD guidelines (Organization of Economic Cooperation and Development) 423 (Acute Toxic Method). The method uses defined doses (5, 50, 300, 2000 mg/kg body weight). The methanol extract of Viscum capitellatum Smith. did not produce lethality and toxic symptoms up to the dose level of 2000 mg/kg.
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The crude extract of the plant showed significant analgesic action compared to the reference drug Diclofenac sodium against acetic acid induced pain in mice. Significant decrease in rat paw inflammation (42.93%) was noted as compared to standard Diclofenac sodium (46.91%). Dose dependent pattern for inhibition of granoloma was observed in sub acute inflammation model. Methanol extract showed inhibition (38.55%) of granuloma formation compared to standard Diclofenac sodium (44.74%). Methanol extract of the plant observed to have a significant immunosuppressive activity (49.68%) in Complete Freund's adjuvant induced arthritis in comparison with standard Diclofenac sodium (58.58%). The methanol extract of plant showed significant decrease in Fasting blood glucose level, which was observed in dose dependent manner. The extract also showed significant changes on lipid profile of diabetic rats. The methanol extract (LC50 27.03) also showed to have more potent in vitro cytotoxic activity than that of reference standard caffeine (LC50 287.71).
The extract found to possess equipotent free radical scavenging activity, nitrous oxide scavenging activity, reducing power and lipid peroxidation activity compared to reference standards, ascorbic acid and BHA.
The antibacterial activity was performed using five gram positive strains and five gram negative strains. Methanol extract showed maximum antibacterial activity against Bacillus subtilis and Proteus vulgaris. The antifungal activity was performed using five strains. The antifungal activity was performed using five fungal strains. Petroleum ether extract showed maximum antifungal activity against Trichoderma viride.
The main problem in validation of ethnomedicinal claims of mistletoes is conversion of similar folk remedic claims to several different mistletoes. Thus it becomes required to typify the individual mistletoes in terms of its morphological and / or phytochemical to authenticate the folk remedy claims and to assign observed pharmacological activities.
The characters such as replacement of epidermis by cuticular epithelium and transverse orientation of stomata to the plant axis are in accordance with other members of Viscaceae. The occurrence of chlorophyll in epidermal cells is a discrete character of mistletoe. The marphoanatomy together with chromatographic data therefore can constitute the basis for identification of these medicinally important species of Viscum.
V. capitellatum contains high amount of phenolics which can be better connected with their antioxidant activity. The extract found to possess equivalent anti-oxidant activity compared to reference standards, ascorbic acid and BHA.
The extract also confirmed significant activity in both acute as well as sub-acute model of inflammation. V. capitellatum is, conversely, effective in late phase of inflammation and gives dose-dependent inhibition.
Due to the presence of flavonoids the methanol extract showed an immunosuppressive activity in Freund's adjuvamt induced arthritis model. Many authors reported the effect of flavonoids e.g. quercetin on arachidonic acid metabolism which indicates that the antioxidant potential is important for inhibition of lipoxygenase.
It also reported that many of phenolic and flavonoid compounds having anti- diabetic activity due to their antioxidant potential. Methanol extract showed significant anti- diabetic activity in Alloxan induced diabetes model.