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The current study evaluates morphologicl and histological characters, Qualitative chemical analysis for plant constituents and TLC fingerprints, fluorescence analysis of powder and Physicochemical characters of different extractives of Butea monosperma flower, Elaeocarpus oblongus fruits and Canarium strictum bark and Plumbago indica root.
4.4.1. Butea monosperma
The detailed histological study of Butea monosperma was carried out in the present study, which helps to identify this species without any confusion. The most convenient and cost effective method of identification of a medicinal herb would be the use of microscopic characteristics; it has been the foundation of conventional Pharmacognosy and remains a fundamental module of the contemporary monograph (Samuelsson, 1999). Flower is a unique character of plant identification, the petal arrangement in this plant plays a major role to identify this plant. Even though Ayurvedic pharmacopoeia has included monograph of this flower, but the elaborate study of the flower microscopic is crucial as flower has been used in traditional medicine. Powder microscopy also plays an important role in pharmacognostical evaluation and sometimes may be an identifying parameter of herbal drugs.
According to quantitative microscopy, the middle part of the standard petal was 500 µm thick; the marginal part of the strand petal was 80 µm thick; the wing petal was 500 µm thick in the middle and 150 µm thick along the margin. The middle part of the keel petal was curved and it was 400 µm thick. The petal has long epidermal trichomes of 400 µm to 1mm length and 10 µm thickness. The pollen grains were triporate with 40 µm diameter in the equatorial plane. The ovary is monocarpellary and the ovules were anatropous and bitegmic. Stamens was diadelphous; pollen grains are spheroidal; about 28µm long and 30µ m broad, pore oval to elongate, 8 to 12.5 µm exine wall surface foveolate (Ayurvedic pharmacopoeia, 1999). These are evaluated in first time in our studies which would stand as unique quality control parameters towards identification of flowers of this plant.
Interestingly, during the present study mineral elements in the dried flower powder were found produce lot of therapeutic effects in the human body. Minerals also responsible of imparting colour to the natural products. Chromium imparts orange colour, whereas Copper given red color and Manganese made orange yellow color (http://www.minsocam.org/msa/collectors_corner/arc/color.htm).
Various literature showed the minerals are producing lots of therapeutic effects in the humans. Therefore the present study has keen to found that the presence of mineral elements in the dried flower powder. Copper deficiency is exemplified by chronic or repeated diarrhoea and low resistant to infection and anaemia. Manganese needed for heamoglobulin development, reproduction and skeleton growth in humans and to cure skin illness, scabies and piles and rheumatism. Zinc is composed of many metalloenzymes and also membrane stabilizer and stimulator of the immune response. Its deficiency leads to loss of appetite and weakens immune function and hair loss, delayed sexual maturation, impotence and hypogonadism in males and eye and skin lesions. Chromium assists carbohydrate metabolism and supports transmission in the neuromuscular system its deficiency causes diabetes in humans (Ariyanathan et al 2010).
4.4.2. Elaeocarpus oblongus
In the view of developing customary monograph for this species, the scanning electron microscope [SEM] studies of leaves of Elaeocarpus oblongus had been performed by Singh HP, Dube VP, 1991. The results shows variation in the cuticular ornamentation compared to stomata and trichomes of E.oblongus with other species. These distinguished characters found to be useful in identifying the members of this species. The cuticular features were aiding in deciding the taxonomic positions of the number of Elaeocarpaceae family (Singh, 1991).
A chemical character of the Elaeocarpus species almost habitually contains either myricetin or gallic acid. Four out of seven species have been found to contain mearnesetin, rare methyl ether of myricetin. The leaf of the Elaeocarpus oblongus contains mearnsetin (flavonoid) 00.015 % and myricetin, 00.19% flavonoid (Lalchand, 1977) The quantitative analysis report showed (Table 3) the presence of primary metabolites such as Carbohydrate in the form of fructose in the fruit. Phenolic compounds, tannin, flavonoids, saponins and sterols are the secondary metabolites in the fruit.
Stem bark of Elaeocarpus oblongus extracts was evaluated biologically for various activities such as antibacterial, antiviral, antifungal and effect on respiration and effect on cardiovascular and effect on isolated ileum and effect on isolated rat uterus and effect on CNS and cross behavior and effect of hypothermia analgesia and supra maximal electroshock seizure pattern test and diuretic and anti inflammatory. It is reported that there was no activity observed in extracts. The ID50 of the stem bark is 1000 mg/kg (Abraham, 1986). While stem bark was not showing any potent biological activity the current study select the much used edible fruits of this species for setting the quality control parameters. The fruits of E. oblongus exhibited potent antioxidant activity towards 1, 1-diphenyl-2- picrylhydrazyl free radical (DPPH·) (EC50 = 0.200 ± 0.76 and 0.250 ± 1.76 mg mL-1), superoxide anion (O2Î‡¯ ) (EC50 = 0.50 ± 0.82 and 0. 50 ± 1.20 mg mL-1), hydroxyl ion (Ë™OH) (EC50 = 0.250 ± 0.22 and 0.500 ± 0.48 mg mL-1) and nitric oxide (NO) (EC50 = 0.250 ± 0.22 and0. 250 ± 0.552 mg mL-1 (Ramakrishnan et al., 2011).
This fruit is susceptible to fungal growth since the fruit contains much moisture content estimated about 12% w/w. In our antifungal studies we have got negative result, 10mg of the alcoholic extract of fruit in DMSO was failed to prevent the growth of Candida albicans in potato dextrose agar medium. However Mycorrhizal type and Dark septate endophyte (DSE) fungal association was not found in the root of Elaeocarpus oblongus.
4.4.3. Canarium strictum
During our Nilgiri Biosphere Reserve (NBR) visit we found that the crimson colour resin was collected from the wounded trunk of the tree was given orally in the form of decoction, twice a day used to treat body pain and arthritis. The bark along with resin was grounded with water and the resultant paste was applied locally for the treatment of psoriasis and skin rashes by baduga tribal community and people of Mizoram as a folkloric medicine (Prabhat kumar Rai, 2010). In eastern Arunachal Pradesh, Khamti tribal were applied melted resin over the skin to reduce the inflammation induced by contact poison of brown hairs of caterpillar (Hui tag, 2007).The resin is important worship item for tribal groups of nilgiris district, particularly baduga they hold 3 to 4 kg in home and they collect resin only from female tree rather than male (Anita Varghese, 2008).
Concerning, anatomical characters of the Canarium L. tree are grown up to 50 m tall. The barks are grayish white6,(Gan lan shu) Outer bark are slim whereas inner bark reddish brown, soft and aromatic with sticky oily exudates of resin. The stems are normally terete. The leaves are pinnate, curved and stipulated. The rachis is terete compressed to channeled puffy base, and bears 5-21 folioles. The folioles are oblique at bottom, whole, dentate or serrate at border, often thick and acuminate at apex. The secondary nerves are arching and joined near margin. The tertiary nerves are reticulate. The inflorescence is an axillary or terminal panicle. The corolla consists of 3 cream colour petals. The androecium contains a whorl of 6 stamens. The disc inside the stamens is 6 lobed. The gynaecium comprised 3 carpels joined into a 3 lobular ovary. The drupes are seated on a persistent inflated calyx and surrounded woody stone (Mogana et al., 2011).
Literature shows that the genus Canarium L. contains terpenes which include monoterpenes, triterpenes and tetraterpenes like carotenoids, sesquiterpenes, cyclohexane and sterols and carboxylic acids, coumarins, furans, lipids and phenolic compounds such as flavonoids, tannins, phenolic acids (Mogana et al., 2011). The cheapest way to identify the plant and to develop monograph is to studies of anatomical characters including macro and microscopical characters are fundamental of pharmacognostical evaluation of crude drugs (Samuelsson, 1999). Optimistically this study provided information helpful to develop official monograph of Canarium strictum Roxb.
4.4.4. Plumbago indica
The TS of the root is nearly cuticular in outline with 5-6 layered cork tissue, consisting of cubical to rectangular cells with their wall light yellow to yellowish brown content forms the major part of the root. The greater part of the cortex consists of thin walled rounded, polygonal tangentially elongate cells with well defined intracellular space. The cells are normally devoid of the starch grains. The innermost rows of cortical cells are nearly regular and rounded.There is no clear demarcation between cortex and bast. The phloem parenchyma cells are very small, thin walled and polygonal. Most of the cells have yellow contents. No mechanical elements are in the bast. The cambium consists of one or two layers. The xylem vessels are arranged mostly in the single file in radial rows and are surrounded by mechanical cells. The medullary rays are multiseriate. The root is triarch and protostelic (Krishnan Nambiar, 2003).
Earlier studies on root shows the presence of elements were Fe (6.47), Mg (0.92), Zn (0.34), Cu (0.22), Co (0.06), Ni (0.01), Na (82), K (113), Ca (18), As (0.0001), Cd (0.007), Pd (0.3116), Hg (0.0053) and the absence of Cr (Ariyanathan et al., 2010). The current study found that the presence of 18.5µg/g chromium in dried root powder. Plumbago rosea root has a major amount of iron therefore it seems to be dark in color similar to dates fruits. Iron is diuretic, demulcent and used to treat diarrhea and croup. Copper deficiency leads to chronic or repeated diarrhea, low resistance to infection and anemia. Manganese needed for hemoglobin development, reproduction and skeleton growth in humans. The highest content of Manganese was reported in all three species of Plumbago zeylanica, Plumbago rosea and Plumbago capsensis has been given to cure skin diseases, scabies and piles and rheumatism. Zinc is constituent of many metalloenzymes and also membrane stabilizer and stimulator of the immune response. Its deficiency develops loss of appetite and impaired immune function. In case of vigorous zinc deficiency causes hair loss, delayed sexual maturation, impotence hypogonadism in males, the eye and skin lesions. The zinc was higher content in Plumbago zeylanica, whereas the amount is equal in Plumbago capsensis, Plumbago rosea which can be cure skin diseases and rheumatism. Chromium plays major role in carbohydrate metabolism and its deficiency leads to Diabetes in humans. Chromium supports transmission in the neuromuscular system. Sodium is influenced in intracellular and extracellular fluid balance and safeguarding of viscosity of blood. Potassium is diuretic, both sodium and potassium facilitated the ionic balance of the human body and sustains tissue excitability. Calcium is essential in the nerve impulse conduction and the mechanism of the neuromuscular system.
The Plumbagin content is high in Plumbago rosea 0.17% compared to other species of Plumbago zeylanica, 0.01% and Plumbago capsensis 0.04%. Crude fiber content largely depends on cellulose and lignin plus some mineral matter. Pesticide residue and Aflatoxin B1B2G1G2 were not detectable range in root powder (Ariyanathan et al., 2010). Acid insoluble ash specified the occurrence of more siliceous matter in the drug. It was found to be 1.55%. The alcohol soluble extractive values reveal the contents of polar phytochemicals such as flavanoids, anthraquinone, glycosides, alkaloids, steroids, triterpenoids present in the plant materials. It was estimated about 5.30% in root. The water soluble extractive values reveal the presence of water soluble materials such as vitamins, amino acids, sugars, carbolic acids, and it was calculated to be 11.21%. As this root contains a rich quantity of iron, manganese and other essential minerals and flavanoids it would be considered as a general tonic in the future.
The current study developed the monograph which includes Microscopical characters, Qualitative chemical analysis for plant constituents and TLC fingerprints of the extracts, fluorescence analysis of powder and Physicochemical characters of Butea monosperma flower, Elaeocarpus oblongus fruits and Canarium strictum bark and Plumbago indica root. These data will be act as a standard data which facilitate to identifying differencing the plant species of the same categories. This is the first report on the physicochemical characters and microscopical characters of Elaeocarpus oblongus fruits and Canarium strictum bark.