PHARMACOGNOSTICAL AND PHARMACOLOGICAL STUDY OF GUDUCHI (TINOSPORA CORDIFOLIA)

Pharmacognosy is the study of medicinal drugs derived from plants or other natural sources

The American Society of Pharmacognosy defines pharmacognosy as "the study of the physical, chemical, biochemical and biological properties of drugs, drug substances or potential drugs or drug substances of natural origin as well as the search for new drugs from natural sources.It is also defined as the study of crude drugs.The word 'pharmacognosy' is derived from the Greek 'pharmacon', a drug' and gignosco, 'to acquire knowledge of'.
It is closely related to both botany and plant chemistry and, both originated from the earlier scientific studies on medicinal plants.With the beginning of the 20th century, the subject had developed mainly on the botanical side, being particularly concerned with the description and identification of drugs, both in the whole state and in powder and with their history, commerce, collection, preparation and storage.Such branches of pharmacognosy are still of fundamental importance, the rapid development in particularly phytochemistry and pharmacology, have enormously expanded the subject.As a result it is now possible to approach the study of medicinal plants from the botanical, phytochemical and pharmacological view points (Trease and Evams, 14th ed., 1997).Undoubtedly, the plant kingdom still holds many species of plants containing substances of medicinal value which have yet to be discovered; large numbers of plants are constantly being screened for their possible pharmacological value.
The plant selected for the present study Tinospora cordifolia, of the family Menispermaceae.
Here an attempt is made to study the plant pharmacognostically in this chapter.The whole plant has taken for study.Guduchi consists of dried, matured pieces of stem of Tinospora cordifolia (Willd.)Miers.(Family-Menispermaceae), a perennial climber found throughout

Species
The 40 species are distributed in tropical Africa, South -East Asia, Indo Malaya region and Australia of which 3 species have been recorded from India.According to Indian medicinal plant, the following species are used medicinally.
Substitute of Guduchi has not been mentioned in any classics.However, in practice fresh stem is more efficacious than dry and is a good substitute for Columba.

Adulteration
The commonest species of Tinospora with which Tinospora cordifolia is likely to be substitute or adulterated are Tinospora sinensis (lour) Merril.(syn.Tinospora malabarica(Mier) and Tinospora crispa (Linn.)are met with and often confused with each other.

Part used
The entire plant stem leaves and roots are used in medicine, preferably in the fresh state.

Collection
The root and stem are collected in hot season when the bitter principle in most abundant and concentrated (Indian pharmacopoeia).It is not official in the British pharmacopoeia or the United state pharmacopoeia.For Sattva preparation fresh stem should be collected at the flowering time when there are no leaves in the stem to get both quantitative and qualitative starch.

Side Effects
No significant information on side effects is available so far.The drug is traditionally considered to be safe in the dosage mentioned.

DESCRIPTION a) Macroscopic
Drug occurs in pieces of varying thickness ranging from 0.6-5 cm in diameter, young stems green with smooth surfaces and swelling at nodes, older ones show a light brown surface marked with warty protuberances due to circular lenticels, transversely smoothened surface shows a radial structure with conspicuous medullary rays traversing porous tissues, taste bitter.

Botanical description
It is a large, glabrous, deciduous climbing shrub.

Stem:
The stem of Tinospora cordifolia is rather succulent with long filiform.Fleshy aerial roots from the branches with a thick, soft, warted bark4 Bark: The bark is creamy white to grey, deeply left spirally, the space in between being spotted with large rosette like lenticels.

Leaf:
The leaves are membranous and cordate at the base.Leaves alternate, on long flexnose petioles, spreading 2-4 inches long, roundish oval, entire, acute at the apex, quite smooth and thin.The leaves have bitter taste and an indistinct odour, when the leaves seen in bulk, they look intensely green.Mature leaves show yellowish to green colour.

Flower:
The flowers are small and yellow or greenish in colour.In. auxiliary and terminal racemes or racemose panicles, the male flowers are clustered and female are usually solitary.

Ranjan et al. World Journal of Pharmacy and Pharmaceutical Sciences
Male flower: Sepals 6 in two rows of 3 each, the outer short & roundish, the inner twice as long, broadly abovate, concave, smooth, petals 6, opposite the sepals and about half length of the inner ones, 3 lobed above with the lateral lobes.
Female flower: Sepals as in the male; petals oblong spasthuatate; stamens reduced to small oblong sepals in front of the petals; carples 3, distinct, opposite the outer sepals.
Fruits: 3 or less usually less by abortion shortly, stalked, subglobose drupes.The drupes are ovoid, glossy, succulent, red and pea sized.Flowers grow during the summer and Fruits during the winter and fruits are fleshy.

Seed:
The seeds are curved and single.

Aerial root
The aerial root is characterized by tetrarch primary structure; cortex is divided in to outer thick walled zone representing the velamen and inner parenchymatous zone containing secretary canals and starch grains are present throughout the parenchyma of the aerial root.

Stem
Transverse section of stem shows outer-most layer of cork, differentiating into outer zone of thick-walled brownish and compressed cells, inner zone of thin walled colourless, tangentially arranged 3-4 rows of cells, cork broken at some places due to opening of lenticels, followed by 5 or more rows of secondary cortex of which the cells of outer rows

Ranjan et al. World Journal of Pharmacy and Pharmaceutical Sciences
smaller than the inner one, just within the opening of lenticels, groups of sclereids consisting of 2-10 cells found in secondary cortex region, outer zone of cortex consists of 3--5 rows of irregularly arranged, tangentially elongated chlorenchymatous cells, cortical cells situated towards inner side, polygonal in shape and filled with plenty of starch grains, simple, ovoid, or irregularly ovoid-elliptical, occasionally compound of 2-4 components, several secretory cells, found scattered in the cortex, pericyclic fibres lignified with wide lumen and pointed ends, associated with a large number of crystal.

Pharmacological activity
Many research works have been carried out on Tinospora cordifolia with different activities i.e. adaptogenic, anti neoplastic, anti diabetic, anti bacterial, miscellaneous.But here some research works have been mentioned which are related to this study.
 Tinospora cordifolia was found to have immunomodulating activity.

Ranjan et al. World Journal of Pharmacy and Pharmaceutical Sciences
Anti-inflammatory activity  The aqueous extract of Tinospora cordifolia showed significant anti inflammatory activity in rats against acute and chronic type of inflammation induced by carrageenin and the activity resembles that of NSAIDS.
 Singh et al. have shown that the aqueous suspension of the alcoholic extract of the stem of T. cordifolia provided protection to liver damage induced by administration of carbon tetrachloride in mice, rats, and rabbits.
 The decoction of Tinospora cordifolia showed anti-inflammatory activity on carrageenin induced hind paw edema in rats.

Hepato-protective activity
A study of hepato-protective activity of T. cordifolia on Kupffer cell function using carbon clearance test as a parameter showed significant improvement in kupffer cell function and a trend towards normalization.Tinospora cordifolia appears to improve surgical outcome, in patient with malignant obstructive jaundice by strengthening the host defenses.Thus the Ayurvedic use of the plant in liver ailments is justified although the Active principle is yet to be isolated and identified.

Anti-stress activity
 The ethanolic extract of Tinospora cordifolia exhibited significant anti stress activity at 100 ml/kg compared with diazepam at 2.5mg/kg.The ethanolic extract of Tinospora cordifolia at a dose 100 mg/kg was shown to induce a marked protective action against 8 hr restraint stress induced ulcerization.
 Sharma and Khosla have reported that the alcoholic extract of Tinospora cordifolia roots possessed normalizing activity against stress induced changes in nor epinephrine, dopamine, 5-HT and 5 hydroxyindoleaceticacidlevels of experimental rats.103

Anti-histaminic activity
 Tinospora cordifolia stem extract significantly decreased bronchospasm induced by 5% histamine aerosol in Guinea pig and permeability in mice.It also reduced the no. of disrupted mast cell in rats.

Preliminary Screening of Phytochemicals
The preliminary phytochemical studies were performed for testing the different chemical groups present the drug.10% (w/v) solution of extract was taken unless otherwise mentioned World Journal of Pharmacy and Pharmaceutical Sciences in solution of Ethanolacetic Acid (4:1)], white spot with blue back ground shows presence of glycoside.

IDENTITY, PURITY AND STRENGTH
Loss on drying at 105C/Moisture content Place about 10 gm of drug sample after accurately weighing it in a tared evaporating dish.
After placing the above said amount of sample in a tared evaporating dish dry at 105 C for 5 hours and weigh.After drying tared evaporating dish cool in desiccator for 30 minutes and then weight.

Determination of Total Ash
About 2.0g of powdered drug was incinerated in a tared silica dish at a temperature not exceeding 450C until free carbon was left, cooled and final weight was taken.The percentage of ash calculated with reference to the air-dried drug (PASF, 1987).

Determination of Acid Insoluble Ash
The ash obtained as above method was boiled for 5 minutes with 25 ml of dilute hydrochloric acid and collected the insoluble matter on an ash-less filter paper, washed with hot water and ignited to constant weight.The percentage of acid-insoluble ash with reference to the air dried drug was calculated (PASF, 1987).

Determination of Sulphated Ash
Heat a silica crucible to redness for 10 minutes; allow cooling in a desiccator and weighing.
Put about 2.0g of powdered drug into the crucible; ignite gently at first, until the sample is thoroughly charred.Cool, moisten the residue with 1 ml of sulphuric acid, heat gently until white fumes are no longer evolved and ignite at 800 ± 25 until all black particles have disappeared.Allow the crucible cool; add few drops of sulphuric acid and heat.Ignite as before, allow to cool and final weight was taken after two successive operation repetitions.

THIN LAYER CHROMATOGRAPHY
The principle underlying the separation of the compounds is their adsorption at the solidliquid interface.For successful separation the compounds of the mixture should show

Ranjan et al. World Journal of Pharmacy and Pharmaceutical Sciences
different degrees of affinity for the solid support (or adsorbent) and the interaction between adsorbent and the component must be reversible.As the adsorbent is washed with the fresh solvent, the various components move down the column and arrange themselves in the order of affinity to the adsorbent.Those with the least affinity move down the column at a faster rate than those with greater affinity.

Materials used
All the TLC plates used for the analysis were prepared with silica gel containing binder.Most frequently used binder in silica gel is calcium sulphate (Silica gel 60 F 254 , Merck).Precoated aluminium sheets were also used which is coated with silica gel 60 F 254 (Merck).

Preparation of TLC plates
Required quantity of silica gel was mixed in a glass mortar to as smooth consistency with the requisite amount of water and the slurry quickly transferred to the spreader.The mixture has been spreaded over the plates in thickness of 0.2mm and was allowed to set into a thin layer.
The plates were transferred carefully to a suitable holder and after 30 minutes, dried and heated at 100-120C for at least one hour.The plates were kept in a desiccates after cooling, until required for further use.The pre coated plates were also activated by heating them for 30 minutes at 100C.

Application of sample
A known quantity of sample was dissolved in a known volume of solvent and the sample applied on pre coated TLC plates.

Selection of Solvent Systems-
The choices of the solvents depend upon the nature of the substances to be separated and also in the material on which the separation is to be achieved.
The solvents system was selected on the basis of trial and error method and by elutropic series.It has been found that combination of two solvents gave better separation than with a single solvent.

Chromatographic development (separation)
Development of the chromatogram is effected after the solvent of the applied sample is completely evaporated.Rectangular glass chambers or twin trough chambers are commonly used for TLC development.
Ranjan* and Dr. Satyendra Kr.Singh MD(Ay) AMO RBSK Bihar.Pharmacognosy is the study of medicinal drugs derived from plants or other natural sources.

Matter in water and alcohol
About 4.0g of coarsely powdered air dried material, was accurately weighed in a glass stoppered conical flask and macerated with 100ml of the solvent specified for the plant

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concerned for 6 hours, shaking frequently, then allowed to stand for 18 hours.Filtered rapidly, taking care not to lose any solvent.The extracted matter was dried at 105C for 6 hours, cooled in a desiccator for 30 minutes and then weighed.The percentage extractable matter was calculated.