Preliminary phytochemical screening and Pharmacogcostic studies of the common weeds, Mimosa pudica Linn., Chromolaena odorata Linn. and Tragia involucrata Linn. collected from Ganjam District, Odisha: A Comparative Study

Debashisha Panda¹*, Santosh Kumar Dash¹ and Gouri Kumar Dash ²

¹College of Pharmaceutical Sciences, Mohuda (Ganjam), Berhampur, Odisha

²Institute of Pharmacy and Technology, Salipur, Cuttack, Odisha

ABSTRACT:

The widespread use of herbal remedies in healthcare preparations, such as those described in ancient texts as ‘Ethnomedicines’, has been traced as the natural products with medicinal properties. Plants, as source bioactive medicinal principles, have continued to play a dominant role in the maintenance of human health since past decades. Keeping pace with the tribal folklore and wide spread use of botanicals in the locality, three plants namely, Mimosa pudica Linn., Chromolaena odorata Linn. and Tragia involucrata Linn. were selected and properly authenticated. The selected plant parts of the above plants were used for comparative studies on presence of various phyto-constituents by preliminary screening methods as well as macroscopic and microscopic features. The powdered drug examination was utilized as a diagnostic tool for the determination of pharmacognostic features including plant description and organoleptic evaluation. The preliminary phytochemical screening of M. pudica root extracts revealed presence of steroids and sterols, triterpenoids, alkaloids, flavonoids, tannins and phenolic substances. Under the same screening procedure, presence of carbohydrates, cardiac glycosides, steroids and sterols, triterpenoids, saponins, flavonoids, tannins and phenolic substances were identified with the leaf extracts of C. odorata and presence of steroids and sterols, triterpenoids, alkaloids, flavonoids, saponins, tannins and phenolic substances with the root extracts of T. involucrata. The powdered drug in various chemical reagents showed characteristic fluorescence at 365 nm of UV light which can play significant role as diagnostic tools in quality control of crude drugs. The monographic analysis of the plant materials was performed according to the Pharmacopeial standards.

KEY WORDS: Solvent extracts, preliminary phytochemical analysis, macroscopic features, powder microscopy, physical characteristics.

INTRODUCTION:

Nature is the paradise of medicinal principles offers to the humanity through plants which act as richest source of phytochemicals since time immemorial. An impressive number of modern drugs have been isolated from the floristic resources; many being tapped basing on their use in the treatises of traditional medicines. Various medicinal plants have been used for years in daily life to combat diseases, world over. The widespread use of herbal remedies in healthcare preparations, such as those described in ancient texts described as ‘Ethnomedicines’, has been traced as the natural products with medicinal properties.

In fact, plants (otherwise termed as botanicals) produce a diverse range of bioactive molecules, making them a rich source of various types of medicaments. Plants, as source bioactive medicinal principles, have continued to play a dominant role in the maintenance of human health since past decades ^[1]. Most of the forest habitats in the state of Odisha are covered by aboriginal tribes out of which khond (Kondha) and Soura/Langia soura are considered ethnically to be most primitive and are found especially in undivided Ganjam, Khandmals and Kalahandi districts. Their healthcare status and medical know-hows over ages interpreted as ‘Traditional healthcare system’ which mostly depends on herbals and the psychosomatic lines of treatment associated with medico religious belief, mysticism and magics, etc. The combination of such healthcare therapy associated with modern allopathic system of treatment may become more affordable by poors and its scientific evaluation needs to be established ^[2,3,4].

Keeping pace with the tribal folklore, three important plant species, namely, Mimosa pudica Linn., Chromolaena odorata Linn. and Tragia involucrata Linn. have been selected which are found have less exploited record and more usable value by the locals. These species are abundantly available and more eco-specific with their wide medicinal uses in the locality.

MATERIALS AND METHOD :

Collection of Plant material

The fresh plant parts of young matured plants of Mimosa pudica Linn.(roots), Chromolaena odorata Linn.(leaves) and Tragia involucrata Linn.(roots) were collected from various locations of Lathi and Mohuda villages, Ganjam district, Odisha and were properly authenticated. After authentication, the specified plant parts were collected in bulk, washed under running tap water to remove adhering dirt followed by rinsing with distilled water. Then, those were separately shade dried and pulverized in a mechanical grinder followed by sieving (sieve no. 40) to obtain coarse powders and kept in air-tight containers separately for future use.

Preparation of Extracts

The dried powdered plants materials were separately extracted successively with various solvents viz. petroleum ether (40⁰–60⁰C), ethyl acetate and methanol in increasing order of polarity using a Soxhlet extractor. The period of extraction was fixed at 48 h for every solvent at every stage of the extraction process. The solvents were purified by distillation prior to extraction ^[5,6].

Preliminary phytochemical analysis of crude extracts of selected plant species

Different extracts obtained from the above extraction processes were analyzed for various phyto-constituents present in these extracts by the method of qualitative phytochemical analysis ^[7,8.9] and results were depicted in Table - 1.

Pharmacognostic studies of selected plant species

I) Macroscopic features

A) Roots of Mimosa pudica Linn.

Colour Greyish brown

Dimensions Length - 7.2 cm to 29.6 cm and Diameter - 0.4 cm to 2.2 cm

Shape Cylindrical, fibrous with nodules

Branching Tap root (primary) with secondary and tertiary branches

Rootlets Present, of true kind, thin and wiry, scattered profusely around the primary root

Direction of Growth Taproot downward, i.e. positively geotropic and secondary roots horizontal with root nodules

Surface Characters Rough

Texture Fibrous

Fracture Hard, woody

Odour Distinctly characteristic

Taste Slightly astringent

B) Leaves of Chromolaena odorata Linn.

Dimensions Length - 5 to 12 cm. & breadth - 3 to 6 cm.

Colour Bright Green

Phyllotaxy Opposite (decussate)

Venation Reticulate

Margin Serrate

Apex Acute to shortly acuminate

Base Cuneate stomatic, dorsiventral, slightly pubescent

Texture Slightly coriaceous

Petiole Up to 1.5 cm long

Taste Pungent

Odour Aromatic

C) Roots of Tragia involucrata Linn.

Colour Light brown

Dimensions Length - 2.2 cm to 9.4 cm &

Diameter - 0.4 cm to 1.2 cm

Shape Woody, hard, Cylindrical, tortuous

Branching Taproot system

Rootlets Present, of true kind, thick and wiry secondary roots

Scattered profusely around the primary root

Direction of Growth Horizontal secondary roots, positively geotropic primary

Surface Characters More or less rough due to presence of secondary roots and root scars

Odour No significant characteristic

Taste No significant characteristic

II) Microscopic features

Powder microscopy of selected plant materials

Roots of Mimosa pudica Linn.

The powdered microscopic characteristics of the roots of M. pudica revealed uniformly shaped polygonal cork cells, oval shaped thick walled phloem fibres, striated walled polygonal cells conforming hard tissues with fibrous sclerenchymatous patches, rhomboid crystals of calcium oxalate and simple or compound starch grains. Cork consists of several layers of orange-brown, thick walled brick-like rectangular cells, polygonal parenchymatous cells relatively narrow, probably are cortical origin. Phloem fibres (secondary/primary) abundant associated with patches of parenchymatous tissues, unlignified or slightly lignified with calcium oxalate prismatic crystals; Vessels seen to be striated, thick, yellow, pitted with reticulately thickened walls associated with sclerenchymatous fibres. Unicellular trichomes, patches of bunches of fibres (bast/wood) are observed (Figure - 1)

Leaves of Chromolaena odorata Linn.

The powdered microscopic characteristics of the leaves of C. odorata revealed the following microscopic characteristics. Fragments of rectangular epidermal cells with an external cuticle layer; anomocytic type stomata encircled by irregular subsidiary cells were seen. Elongated unicellular epidermal trichomes with multicellular base were observed. Well-developed thin, long non-lignified phloem fibers and lignified sclerenchymatous fibers were found in groups or isolated. Fragments of parenchymatous cells and single layered palisade cells with no intercellular space were observed. Starch grains and calcium oxalate crystals not seen, probably might be absent (Figure - 2).

Roots of Tragia involucrata Linn.

The powdered microscopic characteristics of the roots of T .involucrata revealed stratified isodiametric cork cells (few lignified), isolated patches of parenchymatous cells (cortical/phloem), vessels solitary large with numerous bordered pits, fibers and tracheids thick walled. Rosette calcium oxalate crystals and starch grains were also found. (Figure - 3).

Behaviour of powdered drugs with various chemical reagentsThe behaviour of powdered drugs with various chemical reagents were studied under U.V. light (365 nm) after treating with different chemical reagents and reported as in the table - 2 ^[10].

MPR: Mimosa pudica Root; COL: Chromolaena odorata Leaf; TIR: Tragia involucrata Root

Physical characteristics

Determination of Ash Values

The ash remaining following ignition of medicinal plants materials are determined by four different methods which measure total ash, acid-insoluble ash, water-soluble ash and sulphated ash. The procedures for determining ash values are followed as per existing standards ^[11,12,13].

The total ash method is designed to measure the total amount of material remaining after ignition. This includes both “physiological ash”, which is derived from the plant tissue itself, and “non-physiological” ash, which is the residue of the extraneous matter (e.g. sand and soil) adhering to the plant surface.

Acid-insoluble ash is the residue obtained after boiling the total ash with dilute hydrochloric acid, and igniting the remaining insoluble matter. This measures the amount of silica present, especially as sand and silicone earth.

Water-soluble ash is the difference in weight between the total ash and the residue after treatment of the total ash with water. The results of the ash values were reported

Table - 1: Data showing preliminary phytochemical test results of various extracts of selected plant species

Part Used

Plant/s

Extracts

Alkaloids

Carbo-hydrates

Cardiac glycosides

Phenolic compounds and Tannins

Proteins and Amino acids

Gums and Mucilage

Flavonoids

Saponins

Steroids

Triterpe-noids

Mimosa pudica Linn.

Root

Pet.ether ext.

Ethyl acetate ext.

Methanol ext.

Chromolaena odorata Linn.

Leaf

Pet.ether ext.

Ethyl acetate ext.

Methanol ext.

Tragia involucrata Linn.

Root

Pet.ether ext.

Ethyl acetate ext.

Methanol ext.

‘+’: Present and ‘-’: Absent

Table - 2 : Data showing behaviour of powdered drugs with various chemical reagents

Sl. No.

Treatment of powdered drug with various chemical reagents

Fluorescence under Long UV Light ( 365 nm )

MPR

COL

TIR

Untreated Drug powder

Treated with Sod. Hydroxide in Methanol

Treated with Hydrochloric acid

Treated with equal vol. mixture of Nitric acid & water

Treated with Sodium hydroxide solution (5N aqueous solution)

Treated with Picric acid

Treated with Conc. Sulphuric acid

Treated with Ferric Chloride Solution

(5% w/v aqueous solution)

Treated with Iodine solution (N/20)

Brownish

Yellow

Black

Brown

Black

Brown

Dark Green

Green

Black

Greenish Red

Black

Yellowish Brown

Black

Brownish Black

Greenish yellow

Yellowish brown

Pale yellow

Pale Green

Pale Yellow

Pale Green

Blue

Bluish Black

Greenish Black

Method A

A glass stoppered shallow weighing bottle that has been dried under the same condition, was employed in the determination. The bottle was weighed and 1 g of the plant material was transferred in the bottle and covered. The bottle and the contents were accurately weighed. The loaded bottle was placed in the oven by removing the stopper left in the chamber. The sample was dried to constant weight. After the completion of drying, the bottle was promptly closed and then allowed to cool to room temperature in a desiccator before weighing. The bottle and contents were reweighed and reported. The results were tabulated in table - 5.

RESULTS AND DISCUSSION:

The macroscopic and microscopic findings of the selected plant materials have been reported at appropriate places.

The powdered drug examination was utilized as a diagnostic tool for the determination of pharmacognostic features including plant description and organoleptic evaluation. The powder microscopy of the root of M. pudica and T. involucrata revealed presence of ground tissues (parenchyma) associated with phloem, vessels and fibres. Presence of calcium oxalate crystals was found to be existing in cellular environment (Figure - 1 and 3). The powder microscopy of the leaf of C. odorata (Figure - 2) revealed presence glandular trichomes and more mesophyll tissues (more soft tissues and less hard tissues).

The preliminary phytochemical screening of M. pudica root extracts revealed presence of steroids and sterols, triterpenoids, alkaloids, flavonoids, tannins and phenolic substances (Table - 1). Under the same screening procedure, presence of carbohydrates, cardiac glycosides, steroids and sterols, triterpenoids, saponins, flavonoids, tannins and phenolic substances were identified with the leaf extracts of C. odorata (Table - 1) and presence of steroids and sterols, triterpenoids, alkaloids, flavonoids, saponins, tannins and phenolic substances with the root extracts of T. involucrata (Table - 1).

The powdered drug in various chemical reagents showed characteristic fluorescence at 365 nm of UV light (Table 2) which can play significant role as diagnostic tools in quality control of crude drugs.

The monographic analysis of the plant materials was performed according to the Pharmacopeial standards (Table-3, 4 and 5). It may be noted that the plants M. pudica and T. involucrata are official in Ayurvedic Pharmacopeia of India, where the standards for ash values and extractive values have been listed. Our plant materials also confirmed to these parameters listed in the Pharmacopoeia and it was found that all the values were within the pharmacopeia limits. However, for other plant material (C. odorata), obtained experimental values are listed, since there is no pharmacopoeia standards available.

CONCLUSION:

Pharmacognostic standards established for the plants M. pudica, C. odorata and T. involucrata will be useful to the future investigators for pharmacognostical standardization of the plant materials.

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Received on 20.01.2012

Modified on 05.02.2012

Accepted on 12.02.2012

Research Journal of Pharmacognosy and Phytochemistry. 4(2): March-April 2012, 92-96