Pharmacognostical Study and Phytochemical Evaluation of Pterisvittata L.

 

Lakshmi Kanta Kanthal*, Suman Pattanayak, Rimi Mondal, Susmita Roy, Somnath Das, DibyenduGuria, Ujjwal Khatua, Surajit Bera

Haldia Institute of Pharmacy, Haldia, Purba Medinipur, West Bengal, India – 721657.

 

ABSTRACT:

Objective: To evaluate the Pharmacognostical Study and Phytochemical screening of Pterisvittata L. Methods: Physicochemical and pharmacognostical standardization parameters for C. bonplandianum was developed as per the methods provided in World Health Organization (WHO) guidelines. Quantitative estimation of flavonoid total phenolics, tannins and total saponines contents were estimated by UV–spectrophotometry. Total alkaloids content was estimated by gravimetric analysis. Results: Morphological observations revealed that leaves of C. bonplandianum are simple, petiolate, alternate, 3–5cm long, oval to lanceolate in shape, serrated margin with acute apex. The stem is cylindrical, branched, woody, light brown in color, odourless and bitter in taste. Roots are small, dark brown to black in color, cylindrical, odourless and bitter in taste. Histological observations revealed the presence of discontinuous layer of lignified sclerenchymatous cells (stone cells), paracytic stomata, stellate trichomes, xylem with scalariform thickenings and libriformfibres. Qualitative and quantitative study of phytoconstituents revealed the presence of flavonoids (0.57% w/w, equivalent to rutin), phenolics (4.25% w/w, equivalent to tannic acid) tannins (2.15% w/w, equivalent tannic acid) and saponins (7.4% w/w, equivalent to diosgenin) and gravimetric estimation of alkaloids content was found to be 2.5% w/w. Conclusion: The pharmacognostical and phytochemical parameters studied in the present investigation will aid in the identification and will be helpful in maintaining the standard profile of C. Bonplandianum. Methods: Pharmacognostical& Phytochemical screening parameters for Pterisvittata L. Results: Morphological observations of P.vittatarevealed that leaflet comprises mesophyll, epidermis and the vascular bundle. Sclerenchymatous strips are found both in adaxial and abaxial sides in the hypodermal regions of the midrib. Epidermis is covered with thick cuticles and is single-layered with quadrangular cells. Phytochemical screening revealed thepresence of alkaloids, flavonoids, phenolics, tannins and saponins. Different pharmacognostical studies were performed like Fluorescence analysis, extractive values, ash value, loss on drying, foaming index & swelling index. Conclusion: The pharmacognosticaland phytochemical parameters showed in the present investigation will aid in the scientific research will be helpful in maintaining the standard profile of Pterisvittata L.

 

 

 

 

 

NTRODUCTION:

Medicinal plants have traditionally occupied an important position in the socio-cultural, spiritual and medicinal fields of rural and tribal life in Bangladesh.¹ In recent years; the growing demand for plant products has led to a quantum leap in the volume of trade in plant materials within and across the country.²

 

Since the very beginning of human civilization, people have used different parts of plants to treat various diseases. Until now, the plant extract is widely used for the preparation of many drugs in pharmacy. There are many traditional healers known as Kobiraj in the village who depend only on plant parts. Medicines were prepared from different parts of plants using traditional methods. The therapeutic efficacy of plants depends on some bioactive compounds or phytochemicals.³

 

Ferns are an ancient plant group that produces spores rather than seeds. Many stages of the fern reproductive cycle are hidden. The most visible stage of a fern's life cycle produces spores and is called the sporophyte (literally "spore plant").⁴ Fern spores are released from sporangia that cluster into sori.⁵ Thesori are usually found on the underside of the leaves, the shape and arrangement of these sori vary greatly between fern species. In Pterisvittata sori, they are arranged in continuous, copper-colored ridges along both leaf margins.⁶ Up close, the sporangia resemble tiny pearls clustered between threads of yarn.⁷

 

Ethno medicinally, Pterisvittata L. is used for wound healing. This fern is a hyper accumulator of arsenic, but its therapeutic aspect is still unexplored.⁸

 

A literature survey shows that in Malaysia the juice of the young leaves is applied to the clean, unhealthy tongue of young children, while the juice of the rhizome is used for swollen glands in the throat.⁹ In New Guinea the juice is used for ulcers, sores and wounds.¹⁰  Ground dried roots and leaves boiled in sesame oil are used to treat skin diseases in children. P. vittata stands have been proposed to remediate arsenic-contaminated soils, but no field test results are available yet.¹¹

 

So far, a large number of active phytochemical components of this plant have been discovered namely limonene, beta-caryophyllene, beta-pinene, geranial edulinin, ribalinin, isoplatydesmin and so on. Several studies have shown investigation of pharmacological potential.¹²

 

These phytochemicals or secondary metabolites are naturally synthesized in various parts of plants such as leaf, bark, root, fruit, stem, etc.¹³ These chemicals are not used in plant growth but play an important role in plant defence against herbivores and other interspecific defenses.¹⁴ There are possibly more than 25,000 secondary metabolites in plants.¹⁵ These are usually classified based on the pathway of chemical synthesis and are generally of 4 types.¹⁵ They are alkaloids, glycosides, phenols and terpenoids.¹⁶ Most of them are synthesized from some common amino acids such as tyrosine, ornithine, tryptophan, arginine and lysine. These are specific to individual species.¹⁶

 

The  aim  of the  present study  are to  investigate the Histological characters, pharmacognostical parameters presence of qualitative phytochemicals and thin  layer chromatography  (TLC)  of  Pterisvittata L.¹⁷

 

MATERIALS AND METHODS:

Plant material:

The  whole  plant  of  Pterisvittata  was  collected  during  the month  of  April-May,  2022.  The Pterisvittata L. plant were collected from the local areas of ICARE campus, Haldia Institute of Pharmacy, West Bengal, India and were authenticated by Central National  Herbarium, Botanical Survey of India.

 

Preparation of plant extract:

Fresh P.vittata plant material was thoroughly washed under running tap water and dried in the shade for two weeks. The whole plant material was crushed into a coarse powder using a mechanical crusher and passed through a sieve (20#). The coarse powdered drug (5g) was then gradually extracted by a cold maceration process with 100mL of petroleum ether and Ethanol for 72h (frequent shaking for 6 h and left to stand for 18h). At the end of the extraction, the micelle is separated from the matolin by filtration. Subsequently, the micelle is separated from the solvent by the distillation method. Both extracts obtained were filtered, concentrated and stored indesiccators until use.

 

Pharmacognostical evaluation:

Macroscopical, Microscopic and powder evaluation:

Macroscopic evaluation of whole parts of the plant shows leaf, stem and roots of P.vittata was performed according to the methods. Various organoleptic characters such as colour, shape, size, aroma, taste and texture were carefully studied using a simple microscope. For microscopic evaluation, fresh specimens were cut freehand and immersed in a cleaning agent (lactophenol). Sections were dehydrated with various strengths of absolute alcohol and then stained with a mixture of phloroglucinol and conc. HC1 (1:1, v/v). Finally, the stained sections were fixed for histological observations. To study isolated elements, pieces of roots were macerated with concentrated nitric acid, washed with distilled water and finally placed in glycerin for observation. Photographs were taken at various magnifications using a binocular microscope.

 

Determination of physicochemical parameter:

Crude drug powder (whole plant) of P.vittata was used to determine various physico-chemical parameters such as total ash value, acid-insoluble ash value, water-soluble ash value, loss on drying, extraneous matter, extractable matter, foaming index, swelling index. All these parameters were evaluated according to the standard methods of W.H.O. instructions

 

Fluorescence analysis of powdered drug:

Fluorescent powder drug analysis of P.vittata crude powder was performed in daylight, short UV (254nm) and long UV (366nm) according to the method. Fluorescence patterns were obtained after the powdered drug reacted with various chemical agents.

 

Phytochemical evaluation:

Preliminary phytochemical screening was performed for the presence of various phytoconstituents such as alkaloids, carbohydrates, glycosides, saponins, steroids, terpenoids, phenols, flavonoids and proteins were analyzed in all tested extracts viz. petroleum ether extract and ethanol extract of P.vittata.

 

RESULT:

Pharmacognostical evaluation:

Morphological evaluation:

Rhizome short, erect to erect, covered with colored, brownish scales throughout. Leaves pinnate, monomorphic; petiole 5-50cm long, green, scaly when young; lamina oblanceolate.

 

The underground stem is branched and persistent, rhizome and surrounded by brown scales. A few species have permanent leaf bases on the rhizomes. The leaves grow from the upper part of the rhizome with a long spindle.

 

The leaves are unipinnately compound, macrophyllous, growing in an acropetal pattern on the rhizome. Pinnae dissections are not so low compared to Pteridium. In some species, bipinnate leaves, finger-like and spreading leaves are also observed. Leaves that are developed are referred to as leaves. The spindle contains many sessile, coriaceous, lanceolate leaflets arranged in pairs except for the terminal leaflet.

 

Table 1: Macroscopical evaluation of  Pterisvittata L.

 

Microscopical Evaluation:

Rhizome - is oval and when viewed through T.S. is distinguished into cortex, epidermis and stele sections. Epidermis – It is covered with a thick cuticle and is single layered with square cells. Bark – It is divided into four to five layers of sclerenchymatous hypodermis and inner broad parenchymatous areas. These areas have traces of leaves and roots.

 

Stele - This structure varies between species. The meristeles (annulus of vascular fibers) are implanted in the underlying tissue (of a parenchymatous nature). Each of these meristeles is single-layered elliptical, where the endodermis contains Casparian strips within their radial walls. A 1-2 layered pericycle (thin walls) is found surrounding the phloem under the epidermis. The xylem is located principally to the meristem, showing a central protoxylem surrounding both sides of the metaxylem. It includes xylem parenchyma and tracheids. Phloem on the other hand has phloem parenchyma and sieve cells. It completely surrounds the xylem.

 

Leaflet – Leaflet contains mesophyll, epidermis and vascular bundle. The epidermis is a single layer located on the lower and upper surfaces. It is only the lower epidermis that has stomata. The mesophyll can be distinguished into lower spongy spaces and upper palisade or homogeneous. Sclerenchymatous bands are found on both the adaxial and abaxial sides in the hypodermal regions of the midrib. The midrib has an amphicribral vascular bundle that is concentric and surrounds a single layer of pericycle and endodermis. Vascular fibers are implanted in the mesophyll.

 

 

 

 

Fig No. 3: T.S. of  stem of  Pterisvittata L.Fig No. 4 :T.S. of  spores of Pterisvittata L.

 

 

Fig No. 5: T.S. of  root of Pterisvittata L.

 

Table No. 2 :Powder Microscopy

 

Table No. 3 :Proximate value

 

Table No. 4 :Fluorescence analysis of  powder of  Pterisvittata L

 

Table No. 5 :Phytochemical screening test of Ethanolic extract

 

 

Table No. 6 :Phytochemical screening test of petroleum ether extract

 

DISCUSSION:

Present research includes Pharmacognostical and Phytochemical investigations of the whole plant of Pterisvittata L.

 

Pharmacognostical Contributions:

Pterisvittata L. was found to be greenish to reddish brown in color, odorless, simple quill with rough surface. Transverse section and powder microscopy of the whole plant revealed the presence of elongated axillary epidermal cells with raised periclinal and sinusoidal anticlinal walls. Boletuses were hypostomatous with randomly scattered anomocyticstomatal complexes located at the same level as the epidermis. The stomata were large and elliptical. Transverse sections from the central regions of the spindle and stipe revealed V- and U-shaped vascular bundles. In each vascular bundle, the xylem fibers were seahorse-shaped (hippocampus). In contrast, needles had a triangular vascular bundle with a uniform mesophyll organization containing homogeneous lobed parenchyma cells. The indumentum consisted of trichomes and scales that formed different types of robes. Trichomes were borne only on the auricle and scales on the spindle and pith. The roots developed a dense network of long root hairs and the xylem consisted of tracheids with scalariform pits. The Sori were submarginal; continuous along both edges of the pinnae and were covered by a false indusium. Spores were elongated with a short stout pedicel and the annulus was positioned vertically, resulting in transverse dehiscence of the sporangium. Paraphyses were uniseriate, unbranched, septate and found to be mixed with sporangia. The spore of the spherical spores was decorated with a thick reticulum in which the areoles contained round tubercles.

 

Proximate values:

Approximate values ​​for the whole Pterisvittata L. plant are as follows: ethanol-soluble extractive value (0.5%), kerosene-soluble extractive value (0.3%), total ash (12.5%), acid-insoluble ash (3.5%), water soluble ash (7.5%), Loss on drying (0.75%), These values ​​are a criterion for determining the identity and purity of the crude drug. The water and decoction of the plant material were in a ratio of ten tubes, and after shaking the tubes, the foam was measured with a scale. The measured anterior height was less than 1 cm in each tube. Therefore, the foaming index was found to be less than 100. The swelling index was calculated to know how much the plant material can swell when placed in water and also to know that the plant material contains some slimy content. After adding water to the plant material, two readings were taken, an initial reading and a final reading after 3 hours. The swelling index of P.vittata was found to be 1.84.

 

Phytochemical Investigations:

Preliminary phytochemical investigations of petroleum ether and ethanol extract revealed the presence of alkaloids, steroids, terpenoids, tannins, flavonoids, carbohydrates, saponins and phenolic compounds, the absence of proteins and amino acids, glycosides.

 

CONCLUSION:

This study investigated the pharmacognostic parameters and phytoconstituent of the ethanolic and petroleum ether extracts of the whole plant Pterisvittata L. and demonstrated the significant pharmacognostic value and significance of the various phytoconstituents present in this plant. Therefore, Pterisvittata L. can be useful for the treatment of various diseases and can be introduced as a medicinal plant. Different pharmacological activity should be done by the researcher for Pterisvittata L.

 

CONFLICT OF INTEREST:

The authors have no conflicts of interest regarding these investigations.

 

ACKNOWLEDGMENTS:

We have expressed our special thanks to our respected Management of Haldia Institute of Pharmacy, Haldia, West Bengal, India for their motivation and support in every step.

 

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Received on 19.10.2022          Modified on 13.12.2022

Accepted on 07.01.2023     ©A&V Publications All right reserved