Indian Journal of Research in Homeopathy

ORIGINAL ARTICLE
Year
: 2020  |  Volume : 14  |  Issue : 4  |  Page : 260--266

Pharmacognostical studies of Smilax aspera Linn. – A herbal drug


Satish Patel1, B Biswas2, K Rambabu1, EN Sundaram1, Renu Arya3,  
1 Drug Standardisation Unit (H), Hyderabad, Telangana, India
2 Department of Drug Standardization, Dr. Anjali Chatterjee Regional Research Institute for Homoeopathy, Kolkata, West Bengal, India
3 Central Council for Research in Homoeopathy, New Delhi, India

Correspondence Address:
Dr. Satish Patel
Drug Standardisation Unit (H), O.U.B-32, Vikrampuri, Road No. 4, Habsiguda, Hyderabad - 500 007, Telangana
India

Abstract

Background: Smilax aspera L. (sarsaparilla or prickly ivy) is a perennial climber from the family Smilacaceae. Its root and rhizome are used as alterative, demulcent, depurative, diaphoretic, diuretic, stimulant and tonic. Objective: To perform standardization of root and rhizome of S. aspera for authentication and identification of raw drug by pharmacognostical, physiochemical, powder and finish product evaluation. Materials and Methods: Air-dried rhizome and roots were boiled, sectioned and stained for macroscopical and microscopical analysis. For physicochemical studies, rhizome and roots were coarsely powdered and subjected for determination of extractive values, ash values, chemical constituents, weight per millilitre, alcohol content, total solids and loss on drying. Finished product analyses (chromatographic studies, sediments, pH and total solid) were also undertaken. Results: The root was longitudinally wrinkled, about 3 mm in diameter with numerous branching, rootless, tough and flexible. Cortex consisted of 18–20 rows of parenchymatous cells; xylem and phloem were arranged in a radiate manner. Rhizome was wrinkled, hard and brown externally and white or light-yellow internally. The outer cortex consisted of polygonal parenchymatous cells. The findings of physicochemical determination of raw drugs including maximum extractive values in alcohol were 5.67% w/w, 0.1% w/w foreign matter, 8.90% w/w moisture content, 10.60% w/w total ash, etc., and finished product parameter showed pH near to 7, total solid 1.07% w/v and 50% v/v alcohol content. Conclusion: The data represented in this article may be used as distinctive diagnostic characters for proper identification, authentication of raw drug to ensure purity, quality and efficacy of drug S. aspera.



How to cite this article:
Patel S, Biswas B, Rambabu K, Sundaram E N, Arya R. Pharmacognostical studies of Smilax aspera Linn. – A herbal drug.Indian J Res Homoeopathy 2020;14:260-266


How to cite this URL:
Patel S, Biswas B, Rambabu K, Sundaram E N, Arya R. Pharmacognostical studies of Smilax aspera Linn. – A herbal drug. Indian J Res Homoeopathy [serial online] 2020 [cited 2021 Jan 21 ];14:260-266
Available from: https://www.ijrh.org/text.asp?2020/14/4/260/305252


Full Text



 Introduction



Smilacaceae or the Greenbrier family having 350 species in two genera (Smilax and Heterosmilax) is the second-largest family. The genus Smilax is widely distributed in tropical, temperate and subtropical zones and found all over Asia, Europe, Oceania and the USA.[1] Out of 24 species found in India, 4 species, namely Smilax aspera Linn., Smilax perfoliata Lour., Smilax wightii A. DC. and Smilax zeylanica Linn., are available in southern India.[2],[3],[4] These species are commonly known as sarsaparilla and mainly present as climbers, with long, thin thorny stem. The stem branches with the help of tendrils attached to other plants or objects to grow steadily upward. All the parts such as rhizome, roots, stems and leaves of sarsaparilla are used in traditional systems of medicine including Ayurveda; in Homoeopathy, mainly its root and rhizome are used.[1],[5],[6]

S. aspera L., commonly known as sarsaparilla (common smilax, prickly ivy or rough bindweed) belonging to the family Smilacaceae, was formerly included in the monocot family Liliaceae. Smilax comes from ancient Greek language for bindweed. It is an evergreen, perennial climber with a flexible and delicate stem with sharp thorns.[7],[8] Climber stems reach a height of 1–4 m in woodland or scrub habitats and are sheltered in sharp hooks which also lengthen along the base of the midribs of the leaves. Leaves are dark green, 8–10 cm long, petiolated, alternate, tough, leathery heart shaped and toothed. Yellowish or greenish flowers are very fragrant, small and in axillary racemoses,[9] and fruits are 1–3 seeded, red to blue-black coloured berries, 5–10 mm in diameter, rubbery in texture and spherical in shape.[10] The seeds are round and bird dispersed. This species is broadly dispersed across the Mediterranean, Central Africa and Southern Europe through Asia into China, Pakistan, India, Sri Lanka, Nepal and Myanmar.[9],[11]

Principal chemical constituents present in S. aspera fruit are anthocyanins, pelargonidins and cyanidins. The active constituent is pelargonidin 3-O-rutinoside.[12] Root and rhizome mainly contain steroidal saponins, flavonoids and anthocyanins.[10],[12],[13] Primary steroidal saponins are (25S)-5b-spirostane-3b-ol-3-O-a-L-rhamnopyranosyl-(1-2)-b-D-glucopyranosyl-(1-2)-b-D-glucopyranoside, curillin G, asparagoside E, asparoside A, asparoside B, (25S)-5b-furostan-1b, 2b, 3b, 5b, 22a, 26-hexaol-26-O-b-D-glucopyranoside and 26-O-b-D-glucopyranosyl-(25S)-5b-furostan-1b, 3b, 22a, 26-tetraol-1-O-b-D-glucopyranoside.[9],[14],[15] Seed mainly contains different fatty acids.[10] S. aspera has been used in the treatment of syphilis, rheumatism, stomach pain, bloating, leprosy, psoriasis and diabetes and as an antioxidant to diminish the problems of menopause.[1],[9],[10],[16],[17] The root and rhizome of plant are used as alterative, demulcent, depurative, diaphoretic, diuretic, stimulant and tonic[18],[19] and are also used in the treatment of inflammatory disease, rheumatic arthritis, joint paint and oedema.[20],[21]

The present study was, therefore, undertaken to produce standard information for correct identification/authentication of this plant. Even though present modern analytical techniques, for example DNA barcoding, high-performance thin-layer chromatography, high-performance liquid chromatography (HPLC) and infrared, can provide high-quality data for authentication and quality control and quality assurance, here we seek to explore the employment of low-cost yet robust analytical techniques to ensure the quality control and quality assurance of the drug. This information presented in this study may also be consider as the standards ensuring the purity and quality of the drug S. aspera L.

 Materials and Methods



The rhizome and roots were obtained from Centre of Medicinal Plants Research in Homoeopathy (CMPRH), Emerald, Ooty (Tamil Nadu), India. Voucher herbarium sample (Herbarium no. 6511) was prepared and preserved at CMPRH, Emerald, Ooty (Tamil Nadu), India. The pharmacognostic and physicochemical parameters were performed as per the protocols mentioned in Homoeopathic Pharmacopoeia of India (HPI).[22] For physicochemical studies, the rhizome and roots were dried in shade. The dried material was powdered coarsely for the preparation of mother tincture.

The chemicals used in the study were double-distilled water, 37% hydrochloric acid (dilution to 10% was done in-house), strong alcohol, chloroform and methanol. All the chemicals used in this study were of analytical grade.

Macroscopy study

Macroscopic characteristics of root and rhizome were noted on the basis of visual observation of raw drug materials as per the prescribed methods.[23],[24] It included observation of condition, branching, shape, length, diameter, colour, odour, taste, surface texture and fractures.

Microscopy study

The microscopic study was undertaken by taking appropriate section of the plant parts. Rhizome and roots were boiled separately, cut into small pieces and processed for paraffin method of microtomy as per the method described by Johansen and Wallis.[25],[26] Slides of microtome sections cut at 12–15 μm were made, and these slides were further stained with safranin and light green or crystal violet and basic fusion combinations and mounted in Canada balsam and subjected for microscopic observations. Photograph of transverse section of rhizome and roots was taken using Olympus BX53 Research Trinocular Microscope.[27]

Organoleptic characters

Organoleptic characters of powder were evaluated by taking a minute quantity of powder and spread on a white background and visually examined for general appearance, namely nature, colour, odour, taste and texture.

Physicochemical studies

The dried plant material was coarsely powdered and subjected to physicochemical studies which include loss on drying, ash values and extractive values. Mother tincture was prepared following the method described in HPI[22] and subjected for its specific gravity measurement, pH metry, chromatographic profile and ultraviolet (UV) spectroscopy studies.

Thin-layer chromatography

Around 25 mL of mother tincture was heated on a water bath to remove the alcohol. The organics from the aqueous part were extracted using three 25 mL portions of chloroform. The chloroform extract was evaporated to around 2 mL, and thin-layer chromatography was performed using the concentrated chloroform extract using silica gel and chloroform: methanol (90:10) as mobile phase. The spots were detected using UV light of wavelength 365 nm and 254 nm.

 Observations and Results



Pharmacognostic studies

Macroscopy

Macroscopic studies revealed that root was longitudinally wrinkled, about 3 mm in diameter, dark reddish-brown colour and with numerous branching rootless, tough, flexible and not breaking easily even when bent double. Rhizome was also wrinkled, hard, externally brown coloured, internally white or light yellow coloured and gave rise to several roots at different points, fracture short, odourless and taste slightly bitter [Figure 1].{Figure 1}

Microscopic studies

Root

Transection of root was circular in outline. Outermost layer epidermis was single layered; epidermal cells were compactly arranged, polygonal or spherical in shape. Some epidermal cells were modified into root hairs. Epidermis was followed by of 2–4-layered hypodermis. Hypodermal cells were polygonal in shape, tightly packed cells with no intercellular spaces. Hypodermis was followed by multilayered cortex. Cortex consisted of 18–20 rows of parenchymatous cells. These cells were small to large spherical in shape with intercellular space. Endodermis was uniseriate with uniformly thickened walls. Endodermis was followed by a biseriate pericycle layer. Cells of pericycle were smaller in size as compared to endodermis cells. Vascular bundles were radial and polyarch. Xylem and phloem were arranged in radiate manner. Xylem consisted of lignified vessels, with spiral or reticulate thickening and bordered pitted tracheids, fibres lignified, with narrow ends, pitted thickening. Phloem consisted of phloem parenchyma, sieve tubes and companion cells. Large pith was present in the centre composing parenchymatous cells with intercellular space [Figure 2].{Figure 2}

Rhizome

Transection of rhizome showed a somewhat circular outline. The outermost layer was single-layered epidermis. Epidermal cells were closely arranged, rectangular in shape. Epidermis was followed by hypodermal ground tissue. Hypodermal cells were lignified. Hypodermis was followed by the cortical region which is composed of the outer and inner cortex. The outer cortex consisted of polygonal shape parenchymatous cells; cells contain starch grains, tightly arranged without intercellular space. Inner cortical cells were spherical or oval in shape, loosely arranged without intercellular space; bundles of acicular crystals of calcium oxalate were present. Vascular bundle was collateral, conjoint, closed and scattered. A group of sclerenchymatous fibres was present in inner cortex which also partially covered the vascular bundles. Xylem fibres were of different size, lignified, septate, with pits and large lumen. Xylem vessels were present in small and large size, with scalariform thickening and bordered pitted [Figure 3].{Figure 3}

Powder studies

The powder microscopy study showed the presence of pieces of epidermal cells and modified root hairs, polygonal hypodermal cell, pieces of cork with groups of tangentially elongated cells; isolated vessels or tracheids either whole or broken and acicular crystals of calcium oxalate.

Organoleptic characters

Colour: Whitish brownTouch: RoughOdour: CharacteristicTaste: Slightly bitter.

Physicochemical studies

The results of the physicochemical study of the raw drug are described in [Table 1]. The results of physicochemical studies of raw drugs showed 0.1% w/w foreign matter, 8.90% w/w moisture content, 10.60% w/w total ash, etc., and finished product parameters showed pH near to 7, total solid 1.07% w/v and 50% v/v alcohol content. The loss of drying (LOD) was found to be 8.90%.{Table 1}

In [Table 2], the physicochemical data of the mother tincture have been provided. The pH is close to 7. Furthermore, the total solid is 1.07% w/v. This shows that the solvent system is an excellent solvent system for phytochemical extraction.{Table 2}

Preparation of mother tincture

The mother tincture prepared as per [Table 3].{Table 3}

 Discussion and Conclusion



S. aspera is widely used in different medicinal systems for a variety of disorders. The present study demonstrates detailed macroscopic, microscopic, powder studies and physicochemical standards of crude drug and mother tincture of S. aspera. The root is longitudinally wrinkled, about 3 mm in diameter with numerous branching rootless, tough and flexible. Rhizome is also wrinkled, hard and externally brown colour, internally white or light yellow colour. Microscopic studies show the unique arrangements of epidermal, hypodermal, cortex and vascular tissue in root and rhizome of S. aspera. In physicochemical parameters, LOD percentage provides the information regarding the amount of raw wet drug required for the mother tincture preparation. The total ash was found to be around 10%. This value suggests that the metallic component in the raw drug is quite low. The acid-insoluble ash is <5%. This indicates that the amount of silicates in the raw drug is moderate. This is quite expected from the fact that the part for the drug used is exclusively sourced from the underground part of the plant. The pH of the mother tincture is close to 7, indicating that the mother tincture is safe for oral administration. Here, we have shown that even without using high-end instruments this study has been carried out, and we hope, this will pave the path for simple yet robust drug standardisation research. However, considering the new age instrumentation advances in analysis, for example HPLC-mass spectrometry (MS), gas chromatography-MS and nuclear magnetic resonance, we will also seek to explore the use of these modern analytical instruments in the future. This will provide a comparison of our present study with future studies employing modern analytical instruments. We are hopeful that these data may also be considered as pharmacopoeial standards for the drug S. aspera.

Acknowledgement

The authors are thankful to Dr. Anil Khurana, Director General, Central Council for Research in Homoeopathy, New Delhi, for the facilities provided and encouragements. The authors are also thankful to the Survey Officer, CMPRH, Ooty, for providing authentic raw plant drug.

Financial support and sponsorship

Nil.

Conflicts of interest

None declared.

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