|Year : 2022 | Volume
| Issue : 1 | Page : 12-21
Pharmacognostical studies on whole plant of Mimosa pudica L
Harikrishnan Rajalakshmi, Amaranathan Sethuramani
Department of Pharmacognosy, College of Pharmacy, Madurai Medical College, Madurai, Tamil Nadu, India
|Date of Submission||02-May-2022|
|Date of Decision||11-Jul-2022|
|Date of Acceptance||21-Jan-2023|
|Date of Web Publication||18-Apr-2023|
Ms. Harikrishnan Rajalakshmi
Department of Pharmacognosy, College of Pharmacy, Madurai Medical College, Madurai - 625 020, Tamil Nadu
Source of Support: None, Conflict of Interest: None
Background: Mimosa pudica is a creeping, diffuse, prickly undershrub of genus mimosa, native to tropical America which is also scattered along tropical and subtropical parts of India. It is well known as a sensitive plant, shame plant, touch-me-not, humble plant, shyness princess, and sleeping plant by virtue of rapid leaf movements in response to touch.
Aim: The intention of this research was to investigate the morphological, micro anatomical, powder microscopy, proximate, and phytochemical analysis in whole plant of M. pudica.
Materials and Methods: The investigation on macroscopy, microscopy, and physiochemical and phytochemical analyses were implemented using standard methods.
Results: This study divulged unique macroscopical and anatomical characters present in the whole plant of M. pudica. The powder microscopy revealed the presence of trichomes, paracytic stomata, starch grains, prismatic crystals, mucilage, and brownish content. The quantitative microscopy performed in epidermal peelings of leaf quantified the epidermal number, stomatal number, stomatal index, and palisade ratio which determine its quality and purity. The proximate analysis performed was within the specific limits. The phytochemical analysis identifies different phytoconstituents in n-hexane and ethyl acetate extract. Further quantitative estimation disclosed that the total quercetin equivalent in 1 g of ethyl acetate extract of M. pudica was to be 13.39 mg and 27.5 mg of total β-sitosterol equivalent in n-hexane extract of M. pudica.
Conclusion: This research concludes the assessment in whole plant of M. pudica by pharmacognostical, physiochemical, and phytochemical estimation for its identification and confirmation of species epithet from adulterated, substituted, or similar distant group species such as Albizia julibrissin, Acacia dealbata, Neptunia gracilis, and Mimosa tenuiflora.
Keywords: Mimosa pudica, powder microscopy, proximate, quantitative and qualitative analysis, transverse section
|How to cite this article:|
Rajalakshmi H, Sethuramani A. Pharmacognostical studies on whole plant of Mimosa pudica L. J Res Siddha Med 2022;5:12-21
|How to cite this URL:|
Rajalakshmi H, Sethuramani A. Pharmacognostical studies on whole plant of Mimosa pudica L. J Res Siddha Med [serial online] 2022 [cited 2023 Jun 3];5:12-21. Available from: http://www.jrsm.in/text.asp?2022/5/1/12/374334
| Introduction|| |
The genus mimosa consists of about 400–450 species of herbs, shrubs, and trees. The taxonomic character of the genus Mimosa has been analogous with different genera and the species demonstrating with similar pinnate and bipinnate leaves such as Albizia julibrissin, Acacia dealbata, and Neptunia gracilis. Two species in the genus Mimosa are illustrious, Mimosa pudica and Mimosa tenuiflora. The plant M. pudica is a prostate, annual, or perennial belonging to the family fabaceae. The genus Mimosa is derived from the Greek word “mimos” meaning mimic owing to nimble response of an external stimuli, the species pudica is a Latin word meaning bashful or shrinking. Traditionally M. pudica is used in snake bites, hyperglycemic and hypercholesteremic conditions. It also acts as a therapeutic agent against small pox, hemorrhoids, rheumatism, cancer, benign tumor, edema, leprosy, dysentery, vaginal and uterine complaints, inflammations, asthma, and blood disorders. M. pudica is most commonly used as an ornamental plant and as an insect repellant. This current research deals with the comprehensive examination on pharmacognostical and phytochemical investigations.
| Materials and methods|| |
Pharmacognostical assessment is fairly essential to generate assurance about the identification of adulterated and substituted medicinal plants. Therapeutic efficacy of medicinal plants depends upon the quality and quantity of chemical constituents in secondary metabolites; thus, pharmacognostical validation is crucial for the standardization of therapeutic drug. The appraisement of anatomical studies of medicinal plant materials is indispensable. The existent inquiry deals with evaluation of whole plant M. pudica. Macromorplogy and microscopical validation include transverse sections (TSs); microchemistry, powder analysis, and quantitative microscopy were implemented using standard methods. The physicochemical analysis is helpful in judging the identity and purity of the crude drug.
Collection and authentication
The whole plants were collected from Cheranmadevi, Tirunelveli district, Tamil Nadu, India and were authenticated by Dr S. Mutheeswaran M.Sc., M.Phil., Ph.D., Scientist, Centre for Biodiversity and Biotechnology, Xavier Research Foundation, St Xavier’s College, Palayamkottai, Tamil Nadu.
Preparation of plant materials
The whole plants were subjected to shade dried and were powdered in a mixer. The coarse powder was sieved and was stored in a well closed container.
Macro and microscopical evaluation
The whole plants were studied separately for its morphological characters by organoleptic test such as color, odor, size, shape, taste, and texture. Sample was preserved in fixative Formaldehyde Alcohol Acetic Acid, 10%:50%:5% + 35% water for more than 48 hours. The preserved specimens were cut into thin TS using a sharp blade and the sections were stained with safranin. TSs were photographed using Nikon ECLIPSE E200 trinocular microscope attached with Zeiss AxioCam Erc5s digital camera under bright field light. Magnifications were indicated by scale bar. A pinch of the powdered sample of M. pudica was mounted on a microscopic slide with a drop of 50% glycerol. Characters were observed using Nikon ECLIPSE E200 trinocular microscope attached with Zeiss ERc5s digital camera under bright field light. Photomicrographs of diagnostic characters were captured and documented. The fresh leaf samples of M. pudica were boiled with 0.1% chloral hydrate solution for slide preparation. Vein islets, vein termination, epidermal number, stomatal number, stomatal index, and palisade ratio were determined and calculated by observing the slide using microscope.
The powdered drugs were evaluated for loss on drying, foreign matter analysis, ash value, and extractive value using standard methods and instruments.
Phytochemistry is a branch of science which deals with the chemical and structural composition of plant secondary metabolites. The appropriate considerate of phytochemical composition is crucial for scientific validation of existing traditional and ethnomedical claim of medicinal plants. The modern phytochemistry provides various techniques to affirm the identification, separation, and characterization of secondary metabolities.
Extraction of medicinal plants is a process of separating active plant materials or secondary metabolites such as alkaloids, flavonoids, terpenes, saponins, steroids, and glycosides from inert or inactive material using an appropriate solvent and standard extraction procedure. Phytosterols are usually extracted by Soxhlet extraction (hot continuous extraction) and maceration (single continuous extraction). Maceration techniques are preferable for the extraction of whole plant of M. pudica because of its coarse nature of plant powder by using n-hexane and ethyl acetate as solvent. Extraction of plant materials is based on the selection of solvent, temperature, particle size, and pH, which is the important feature of phytoconstituents from medicinal plants. Retrospective data considered the health promoting effect of phytosterols and the techniques for the extraction. The extraction and analysis process of phytosterols are complex and have not been still established. Selection of solvent should be conscripted for the extraction of targeted compounds using maceration or Soxhlet extraction. Soxhlet extraction is time consuming and involves large amounts of organic solvents which are health and environmental hazards and not suitable for thermolabile substances, thus maceration method is considered as a choice for extraction. The yield and composition of the extract vary because of the use of different extracting solvents. The most widely used solvents for the extraction of phytosterols are n-hexane, petroleum ether, ethanol, and methylene chloride. Phytosterols are lipophilic and are soluble in non-polar solvents. Reminiscing survey of disputes, maceration method and n-hexane solvent were pegged. The qualitative and quantitative phytochemical screening of ethyl acetate and n-hexane extract of M. pudica leaves were performed with the aid of standard procedures and references.
| Results|| |
The morphology [Figure 1] and organoleptic characters of whole plant of M. pudica were recapitulated in [Table 1]. Morphological features of whole plant of M. pudica Linn. divulged that the leaves are digitatively arranged, disitichous, bipinnately compound, hairy pinnae (1–2 pairs) with prickly petiole, stipulate, linear lanceolate leaflet (24–40 pairs) without stalks; leathery, measuring 0.6–1.2 cm long and 0.3–0.4 cm broad with obliquely rounded to truncate base; apiculate apex with entire to ciliate margin. The stems are erect, hispid with fibrous bark. Flowers are lilac pink, bisexual, glabrous, campanulate calyx, corolla four lobes, puberculous, crenate toward base, globose head, axillary, monadelphous, sessile ovary with numerous ovules, filiform style, and pedonculate glomerule inflorescence. The fruit/pod with 30 per capitulum, lance shaped. Oval or elliptical brown to gray four-seeded measures about 0 to 0.3 long and 2.5 mm broad, compressed, central ring on surface. The roots are slightly woody or wrinkled, tapering taproot repentant with secondary and tertiary branches.
Transverse sectional of root
Transverse sectional of root was represented in [Figure 2]. The Cork exhibits thick, narrow, corrugated, tangentially elongated rectangular walled with 5–12 layers, with a few outer layers crushed or exfoliated. The cortex cells with 6–8 layered, oval, or narrow in upper layer and rectangular and wide in lower layer contains intercellular spaces with fibers and crystals scattering in outermost layer. Medullary rays are wider toward cortex and narrow toward center. Vascular bundles consist of secondary phloem with sieve elements, crystal fibers, and phloem parenchyma traversed by phloem rays in single or groups arranged in tangential bands; thick walled crystal fibers with single or two to four prismatic crystals of calcium oxalate; uni- to multiseriate phloem rays; secondary xylem consists of usual elements traversed by xylem rays; vessels and thick walled parenchyma scattered throughout secondary xylem; uni- to biseriate xylem rays, rarely multiseriate, wider toward secondary phloem and narrow toward center. Secretory canals, simple and compound starch grains are seen. Besides, TS of root under polarizer was focused and depicted in [Figure 3].
|Figure 2: TS of Mimosa pudica root Ck, cork; Ct, cortex; MR, medullary ray; Pa, parenchyma; PCr, prismatic crystal; PF, phloem fibre; SG, starch grain; Ve, vessel; XF, xylem fibre|
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Transverse section of stem
TS of stem was shown in [Figure 4]. Mature stem shows four to eight layered, exfoliated cork of tangentially elongated cell filled with reddish brown contents; below the cork a single layer of collenchyma is present; secondary cortex wide consisting of large, moderately thick walled, tangentially elongated to oval parenchymatous cells, filled with reddish brown contents. A few cells contain prismatic crystals of calcium oxalate, a number of lignified fibers in single or groups scattered throughout the cortex. Pericycle with composite ring of sclerenchymatous fibers, the fibers often being mucilaginous or unlignified. Secondary phloem consisting of usual elements with two to five transversely arranged strips of fibers occur alternating with narrow strips of sieve elements and parenchyma; crystal fibers elongated, thick-walled, containing single crystal of calcium oxalate in each chamber; thick walled radially elongated phloem rays, loosely arranged unlignified thick walled phloem fibers; secondary xylem with usual elements traversed by xylem rays, vessels, tracheids, fibers, radially elongated xylem rays, polygonal pith contains parenchymatous cells with intracellular spaces. An enlarged portion of TS of stem was shown in [Figure 5].
|Figure 4: TS of Mimosa pudica stem Ck, cork; Ct, cortex; MR, medullary ray; Pa, parenchyma; PCr, prismatic crystal; Per, pericycle; PF, phloem fibre; Pi, pith; RBC, reddish brown content; SG, starch grain; Ve, vein; XF, xylem fibre|
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Transverse section of petiole
TS of petiole [Figure 6] is oblong, lunette with two inconspicuous winged projections on the upper region. Many reticulate lacunae were found on both sides of cortex of adaxial petiole, whose parenchyma cells were big and irregular, usually called motor tissue, but the reticulate lacuna in the abaxial side was not as that of adaxial side. The adaxial part of the phloem is with/without a few under developed fibers, while the abaxial part of the phloem was with well-developed fibers. The vascular cylinder compost of four bicollateral vascular bundles associated with arc shaped pericycle. Few layered of thick walled libriform fibers forming a sheath around the vascular cylinder, detailed TS shows single layered epidermis covered with thin cuticle and few multicellular trichomes; parenchymatous cortex with few cavities; sclerenchymatous pericycle band encircles the stellar region containing four vascular bundles, the two large bundles are arranged opposite to each other and the two smaller bundle are arranged horizontally opposite; centrally located parenchymatous pith is present.
|Figure 6: TS of petiole of Mimosa pudica E, epidermis; Hyp, Hypodemis; Per, pericycle; Pa, parenchyma; PCr, prismatic crystal; Ph, phloem; T, trichome; VB, vascular bundle; Ve, vessel; Xy, xylem|
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Transverse section of leaflet
TS of leaflet is dorsiventral and amphistomatic; the TS passing through the midrib is marginally elevated at lower side and comparatively flat on the upper side; single layered upper and lower epidermis embedded with stomata and covered with thin cuticle; a layer of palisade is seen underneath the upper epidermis followed by two layers of spongy parenchyma; a single meistele in the center surrounded by sclerenchymatous band of pericycle. The lamina shows both upper and lower epidermis, single layered palisade is seen following which two to three layers of spongy parenchyma is present; single mucilage canal is seen; vascular bundle with normal elements, prismatic crystals and few veins are present in spongy parenchyma. The TS of lamina through midrib, leaflet with midrib enlarged, lamina with mucilage cavity, enlarged lamina were shown in [Figure 7][Figure 8][Figure 9][Figure 10].
|Figure 10: Enlarged lamina Cu, cuticle; LE, lower epidermis; MuC, mucilage cavity; Pal, palisade; PCr, prismatic crystal; Per, pericycle; Ph, phloem; SP, spongy parenchyma; St, stomata; T, trichome; UE, upper epidermis; VB, vascular bundle; Ve, vein; Xy, xylem|
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The powder microscopical studies identify the presence of trichomes, cork cells, paracytic stomata, mesophyll cells, pitted parenchyma, pitted vessels, bordered pitted vessels, crystal fibers, prismatic crystal, brownish content, and simple and compound starch grains. These characters are shown in [Figure 11].
The values obtained with respect to quantitative analysis are tabulated in [Table 2]. It reveals that the leaf showed paracytic stomata on both surfaces with frequency of stomata on the upper surface [Figure 12] is less as compared to that of the lower surface [Figure 13]. The respective epidermal numbers of adaxial and abaxial surface ranged 680–760/mm–2 and 830–940/mm–2, whereas the stomatal number of adaxial and abaxial surface are 146–160/mm–2 and 210–240/mm–2 and the stomatal index of adaxial and abaxial surface are 17.52/mm–2 and 20.27/mm–2, respectively. Palisade ratio of the leaves was found to be in the range 42–60/mm–2. The vein islet number and vein termination number [Figure 14] of the leaf were found to be 28 and 42, respectively. Physiochemical parameters are used to identify the purity and quality of powdered drug. The organoleptic characters of whole plant powder were identified and given in [Table 3]. The powdered drugs were evaluated for loss on drying, foreign matter analysis, ash value, and extractive value. The physiochemical constants of whole plant of M. pudica were analyzed and tabulated in [Table 4]. The organoleptic characters of n-hexane extract of M. pudica (HEMP) and ethyl acetate extract of M. pudica (EAEMP) are given in [Table 5].
Preliminary phytochemical tests were observed and are tabulated in [Table 6]. From the above accentuation different chemical compounds such as carbohydrates, alkaloids terpenoids, flavonoids, saponins, gums, and mucilage were detected in ethyl acetate extract of M. pudica, whereas sterols, glycoside, tannins, and phenol are absent. The n-hexane extract of M. pudica leaves indicates the presence of carbohydrates, alkaloids, sterols, terpenoid, saponins, gums, and mucilage, whereas glycoside, flavonoids, phenol, and tannins, are absent in n-hexane extract of M. pudica.
|Table 6: Preliminary phytochemical screening of hexane and ethyl acetate extract|
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The quantitative estimation was performed by using UV spectroscopy to detect the amount of quercetin and β-sitosterol equivalent. The standard curve for quercetin and β-sitosterol was plotted in [Figure 15] and [Figure 16]. The linear regression equation for quercetin was found to be y = 0.0005x + 0.0233 and the correlation coefficient was found to be 0.9746. The amount of total flavonoid content present in the extract in terms of mg quercetin/g of extract was found to be 13.39 mg/g by using the linear equation. The linear regression equation for β-sitosterol was found to be y = 0.0008x – 0.042 and the correlation coefficient was found to be 0.982. The amount of total steroid and flavonoid content present in the extract in terms of mg/g was calculated by using the linear equation. Therapeutic efficacy of medicinal plants is due to the quality and quantity of biologically active compound present in the crude drug. Quantitative phytochemical screening of whole plant of M. pudica disclosed that the total quercetin equivalent present in 1 g of ethyl acetate extract of M. pudica was found to be 13.39 mg, whereas total β sitosterol equivalent in n-hexane extract of M. pudica was found to be 27.5 mg. The phytosterol content is higher than that of flavonoids content; instead both the flavonoids and phytosterol are affirming the therapeutic efficacy of the M. pudica.
| Discussion|| |
The consequences of preceding literature stated that the leaf surface is smooth in adaxial region and bristle like appearance in abaxial region; digitatively arranged, bipinnately compound, and hairy margin; rachis enclosed with ascending bristles; linear glabrous leaflet weakly bristle stem, sparsely prickly fibrous bark; bisexual flower, campanulate calyx, lance shaped pod; tapering root. Our findings accommodate a couple of freakish information regarding morphological variation of this taxon such as white wax-like substances on leaf surface that correlates with statement. Obtuse to truncate base; apiculate apex; ciliate to entire margin; hispidulous erect stem, weak bristly peduncled flowers; pinkish red especially on the inner faces of the lobes during anthesis; branched taproot with longitudinal rough surface. This crucial feature provides referential information for correct identification and helps in checking adulterants and substituent’s (N. gracilis and A. julibrissin). These observations will help in differentiating the whole plant of M. pudica from closely related species such as Mimosa pigra, Mimosa hamata, and Mimosa himalayan of the same genus and family. Metcalfe and Chalk stated that anatomical characters are delicious to differentiate genus and species. The literature, revealed similarities with statement of literature that describes root with 5–12 layered cork, secondary phloem and xylem consist of thick walled parenchymatous cell with both simple and compound starch grains; tannins in secondary cortex. Stem with reddish brown contents; uniseriate to multiseriate phloem rays; uniseriate to biseriate xylem rays; vessels, tracheids and fibres; polygonal pith with intracellular parenchymatous cells; dorsiventral leaf with thin cuticle, four bicollateral vascular bundles. An attempt has been made by, on investigation of anatomy of petiole confessed ring of sclerenchymatous sheath, many reticulate lacuna, libriform fibers, few tracheary elements which are stretched and crushed are discernible when their lignified wall is stained. These above premeditated reports contextually similar to our statement and observations recorded in disparate literature. The diverse phylogenetic nature of the medicinal plant is closely related to nature and is mostly affected due to environmental alterations. Thus, reiterated swotting of medicinal plant is obligatory. The results of our findings are similar to that of the observations recorded in different literature. Our studies exclusively identified leaves with paracytic stomata, smaller vein, presence of glands, mucilage, secretory elements and canals scattered in root; that correlates with the existing literature of Metcalfe and Chalk. A single layer of collenchyma cells in stem portion was seen that were similar with the findings. Though Mammiform papillae is an important anatomical character in genus of Mimosa, they were not identified in species of M. pudica in our studies. Further powder and quantitative microscopy confer its identity and purity. Metcalfe and Chalk stated that anatomical characters are delicious to differentiate genus and species. With the worldwide increasing popularity and acceptance of herbal medicines the classical tool like quantitative microscopy is needed for purity assessment and quality control of plant products. Based on meticulous study the epidermal number and size in the abaxial surface is higher than the adaxial surface. The leaves are basically amphistomatic in M. pudica, since stomata are present on both side of the leaves. Based on the investigational reports, the quantitative constants were predicted significantly and they are considered as valued taxonomical facts of whole plant of M. pudica. Fluorescence studies of powder and extracts help to identify particular drug by the use of estimates of intensity of fluorescence. The comparison of the unknown should be made with a sample of known identity. It is also one of the reliable pharmacognostical aids in the identification of authentic samples and recognizing of adulterants. The shelf life of medicinal plants depends on moisture temperature and other environmental factors. It was argued that plant materials with a level of moisture higher than 8% favor the invasion of insects and moisture content of plant materials higher than 15% stands the risk of contamination with bacteria and fungi. The outcome of this research reveals that the moisture content of the dried samples of the whole plant M. pudica (3.30 + 0.03%) moisture and could be prescribes with little risk of insects and microorganism invasion, have increasing its shelf life. The level of ash contains of the plant suggested the availability of inorganic constituents which is subsequently confirmed by the considerably high content of mineral composition. The high level of ash obtained during incineration reveals the presence of high mineral composition in this plant.
Various medicinal properties have been attributed to natural resources. Tremendously medicinal plants constitute the main source of new pharmaceuticals and health care products. Extraction and phytochemical analysis of several active components from these medicinal plants have given birth to some high activity profile drugs. The use of traditional medicine is wide spread in India. A growing body of evidence indicates that secondary metabolities play a vital role in human health and may be nutritionally important. It is believed that crude extract from medicinal plants was highly biologically active isolated compound due to their synergetic effect. The present study revealed the presence of numerous therapeutically effective secondary metabolities including carbohydrates, proteins, alkaloids terpenoids, flavonoids, saponins, sterol, terpenoid, saponins, etc. These metabolites might make the plant useful in curing different illness. Innumerable traditional and ethnomedical claims (in the treatment of boils, glandular swellings, Rheumatoid arthritis, and fibroadenoma) of M. pudica were unproven scientifically so interrogation of qualitative and quantitative analysis galvanize in further research in proving ethnomedical claims of M. pudica Linn.
| Conclusion|| |
This research concludes the nature of pharmacognostical, physiochemical, and phytochemical characters of M. pudica for its identification and to determine its quality and purity from genetically distant species for its therapeutic importance. Therapeutic efficacy of plant is generally eminent by the nature of secondary metabolites. At present free phytosterols and flavonoids exerted various sources of fortified foods, medicine, and dietary supplements. These qualitative and quantitative analyses play a critical role in the efficacy of therapeutic action of M. pudica.
The research activities and manuscript writing were contributed by H. Rajalakshmi. Reviewing and revising the final version of the manuscript were contributed by Dr. A. Sethuramani.
Financial support and sponsorship
Conflicts of interest
There are no conflicts of interest.
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[Figure 1], [Figure 2], [Figure 3], [Figure 4], [Figure 5], [Figure 6], [Figure 7], [Figure 8], [Figure 9], [Figure 10], [Figure 11], [Figure 12], [Figure 13], [Figure 14], [Figure 15], [Figure 16]
[Table 1], [Table 2], [Table 3], [Table 4], [Table 5], [Table 6]