|Year : 2012 | Volume
| Issue : 2 | Page : 107-111
Pharmacognostic and phytochemical studies on Ficus Microcarpa L. fil
Mohan G Kalaskar, Sanjay J Surana
Department of Pharmacognosy, R. C. Patel Institute of Pharmaceutical Education and Research, Shirpur, Dhule, Maharashtra, India
|Date of Web Publication||20-Sep-2013|
Mohan G Kalaskar
Department of Pharmacognosy, R. C. Patel Institute of Pharmaceutical Education and Research, Shirpur, Dhule - 425 405, Maharashtra
Source of Support: None, Conflict of Interest: None
Background: Ficus microcarpa L. fil. (Syn: Ficus retusa) (Moraceae) is well-known traditional medicinal plant. The bark is used for diverse health ailments in traditional and folklore remedies.
Aims: The present study was undertaken to lay down pharmacognostical and phytochemical standards.
Materials and Methods: Pharmacognostic studies on fresh, dried and powdered bark was carried out to determine it's morphological, anatomical, and phytochemical diagnostic features. Furthermore, major phytoconstituents were identified from the extracts with the help of high performance liquid chromatography (HPLC) study.
Results: The morphology showed to be soft, brittle, rough, shallow vertical, irregularly oriented fissures, curved surface; with splintering, laminated fracture. Microscopically F. microcarpa showed all general characteristics of bark with some distinct differentiation. The phellem is thin and even, phelloderm few cell and rectangular and followed by smaller sclerides, the phloem rays are broad, multi-serrate and showed the scattered bundles of sclerides. The fluorescence and physicochemical standards for bark were established. HPLC analysis showed the predominant presence of therapeutically important phytoconstituents such as oleanolic, betulinic acid, lupeol, β-sitosterol, catechin, and gallic acid.
Conclusion: The bark of F. microcarpa considered equivalent to other Ficus species, such as Ficus virens, Ficus infectoria, Ficus arnottiana, Ficus lacor, and Ficus talboti. However, there is no pharmacognostical and phytochemical reports on F. microcarpa to authenticate and differentiate it from similar species. Present work has described pharmacognostical and phytochemical characteristics of F. microcarpa and diagnostic features to differentiate it.
Keywords: Ficus microcarpa, Ficus retusa, pharmacognostic, phytochemical study
|How to cite this article:|
Kalaskar MG, Surana SJ. Pharmacognostic and phytochemical studies on Ficus Microcarpa L. fil. Ancient Sci Life 2012;32:107-11
| Introduction|| |
Ficus microcarpa L. fil. (Syn: Ficus retusa) (Moraceae) is well-known traditional medicinal plant known commonly known as Indian Laurel, distributed across the hotter parts of India. Its dried leaves, aerial roots, and bark are used in diverse ailments in traditional and folklore remedies. The phytochemistry of the plant is well-studied and the literature supports the presence of triterpenoids, fatty alcohol, steroids, coumarin, flavane, 4-hydroxybenzoates, and a carotenoid-like compound in Ficus microcarpa bark (FMB). The antioxidant and antibacterial potential plant phenolics such as protocatechuic acid, catechol, p-vinylguaiacol, syringol, p-propylphenol, vanillin, and syringaldehyde have been identified in FMB. The latex of the plant has been reported to contain chitinase, giving rise to antifungal properties. Liu et al., reported antitussive and expectorant potential of F. microcarpa. Traditionally, the bark has a reputation of being efficient in the treatment of diabetes, ulcers, burning sensation, hemorrhages, leprosy, itching, liver disease, and toothache. Despite extensive traditional utilization and source of important phytoconstituents, the present plant has not been well standardized for pharmacognosy and phytochemistry. The present study was undertaken to lay down pharmacognostical and phytochemical standards for F. microcarpa.
| Materials and Methods|| |
FMB was collected from Lonawala, Maharashtra, in August 2009 and authenticated by the Botany Department of SSVPS College, Dhule, Maharashtra. The bark was washed, cleaned, shade dried, powdered, and passed through a 40-mesh sieve. It was stored in a tightly closed container. A voucher specimen R C Patel (it is the college code for herbarium library) (RCP-42) was authenticated and deposited in Pharmacognosy Department.
The bark was separated from other parts, washed, cleaned, and dried for further use. The following macroscopic characters of the fresh leaves were noted: Color, odor, taste, size and shape, touch, texture, and fracture [Figure 1].
|Figure 1: Ficus microcarpa: (a) External features of bark. (b) Transverse section of bark. (c) Enlarged portions. Ph = Phellem, Pd = Phelloderm, Sc = Sclereids, LF = Lignified fibers, SSc = Scattered Sclereids, MR = Medulary rays and Phl F-Phloem fiber; Phl P-Phleom Parenchyma|
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The free hand thin transverse sections of the fresh bark taken and treated with the different staining agent and observed for the general and specific microscopic characteristic [Figure 1]. Furthermore, small quantity of the powdered leaves was cleared, mounted, and observed for diagnostic powder characteristics [Figure 2]. Photomicrographs of the microscopical sections were taken with the help of Motic photomicroscope provided with Motic Images plus 2.0 software.
|Figure 2: Powder microscopy of Ficus microcarpa bark. (a) Calcium oxalate crystal. (b) Phloem fiber. (c) Stone cell/sclereids. (d) Lignified fibers|
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Physicochemical and fluorescence analysis
Bark powder was analyzed for their fluorescence analysis according to Kalaskar et al. The dried bark material was analyzed under visible light, short, and long ultra violet (UV) light after treatment with various acids, alkalis, other reagents [Table 1] and [Table 2]. Physicochemical parameters such as a percentage of ash, extractive values, foreign matter content, and loss on drying were determined in triplicates as per Kalaskar and Surana. The mean and standard deviation (±SD) from triplicate samples were summarized in [Table 3].
|Table 2: Behavior of Ficus microcarpa bark powder with different chemical reagents|
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The FMB extracted with four successive solvent to get different polarity solvents, to screen phytoconstituents based on their polarity. These extracts were evaluated for qualitative phytochemical analysis. Furthermore, the presence of triterpenoids, steroids, and polyphenols in FMB extract were estimated qualitatively by HPLC, as per the method described by Pawar and Surana, with some modification. A Shimadzu HPLC system with LC-10AT, UV detector (Spectra System UV 1000), and Luna C18 reverse-phase column (250 mm × 4.6 mm, i.d. particle size 5 μ) in an isocratic mode was used with the specification given in [Table 4].
| Results|| |
The bark has a thickness of about 4-7 mm and grayish to dark brown outside and light brown inside. It is soft, brittle, with a rough surface with shallow vertical, irregularly oriented fissures, and curved. The fracture was found to be splintery and laminated [Figure 1].
Bark differentiated into outer bark as periderm and inner bark or secondary phloem. Periderm well differentiated into phellem (Ph) and phelloderm (Pd). Ph zone is thin and even in transaction. Ph cells are organized into thin tangential compressed cells, rectangular, and the older layers exfoliate. The periderm consists of continuous bands of sequential periderm, and originates from the secondary phloem. Ph cells are homogeneous thin walled rectangular and suberized while Pd is wide and distinct consist of 5-7 layers if containing chloroplast or tannin. Pd cells (23.87 μ × 3.45 μ) are turned into cubical sclereids (Sc) (49.51 μ × 21.24 μ) arranged 2-5 layers, lignified [Figure 1].
Secondary phloem is differentiated into inner intact non-collapsed zone and outer collapsed phloem zone. The collapsed phloem zone consists of wide dilated modular rays, few phloem fibers (17.70 μ), lignified fibers (25.75 μ), and cubical Sc are scattered in this region [Figure 1] In the non-collapsed zone phloem elements occur in small clusters and consist of phloem fibers, sieve tube members, companion cells, and phloem parenchyma. The phloem rays are uniseriate or multi-seriate. They are homocellular or heterocellular. Laticifers are found in the inner bark. Starch and prismatic calcium oxalate is abundant in most of the axial parenchyma cells.
Powder drug analysis
The microscopical powder analysis showed the presence of lignified phloem fibers, sclerides in the form of stone cells, rhomboidal prisms of calcium oxalate and abundant starch grain in parenchymatous cells observed [Figure 2].
The fluorescence analysis and behaviors of powder with different chemical reagents were studied [Table 1] and [Table 2]. The physicochemical standards are important to check the quality, purity, and adulteration of given crude drug. The foreign matter, loss on drying (LOD), ash, and extractive values were determined and summarized in [Table 3].
The preliminary phytochemical analysis of bark extracts showed the medicinally potential constituents include carbohydrates, tannins, flavonoids, steroids, and triterpenoids. The HPLC study of successive FMB extracts identified predominant phytoconstituents such as triterpenoids and steroids such as oleanolic acid (OA), betulinic acid (BA), lupeol, and β-sitosterol in pet-ether extract. Triterpenoids such as OA, BA and polyphenol, catechin (Cat) were identified in ethyl acetate extract. The ethanol extract showed the presence of polyphenols, Cat, and gallic acid (GA) while phenolic acid, GA was identified in aqueous extract [Table 4].
| Discussion|| |
The bark is one of the important medicinal plant parts, used traditionally for therapeutic purposes. The bark is very complex in structure and has the potential of containing many primary and secondary metabolites. The complex structure of the bark can be utilized for correct identification to maintain the quality and purity of the drug. The different Ficus species are utilized extensively in Indian traditional medicinal systems. Among these species F. microcarpa having credentials in treatment of diabetes, ulcers, burning sensations, hemorrhages, leprosy, itching, liver disease, and toothache. The FMBs considered equivalent with other species such as Ficus virens, Ficus infectoria, Ficus arnottiana, Ficus lacor and Ficus talboti.,, Hence, it is always a problem to identify the correct species without sets of standards.
Thus, the present work was taken up with an objective to lay down detail pharmacognostical and phytochemical standards that will contribute significantly to quality control of medicinally useful F. microcarpa.
Microscopy of F. microcarpa showed all general characteristics of bark with some distinct differentiation. The bark has yellowish brown to reddish brown color with very few distinct irregular shallow fissures. Microscopically the Ph is thin and even, which is not so in other species of Ficus. Furthermore, the Pd few cell and rectangular and followed by sclerides as stone cell layer. The sclerids are smaller when compared with other species. In the upper phloem region, the phloem rays are broad, multiserrate and showed the scattered bundles of sclerides. The phloem fibers are distinct and solitary, lignified. The phloem rays and parenchyma showed the presence of starch and rhomboidal calcium oxalate crystals.
The ash values of a drug give an idea of the earthy matter or inorganic composition and other impurities present along with the drug. Extractive values are useful for determination of exhausted or adulterated drug. Thus ash, extractive values, fluorescence analysis will be helpful in the identification and authentication of plant material. The preliminary phytochemical screening showed the presence of different phytoconstituents groups such as steroids, triterpenoids, flavonoids, polyphenolic, and tannin. HPLC analysis showed the predominant presence of therapeutically important phytoconstituents such as oleanolic, BA, lupeol, β-sitosterol, Cat, and GA.
The present study provides in-depth microscopical features, and identification of biologically active phytoconstituents, which also provide pharmacopoeia standards for easy identification of the F. microcarpa and hence differentiating it from closely related species.
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[Figure 1], [Figure 2]
[Table 1], [Table 2], [Table 3], [Table 4]