|Year : 2013 | Volume
| Issue : 4 | Page : 153-157
Standardisation of Ficus carica L.: In homoeopathic perspective
P Subramanian1, P Padma Rao2, T Sheshashena Reddy1, P Sudhakar2, PR Reddy3, Santhosh Kr. Sharma4, D Suresh Baburaj4
1 Chemistry Section, Drug Standardisation Unit, Hyderabad, India
2 Pharmacognosy Section, Drug Standardisation Unit, Hyderabad, India
3 Department of Botany, Osmania University, Hyderabad, India
4 Drug Standardisation Section, Central Council for Research in Homoeopathy, New Delhi, India
|Date of Submission||27-Jun-2013|
|Date of Acceptance||11-Dec-2013|
|Date of Web Publication||26-Dec-2013|
Chemistry Section, Drug Standardisation Unit, O.U.B 32, Road No. 4, Habsiguda, Hyderabad - 07
Source of Support: None, Conflict of Interest: None
Background: Ficus carica L. known as 'Common Fig' in English and 'Anjir' in Hindi, is a well known tree belonging to the family Moraceae. The unripe fruits are used as medicine in Homoeopathy.
Objective: The pharmacognostic and physico-chemical studies are carried out to facilitate use of correct species and standard raw materials.
Material and Methods: Pharmacognostic studies on unripe fruits of authentic samples of Ficus carica L. have been carried out; Physico-chemical parameters of raw drug viz., extractive value, ash values, formulation, besides weight per mL, total solids, alcohol content along with High Performance Thin Layer Chromatography (HPTLC) and Ultra violet-visible studies have been worked out for mother tincture.
Results: The unripe fruit is a syconium, globose, light green and externally tomentose. Stomata are anomocytic or actinocytic. Trichomes are unicellular conical. The 1-layered epidermis in transection (T.S.) is marked by hair bases with conspicuously large encircling cells. Inner cortex is aerenchymatous and possesses laticiferous tubes. Vascular bundles are present in the cortex towards inside. In T.S. inner fruit wall possess female flowers. Physico-chemical properties and HPTLC values of this plant have been standardised.
Conclusion: The powder microscopic features and organoleptic characters along with anatomical and physico-chemical studies are diagnostic to establish the standards for the drug.
Keywords: Ficus carica L., Figs, High performance thin layer chromatography, Syconium, Laticifer, Pharmacognosy
|How to cite this article:|
Subramanian P, Rao P P, Reddy T S, Sudhakar P, Reddy P R, Sharma SK, Baburaj D S. Standardisation of Ficus carica L.: In homoeopathic perspective. Indian J Res Homoeopathy 2013;7:153-7
|How to cite this URL:|
Subramanian P, Rao P P, Reddy T S, Sudhakar P, Reddy P R, Sharma SK, Baburaj D S. Standardisation of Ficus carica L.: In homoeopathic perspective. Indian J Res Homoeopathy [serial online] 2013 [cited 2019 Sep 17];7:153-7. Available from: http://www.ijrh.org/text.asp?2013/7/4/153/123387
| Introduction|| |
Ficus carica L. popularly known as 'Common Fig' in English and 'Anjir' in Hindi is a tree of moderate size, belonging to the family Moraceae. It is native to Western Asia and is cultivated in many temperate and sub tropical regions.  The unripe fruits are used as medicine in Homoeopathy. Medicinally, it is useful as aperient, emollient, demulcent, nutritive, for removal of stones in kidney or bladder, cures piles, gout, disorders of liver and removal of warts. ,, In general, figs are easy to digest, rich in vitamins, fibre and minerals.  Chemically the plant contains some bioactive compounds such as psoralen, bergapten, chrysin, galangin, anthocyanin, β-amyrin, quercetin, fucosterol, nicotinic acid, riboflavin, ficusin, germacrene D, agelicin, linalool, linalool oxides, cinnamic aldehyde and lupeol acetate. ,, They are good source of flavonoids and polyphenols  and β-carotenes.  It is reported to have anti oxidant, antibacterial, hypoglycaemic, cancer suppressive, hypotriglyceridaemic and anthelmintic properties. 
The pharmacognostic studies on fruits of Ficus carica L. in general is rare  and on unripe fruit in particular is absent. Considering the medicinal importance of the drug and absence of earlier pharmacognostic studies on unripe fruits, the present study is undertaken.
| Material and Methods|| |
The unripe fruits of Ficus carica L. was supplied by Survey of Medicinal Plants and Collection Unit, Nilgiris, Tamil Nadu, under CCRH. The fruits were slightly boiled and fixed in F.A.A. (Formaldehyde - Acetic acid - Alcohol) and processed for microtomy (Paraffin method), sectioned, stained and permanent slides prepared as per method of Johansen.  The surface peels of the fruit were obtained by gently scraping with razor blade. The powder microscopy characters were studied by boiling the powdered drug in distilled water, stained in saffranine and mounted with glycerine. Photomicrography was done with Olympus BX 53 trinocular research microscope attached with Sony digital camera.
The air-dried unripe fruits were coarsely powdered to 10/44 (sieve size) and subjected to determination of moisture content (loss on drying at 105°C), total ash, water soluble ash, acid insoluble ash, extractability in different solvents, physico-chemical constants; UV aspects of mother tincture, including its preparation by percolation method given in Homoeopathic Pharmacopeia of India  .
High Performance Thin Layer Chromatography (HPTLC) Analysis
25 mL of mother tincture was evaporated on water bath to remove alcohol. The residue was extracted with 3 × 25 mL chloroform. Concentrated chloroform extract was used for the HPTLC study. The concentrated chloroform extract was spotted in the form of band of width 4 mm with a Camag microlitre syringe on pre-coated Silica gel Aluminum plate 60F-254, (5 × 10 cm with 0.25 mm thickness; Merck, Darmstadt, Germany) using a Linomat IV sample applicator (Camag, Muttenz, Switzerland, supplied by Anchrom technologists, Mumbai). A constant application rate of 6 mL/sec was employed. The slit dimension was kept at 4 × 0.45 mm and 20 mm/sec scanning speed was employed. The mobile phase consisted of Chloroform: Methanol (9:1 v/v) and 10 mL of mobile phase was used for chromatography. Linear ascending development was carried out in a 10 × 10 cm twin trough glass chamber (Camag, Muttenz, Switzerland) saturated with the mobile phase at room temperature (25 ± 2° C) for 20 minutes. The length of the chromatogram run was 8 cm and subsequent to the development, the TLC plates were dried in a current of air with the help of hot air dryer in a wooden chamber with adequate ventilation. Densitometric scanning was performed (Camag TLC scanner III) at 254 nm and 366 nm by reflectance scanning and operated by Wincats software (Camag) present in the system. ,,
| Observations and Results|| |
Unripe fruit is syconium, produced into a globose, clavate structure, 2-8 cm in diameter, bearing pistillate flowers, pale to light green, externally tomentose, with a pore over the top at centre. Plant photograph with fruit branch is given as [Figure 1].1.
The epidermal cells over the surface polygonal isodiametric, 5-6 sided, sides thin, straight to curved, cytoplasm slightly dense. Stomata are anomocytic or actinocytic. Trichomes are unicellular conical hair; walls thick, contents dense, surface verrucose [Figure 2].2 Distribution: Common, all over the surface.
|Figure 2: (1) T.S. of receptacle X 33, (2) Fruit epidermis (Surface) X 59, (3) T.S.of receptacle showing L.S. of female flower& floral appendeges X 55|
Click here to view
In T.S. the outer most epidermis is 1-layered, cells tabular, radially long, wall thin, contents dense, often interspersed with conical hair and stomata [Figure 2].1 and [Figure 3].3. The epidermis is marked with large hair bases having surrounding cells projecting above the surface [Figure 2].1 and [Figure 3].3. Hypodermal cortex is 8-10 layered with peripheral 2-3 layers with smaller cells which are tabular, arranged in rows and often containing sphaeraphides [Figure 3].3 and [Figure 4].2. The inner cortex is up to 40 layers, aerenchymatous, having oval to spherical and elongated cells with thin walls and dense contents [Figure 2].1, [Figure 3] and [Figure 3].1, [Figure 2]. The cortex is interspersed with laticiferous canals which are unbranched [Figure 3].3 and [Figure 3].1. The vascular bundles arranged in the form of a ring frequently occur towards inside [Figure 4].3. The vascular bundles are endarch, conjoint, collateral and capped by sclerenchyma. Some bundles occur dispersed irregularly in the aerenchyma or middle cortex [Figure 3].1 and [Figure 2]. The innermost layer of receptacle consists of inner epidermis covered with unicellular conical hairs over its surface [Figure 2].1, [Figure 3] and [Figure 3].1 The inner wall of receptacle possesses female flowers  [Figure 2].3.
|Figure 3: (1) T.S. of receptacle (inner most side) X 123, (2) T.S. of receptacle (median region) X 107, (3) T.S. of receptacle (peripheral) X 113|
Click here to view
|Figure 4: (1) T.S. receptacle at peripheral with latex tube X 401, (2) T.S. of receptacle at hypodermal region with sphaeraphides X 329|
Click here to view
The flowering stalk arises from thalamus. The female flowers consist of outermost tepals in L.S. having 1-layered epithelium like epidermis and underlying mesophyll [Figure 2].2. The mesophyll is 3-4 layered and with closely packed cells [Figure 2].3. The central gynoecium is a flask - shaped structure consisting of epidermis and underlying parenchyma with a vascular bundle at its base [Figure 2].3. Apically the gynoecium is narrowed into a stalk covered by a glandular epithelium.
The fruit in T.S. show many floral appendages mostly the tepals and central area of the female flowers in various planes.
- Pieces of thalamus with epidermis, hypodermis and cortex
- Numerous fragments of conical hair either whole or broken
- Pieces of cortex with latex tubes
- Fragments of stalk of gynoecium with dense contents
- Pieces of ovary with an ovule
- Pieces of tracheary tissue showing vessels/tracheids with helical thickenings
- Pieces of epidermis with anomocytic stomata and conical hair
- Pieces of black tanniniferous masses.
Colour - Creamish yellow
Touch - Slightly coarse
Odour - Not characteristic
Taste - Not characteristic.
The determined data under the physico-chemical study for the raw drug is summarised in [Table 1] and that of mother tincture preparation and its standardisation in [Table 2] and [Table 3] respectively.
Qualitative Phytochemical Tests
Moisture content reveals the presence of water in the plant and also some volatile organic matter. Results of physico-chemical studies are summarised in [Table 1], [Table 2] and [Table 3].
HPTLC Finger Printing
The profile of chromatographic separation scanned at 254 nm, reveals 10 spots [Figure 5] out of which 9, 7 and 5 spots possess maximum composition with R f at 0.73, 0.59 and 0.45 respectively. While, chromatogram scanned at 366 nm, revealed 6 spots with 4, 5 and 6 spots showing maximum composition at R f 0.86, 0.70 and 0.60 respectively. It is evident from the data that these are characteristic for the studied drug, which will help in identification and authentication of the mother tincture. This is considered as valuable standard in pharmacopoeia. At 254 nm, ten spots appear at R f 0.17, 0.21, 0.28, 0.37, 0.45, 0.54, 0.59, 0.66, 0.73 and 0.85 (all are in brown colour) [Figure 5] with various concentrations while at 366 nm, five spots at R f , 0.23 (red), 0.43 (light yellow), 0.60 (red), 0.70 (blue) and 0.86 (red).
|Figure 5: HPTLC finger printing of Ficus carica mother tincture chloroform extract scanned at 254 nm|
Click here to view
| Discussion|| |
The unripe fruit is green  which is presently confirmed. The fruit is a syconium, produced into a globose or pear-shaped structure, 4-8 cm bearing pistillate flowers as reported earlier  and externally tomentose.
The epidermal surface of the fruit consists of polygonal isodiametric cells with straight to curved anticlinal walls as reported earlier.  Stomata are reported as anomocytic  and is presently confirmed, besides a few actinocytic types.
The epidermis in T.S. is single layered with oval to barrel-shaped cells;  however, it is 1-layered with tabular and radially elongated cells interspersed with conical hairs. Further, the epidermis is intermittently marked with large encircling cells around hair bases projecting over the surface.
Hypodermal region is earlier reported as 3-5 layered collenchymatous tissue but it is 8-10 layered parenchyma tissue with peripheral 2-3 layers having closely packed tabular cells often containing sphaeraphides as was also found.  The mesocarp region is reported as having large, thin walled oval to polygonal or squarish parenchymatous cells without intercellular spaces.  But presently the inner cortex is extensive, aerenchymatous with cells possessing dense contents.
The cortex also possesses laticifers which are tabular, elongated with few branched  which is also presently found except they are unbranched. Vascular bundles occur dispersed in the cortex  In L.S. vessels were found to possess helical or reticulate thickenings which is presently confirmed. Besides the receptacle on its inner side possess female flowers  cut in various planes.
The identification of the active principles and investigation of mother tinctures are quintessential in order to ensure safe, effective and constant activity. Chemical profiling is a versatile technique and can be made to good use in standardisation. Pharmacopoeia largely relies on chemical assay and physical constants for routine standardisation. HPTLC finger printing is a modern, precise and accurate tool to authenticate and characterise the drug, which in turn ensures the quality, purity and guarantees uniformity and reproducibility. Thus, the developed chromatogram will be specific with the selected solvent system and R f value. In terms of area and height of the peak, help in assigning the identity and specificity. UV-visible study of the mother tincture exhibits two prominent peaks, which serve as characteristic standards. Physico-chemical analysis of Ficus carica presented in [Table 1], [Table 2] and [Table 3], helps substantiate the standards for the drug.
| Conclusion|| |
The determined physico-chemical data, spectrophotometric profile along with macro and microscopical characters and methodology employed in the study will help in not only identification, authenticity of the drug but also to ensure batch- to- batch consistency of the products manufactured by the pharmaceutical companies and provide quality medicines to the consumers.
| References|| |
|1.||Anon., Wealth of India, Raw materials, Vol. 4. New Delhi: Publications and Information Directorate, Council of Scientific and Industrial Research, New Delhi, 1980, p. 26-7. |
|2.||Kirtikar KR, Basu BD. Indian Medicinal Plants. Vol. 3. Dehradun: Bishen Singh Mahendrapal Singh. 1980: p. 2329-31. |
|3.||Nadkarni AK. Indian Materia Medica (Revised). Vol. 1. Bombay: Bombay Popular Prakashan. 1976: p. 545-47. |
|4.||Gibernau M, Buser HR, Jurg JE and McKey MH. Volatile compounds from extracts of figs of Ficus carica L. Phytochemistry. 1997;46:241-44. |
|5.||Kazemipoor M, Aorestani, Ansari M. Extraction and determination of biomarkers in Ficus carica L. Leaves from various species and different cultivars by HPLC. J. Liq. Chromatogr. Related Technol. 2012;35:2831-44. |
|6.||Vinson JA, Zubik L, Bose P, Samman N, Proch John. Dried fruits: Excellent in vitro and in vivo antioxidants. J. Am. Coll. Nutr. 2005;24:44-50. |
|7.||Ross JA, Kasum CM. Dietary flavonoids Bioavailability, metabolic effects and safety. Annu Rev Nutr. 2002;22:19-34. |
|8.||Solomon A, Golubowicz S, Yablowicz Z, Grossman S, Bergman M, Gottlieb HE et al., Antioxidant activities and anthocyanin content of fresh fruits of common fig (Ficus carica L.). J Agric Food Chem. 2006;54:7717-23. |
|9.||Anon., Standardisation of Single Drugs of Unani Medicine, Part I, New Delhi: CCRUM, Ministry of Health Family Welfare, Government of India. 1987, p. 7-11. |
|10.||Johansen DA. Plant Microtechnique. New York: Mcgraw Hill Book Co. 1940. |
|11.||Anon., Homoeopathic Pharmacopoeia of India. Vol. 1. New Delhi: Controller of Publications, Ministry of Health and Family Welfare; 1971. |
|12.||Sethi PD. High Performance Thin Layer Chromatography. New Delhi: CBS Publishers and Distributors, 1996. |
|13.||Wagner H, Bladt S, Zgainski EM, Plant Drug Analysis, A Thin Layer Chromatography Atlas. Heidelberg: Springer-Verlag, 1996. |
|14.||Stahl E. Thin Layer Chromatography, A Laboratory Hand Book. Berlin: Springer- Verlag, 1969. |
[Figure 1], [Figure 2], [Figure 3], [Figure 4], [Figure 5]
[Table 1], [Table 2], [Table 3]