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PURPOSE: The Ficus reticulata fruit mucilage is an edible fruit and its pulp is rich in mucilage. But there are no reports on isolation and characterization of Ficus reticulata fruit mucilage. Hence, the present study was designed to isolate, purify and characterization of Ficus reticulata fruit mucilage. METHODS: The Ficus reticulata fruit mucilage was extracted, purified and identified by official methods. The isolated mucilage was characterized for physical, chemical and flow properties. The mucilage was further characterized by Fourier transform Infrared Spectroscopy. RESULTS: The Ficus reticulata fruits gave sufficient quantity of mucilage and it has good physical, chemical and flow properties. The Fourier transform Infrared Spectroscopic study revealed that the mucilage has characteristic peaks. CONCLUSIONS: It was concluded that the Ficus reticulata fruit mucilage can be used as matrix forming material in pharmaceutical dosage forms.
Key words: Ficus reticulata; mucilage; isolation; characterization;
Ficus reticulata is laticiferous tree grows up to 12.2 m tall with reddish grey bark, alternate leaves, stipules ovate-Ianceolate, pubescent, 1.25 to 2.5 cm long, petioles 2.5 to 5.0 cm long. Fruits borne in clusters on the main trunk and leafless short branches, sub globose or pyriform, 2.5 to 5.0 cm in diameter, they become red when ripe (1). The taxonomical classification of Ficus reticulata was shown in Table 1. The dried uncooked Ficus reticulata fruits give 275kcal (1151.4 kJ) of energy per 100 g (3.5 oz) (2). The nutritional value of Ficus reticulata fruits was shown in Table 2. These fruits were already proved its medicinal values (3-6) in relieving fever, Pain and Inflammation; Wound healing, Blood purifying and urinary problems. More over the fruits were used in conditions like Impotence and decreased sperm count. Presently the use of natural gums and mucilages is gaining importance in pharmaceutical formulations as an excipient in dosage forms. Synthetic polymers have certain drawbacks viz., high cost, non-renewable sources, side effects, toxicity, causing environmental pollution during their synthesis, non-biodegradable ( where as biodegradable synthetic polymers are costlier) and less patient compliance (7, 8) etc. While natural plant based materials are economical, devoid of side effects, biocompatible, biodegradable, renewable source, environmental-friendly processing and better patient compliance (9-13). Mucilages are polysaccharide complexes formed from sugar and uronic acid units. Mucilages form slimy masses in water, are typically heterogeneous in composition. Upon hydrolysis mucilages gives Arabinose, Galactose, Glucose, Mannose, Xylose and various Uronic acids (14). Mucilages are obtained mainly from fruits have an added advantage of wide acceptability by the patients. The prospects of natural polymers are brighter but even here extensive testing will be required. In present study the fruits of Ficus reticulata were selected for the isolation and purification of mucilage. However there are no reports on isolation, purification and characterization of Ficus reticulata fruit mucilage. Hence, the present study was planned to isolate and characterize mucilage of Ficus reticulata fruits. The data so obtained will be a standardizing parameter for future research work.
MATERIALS AND METHODS
The fresh Ficus reticulata fruits were collected from plants growing in and around Anantapur, India. The plant and fruits were authenticated at the Botany Department, Sri Krishnadevaraya University, Anantapur, India. Ethanol (95%), Acetone, trichloro acetic acid, sodium hydroxide and diethyl ether were procured from SD Fine chemicals (Mumbai, India). All other chemicals used were of AR grade and double distilled water was used throughout the experiments.
Extraction of mucilage
The fresh Ficus reticulata fruits were collected and washed with water. The fruits were opened and seeds were removed. The fruits were crushed and soaked in water for 5-6 h, boiled for 30 min and left to stand for 1 h to allow complete release of the mucilage into the water. The mucilage was extracted using a multi-layer muslin cloth bag to remove the marc from the solution. Acetone (in the volumes of three times to the volume of filtrate) was added to precipitate the mucilage. The precipitated mucilage was separated, dried in an oven at 40Â°C, collected, ground, passed through a # 80 sieve and stored in desiccator at 30Â°C & 45% relative humidity till use (15-17).
Purification of the Mucilage
The crude mucilage (1 %) was homogenized (Potter S homogenizer, Sartorius AG, Germany) with cold dilute trichloro acetic acid solution (5%). The solution was centrifuged (3500 r/min for 20 min), neutralized with sodium hydroxide by drop wise addition and then dialyzed for 30 h against distilled water. The mucilage was precipitated with ethanol (in the quantities of three times the volumes) and washed successively with ethanol, acetone and diethyl ether. The mucilage so obtained was dried under vacuum (less than 1 Torr at 25Â°C for 12 h). The so obtained mucilage was passed through a # 80 sieve and stored in desiccator at 30Â°C & 45% relative humidity till use (18, 19).
Characterization of Mucilage
The collected mucilage was evaluated for physical characteristics (20, 21) viz., Appearance, Odour, Solubility, percentage yield, average particle size, swelling ratio, weight loss on drying, pH, Charring, density and bio burden. All these values were tested in triplicate.
The extracted mucilage was tested for chemical characteristics for identification, test for Carbohydrate, test for Tannins, test for chloride, test for sulphate and test for Uronic acid. The mucilage was also tested for unwanted chemicals (20, 21) viz., foreign matter, heavy metal and Arsenic.
The dried Ficus reticulata fruit mucilage was tested for the flow properties22 viz., Angle of repose, Bulk densities, compressibility index and Hausner's ratio. All these evaluations were carried out as per procedures describe in official books. All these experiments were conducted for five times.
Fourier Transform Infrared (FTIR) Spectroscopy
FTIR spectrums of dried mucilage were recorded on samples prepared in potassium bromide disks using FTIR spectrophotometer (Shimadzu 1601 PC, Tokyo, Japan). Samples were prepared in KBr disks. The scanning range was 500 to 4000 cm-1.
RESULTS AND DISCUSSION
The extracted mucilage was yellow in colour with a characteristic odour and soluble in water produces hage viscous solution. The fruits gave 28 Â±2.457 g of yield per kg. The Average particle size of dried mucilage was 159.32Â±9.543 Âµm. The weight loss on drying was 2.58Â±0.159 and the percentage of swelling was 47.0Â±2.575%. The dried mucilage was melted and charred at and above 152Â±6.8510C. The density of 1.0% w/v solution was 1.058Â±0.018 and a pH of 7.1Â±0.111. The mucilage has very negligible bio burden. All these values were shown in Table 3.
The mucilage gave positive test for carbohydrates and uronic acid (common for all mucilages) and negative test for tannins, chlorides and sulphates. The amount of foreign matter was negligible. The heavy metal concentration was also found to be within the limits. All these values were shown in Table 4.
The dried Ficus reticulata fruit mucilage has excellent flow properties as the Angle of repose was 25.77Â±1.680. The Bulk density values were considered for calculating compressibility index and Hausner's ratio, which were 20.8Â±0.04% and 1.26Â±0.03 respectively. All these values were shown in Table 5.
The FTIR spectrum of Ficus reticulata fruit mucilage showed sharp and characteristic peaks at 3155.3, 2935.5, 2885.3, 2098.4, 1635.5, 1234.4, 1110.9, 991.3 and 852.5cm-1. The FTIR spectral values and probable bonds present were represented in Table 6 and shown in Figure 1.
This study revealed that Ficus reticulata fruit mucilage has good physicochemical characteristics with good flow properties. The mucilage can be used as binder in tablet formulations and as matrix forming material in pharmaceutical dosage forms.
Table 1: Taxonomical Classification of Ficus reticulata
Ficus L. - fig
Table 2: Composition of 100 g of dried fruits of Ficus reticulata
Dietary fiber Â
5.6 g Â
Thiamin (Vitamin B1)
0.040 mg Â
Riboflavin (Vitamin B2)
0.050 mg Â
Niacin (Vitamin B3)
0.324 mg Â
Patothenic acid (VitaminB5) Â
Vitamin (Vitamin B6)
Folic acid (Vitamin B9) Â
540 mg Â
Table 3: Physical characterization of Ficus reticulata fruit mucilage
Slowly soluble in water produces hage viscous solution
*Percent yield (g /kg)
*Average particle size (Âµm)
*Weight loss on drying (mg)
*Swelling Index (%)
*Density of liquid (1.0% w/v)
*Microbial count (cfu/g)
Bacteria: 6 ; Fungi: 2
Cfu = Colony forming units; *= experiments were conducted in triplicates (n=3)
Table 4: Chemical characterization of Ficus reticulata fruit mucilage
Mounted in 96% ethanol
Transparent angular masses
Mounted in ruthenium red
Particles stained red
Mounted in Iodine solution
Particles stained blue
Mollish test (for Carbohydrates)
Ferric chloride test (for Tannins)
Silver-nitrate test (for chlorides)
Barium chloride test (for Sulphates)
Test for Uronic acid
Test for foreign matter (%)
Test for heavy metal (lead)
Test for Arsenic
NMT = Not more than
Table 5: Flow properties of Ficus reticulata fruit mucilage
Angle of repose (qÂ°)
Loose Bulk density (g/cm3)
Tapped bulk density(g/cm3)
Carr's Index (%)
Number of trials (n)=5
Table 6: FTIR spectral data of Ficus reticulata fruit mucilage
Specific type of bond
Nâ”€H bond ( ammonium ions)
Câ”€H bond (alkyl, methyl)
Nâ”€H bond ( ammonium ions)
Câ”€N bond (any)
Câ•N bond (any)
Nâ”€H (primary amine)
Câ”€O bond (ethers, aromatic)
Câ”€O bond (alcohols, secondary)
Câ”€H bond (vinyl, mono substituted alkenes)
Câ”€H bond (aromatic, para-di substituted Benzene)