PHARMA SCIENCE MONITOR

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1 PHARMA SCIENCE MONITOR AN INTERNATIONAL JOURNAL OF PHARMACEUTICAL SCIENCES NOVEL APPROACH FOR ISOLATION OF MUCILAGE FROM LUFFA ACUTANGULA FRUITS AND SCREENING OF ITS BINDING PROPERTY Neha H. Ravalji*, Biren. N. Shah, D. C. Modi and V. B. Lambole Vidyabharti Trust College of Pharmacy, Umrakh, Gujarat, India. ABSTRACT A microwave assisted extraction technique was developed to optimize the extraction of mucilage from the fruits of Luffa acutangula plant. The plant has been extracted by conventional and microwave assisted methods for the isolation of mucilage. Microwave extraction at 640 W intensity and 5 min heating duration increase 23.07% in the yield of mucilage when compared to 1 h conventional heating method. The products obtained by both the methods were of similar nature chemically. The developed microwave procedure can be used successfully in commercial and routine laboratory isolation of mucilage. The tablets prepared using mucilage of Luffa acutangula having uniform length and thickness. Disintegration time of tablets increased with increase in concentration of tablets. Friability of tablets decreased with increase in binder concentration. Keywords: Microwave, Mucilage, Luffa acutangula, Binder. INTRODUCTION Many extraction techniques have been used for obtaining bioactive compounds from plant origins nowadays. Conventional extraction is usually performed at reflux temperature for several hours, and this method is very time consuming and requires relatively large quantities of solvents. [1] Over the last decade, the demand for new extraction techniques has encouraged the development of alternative extraction techniques [2]. Microwave-assisted extraction is the simplest and the most economical technique for extraction of many plant derived compounds [2]. Microwave assisted extraction can reduce both the extraction time and solvent consumption compared to conventional methods, as demonstrated by the drastically accelerated removal of a variety of compounds from solid matrices. Furthermore, it has the potential to improve extraction quality [3]. Many studies have been undertaken to investigate bioactive compounds of plants deal with microwave-assisted extraction [4]. A microwave is a form of electromagnetic IC Value

2 energy, which falls at the lower end of the electromagnetic spectrum and is defined in a measurement of frequency as 300 to 300,000 Megahertz, corresponding to wavelengths of 1 cm to 1 m [5]. Recently, microwave energy is being used for extraction of phytoconstituents from plants. Microwave extraction follows the same principle as maceration or percolation, but the speed of breaking up of plant cells and plant tissues is much higher. This reduced processing time is an economic advantage and also there is less risk of decomposition or disintegration and oxidation of the valuable plant constituents [6]. Microwave-assisted extraction system for biologically active compounds has many advantages over other conventional extraction methods [7]. The main advantages of microwave assisted extraction over the conventional extraction techniques are reduced solvent consumption, shorter operational times, moderately high recoveries, good reproducibility and minimal sample manipulation for extraction process [8]. In order to reduce the duration involved in their extraction and study the yield, mucilage from the fruits of Luffa acutangula plant was isolated in the present study using microwave method. Mucilage and gums have been known since ancient times for their medicinal uses [9]. In the modern era also they are widely used in the pharmaceutical industries as thickeners, water retention agents, emulsion stabilizers, suspending agents, binders and film formers 10. Binders and suspending agents from natural sources do hold advantages over the synthetic excipients, because of its inertness, cost, and availability [11]. The high cost of imitation polymer and ecological pollution by chemical diligence has made the scientist in budding country to enter into era, in which plant products serve as alternative to synthetic products because of local accessibility, environmental gracious nature, subordinate prices and nontoxic compared to imported synthetic products [9].In recent years, plant derived polymers have evoked tremendous interest due to their diverse pharmaceutical applications such as diluent, binder, disintegrant in tablets, thickeners in oral liquids, protective colloids in suspensions, gelling agents in gels and bases in suppository; they are also used in cosmetics, textiles, paints and paper-making [12]. IC Value

3 These polymers such as natural gums and mucilage are biocompatible, cheap and easily available and are preferred to semi synthetic and synthetic excipients because of their lack of toxicity, low cost, availability, soothing action and non irritant nature [12]. Scientific and technically mucilage is a naturally occurring, high-molecular-weight (,000 and up), organic plant product [13]. Mucilages are most commonly used adjuvant in pharmaceutical preparations [12]. They consist of sugar and uronic acid units [14]. A part from its use in finished medicines, newer uses have been found in the preparation of cosmetics, textiles and paint paper. Hence the demand for these substances is increasing and new sources are getting tapped [10]. Mucilages of different origins are also used in conventional dosage forms of various drugs for their binding, thickening, stabilizing and humidifying properties in medicine [15]. Binders are agents used to impart cohesive qualities to the powdered material during the production of tablet. With the increase in demand for natural mucilages, it has become necessary to isolate and evaluate the newer sources of mucilages to meet the needs [11]. MATERIALS AND METHODS Collection and Authentication: The fruits of Luffa acutangula (Cucurbitaceae) plant were used for the isolation of mucilage. These materials were purchased from the local market of Mandvi, Gujarat and authenticated by Botanist from Science College Bardoli. Isolation of Mucilage by Conventional Procedure: Luffa acutangula fruits (5 g) were powdered for 5 min in a mechanical blender and soaked in distilled water (150 ml) for 24 h in a RB flask. It was boiled for 1 h under reflux with occasional stirring and kept aside for 2 h for the release of mucilage into water. The material was filtered through a muslin bag and hot distilled water (25 ml) was added through the sides of the marc and squeezed well in order to remove the mucilage completely. Equal volume of ethanol was added to the filtrate to precipitate the mucilage and kept inside a refrigerator for one day for effective settling. It was filtered and dried completely in an incubator at 37º C, powdered and weighed. It was subjected to chemical tests to confirm its identity [16],[17]. IC Value

4 Isolation of Mucilage by Microwave Procedure: Luffa acutangula fruits (5 g) were powdered in a mechanical blender for 5 min and soaked in distilled water (150 ml) for 24 h in a 1000 ml beaker. It was kept in a microwave oven (LG Grill Intello wave System, Model No. MG-2381LE) along with a glass tube inside to prevent bumping. It was subjected to microwave irradiation at 640 W intensity for 5 min. The beaker was removed from the oven and kept aside for 2 h for the release of mucilage into water. It was processed in a similar way as explained in the conventional procedure, weighed and chemical tests were carried out. The experiment was repeated several times using various intensities and different durations as shown in Table 1. In each case, the yield was calculated. Physiochemical characteristics of mucilage The physicochemical properties such as swelling index, ash value, identification test, state, colour, odour were determined. The ph of the mucilage was determined using a digital ph meter. The swelling index is the volume in milliliter occupied by 1 g of a material, including any adhering mucilage, after it has swollen in aqueous liquid for 4 h [18]. Results are shown in table 2. Preparation of binder solution The binder solution was prepared by dissolving the mucilage of Luffa acutangula fruits in water at 2, 4, 6, 8, and 10 % w/v concentration. Standard binder (starch) was prepared by dispersing starch powder in 20 ml of distilled water and adding boiled water whilst stirring with a glass rod to make up to100 ml at 2, 4, 6, 8 and 10% w/v concentration [19]. Composition of each tablet shown in table 3. Preparation and evaluation of granules The granules were prepared by wet granulation method. Paracetamol was used as a model drug to formulate granules. Lactose used as diluents, magnesium stearate and talc as lubricant respectively. The drug and lactose were mixed thoroughly and a sufficient volume of 2, 4, 6, 8 and 10 % w/w of mucilage of Luffa acutangula was added slowly to the powder blend and cohesive wet mass was prepared. For standard used 2, 4, 6, 8 and 10%w/w of starch as a binder. The batch size was. The wet mass was then sieved through sieve number. 10 and dried at not more than 60 0 C in hot air oven up to LOD NMT 3%. The dried granules were re-sieved through sieve number 20. The IC Value

5 prepared granules were then evaluated for percentage of fines, particle size and flow properties (by measuring angle of repose). The bulk and tapped densities were determined. Compressibility index of the granules was determined by Carr s compressibility index [20]. Results are shown in table 4. Preparation and evaluation of tablets The granules were compressed into tablets using single punch tablet machine. The batch size of 70 tablets was prepared. The prepared tablets were evaluated for hardness, friability, disintegration time, thickness, weight variation and dissolution. Monsanto hardness tester/ Pfizer type and Roche friabilator were used to test hardness and friability respectively. The disintegration time was determined using disintegration apparatus and distilled water was used as medium. The dissolution was determined using dissolution apparatus USP 2 nd type [20],[21]. Results are shown in table 5 and 6. RESULTS AND DISCUSSION TABLE 1: STANDARDIZATION OF MICROWAVE METHOD FOR THE ISOLATION OF MUCILAGE FROM LUFFA ACUTANGULA FRUIT. Conventional Method Sr. No. Duration Yield (g) Intensity (W) Microwave Method Duration Yield (g) Percentage Increase in yield * 1 1 h *In comparison to conventional method Using Luffa acutangula fruits for the isolation of mucilage, microwave standardization was carried out at several intensities and durations (Tables 1). Using 640 W intensity and 5 min heating duration, % increase in the yield of mucilage compared to the conventional heating method. IC Value

6 The mucilage isolated by both the methods was found to be identical in nature and gave positive results for all the chemical tests performed. The chemical tests that were conducted are: Ruthenium red test and Molisch test. Thus, the selected plant source produced high yield of mucilage in shorter duration by the developed microwave method when compared to conventional heating. TABLE 2: PHYSICO-CHEMICAL PARAMETERS OF MUCILAGE Parameters L.acutangula State Odour Taste Color Identification : Mounted in ruthenium red Amorphous powder Odourless Tasteless Brown Particles stained red Mounted in iodine solution Particles stained blue ph (1% w/w) 6.7 Ash value (% w/w) 80.5 Solubility Slightly soluble in water, insoluble in alcohol, chloroform and acetone Swelling index 29 Results showed that mucilage isolated from the Luffa acutangula fruits are amorphous in state, odourless, taste less, brown in colour. Mucilages gave positive chemical test, ph is 6.7, and ash value and swelling index was found to be 80.5% and 29 respectively. IC Value

7 TABLE 3: COMPOSITION OF TABLETS CONTAINING LUFFA ACUTANGULA MUCILAGE AS BINDER Ingredients LA 1 LA II LA III LA IV LA V S I S II S III S IV S V Paracetamol Lactose LA mucilage 2% 4% 6% 8% 10% Starch % 4% 6% 8% 10% Magnesium stearate Talc TABLE 4: EVALUATION OF GRANULES Batch Angle of Bulk Tap density Carr s Hausner s repose density index ratio LA LA 1I LA 1II LA 1V LA V S S 1I S II S 1V S V Results showed that angle of repose for all the granules are found in range from 28 to 30 are good in flow property, 30 to 40 are in passable flow properties and granules of batch LA III and SV having very poor flow properties. Carr s index between the range of having good flow property, having passable flow property, having poor flow properties and above 40 having very poor flow properties was found in batch SV. Hausner s ratio of granules of all the batches having value greater than 1.25 which indicates granules having fair to poor flow properties. IC Value

8 TABLE 5: THICKNESS, BROADNESS, LENGTH, WEIGHT VARIATION, FRIABILITY, DISINTEGRATION TIME AND HARDNESS OF TABLETS Batch Thickness * Broadness * Length* Weight variatio n Friabilit y (%) Disintegratio n time Hardness * LA I 5±0.0 5±0.0 1± ± ± LA II 5±0.0 5±0.0 1± ± ± LA III 5±0.0 5±0.0 1± ± ± LA IV 5±0.0 5±0.0 1± ± ± LA V 5±0.0 5±0.0 1± ± ± S I 5±0.0 5±0.0 1± ± ± S II 5±0.0 5±0.0 1± ± ± S III 5±0.0 5±0.0 1± ± ± S IV 5±0.0 5±0.0 1± ± ± S V 5±0.0 5±0.0 1± ± ± *Average of six reading Tablets prepared in each batches were evaluated for its length, thickness and broadness. Length of tablets was found to be 1±0.0 for all the batches, and thickness and broadness are 5±0.0 and 5±0.0 respectively. Hardness of tablets prepared using LA mucilage was found in range between 4.25 to 6.08 while tablets prepared using starch as binder having hardness between 3.16 to 4.48, hardness of the tablets increased with increase in concentration of binder. Friability of tablets prepared using LA mucilage was found in range between 0.88 to 0.57 while tablets prepared using starch as binder having friability between 0.98 to 0.41, friability of the tablets decreased with increase in concentration of binder. Disintegration time of tablets also increased with increase in concentration time of binders. Percentage cumulative release of drug decreased with increase in concentration of binders. IC Value

9 Batch TABLE 6: DISSOLUTION OF TABLETS Percentage cumulative drug release illustrate in below table LA I LA II LA III LA IV LA V S I S II S III S IV S V Series2-2%, Series3-4%, Series4-6%, Series5-8%, Series6-10% Figure 1 In vitro drug release of paracetamol tablets prepared using Luffa acutangula mucilage as binder. IC Value

10 Series2-2%, Series3-4%, Series4-6%, Series5-8%, Series6-10% Figure 2 In vitro drug release of paracetamol tablets prepared using starch as binder. CONCLUSION In the present study, a microwave assisted method for rapid extraction of mucilage from Luffa acutangula fruits has been optimized. The developed method improved the yield of mucilage from the selected source. In conclusion, the developed microwave procedure can be used successfully in commercial and routine laboratory isolation of mucilage. It can be observed from the results that tablets prepared using the Luffa acutangula mucilage as binder showed significant hardness and friability. The increased concentration of mucilage showed small retardation in drug release from tablet. Thus on the basis of the study it can be concluded that the mucilage obtained from Luffa acutangula possesses significant binding properties. So it can be used as tablet binder in pharmaceutical formulations. ACKNOWLEDGEMENT I would like to give my sincere thanks to my honorific guide, Mr. Biren N. Shah, Ph.d, Associate professor, Department of Pharmacognosy and Phytochemistry, Vidyabharti Trust College of Pharmacy, Umrakh for giving continuous guidance, direction and everlasting support enabled to me to complete my work. Without their help I could not have finished my dissertation successfully. I am also thankful to Dr. B.R. Patel for the authentification of plant. IC Value

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