B.Ushasri et al. / IJDFR volume 2 Issue 6, Nov.-Dec.

2011
Available online at www.ordonearresearchlibrary.org Research Article ISSN 2229-5054

INTERNATIONAL JOURNAL OF DRUG FORMULATION AND RESEARCH

EVALUATION OF COCCINIA INDICA MUCILAGE AS SUSPENDING AGENT IN PARACETAMOL SUSPENSION

B.Ushasri, M.Kiranmai, Mohammed Ibrahim Department of Pharmacology and Biotechnology, Nizam Institute of Pharmacy, Deshmukhi, Pochampally (Mandal), Near Ramoji Film City, Nalgonda 508284, Andhra Pradesh & Asian Institute of Advance Research, Hyderabad 500058, Andhra Pradesh, India.
Received: 14 Sep. 2011; Revised: 1 Oct. 2011; Accepted: 16 Nov. 2011; Available online: 5 Dec.. 2011

ABSTRACT
The purpose of this study is to find a cheap and effective natural adjuvant that can be used as a useful alternative for the formulation of pharmaceutical suspensions. The mucilage from the fruits of Coccinia indica was subjected to preformulation study for evaluation of its safety and suitability for use as suspending agent. The mucilage extracted is devoid of toxicity. Suspensions of paracetamol were prepared and compared with different concentrations (1% and 2% w/v) of Coccinia indica mucilage and tragacanth gum. Their sedimentation profile, redispersibility, degree of flocculation and rheological behavior were compared. The mucilage was found to be a comparable suspending agent with tragacanth. Studies indicate that the mucilage of Coccinia indica may be used as a pharmaceutical adjuvant and as a suspending agent at 2%w/v, depending on its suspending ability and the stability.

Key words: Coccinia indica, suspending agent, mucilage, sedimentation volume, degree of flocculation

INTRODUCTION Mother Nature has gifted India with great variety of flora and fauna for centuries. Man has made effective use of materials of natural origin in the medical and pharmaceutical field. Natural materials have advantages over synthetic materials because they are non-toxic, less expensive, and freely available1-3. These are generally normal products of metabolism formed within the cell (intracellular formation) and/or produced without injury to plant. Mucilages are plant hydrocolloids and they are also translucent amorphous substances and polymers of a monosaccharide 4-5. Advantages of mucilages in pharmaceutical sciences: Biodegradable, biocompatible, non-toxic, low cost, environmental friendly processing, local availability, better patient tolerance, public acceptance and edible sources6. Mucilages have a variety of application in pharmacy Girish and Jani et.al reported the pharmaceutical applications of natural gums and mucilages7-9. A pharmaceutical suspension like other disperse systems is
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B.Ushasri et al. / IJDFR volume 2 Issue 6, Nov.-Dec.2011 thermodynamically unstable, thus, making it necessary to include in the dosage form, a stabilizer or suspending agent which reduces the rate of settling and permits easy redispersion of any settled particulate matter both by protective colloidal action and by increasing the consistency of suspending medium 10. Plant mucilages are pharmaceutically important polysaccharide with wide range of application11-14 such as thickening, binding, disintegrating, suspending, stabilizing, emulsifying, gelling and altering release properties of mucilage because of its colloidal nature and viscosity can be used to suspend insoluble substances in liquids and help in preventing sedimentation15,16. Coccinia indica, cucurbitaceae is a perennial creeping herb with a long tapering tuberous roots and grows abundantly in India17. Various parts of Coccinia indica (CI) plant has been used as traditional use in India from time immemorial. The medicinal utilities have been described especially for leaf, fruit, root and stem. Anti- inflammatory, antipyretic, analgesic18, antinuclear and antioxidant effect19, antimicrobial activity20- 21, laruictfidal efficiency22, mutagenic effect23, antistress and free radical scavenging24, antidiabetic25-28, antihyperglycemic and hypolipidimic29. Varsha patel reported Coccinia indica fruit mucilage extract as coagulant-floculant for turbid water treatment30. The present study is an attempt to extract and investigate the phytochemical and pharmaceutical properties of Coccinia indica mucilage to assess its suitability as a suspending agent in the pharmaceutical formulation. MATERIALS AND METHODS Fruits of Coccinia indica W&A were purchased from local market. The plant material was identified and authenticated taxonomically at Osmania University. A voucher specimen of the collected sample was deposited in the institutional herbarium for future reference. Paracetamol, all other ingredients and solvents were purchased from Merck(Mumbai ) EXTRACTION OF MUCILAGE31,32 To obtain Coccinia indica mucilage extract (also known as kundru mucilage ), the fruits were thoroughly washed with water, cut into pieces and then soaked in distilled water overnight then the mucilaginous extract was filtered by addition of alcohol. The precipitate obtained was washed with acetone 2-3 times and finally dried by keeping in oven at 400C for 24 hrs. The resultant extract was then used in the experiment. PHYTOCHEMICAL SCREENING Preliminary tests like test for carbohydrates, alkaloids, glycosides, tannins, saponins flavonoids, terpenoids and mucilage were performed to confirm the nature of mucilage obtained33-34. ACUTE TOXICITY STUDIES35 Toxicity studies were carried out according to the method of Knudsen & Curtis. The animals used in the toxicity studies were sanctioned by the institute animal Ethics committee. The male albino rats of wistar stream weighing 160-200g. Animals were divided into different groups comprising of six animals each. The control group received normal saline 20ml/kg i.p. the other group received 500,1000,2000,3000 and 4000 mg/kg of

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gum suspension in normal saline orally. The animals were observed continusally for the behavioral change for the first 4hrs and then observed for mortality if any for 48hrs. PREPARATION OF SUSPENSION 1.0 g of compound tragacanth powder and 10 g of paracetamol were triturated together with 20ml of sugar syrup to form a smooth paste. 2ml of benzoic acid solution and 1ml of amaranth solution were added gradually with constant stirring and then mixed with 50 ml of chloroform water double strength. Now make up the volume to 100 ml and transfer to an amber coloured bottle and shake vigorously for 2min. Thus making 1.0% w/v of the gum in the preparation. The same procedure was repeated with 2 g of tragacanth to prepare 2.0% w/v of suspension. The above procedure was repeated with Coccinia indica mucilage in place of compound tragacanth powder in order to prepare 1.0% and 2.0%w/v mucilage suspensions. Prepared suspension is subjected for evaluation.36 EVALUATION OF SUSPENSION37-39 Sedimentation volume The rate of separation of the suspension were determined by keeping 50ml portion of each suspension in stopered measuring cylinder and stored undisturbed at room temperature. The separation of clear liquid was noted at interval of 5 days up to 50days. The sedimentation volume (F) was then calculated using the following equation
F= Vu/Vo

Where Vu is the ultimate volume of the sediment and Vo is the original volume of the suspension Degree of flocculation Degree of flocculation was determined by following equation
= F/F

Where F is the ultimate sedimentation volume in the flocculated suspension and F is the ultimate sedimentation volume in the deflocculated suspension. Redispersion Fixed volume of each suspension (50ml) was kept in calibrated tubes which were stored at room temperature for various time intervals at regular interval of 5 days up to 50 days, one tube was removed and shaken vigorously to redistribute the sediment and the presence of deposit if any was recorded Rheological study The rheological behavior of the suspensions prepared with gum tragacanth and Coccinia indica mucilage were studied using Brookfield synchroelectric viscometer, spindle number 1 of low viscosity type with great speed

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B.Ushasri et al. / IJDFR volume 2 Issue 6, Nov.-Dec.2011 ranking 0.3 to 0.6 rpm. The dial reading for both up-curve and down curve was recorded and the experiment was repeated for 3 times. Using this observation, the rate of shear was calculated. The results were recorded. Particle size analysis Determination of particle size as mean diameter is based on direct observation under microscope with eye-piece micrometer. After calibrating the eye-piece micrometer, little of the prepared suspension is mounted on the glass slide. The mounted material is placed on mechanical stage to observe through a microscope the particle diameter (breadth) is measured and recorded for 300 particles. The data is represented as size frequency distribution curve and average particle size is calculated and it is determined by using the following formula:

Average particle size= N.X N

RESULTS AND DISCUSSION The average yield of mucilage obtained from coccinia indica fruit was 2.2 g/kg.The obtained mucilage is partially soluble in water and easily soluble in acetone. Phytochemical tests were carried out on coccinia indica mucilage confirmed the absence of alkaloids, glycosides, tannins, saponins,flavonoids,terpenoids.on treatment of mucilage with ruthenium red, it showed red colour confirming the obtained product as mucilage. A violet ring was formed at the junction of two liquids on reaction with molischs reagent indicating the presence of carbohydrates. Mucilage could not reduce fehlings solution so the sugars present were non-reducing sugars. The results of phytochemical screening of mucilage are summarized in Table1.

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B.Ushasri et al. / IJDFR volume 2 Issue 6, Nov.-Dec.2011

Table1: phytochemical screening of mucilage of coccinia indica

S.NO

Test

Observation

1 2 3 4 5 6 7 8

Test for carbohydrates (molisch,s test) Test for alkaloids (mayers test) Test for tannins (lead acetate test) Test for glycosides (keller kelliani test) Test for mucilage (rhuthenium red test) Test for flavonoids (ferric chloride test) Test for reducing sugars (felhing,s test) Test for saponins (foam test)

+ + -

To determine the safety level of coccinia indica mucilage, acute toxicity studies were carried out. In this study, the mucilage revealed no behavioral changes for first few hours and no mortality, no toxic syndromes were observed at the dose level of 4000mg/kg body weight after 24hrs indicating the safety of mucilage. To evaluate the suspending properties of mucilage, suspensions were prepared with fixed concentration of paracetamol but with varying concentration of test mucilage (1% and 2%) as well as traditional suspending agent such as tragacanth. The sedimentation volume profiles of suspension prepared with coccinia indica mucilage and tragacanth were shown in figure1 and 2 respectively.

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B.Ushasri et al. / IJDFR volume 2 Issue 6, Nov.-Dec.2011

1.2 sedimentation volume 1 0.8 0.6 0.4 0.2 0 0 5 10 15 20 25 30 35 40 45 50 Time in days Coccinia mucilage, 1% Coccinia mucilage, 2%

Figure 1:Sedimentation profile of coccinia indica mucilage with different concentrations

1.2 sedimentation volume 1 0.8 0.6 0.4 0.2 0 0 5 10 15 20 25 30 35 40 45 50 Time in days Tragacanth, 1% Tragacanth, 2%

Figure 2:Sedimentation profile of tragacanth mucilage with different concentration

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B.Ushasri et al. / IJDFR volume 2 Issue 6, Nov.-Dec.2011

The dispersed particles sediment at a faster rate in suspension containing 1% w/v of CI mucilage when compared with 2%w/v CI throughout 50 days as shown in figure1 and the suspension containing tragacanth 1%w/v sediment at a slower rate when compared to 2%w/v tragacanth through out 50 days as shown in figure 2. The sedimentation volume provides only a qualitative account of flocculation. The degree of flocculation () is more useful parameter, which is the ratio of ultimate sedimentation volume in the flocculated and deflocculated system. A comparison of values (Table 2) of suspension prepared with CI mucilage and gum tragacanth shows slight higher values at the 2 % w/v and 1 %w/v level for CI mucilage and tragacanth. Suspensions with CI mucilage and tragacanth were found to be redispersible irrespective of their concentration.

Table2: Degree of flocculation () of various suspe4nding agents Suspending agent Concentration(%w/v) Degree of flocculation()

Tragacanth

1% 2%

1.730.18 0.830.12 1.130.10 1.460.09

Mucilage of coccinia indica

1% 2%

The rheological behavior of the suspensions prepared with CI mucilage and tragacanth revealed that their flow rate decreases with increase in viscosity, which is an essential property in the formulation of suspension. Particle size analysis result was found to be 35.3. Viscosity was found to be directly proportional and flow rate is inversely proportional to the concentration. All the formulations were found to obey the Stokes law when subjected to particle size analysis. The results are given in table 3.

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B.Ushasri et al. / IJDFR volume 2 Issue 6, Nov.-Dec.2011

Table3: Effect of type and concentration of suspending agents on the flow rate and viscosity of paracetamol suspensions.

Suspending agent

Concentration(%w/v)

Flow rate (ml/s-1)

Viscosity (poise)

Coccinia indica mucilage

1 2

0.80 0.63

2.22 3.25

Tragacanth

1 2

1.28 1.04

1.14 1.38

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B.Ushasri et al. / IJDFR volume 2 Issue 6, Nov.-Dec.2011

CONCLUSION From the present study, it can be concluded that like compound tragacanth, CI mucilage has the potential as a suspending agent even at lower concentration(2 %) and can be used as a pharmaceutical excipient. This research provides information which could trigger further research in the direction of using CI mucilage in tablet formulation. AKNOWLEDGEMENT The present study is carried out in the Nizam institute of pharmacy and the authors are very much thankful to Mr.Mohammed Jafer, chairman, Nizam group of institutions, Deshmukhi for providing all necessary facilities in order to complete this study. REFERENCES 1. Nakano.M, Nakamura.Y, Juni K, Sustained release of sulfamethizole from agar beads after oral

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