RESEARCH ARTICLE ISSN 2278 – 2818

EXTRACTION, ISOLATION & EVALUATION OF GELLING POTENTIALS OF HIBISCUS CANNABINUS SEED MUCILAGE G. S. Palshikar1†, P. P. Parekh1 and B. M. Firake1 ABSTRACT The present study was undertaken with an objective to find out the gelling potentials of mucilage obtained from plant Hibiscus cannabinus. The mucilage was extracted by using water as solvent and precipitated using ethanol as non-solvent. Physical characteristics such as solubility, swelling index, loss on drying, and pH were studied. Diclofenac sodium was used as model drug for formulation of gels. Six batches of drug loaded gels with concentration of mucilage ranging from 5, 6, 7, 8, 9 and 10% were formulated by using glycerin as plasticizer and methyl paraben as preservative. The pH, viscosity, and in vitro diffusion profiles were studied. The gels prepared with 8% of mucilage were found to be ideal and comparable with a commercial preparation. KEYWORDS: Hibiscus cannabinus, Gel, Mucilage. INTRODUCTION

Preparation of Gel

Natural sources are being utilized in healthcare system throughout the world since recorded history of mankind. According to WHO, around 80% of the population in developing countries relies on alternative and traditional medicines for primary health care [1]. In most of the traditional and alternative systems of medicines, natural sources so much, so plant derived materials form dominating portion. In India, traditional system of medicines like ayurveda, siddha and unani system of medicines highlighted use of plant derived materials in health care with combination of other natural sources. Number of pharmaceutical excipients, which are major raw material for any pharmaceutical company and forms major portion of most of the dosage form, are obtained from natural sources.

Diclofenac used as a model drug to make gel formulation of topical therapy. The gift sample was procured from Emcure Pharmaceuticals Pvt. Ltd. The formula used to prepare gel is given in Table 1. In this formula Diclofenac is used as an anti-inflammatory drug, glycerin act as antimicrobial agent, polyethylene glycol act as a nonirritant nature, methyl paraben act as a preservative and mucilage act as a thickening agent. In above formula mucilage concentration was gradually increased. Procedure Gel was prepared by using different concentrations of mucilage, Diclofenac, methyl paraben, glycerin, polyethylene glycol and sufficient quantity of purified water was added as per mentioned in I.P. Diclofenac is used as an anti-inflammatory drug. diclofenac gel was prepared and stored in cool place until further use [6]. For comparison purpose standard marketed formulation of diclofenac gel (Omnigel) was used and results were compared with test formulation.

Excipients are the additives used to convert active pharmaceutical ingredients into pharmaceutical dosage form suitable for administration to patients [2]. Plant products serve as an alternative to synthetic products because of local accessibility, environment friendly nature and lower prices compared to imported synthetic products. Many products from natural sources like plant exudates, gums, mucilage, and starches are utilized for preparation of pharmaceutical dosage forms like tablets, syrups, suspensions, emulsions, ointments and sustained drug release systems. Dosage form, irrespective of its final structure and nature, is a combination of the drug components and an assortment of different kinds of non-drug components, collectively known as additives.

RESULTS AND DISCUSSION A) Viscosity study The viscosity of sample prepared with different concentrations, was checked. Viscosity was taken with the help of Brookfield viscometer using T bar spindle No. 7at 5 rpm under constant temperatures (10-15°C) and results of standard and H. cannabinus mucilage as a gelling agent is summarized in Table 2 and Figure 1.

These additives play important role in pharmaceutical industry. Drugs are rarely administered in pure forms; generally they are admixed with various kinds of adjuvants, which lead to formation of dosage forms. These non-drug components are collectively called as additives or excipient. These can be used in natural from or after modification and derivatisation [3].

The viscosity of gel prepared with H. cannabinus mucilage as a gelling agent showed viscosity 2800 cps on day 1 at 5% concentration, 5310 cps at 8% concentration and 6000 cps at 10% concentration. On day 25, 8% concentration gave gel with viscosity of 5080 cps which was more stable than other gels. In this case, the viscosity of gel sample showed constant results for a 25 days period of time interval. Different concentrations of gelling agent showed results in acceptable ranges.

Gels are an excellent formulation for several routes of administration. They are useful as liquid formulations in oral, topical, vaginal, and rectal administration. Gels can be clear formulations when all of the particles completely dissolve in the dispersing medium. But this doesn't occur in all gels, and some are therefore turbid. Clear gels are preferred by patients. Clarity also increases the beauty and usefulness of any formulation [4]. Gelling agents for pharmaceutical and cosmetic use should be inert, safe and nonreactive with other formulation components [5].

B) pH study The pH of sample of different concentrations was observed by using digital pH meter. The effect of ageing on pH of standard and different concentration H. cannabinus mucilage is summarized in Table 3 and Figure 2 [7].

MATERIALS AND METHODS The plant material was collected from Naik seeds and Fertilizers, Pune. The botanical identity of plant was confirmed at Agharkar Research Institute, Pune. From the seeds the mucilage was isolated by extraction and precipitation by acetone.

As the preparation is for the sake of topical application, the pH must be in neutral form. In this case the gel preparation prepared by using concentration of 6, 7, 8% gelling agent showed pH range in between 6 to 7. The 8% concentration showed pH of 7.0 on day one and pH of 7.2 on day 25. There observed a stable pH range in this form. In this case, all the concentrations of gelling agent show results within an acceptable limit.

For evaluation of gelling properties of isolated mucilage, topical gel formulation was prepared using model drug Diclofenac. Also various evaluation properties of Gel were checked and results were compared with standard marketed preparation of diclofenac topical gel (Omnigel).

Appearance All the gel preparations prepared showed accepted results of appearance. The standard marketed preparation also showed translucent appearance.

1J.S.P.M.’s

Jayawantrao Sawant College of Pharmacy & Research, Hadapsar, Pune, Maharashtra. †Corresponding author: [email protected]

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Palshikar et al.

It showed that all these preparations showed appearance within an acceptable limit [8].

The percent drug release decreased as the concentration of gelling agent increased. At 5% concentration H. cannabinus mucilage showed complete drug release within 4hr. It showed 78.210% drug release in 1 hr and 99.545% drug release in 4hr at 1% concentration. It showed 42.815% drug release in 1hr. and 99.025% drug release in 8 hr. at 8% concentration. Gel showed 27.825% drug release in 1 hr and 81.335% drug release in 8 hr. at 10% concentration. The comparison was made with standard marketed gel which showed 43.712% drug release in 1hr. and 99.314 % drug release in 8 hrs time period. The data from standard gel formulation matched with test gel containing 8% H. cannabinus mucilage as a gelling agent.

Table 1. Specifications for gel as per U.S.P. Name of ingredient Diclofenac Glycerin Polyethylene glycol Methyl paraben Mucilage Purified water

Quantity % 1 15 10 0.1 55 q.s.

Viscosity-H.cannabinus mucilage

Table 2. The effect of aging on the viscosity of gel using H. Cannabinus mucilage as the gelling agent Viscosity (cp)

7000

Days

% Conc.

1 2800 ±10 3870 ±10 4500 ±10 5310 ±30 5890 ±10 6000 ±10 5200 ±10

5 6 7 8 9 10 Standard

5 2890 ±20 3700 ±10 4610 ±10 5290 ±50 5670 ±50 6100 ±40 5290 ±20

10 2730 ±10 3940 ±30 4530 ±50 6260 ±30 5930 ±10 6050 ±20 5640 ±10

15 2610 ±10 3870 ±10 4610 ±10 5790 ±30 6170 ±50 6040 ±10 5890 ±40

20 2580 ±50 4100 ±50 4540 ±20 5090 ±10 5810 ±20 6310 ±20 5800 ±10

25 2470 ±10 4000 ±20 4780 ±10 5080 ±10 5830 ±20 6020 ±10 5200 ±10

5 7.1±0.5 7.5±0.1 6.9±0.3 7.4±0.1 6.8±0.5 7.2±0.5 7.1±0.1

10 6.9±0.5 6.8±0.5 7.1±0.3 7.6±0.2 7.2±0.2 6.9±0.5 7.0±0.1

7% 8%

3000

9%

2000

10% St.

0 0

H.c. Mucila ge

8

pH

7

5%

6

6%

5

7%

4

8%

3

9%

2

10%

1

St.

0

15 6.8±0.5 6.5±0.5 6.4±0.5 7.3±0.2 7.1±0.1 6.9±0.1 7.2±0.2

20 6.4±0.5 6.8±0.5 6.9±0.5 6.9±0.5 7.1±0.5 7.2±0.2 7.2±0.1

25 6.9±0.1 7.1±0.5 7.3±0.2 7.2±0.2 6.9±0.5 6.7±0.5 7.2±0.2

0

10

7

8

9

20

30

Days

Figure 2. Effect of ageing on pH of different concentrations of H. Cannabinus mucilage and standard Gel Diffusion profile 120 5%

100

Time (hr)

6

30

pH- H.cannabinus mucilage

Table 4. The appearance of gel at different concentrations of the H. Cannabinus mucilage 5

20

Figure 1. Effect of ageing on viscosities of different concentrations of H. Cannabinus mucilage and standard

Each value is the mean ± S.D. (n=3)

Conc. %

10 Days

Days 1 6.7±0.4 6.3±0.5 7.1±0.1 7.0±0.3 6.9±0.2 7.2±0.5 7.2±0.1

6%

1000

Table 3. The pH study of different concentrations of H. Cannabinus mucilage

5 6 7 8 9 10 Standard

5%

5000 4000

Each value is the mean ± S.D. (n=3)

% Conc.

6000

10

6%

80

7%

60

8%

40

9% 10%

Transluc ent

Transluc ent

Transluc ent

Transluc ent

Transluc ent

20

Transluc ent

St.

0 0

Each value is the mean ± S.D. (n=3) (H.c- H. cannabinus)

5

10

Concentration

Each value is the mean ± S.D. (n=3)

Figure 3. Comparative diffusion profile for gel

Table 5. The comparative diffusion study for gel % Conc. 0 5

0.000

6

0.000

7

0.000

8

0.000

9

0.000

10

0.000

Standard

0.000

1 78.210 ±0.550 62.625 ±1.200 50.265 ±0.800 42.815 ±0.500 33.935 ±0.500 27.825 ±1.250 43.712 ±0.100

2 85.730 ±0.850 66.235 ±0.900 56.615 ±0.200 47.335 ±1.450 36.445 ±1.100 36.945 ±0.550 46.123 ±0.100

3 95.345 ±0.300 75.955 ±0.600 67.465 ±0.650 57.465 ±1.800 49.645 ±0.700 42.125 ±1.650 58.537 ±0.100

Time (hr) 4 99.545 ±0.500 87.135 ±0.500 72.825 ±0.500 71.335 ±0.750 56.335 ±0.950 53.335 ±0.500 72.317 ±0.100

Comparative Diffusion Study

6

7

8

92.865 ±0.700 84.355 ±1.100 81.125 0.500 62.865 ±1.450 61.745 ±0.350 83.712 ±0.100

99.055 ±0.500 99.225 ±0.600 86.625 ±0.500 71.125 ±1,850 68.965 ±0.565 87.831 ±0.100

90.735 ±0.300 76.465 ±0.800 76.335 ±0.875 92.613 ±0.100

99.025 ±0.450 85.785 ±0.550 81.335 ±0.750 99.314 ±0.100

CONCLUSION

The diffusion study was observed for 8 hr. time period with different concentration of mucilage. The observations are summarized in Table 5 and Figure 3.

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The property of Hibiscus Cannabinus mucilage as a gelling agent in topical gel formulation was studied. For study the concentration of mucilage from 5 to 10% was used to prepare gel formulation. In this diclofenac

42

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sodium was used as a model drug having anti-inflammatory activity. Different evaluation tests were performed and results compared with standard marketed topical gel preparation containing diclofenac as a drug (Omnigel). The gel formulation with 8% concentration of mucilage as a gelling agent gets matched with results from standard preparation. So the H. cannabinus mucilage can also acts as gelling agent. ACKNOWLEDGEMENT Authors wish to thank Dr. D.Y. Patil Institute of Pharmaceutical Sciences and Research, Pimpri, Pune for providing necessary facilities to perform this research work. Also thankful to Dr. M. J. Patil, Principal M. M. M.’s College of Pharmacy, Kalewadi, Pune for his valuable guidance. REFERENCES 1. 2. 3. 4. 5. 6. 7. 8.

The Wealth of India. Raw materials, Vol. 8, Council of science and Industrial Research, New Delhi, 2003, 77-78. Editor(s), compiler(s) as author: Kibbe AH editors. Handbook of pharmaceutical excipients, London (UK), pharmaceutical press; 2000. Kapoor V P, Joshi H, and Chaubey M. Application of seed gums in pharmaceutical formulations. Published by Photochemistry division, National Botanical Research Institute, Lucknow, India, 1992; 42. Schmolka P, Cosmet. Toilet. 1984; 99(11): 69. Chang, I.B., Cosmet. Perfum. 1977; 92 (7): 25. Panda D S, Swain SIS, Kanungo S K, and Gupta R. Preparation and Evaluation of Gels from Gum of Moringa oleifera. Ind. J of Pharm Sci. 2006; 68 (6): 777-780. Indian Pharmacopoeia, Government of India, Vol. 2, Ministry of Health and Family Welfare, Controller of Publications, New Delhi, 1996: A-53, 54, A-95, A-97, A-109. Aulton M E, Pharmaceutics: The Science of Dosage Form Design, Churchill Livingstone 1988: 133-135.

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