Research J. Pharm. and Tech. 1(4): Oct.-Dec.

2008, ,

ISSN 0974-3618

www.rjptonline.org

RESEARCH ARTICLE

Pharmaceutical and Biological Evaluation of Formulated Cream of Methanolic Extract of Acacia Nilotica Leaves
AA Baravkar1*, RN Kale1, RN Patil1 and SD Sawant2
1

SVPMs College of Pharmacy, Malegaon (Bk), Tal-Baramati, Dist-Pune, M.S., India. 2 STESs SKN College of Pharmacy, Kondhwa (Bk), Pune, M.S., India. *Corresponding Author E-mail: atul200678@rediffmail.com

ABSTRACT
A cream formulation of the methanolic extract of the leaves of plant Acacia nilotica (Fabaceae) was evaluated pharmaceutically as well as biologically for its wound healing property made by excision wound model. The methanolic extract of leaves was obtained by Soxhlet extraction and its medicated as well as non-medicated cream formulation is developed using bees wax, liquid paraffin, borax and water. Commercially available Vicco turmeric cream was used as a standard for this study. All three formulations were evaluated for various pharmaceutical parameters such as Rheological properties, stability, pH and external characters as well as biologically for its wound healing activity on excision wound made on albino rats. Albino rats were differentiated into control group, non-medicated cream group (group of rats with nonmedicated cream) and medicated cream group (group of rats with medicated cream), standard (vicco turmeric cream). Results were found by comparing the wound contraction (in mm2 and in %) as well as period of epithalisation of medicated cream with that of control group, non-medicated cream group and vicco turmeric cream (as standard) and was found faster for medicated cream than control group and non medicated one.

KEY WORDS: INTRODUCTION:

Cream, Wound, Polyphenols, Antioxidants. cooling, styptic, emollient, anthelmintic, aphrodisiac, diuretic, expectorant, emetic, nutritive, in hemorrhage, wound ulcers, leprosy, leucoderma, skin diseases, and seminal weakness. Gum is used as astringent, emollient, liver tonic, antipyretic and antiasthmatic2. Scientific Classifiaction1 2 3 4 Kingdom Division Class Order Plantae Magnoli ophyta Magnoli ophyta Fabales 5 6 7 8 Family Subfamily Genus Species Fabaceae Mimosoideae Acacia Acacia

Plant kingdom consists of number of medicinal plants from which Acacia nilotica leaves methanolic extract as cream formulation was investigated on albino rats for its effect on healing of excision wound using Vicco-turmeric cream as standard1. Acacia nilotica is a tree 5-20 m high with a dense spherical crown, stems and branches usually dark to black colored, fissured bark, grey-pinkish slash, exuding a reddish low quality gum. The tree has thin, straight, light, grey spines in axillary pairs, usually in 3 to 12 pairs, 5 to 7.5 cm long in young trees, mature trees commonly without thorns. The leaves are bipinnate, with 3-6 pairs of pinnulae and 1030 pairs of leaflets each, rachis with a gland at the bottom of the last pair of pinnulae. Flowers in globulous heads 1.21.5 cm in diameter of a bright golden-yellow color set up either axillary or whorly on peduncles 2-3 cm long located at the end of the branches. Pods are strongly constricted, hairy, white-grey, thick. Bark is used as astringent, acrid,

Received on 27.09.2008 Accepted on 10.11.2008

Modified on 10.10.2008 RJPT All right reserved

Research J. Pharm. and Tech. 1(4): Oct.-Dec. 2008;Page 480-483

A wound is the result of physical disruption of the skin, one of the major obstacles to the establishment of infections by bacterial pathogens in internal tissues, which leads to loss or breaking of cellular and anatomic or functional continuity of living tissue3. Wound healing is a process that is fundamentally a connective tissue response. Initial step of this process involves an acute inflammatory response followed by synthesis of collagen and other extracellular that are later remodeled to form scar4. Excision wound model is one of the important model among the others5.

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The leaves of Acacia nilotica were collected from local areas of Baramati city and authenticated in Department of Botany, SPM Science College, Shardanagar, Tal-Baramati, Dist-Pune. Leaves were shade dried and pulverized and subjected to extraction.

MATERIALS AND METHODS:

Methanolic extract was obtained by subjecting dried and coarsely powdered leaves to Soxhlet extraction process6 and extract was concentrated and dried. Extract was subjected to preliminary phytochemical evaluation and chemical tests7. Two types cream formulation were formulated using following formula1. Non-medicated Cream Formulation-

control group, non-medicated cream group, medicated cream group, standard (vicco-turmeric) group. Full thickness excision wounds of 300 mm2 diameter were inflicted on starved animals after shaving the area to be operated under light chloroform anaesthesia, then cleaned with normal saline and medicated, non-medicated and vicco-turmeric cream were applied immediately. Animals were inspected for 15 days, once in 3 days and healing was assessed based on physical parameters namely wound contraction (in mm2 & in %) and period of epithelisation. Tracing of raw wound area on transparent polythene paper was done on 0th, 3rd, 6th, 9th, 12th, 15th post wounding day. The time taken for complete epithalisation was noted by recording the days required for fall of scab leaving no raw wound behind. Results are reported in MeanSEM and the data was analyzed using unpaired Studentst test.

Sr. No. 1 2 3 4 5

Ingredient Bees Wax Liquid Paraffin Borax Water Perfume

Quantity 16 gm 50 gm 0.8 gm 33.2 ml Q.S

RESULTS AND DISCUSSION:

In the present experimental study, an excision wound model is considered for evaluation of wound healing. In this study, the wound is treated with 10 % medicated cream formulation and commercial vicco-turmeric cream and the results shows complete wound healing on around 21st and 26th day respectively. These results are compared with control group as well as with non-medicated cream group which took more that 38 and 29 days for wound healing respectively. In Excision wound study, from the observations it is clear that, medicated cream formulation showed better and faster healing of wound as compared to control and non-medicated cream formulation. During initial days i.e. from day 0, there was not much difference in healing of wounds in all four groups. But after 6th day, healing process was faster with group 3 i.e. wound treated with medicated cream formulation. If the results are compared with group 1 and 2, group 3 shows faster wound contraction. During day 12-15, healing progress of medicated cream formulation was clearly seen in comparison with group 1 and 2. On comparison with group 4 (treated with vicco-turmeric cream), wound healing property of medicated cream formulation is less effective.
Fig.1 Percentage reduction of wound contraction in mice.

2. Medicated Cream Formulation-

Sr. No. Ingredients Quantity 1 Dried Extract 10 gm 2 Bees Wax 16 gm 3 Liquid Paraffin 50 gm 4 Borax 0.8 gm 5 Water 33.2 ml 6 Perfume Q.S 10 % aqueous drug extract was used after Phase-Solubility technique. Pharmaceutical Evaluation of Developed Cream Formulation: A developed cream formulation is evaluated pharmaceutically for various parameters such as Rheological properties, stability, pH and external characters. Rheological properties like apparent viscosity using Brookfield viscometer8 at 2.5 rpm was found to be 27x106 cPs, sreadability9 was 51 sec., extrudability9 was 582 gms and flow Index was 0.23. Stability10 was determined by exposing the formulation to various temperatures such as 4C, 27C & 37C for specified period and no change occurred in properties of formulation which shows that the formulation is stable. pH of the formulation was determined using digital pH meter11 and was found to be 7.2. External characters of developed cream formulation was also find out, such as colour (dark yellow), odour (agreeable), smoothness (adequate), grittiness (absent). Biological Evaluation of Developed Cream Formulation12: It was done using excision wound model. The study protocol was approved by Institutional Animal Ethics Committee. Healthy 24 albino rats weighing between 150250 gms of same age, which are divided in 4 groups such as

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Table no. 1 Values are in Mean SEM, where n=No. of animals. *-p<0.05 Vs control & standard. Wound contraction in mm2 and in % Post wounding day Control (n=6) Non-medicated Medicated cream formulation cream formulation 0 32309(00) 32810(00) 33308(00) 3 28512(11.77) 27909(14.94) 24814(25.53) 6 24007(25.7) 23211(29.27) 19809(40.55) 9 20108(37.78) 19112(41.77) 11511(64.47) 12 166.10(48.01) 15207(53.76) 8807(73.58) 15 12111(62.54) 11608(64.64) 3006(91) Epithelization period (Days) 382.11 29.430.32 26.651.54 Values in parenthesis indicate % wound contraction.

Vicco-Turmeric cream (standard) 32108(00) 23010(28.35) 15806(50.78) 7408(79.59) 2511(92.22) 0 (100) 21.231.88

CONCLUSION:
Results clearly show that medicated cream formulation of methanolic extract of Acacia nilotica leaves has satisfactory results Parmaceutically as well as biologically and promotes fast wound healing than control and nonmedicated group. The activity may be mainly due to free radical scavenging and antioxidant activity of the polyphenols13,14 present in the leaf extract, which are known to reduce lipid peroxidation an important process for burns, wounds, skin ulcers, thereby reduce cell necrosis.

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FUTURE SCOPE:
Cream formulation of extract of Acacia nilotica leaves showed wound healing action, will essential to investigate the clinical trials on human volunteers and to bring the acacia cream formulation as one of the cheap, cost effective and patient affordable cream in the market for healing of wounds, burns and skin ulcers.

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Kokate C K. Plant constituents-Preliminary phytochemical evaluation. Practical Pharmacognosy. Vallabh Prakashan, New Delhi. 1999; 4th ed: pp. 107-111. Alfred Martin. Rheology. Physical Pharmacy. Lippincott Williams and Wilkins, USA. 2003; 4th ed: pp. 465-467. Venkata R M, et al. Comparative Evaluation of Transdermal Formulations of Norfloxacin With Silver Sulfadiazine Cream, USP, for Burn Wound Healing Property. J Am Pharm Ass.1956; 45: 212. Kulkarni G T, Gowthamarajan B, Suresh B. Stability testing of pharmaceutical products-An overview. Indian J Pharm Educ. 2004; 38(4): 194-198. David T Plummer. pH and buffer solutions. An introduction to Practical Biochemistry. Tata McGraw Hill Publications, New Delhi. 1988; 3rd ed: pp. 39-41. Manjunatha B K et al. Wound healing activity of Lucus hirta. Ind J Pharm Sci, 2006; 68 (3): 380-384. Augustin S, Ian T J, Mike S. Polyphenols- antioxidants and beyond. Am J Clin Nutr. 2005; 215S7S: 81. Mooldun M E, Kritchevsly S B. Flavonoid and heart diseases. Brit Med J. 1996; 312: 458-459.

ACKNOWLEDGEMENTS:
Authors are thankful to Mr. Anandrao Pisal, Managing Director, Shivnagar Vidya Prasarak Mandal, Malegaon (Bk) for providing necessary facilities through Principal and to Prof. R. B. Deshmukh, Head of Department, Botany, SPM Science College, Sharadanagar, for providing plant authentication report.

REFERENCES:
1. Hukkeri V I et al. Antipyretic and Wound Healing Activities of Moringa oleifera Lam. in Rats. Ind J Pharm Sci. 2006; 68(1): 124-126. Vaidyaratnam P S. Variers Arya Vaidya Sala: Acacia nilotica. Indian Medicinal Plants. Edited by Warrier PK, Nambiar VPK, Ramankutty C. Orient Longman Ltd, Hyderabad. 1996; Vol-I: pp. 26. Patil M B et al. Antimicrobial and wound healing activities of leaves of Alternanthera Sessilis. Linn. Int J Green Chem. 2008: 2(3): 141-144. Chithra P, Sajithal GB, Chandra KG. Influence of Aloe vera on collagen turnover in healing of dermal wounds in rats. Indian J. Exp. Biol. 1998; 36: 896. Hunt T K et al. Advances in skin and wound care. Int Ann Surg. 1969; 170: 633-641. William Charles Evans. Introduction and general methods. Trease and Evans Pharmacognosy. Bailliere Tindall publisher, London. 1992; ELBS 13th ed: pp. 248- 249.

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