Objective: The present study deals with topical formulations of a bioactive extract of Tinospora cordifolia and its evaluation.Methods: Dried, powdered stem was extracted with chloroform using ultra-sonication method for 3 hrs. Topical formulations like gels containingchloroform extract were formulated using various gelling agents. These gels were evaluated for physicochemical parameters, viscosity, spreadaibility,and antimicrobial activity.Results: A topical gel was successfully formulated containing bioactive chloroform extract of T. cordifolia. The gel was very effective as antimicrobialformulations.Conclusion: These kinds of formulations can be very promising wound healing medicines with ease of use and no side effects.Keywords: Topical gel, Tinospora Cordifolia, Anitmictobia

D DORLE, ABHISHEK

K NAGARE, SUJIT

R HYAM, SUPRIYA

S SWAMI, KUMAR

English

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Vol 8, Issue 6, 2015 ISSN - 0974-2441DESIGN AND EVALUATION OF NOVEL TOPICAL GEL OF TINOSPORA CORDIFOLIA AS ANTIMICROBIAL AGENTABHISHEK D DORLE, KUMAR S SWAMI, SUJIT K NAGARE, SUPRIYA R HYAM*Department of Pharmaceutical Analysis,  Indira Institute of Pharmacy, Sadavali, Devrukh, Ratnagiri - 415 804, Maharashtra, India. Email: supriyahyam@gmail.comReceived: 16 August 2015, Revised and Accepted: 30 September 2015ABSTRACTObjective: The present study deals with topical formulations of a bioactive extract of Tinospora cordifolia and its evaluation.Methods: Dried, powdered stem was extracted with chloroform using ultra-sonication method for 3 hrs. Topical formulations like gels containing chloroform extract were formulated using various gelling agents. These gels were evaluated for physicochemical parameters, viscosity, spreadaibility, and antimicrobial activity.Results: A topical gel was successfully formulated containing bioactive chloroform extract of T. cordifolia. The gel was very effective as antimicrobial formulations.Conclusion: These kinds of formulations can be very promising wound healing medicines with ease of use and no side effects.Keywords: Topical gel, Tinospora Cordifolia, AnitmictobialINTRODUCTIONHerbal medicines have been always in demand because of their potentials and minimal side effects, and thus the research on plant-based medicines for their activity and formulations is constantly in interest. For topical treatment of dermatological disease as well as skin care, a wide variety of vehicles ranging from solids to semisolids and liquid preparations is available to clinicians and patients. Within the major group of semisolid preparations, the use of transparent gels has expanded both in cosmetics and in pharmaceutical preparations. Gels are defined as semi-rigid systems in which movement of the dispersing medium is restricted by an interlacing three dimensional network of particles or solvated macromolecules of the dispersed phase. Gels have better potential as a vehicle to administer drug topically in comparison to ointment, because they are non-sticky, require low energy during formulation, are stable and have esthetic value [1].Traditionally, Tinospora cordifolia (Willd.) Hook. F. and Thomson (Menispermaceae) commonly known as rasayana plant, Guduchi, is one of the commonly used medicinal plants in India for curing ailments ranging from common cold, skin diseases, and dental infections to major disorders such as diabetes, hypertension, jaundice, and rheumatism, and its rejuvenating property is well-reported in Ayurvedic and other ancient literature. This plant is used as stomachic, antipyretic, analgesic, anti-inflammatory, anti-diarrheal, and in fracture of bones in animals [2-6].In past, many herbal gels have been formulated but gel containing T. cordifolia extract as an antimicrobial agent has not been explored. Thus, the present study deals with the investigation of antimicrobial activity of gel containing bioactive extract of T. cordifolia commonly known as Gulvel, Guduchi.METHODSSolvents and chemicalsGuduchi stem was collected from the local region of Devrukh, Ratnagiri. All chemicals and reagents used were of analytical grade and purchased from Rankem and S. D. Fine Chemicals, India.Preparation of extractGuduchi stems cleaned (with 70% ethanol), shade dried, and powdered. Dried, powdered stem was extracted with chloroform using ultra- sonication method for 3 hrs.Formulation of gel [7-12]Weighed amount of Carbopol 940P was soaked in distilled water overnight. The bioactive extract was accurately weighed and uniformly suspended in ethanol. The suspension is then added to the soaked Carbopol. The mixture was stirred using an overhead stirrer for around 1-1.5 hrs; until a uniform suspension was obtained. Care was taken to prevent air entrapment during stirring. This was followed by neutralization of the gelling agent by NaOH or 50% triethanolamine; pH was adjusted to 7.3-7.5. The composition of gel formulations is given in Table 1.Evaluation of gel [7-12]Formulations were evaluated for various parameters such as appearance, color, pH, viscosity, homogeneity, spreadability, extrudability, extract content, uniformity of extract content.ViscosityThe viscosity of the formulations was checked using a Brookfield Viscometer (DV-I PRIME, USA).HomogeneityHomogeneity of formulated gels was examined by visual inspection for the presence of any aggregates.SpreadabilitySpreadability is expressed in terms of time in seconds taken by two slides to slip off from the gel when placed in between the slides under the direction of a certain load. The excess amount of sample was placed between the two glass slides, and a definite amount of weight was placed on these glass slides to compress the glass slides of uniform thickness. A weight of 70 g was added and the time required to separate the two slides was noted. Spreadability was calculated using the formula S = ML/T, where, M = wt tied to upper slide, L = length of glass slides, T = time taken to separate the slides.Research ArticleAsian J Pharm Clin Res, Vol 8, Issue 6, 2015, 237-239 Dorle et al. 238Primary dermal irritation index (PDII)This test is done by applying the formulated gel onto the skin and then observed for any reversible damage to the skin within 4 hrs. Based on their PDII score, the formulation can be graded as irritating or non-irritating.Microbial contaminationMicrobial contamination of gel with bacteria and moulds, respectively, was determined by spreading a thin loop full of the material withdrawn from the depth of the bulk product on a nutrient and Sabouraud agar, and incubating for 24-48 hrs, at 37°C.To assess the degree of contamination, 1 g of material was dispersed in 4 ml of sterile Ringer solution, containing 0.25% Tween 80. Appropriate dilutions were made in the same dispersion vehicle and 0.5 ml was plated out on the appropriate solid medium using the surface viable method. Emergent colonies were counted after necessary incubation.Antimicrobial evaluation [13-17]Test organismThe microbial cultures (Staphylococcus aureus, Bacillus subtilis, Escherichia coli, and Pseudomonas aeruginosa) were procured from National Center for Industrial Microorganisms (NCIM), Pune, India.Antibacterial activityIn-vitro antibacterial activity was evaluated using the agar well diffusion technique. Muller-Hinton agar was used as the medium. The sterile agar was inoculated with the bacteria culture for 48 hrs, at 37°C. Wells were bored by using a sterile borer, and standard formulations (1000 μg/ml was prepared by dissolving the test sample in methanol and the solvent control) were placed into them. Plates were kept for 2 hrs in the refrigerator to enable pre-diffusion of the extracts into the agar. Next, the plates were incubated overnight (24 hrs) at 37°C.StabilityThe stability studies were carried out for all the formulations. The formulations were kept at two different temperatures 4 ± 2°C and 30 ± 2°C, 65 RH, for 3 months. The pH and the viscosity of the formulations, which were determined after 3 months, were compared with the initial pH and viscosity.Statistical analysisAll experimental measurements were carried out in triplicate and were expressed as an average of three analyzes ± standard deviation. Statistical analyzes were performed by the t-test.RESULTS AND DISCUSSIONEvaluations of formulationsThe aqueous extract of guduchi was formulated into a gel into different combination of excipients; formulation E did not show a considerable change in characters such as color, odor, and consistency, and there was no phase separation observed during the course of the study.The results of pH, spreadability, and viscosity of the formulations are recorded in Table 2. The result depicted that formulation E is compatible with the skin, well viscous to spread and to retain onto the skin. The extrusion from the tube and spreadability of the topical formulation is important during the application, as also patient acceptance. The formulation E showed acceptable spreadability along with good extrusion. As, the formulation E was found to be the best among all the combination; it was carried forward for microbial contamination test and antimicrobial study.Microbial contaminationThe microbial contamination of the gel E after 24 hrs was found to be 1.22 colony forming a unit (CFU) and 1.71 CFU for fungi, at a temperature of 37°C, and 2.99 and 2.50 CFU for bacteria, at the end of 5 days.Antimicrobial activityThe antimicrobial activities (by measuring the zone of inhibition) of the formulation E was compared with the marketed formulation.Table 3 shows the antimicrobial activity of the formulation E. The Table 3 represents that the formulations had antimicrobial activity against both all four bacteria.Stability studiesThe stability studies of formulation E showed that it was most stable during the study period as this formulation did not show any physical instability and the pH before and after the study did not show any significant change.DISCUSSIONUntil date, guduchi gel has not been formulated as an antimicrobial agent. T. cordifolia is well-known for its medicinal effects. Thus, this work uplifts one more arena of guduchi and has successfully formulated “formulation E.” The formulation E showed acceptable physical Table 1: Combination of ingredients for formulation of gelIngredients Formulation codeA B C D EAqueous extract 5% w/w 5% w/w 5% w/w 5% w/w 5% w/wCarbopol 940P 1.5% w/w 1.5% w/w 2% w/w 1.5% w/w 2% w/wTween 80 - 5%v/w 5% v/w 5% w/w 5% v/wFlavor q. s q. s q. s q. s q. sWater q. s 100 g 100 g 100 g 100 g 100 g50% triethanolamine - - - pH 7.3-7.5 pH 7.3-7.510% sodium hydroxide pH 7.3-7.5 pH 7.3-7.5 pH 7.3-7.5 - -Table 2: Evaluation parameters of formulated gelSr. no Parameters Observations1. Appearance Lustrous2. Color Pale yellow - white3. pH 7.3-7.54. Viscosity 6015±171 cps5. Spreadability 6.96±1.56 secondsAll the experiments were performed in triplicatesTable 3: Antimicrobial study of formulation gel EOrganisms Zone of inhibition (mm) (for containing 50 mg extract/well)S. aureus 9.5±1.2B. subtilis 10.2±2.01E. coli 8.9±0.99P. aeruginosa 10.5±1.11S. aureus: Staphylococcus aureus, B. subtilis: Bacillus subtilis, E. coli: Escherichia coli, P. aeruginosa: Pseudomonas aeruginosa, All the experiments were performed in triplicatesAsian J Pharm Clin Res, Vol 8, Issue 6, 2015, 237-239 Dorle et al. 239properties, and hence, was compatible with the skin. The in-vitro antimicrobial activity showed maximum activity against B. subtilis and P. aeruginosa, showing promising prospects as an antibacterial gel for open wounds. In addition, the formulation E passed the short-term stability, indicating the physical and chemical stability of the product. Hence, the formulation E of the aqueous extract of T. cordifolia is safe and efficient carriers, with potent antimicrobial activity.CONCULSIONFrom the present study, it was concluded gel formulation of the aqueous extract of T. cordifolia was successfully formulated. Thus, this study proves to be beneficial as the formulation can be put forward for extensive evaluation to make it as a marketed product.REFERENCES1. Yamauchi A. Gels: Introduction. In: Osade Y, Kajiwara K, editors. Gels Handbook. Vol. 1. San Diego: The Fundamentals, Academic Press; 2001. p. 4-12.2. Rana V, Thakur K, Sood R, Sharma V, Sharma TR. Genetic diversity analysis of Tinospora cordifolia germplasm collected from northwestern Himalayan region of India. J Genet 2012;91(1):99-103.3. Parthipan M, Aravindhan V, Rajendran A. Medico-botanical study of Yercaud hills in the Eastern Ghats of Tamil Nadu, India. Anc Sci Life 2011;30(4):104-9.4. Upadhyay AK, Kumar K, Kumar A, Mishra HS. Tinospora cordifolia (Willd.) Hook. f. and Thoms. (Guduchi)-validation of the Ayurvedic pharmacology through experimental and clinical studies. Int J Ayurveda Res 2010;1:112-21.5. Sharma U, Bala M, Kumar N, Singh B, Munshi RK, Bhalerao S. Immunomodulatory active compounds from Tinospora cordifolia. J Ethnopharmacol 2012;141(3):918-26.6. Patel MB, Mishra S. Hypoglycemic activity of alkaloidal fraction of Tinospora cordifolia. Phytomedicine 2011;18(12):1045-52.7. Umachigi SP, Jayaveera K, Ashok Kumar CK. Evaluation of wound healing potential of poly herbal formulation. Pharmacol Online 2009;3:505-12.8. Panda P, Ghosh A. Formulation and Evaluation of topical dosage form of Eupatorium odoratum Linn. and their wound healing activity. Int J Pharm Biol Sci 2010;2:1-10.9. Mohamed MI. Optimization of chlorphenesin emulgel formulation. AAPS J 2004;6(3):e26.10. El Gendy AM, Jun HW, Kassem AA. In vitro release studies of flurbiprofen from different topical formulations. Drug Dev Ind Pharm 2002;28(7):823-31.11. Shivhar UD, Jain KB, Mathur VB. Formulation development and evaluation of diclofenac sodium gel using water soluble Poly acrylamide polymer. Dig J Nanomater Biostruct 2009;4:285-90.12. Gupta GD, Gound RS. Release rate of Nimesulide from different gellants. Indian J Pharm Sci 1999;61:229-34.13. Modarresi Chahardehi A, Ibrahim D, Fariza Sulaiman S. Antioxidant, antimicrobial activity and toxicity test of Pilea microphylla. Int J Microbiol 2010;2010:826830.14. Ofori-Kwakye K, Kwapong AA, Adu F. Antimicrobial activity of extracts and topical products of the stem bark of Spathodea campanulata for wound healing. Afr J Tradit Complement Altern Med 2009;6(2):168-74.15. Olila D, Olwa-Odyek, Opuda-Asibo J. Antibacterial and antifungal activities of extracts of Zanthoxylum chalybeum and Warburgia ugandensis, Ugandan medicinal plants. Afr Health Sci 2001;1(2):66-72.16. El Maghraby GM, Barry BW, Williams AC. Liposomes and skin: From drug delivery to model membranes. Eur J Pharm Sci 2008;34(4-5):203-22.17. Bhalodia NR, Shukla VJ. Antibacterial and antifungal activities from leaf extracts of Cassia 415847 l.: An ethnomedicinal plant. J Adv Pharm Technol Res 2011;2(2):104-9.

DESIGN AND EVALUATION OF NOVEL TOPICAL GEL OF TINOSPORA CORDIFOLIA AS ANTIMICROBIAL AGENT

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DESIGN AND EVALUATION OF NOVEL TOPICAL GEL OF TINOSPORA CORDIFOLIA AS ANTIMICROBIAL AGENT

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