Formulation and Evaluation of Herbal Anti-bacterial gel containing Ethanolic extracts of Rosa indica and Tagetes minuta

 

Sandhya Mandadi*, K. Prathyusha, M. Rani, K. Sridivya Goud, Kalivemula Divya Reddy, M. Venkata Swamy

Vishnu Institute of Pharmaceutical Education and Research, Vishnupur, Narsapur, Medak District– 502313, Telangana, India

 

ABSTRACT:

The present research has been undertaken with the aim to formulate and evaluate herbal gel containing ethanolic extracts of Rosa indica and Tagetes minuta. The antibacterial activity and phytochemical screening was performed for ethanolic extract. The gel preparation was designed by utilizing ethanol extract of Rosa indica and Tagetes minuta petals in varied concentrations (1.0%, 2.0%, 4.0%) and evaluated using physiological measurements. The gel was formulated by using polymer carbopol 934. Rosa indica and Tagetes minuta petals, methyl paraben, propyl paraben and required amounts of distilled water. The antibacterial activity of the gel was evaluated by agar well diffusion method by using microbial strains are E.coli and Staphylococcus aureus. The physicochemical parameters (pH, viscosity, spreadability and homogeneity) was determined. The results showed that formulation containing 4% concentration of gel containing ethanolic extracts of Rosa indica and Tagetes minuta have better antibacterial activity than other formulations.

 

 

 

INTRODUCTION:

For many decades treatment of the acute illness or a chronic illness health problem has been largely accomplished by delivery of medication to patients exploitation numerous pharmaceutical medicines forms, including tablets, capsules, pills, suppositories, creams, gels, ointments, liquids, aerosols and injectables as drug carriers. Delivery of drugs to the skin is an effective and targeted therapy for local dermatological disorders⁽¹⁾. Gel based formulation makes the drug molecules a lot of simply removable from the system than cream and ointment. Gel base formulations for the skin use have plenty favorable properties like being thixotropic, easily spreadable, grease less, easily removable, emollient, compatible with several excipients and water-soluble or miscible. Gels⁽²⁾ were superiorproducts for different routes of administration. Gel is clear formulations once all of the particles fully dissolve within the dispersing phase. Gels formulatedwith thesecompounds (named as gelling agents) that leads to a superiorchain linking or aggregation when hydrated and discrete in the dispersing medium or when dissolved in the dispersing medium. This chain-linking or aggregation of the dispersed particles can alter the glueyness of the dispersing phase. The movement of the dispersing phase is restricted by the dispersed particles, and therefore the glueynessis raised. If the gel includestiny discrete particles, it is called a two-phase system. The gel without tiny discrete particles, it is termed as a one-phase system. The two-phase systems will be thixotropic (means semisolid without agitation but liquefy when agitation). The United state pharmacopeia states that gels are semisolids, being either suspensions of tiny inorganic particles or big organic molecules interpenetrated with liquid.

 

Classification of Gels:

1. On the basis of the nature of their colloidal phase, gels are divide into: Inorganic gels: Bentonite magma is an example of an organic gel. Organic gels: Contain organic polymers as the primary gel forming agent. Examples cellulose derivatives, or polyethylene and its copolymers.2. They can also be classified on the basis of the nature of the solvent: Hydrogels: They are water based aqueous gels that contain an insoluble polymer. They usually have a higher solvent concentration. Organogels: They contain a non-aqueous solvent as a continuous phase. Examples being, plastibase (low molecular weight polyethylene dissolved in mineral oil and shock cooled) and dispersion of metallic stearate in oil. Xerogels: These are solid gels with low concentration. These are often produced by evaporation of solvent leaving a gel framework behind.

 

MATERIALS:

rosa indica, tagetes minuta, carbapol 934, tri ethanol amine, propylene glycol

 

METHODS:

Method of extraction: Maceration method:

Plant Material was pulverized into small powder and placed into a closed vessel. The required quantity of ethanol was added. Then cover the outer part of the vessel with aluminum foil. Then allow to stand for seven days with shaking occasionally. Then filter the liquid, Solid residue obtained. Collect as much as filtrate. remaining filter and pressed liquids mixed clarified by filtration. Then evaporate and collect the concentrated product.

 

Fig 1: Maceration method

 

PHYTOCHEMICAL SCREENING:

TEST	EXPERIMENT
Alkaloids (wagners test)	Ext + wagners reagent
Phenols (ferric chloride test)	Ext + ferric chloride soln
Tannins (Braymers test)	Ext + alc. ferric chloride soln
Flavonoids (Alkaline reagent test)	Ext + NaOH
Proteins and amino acids (ninhydrin test)	Ext + ninhydrin solution
Sterols and triterpenoids (liebermann-burchards test)	Ext + acetic anhydride + sulphuric acid
Carbohydrates (Molisch test)	Ext + Molisch reagent + sulphuric acid
Saponins (Froth test)	Ext + distilled water + shaken
Fixed oils (filter paper test)	Ext + pressed b/w filter paper

 

PROCEDURE FOR ANTIBACTERIAL ACTIVITY:

Anti-bacterial activity of ethanolic extract of Rosa indica and Tagetes minuta by agar diffusion method Showing inhibition zones against the gram positive and gram negative bacterial strains. In this well-known procedure, agar plates are inoculated with a standardized inoculums of the test microorganisms namely E.coli and Staphylococcus aureus. Then, the cavities using borer are made containing the plant extracts (Rosa indica and Tagetes minuta), are placed on the agar surface. The Petri dishes are incubated under suitable conditions. Generally, antibacterial agent diffuses into the agar and inhibits germination and growth of the test microorganism.

 

FORMULATION OF GEL:

PREPARATION OF GEL WITH CARBOPOL⁽³⁾:

Carbopol 934 was taken in a beaker and dispersed in 50 ml of distilled water. Swell the Carbopol for half an hour, stirring should be done using mechanical/lab stirrer at 1200rpm for 30 min. In one beaker take 5ml of propylene glycol add Extract. 5ml propylene glycol in another beaker add propyl paraben and methyl paraben. Carbopol dispersed, extract, preservatives solutions were added and volume made upto 100ml. Triethanolamine was added drop wise. gel at required consistency is obtained.

EVALUATION OF GEL FORMULATIONS:

Determination of pH:

The pH value of gel formulation was determined by using a pH meter.

 

Homogeneity⁽⁴⁾:

All developed gels were tested for homogeneity by visual observation.

Physical appearance:

The physical appearance of the gel was checked visually such as the color, odor, consistency and greasiness. They appear light golden brown and opaque, free from grittiness and homogenously dispersed.

 

Composition of Different Gels:

Table 1: Composition of Different Gels

FORMULATION	CARBAPOL	EXTRACT (Rosa indica and Tagetes minuta)	PROPYLENE GLYCOL	METHYL PARABEN	PROPYL PARABEN	WATER
1%gel	4g in 50 ml of water	1g	10ml	0.2g	0.2g	q.s to 100 ml
2%gel	4g in 50 ml of water	2g	10ml	0.2g	0.2g	q.s to 100 ml
3%gel	4g in 50 ml of water	4g	10ml	0.2g	0.2g	q.s to 100 ml

 

 

Viscosity:

The measurement of viscosity of the prepared gel was done with Brookfield viscometer. The reading was taken at 100 rpm.

 

Spreadability:

The spread ability of gel formulations was determined by measuring the spreading diameter of 1g of gel between two horizontal plates.

RESULTS AND DISCUSSION:

PHYTOCHEMICAL SCREENING:

Phytochemical screening of crude extract of Rosa indica and Tagetes minuta using ethanol solvent. Presence or Absence of the chemical constituents is depicted below:

 

 

Table 2: Phytochemical screening of crude extract

PHYTOCHEMICALS	OBSERVATION	INFERENCE
Alkaloids	Reddish brown ppt	Present
Phenols	Deep blue colour	Present
Tannins	Blue/green colour not observed	Absent
Flavanoids	Yellow to colourless	Present
Proteins and aminoacids	Violet colour not observed	Absent
Sterols and triterpenoids	Red color (lowerlayer)sterols	Present
	Yellow colour (lower layer)-triterpenoids
Carbohydrates	Dull violet colour at interphase	Present
Saponins	Froth formation	Present
Fixed oils	oil stain on paper	Present

 

ANTIBACTERIAL ACTIVITY:

Anti-bacterial activity of ethanolic extracts of Rosa indica and Tagetes minuta by agar well diffusion method showing inhibition zones against the gram positive and gram negative bacterial strains.

 

Fig 2: Antibacterial activity ofExtract againstE.coli and S.aureus

  

Fig 3: Antibacterial activity ofStandard against S.aureus and E.coli

 

Fig 4: Antibacterial activityBlank against S.aureus and E.coli

 

Table 3: Inhibition zones Diameter of standard (Gentamicin), blank (Ethanol) and extract (Rosa indica and Tagetes minuta)

Microbial cultures	Standard	Blank	Extract (Rosa indica and Tagetes minuta)
Staphylococcus aureus	15 mm	0 mm	14.8 mm
E.coli	14.7 mm	0 mm	14 mm

 

 

Table 4: Different composition of gels showing inhibitory zones

CULTURE	1%	2%	4%
E.Coli (Diameter of inhibition zones)	10mm	12mm	14mm
Staphylococcus aureus (Diameter of inhibition zones)	11mm	13mm	16mm

 

Figure 5: Antibacterial activity shown by different concentration of gels.

 

Table 5: Evaluation parameters of different gels

Formulation	pH	Appearance	Homogeneity	Spreadability after 1 min	viscosity
1%	6.1	Light golden yellow	Good	45 mm	4200
2%	5.5	Golden brown	Good	41mm	4486
4%	5.1	Dark golden brown	Good	39mm	4500

Figure 6: Graphical representation of different composition of gels showing inhibitory zones

 

SUMMARY AND CONCLUSION:

Based on literature and current investigation it has been found that the ethanolic extract obtained from flower petals of plants Rosa indica and Tagetes minuta possess significant antibacterial activity. From the above experimental results, it has been concluded that the formulation 4% gel shows best in appearance and shows better antibacterial effect than other gel formulations.

 

REFERENCES:

1.      Raquel Petrilli, Renata Fonseca Vianna Lopez, " Physical methods for topical skin drug delivery: concepts and applications" Braz. J. Pharm. Sci. vol.54 Nov 08, 2018.

2.      Sarode S.M, Bendale A.R, Mittal M, Chaudhri M.A, Kale M.K, Vidyasagar G, Shrivastava B, "Preparation and Evaluation of Anti-Inflammatory Transdermal Drug Delivery System" Asian Journal of Research in Chemistry Year: 2011, Volume: 4, Issue: 5, 791-794.

3.      Deepak p pawar, Prashant B Shamkuwar, "Formulation and evaluation of herbal gel containing Lantana camara leaves extract" Asian J Pharm Clin Res, Vol 6, Issue 3, 2013, 122-124.

4.      U.D. Shivhare, K. B. Jain, V. B. Mathur, K. P. Bhusari, . A. Roy "Formulation development and evaluation of diclofenac sodium gel using water soluble polyacrylamide polymer" Digest Journal of Nanomaterials and Biostructures Vol. 4, No.2, June 2009, p. 285 – 290.

 

Received on 09.04.2020         Modified on 19.04.2020

Accepted on 04.05.2020     ©AandV Publications All right reserved