FORMULATION AND DEVELOPMENT OF QUERCETIN AND MONOAMMONIUM GLYCYRRHIZINATE LOADED GEL FOR THE TREATMENT OF MOUTH ULCER DISEASE

FORMULATION AND DEVELOPMENT OF QUERCETIN AND MONOAMMONIUM GLYCYRRHIZINATE LOADED GEL FOR THE TREATMENT OF MOUTH ULCER DISEASE

Salunkhe Akshay Kumar Shivaji ^* and R. Vaidya Vaibhav

Department of Pharmaceutics, Dr. D. Y. Patil College of Pharmacy, Akurdi, Pune - 411044, Maharashtra, India.

ABSTRACT: Diagnosis of oral ulcerative lesions might be quite challenging. Oral ulcerative lesions were categorized into three major groups as acute, chronic, and recurrent ulcers. Helicobacter pylori (H. pylori) have been found in the oral cavity and stomach, and its infection is one of the most frequent worldwide. There is a close relation between H. pylori infection in the oral cavity and the stomach. H. pylori etiologic agent in recurrent apthous stomatitis, Canker sores. Quercetin inhibits the H. Pylori. Monoammonium glycyrrhizinate (MAG) was recog­nized as a compound possessing antimicrobial properties; MAG tested many researchers using the agar diffusion method which, show significant antibacterial activities against gram-positive (Bacillus subtilis, Stap Hylococcus aureus) as well as gram-negative bacteria (Escherichia coli, Pseu­domonas aeruginosa). The objective of the present investigation was to formulate and evaluate gel for mouth ulcer treatment of Quercetin and MAG. The concentration of carbopol p940 and propylene glycol were optimized using a central composite design for two factors. Formulations were evaluated for various parameters such as pH, viscosity, spreadability, drug content, cumulative drug diffusion, Texture analyzer, ex-vivo permeation, and mucoadhesion. The developed formulation was stable, safe, and effective for the treatment of mouth ulcer.

Mouth ulcers RAS, Quercetin, Monoammonium Glycyrrhizinate

INTRODUCTION: Ulcer means the break in the continuity of the epithelium brought about by molecular necrosis. Ulcers are most commonly occur in the oral region, for which the patient seeks help from their physician/dental surgeon. The presenting symptoms are usually redness, burning sensation and/or pain Fig. 1. They can present in any part of the oral cavity, but it becomes painful if it occurs in the movable area ¹. Mouth ulcers are very common and are usually occur due to trauma, such as from ill-fitting dentures, fractured teeth, or fillings Fig. 2.

However, patients with an ulcer of over 3 week’s duration should be referred for biopsy or other investigations to exclude malignancy or other serious conditions such as chronic infections. Ulcers related to trauma usually resolve within in a week after removal of the cause to maintain good oral hygiene ². MAG possesses a wide range of pharmacological and biological activities. It possesses anti-inflammatory activity, anti-ulcer, anti-bacterial, anti-allergic, antidote, antioxidant, antiviral, anti-tumour, anticonvulsant activity.

Drugs show anti-ulcer, anti-inflammatory, anti-bacterial, anti-viral, which play a major role in curing mouth ulcers, but the major reason for a section of this drug is its sweet test which is mainly not seen in other drugs, so this property increases patient compliance ^3-7. Quercetin chelates ions of transition metals such as iron which can initiate the formation of oxygen free radicals and inhibits xanthine oxidase activity and nitric oxide-induced radical damage, all of which have been implicated in the etiopathogenesis of recurrent aphthous stomatitis. In addition, quercetin also inhibits the enzymes cyclooxygenase and lipooxygenase which catalyzes the conversion of arachidonic acid to its metabolites, suggesting it to be a potent anti-inflammatory agent Relative Efficacy of Quercetin ^8-12.

FIG. 1: APTHOUS MAJOR ULCER IN LOWER LABIALMUCOSA

FIG. 2: TRAUMA ULCER DURING TOOTH BRUSHING, FLOSSING

Topical Drug Delivery: Topical drug delivery systems are localized drug delivery systems for local delivery of therapeutic agents via skin to treat the cutaneous disorder. These systems are generally used for local skin infections. The formulations are available in different forms, like from solid through semisolid to liquid.

If the drug substance in the solution has a favorable lipid/water partition coefficient and if it is a non-electrolyte, then drug absorption is enhanced via the skin. Dermatological products have various formulations and range in consistency though the most popular derma products are semisolid dosage forms ^13-14.

Advantages of Topical Drug Delivery Systems:

1. Avoidance of primary pass metabolism.
2. Convenient to use and easy to apply.
3. Easily to terminate the medications.
4. Drug delivered selectively to a specific site.
5. The gastro-intestinal incompatibility will be avoided.
6. Provides drug utilization with short biological half-life and narrow therapeutic window.
7. It provides effectiveness in low doses and by continuous drug input.
8. Avoids fluctuation in drug levels and risks.
9. A large area of application compared to other routes.
10. Drug delivery at a specific site

Disadvantages of Topical Drug Delivery Systems:

1. Possibility of local skin irritation at the site of application.
2. Contact dermatitis due to some drugs may occur.
3. Some drugs with poor permeability are difficult to penetrate via the skin.
4. Drugs with larger particle sizes are difficult to penetrate.
5. Possibility of allergenic reactions.
6. Drugs with a very small plasma concentration can be used for action

Types of Ulcer:

1. Acute Ulcer:

• Traumatic Ulcers
• Necrotizing Sialometaplasia
• Primary Herpetic Gingivostomatitis
• Varicella-Zoster Virus Infection
• Erythema multiforme
• Recurrent aphthous stomatitis [RAS]
• Behçet Disease [BD (Behçet Syndrome)]

2. Chronic Ulcer:

• Sustained traumatic ulcers (Decubitus ulcer)
• Squamous cell carcinoma
• Traumatic ulcerative granuloma (EosinopHilic ulcer of the tongue)

3. Pemphigus and Pemphigoid:

• Mucormycosis
• Tuberculous ulcers
• SypHilitic ulcers

MATERIALS AND METHODS:  

Materials:

Drugs:

• Quercetin was obtained from Research Lab Fine Chem. Industries, Mumbai.
• Mono-ammonium Glycyrrhizinate was pur-chased from SV Agro Mumbai.

Chemicals: Carbopol 934, Propylene glycol, Sodium benzoate

Solvents: Ethanol, Water.

Method:

Preformulation Studies: ^{13, 15-17}

The aim of Preformulation studies is as follow:

• To check physiochemical characterization of a new drug.
• To establish compatibility with different excipients.

Description: The sample was evaluated visually for appearance, color, and odor.

UV-visible Spectroscopy:

Calibration Curve of Curcumin:

Preparation Stock Solution of Quercetin: Ethanol AR: 10 mg pure Quercetin was dissolved in 100 ml ethanol to get a 100 ug/ml stock solution. Prepare concentration of 2 μg/ml, 4 μg/ml, 6 μg/ml, 8 μg/ml and 10 μg/ml respectively. Then measured absorbance of prepared dilutions at the respective wavelength.

Preparation Stock Solution of Quercetin: In pH 6.8 Phosphate Buffer 10 mg pure Quercetin was dissolved in 10 ml ethanol, then volume makeup upto 100 ml with pH 6.8 Phosphate Buffer, to get a 100 ug/ml stock solution. Prepare concentration of 2 μg/ml, 4 μg/ml, 6 μg/ml, 8 μg/ml and 10 μg/ml respectively. Then measured absorbance of prepared dilutions at the respective wavelength.

Preparation Stock Solution of Monoammonium Glycyrrhizinate:

Preparation Stock Solution of Monoammonium Glycyrrhizinate: Ethanol AR 10 mg pure MAG was dissolved in 100 ml ethanol to get a 100 ug/ml stock solution. Prepared dilutions of 5 μg/ml, 10 μg/ml, 15 μg/ml, 20 μg/ml, 25 μg/ml, 30 μg/ml, 35 μg/ml and 40 μg/ml respectively. Then measured absorbance of prepared dilutions at the respective wavelength.

Preparation Stock Solution of MAG: In pH 6.8 Phosphate buffer, 10 mg pure drug of MAG was dissolved in 10 ml ethanol, then volume makeup upto 100  ml with pH 6.8 phosphate buffer, to get a 100 ug/ml stock solution. Prepare dilutions of 5 μg/ml, 10 μg/ml, 15 μg/ml, 20 μg/ml, 25 μg/ml, 30 μg/ml 35 μg/ml and 40 μg/ml respectively. Then measured absorbance of prepared dilutions at the respective wavelength.

Formulation Design:

Formulation of Gel by Different Concentrations:

Preparation of optimization of Formulation: ¹⁸ Prepare Gel with different concentrations of Gelling agent and penetration enhancer by using design expert. Carbopol in the range of 0.5-2% and propylene glycol in the range 3-7% Table 1.

TABLE 1: OPTIMIZING DESIGN VALUES OF CARBOPOL P940 AND PROPYLENE GLYCOL

Procedure for Formulation of Gel: ¹⁶

• Gel formulations were prepared using carbopol P940 as gelling agents and Propylene Glycol as penetration enhancer.
• The carbopol P940 was mixed with Propylene Glycol in a beaker heated at 70 ºC and
• Quercetin and MAG in a suitable solvent (ethanol) were added to the is dispersion.
• The preservative sodium benzoate was dissolved in water using heat the solution was left to cool and then warmed to about 70 ºC with vigorously stirring using an electric stirrer.
• Resulting solution added to the above solution with stirring until a gel was formed Table 2.

TABLE 2: FORMULATION TABLE OF GEL WITH DIFFERENT CONCENTRATION OF CARBOPOL P340 ANDPG

Evaluation of Gel: ^19-22

1. pH
2. Spreadability test.
3. Drug content test.
4. In-vitro drug diffusion study studies.

Finalization of Batch: Finalization of batch depends on various evaluations of all nine (F1-F9) batches.

From evaluation parameters which batch fulfill all evaluation parameters, this batch selected as optimized batch.

Evaluation of Optimized Gel: ^23-29s

1. Viscosity Table 13.
2. Texture analyzer Table 12, Fig. 6.
3. Ex-vivo drug permeation Table 14, 3.
4. Mucoadhesive strength determination Table 15.
5. Antimicrobial study Table 16, Fig. 7.
6. Stability testing Table 17, 18.
7. pH
8. Spreadability
9. Percentage drug content

RESULTS:

Physical Properties of Drug:

TABLE 3: APPEARANCE OF QUERCETIN AND MONOAMMONIUM GLYCYRRHIZINATE

UV-visible Spectroscopy:

Summary of Validation Parameter:

TABLE 4: SUMMARY OF VALIDATION PARAMETER OF QUERCETIN AND MAG IN ETHANOL (AR)

TABLE 5: SUMMARY OF VALIDATION PARAMETER OF QUERCETIN AND MAG IN pH 6.8 PHOSPHATE BUFFER

TABLE 6: pH OF DIFFERENT FORMULATION OF GEL

TABLE 7: SPREADABILITY OF DIFFERENT FORMULATION OF GEL

FIG. 3D AND 2D: PLOT FOR EFFECT OF CONCENTRATION OF CARBOPOL AND CONC. OF PG ON SPREADABILITY

TABLE 8: PERCENTAGE DRUG CONTENT OF QUERCETIN FOR DIFFERENT FORMULATION OF GEL

TABLE 9: PERCENTAGE DRUG CONTENT OF MAG FOR DIFFERENT FORMULATION OF GEL

TABLE 10: % DRUG RELEASE OF QUERCETIN FROM DIFFERENT GEL FORMULATION FOR DIFFERENT TIME INTERVAL

GRAPH 1: %DRUG RELEASE OF QUERCETIN FROM DIFFERENT GEL FORMULATION FOR DIFFERENT TIME INTERVAL

TABLE 11: % DRUG RELEASE OF MAG FROM DIFFERENT GEL FORMULATION FOR DIFFERENT TIME INTERVAL

GRAPH 2: %DRUG RELEASE OF MAG FROM DIFFERENT GEL FORMULATION FOR DIFFERENT TIME INTERVAL

FIG. 3: 3D AND 2D PLOT FOR EFFECT OF CONC. OF CARBOPOL AND CONC. OF PG ON DRUG DIFFUSION OF QUERCETIN

FIG. 4-3D AND 2D: PLOT FOR EFFECT OF CONC. OF CARBOPOL AND CONC. OF PG ON DRUG DIFFUSION OF MAG

Finalization of Batch: Selection of final batch depends on various evaluation parameters e.g., spred ability, in-vitro diffusion study.

After evaluation of the above parameters, we selected “F2” formulation, which shown the ideal property for gel.

Evaluation of Optimized Gel:

Texture Analyzer:

TABLE 12:  TEXTURE ANALYZER FOR OPTIMIZED FORMULATION

FIG. 6: TEXTURE ANALYZER OF OPTIMIZED FORMULATION (TIME (S) VS LOAD (G)

Viscosity:

TABLE 13: VISCOSITY OF OPTIMIZED FORMU-LATION

Ex-vivo Drug Permeation Table 14, Graph 3:

TABLE 14: EX-VIVO PERMEATION OF QUERCETIN AND MAG THROUGH GOAT MEMBRANE OF OPTIMIZED FORMULATION

GRAPH 3: EX-VIVO PERMEATION OF QUERCETIN AND MAG THROUGH GOAT BUCCAL MUCOSA OF OPTIMIZED FORMULATION

Mucoadhesive Strength Determination Table 15:

TABLE 15: DETERMINATION OF MUCOADHESIVE STRENGTH OF OPTIMIZED FORMULATION (F2)

Antimicrobial Study:

TABLE 16: DETERMINATION OF ANTIMICROBIAL ACTIVITY OF OPTIMIZED FORMULATION

FIG. 7: DETERMINATION OF ANTIMICROBIAL ACTIVITY OF OPTIMIZED FORMULATION

Stability Testing Table 17, 18:

pH:

Table 17: pH of Optimized Gel (F2) for Stability Study at 5 ºC and 25 ºC

Spreadability:

TABLE 18: SPREADABILITY OF OPTIMIZED GEL (F2) FOR STABILITY STUDY AT 5 ºC AND 25 ºC

Percentage Drug Content:

TABLE 19: % DRUG CONTENT OF OPTIMIZED GEL (F2) FOR STABILITY STUDY AT 5 ºC AND 25 ºC

SUMMARY: The present work describes a study on “Formulation Development and Evaluation studies of Quercetin and MAG gels for the treatment of Mouth ulcer”. The polymers, namely Carbopol-940, Propylene Glycol were used for the formulation of gels and studied for their drug release from the gel formulations. It is evident from the IR spectrum that all the polymers used in the gel formulations were compatible with the drug Quercetin and MAG. The calibration curve of Quercetin and MAG was performed in ethanol and Phosphate Buffer pH 6.8 which can be used for drug quantification in diffusion analysis. Different formulations of gel were prepared by using a varying concentration of Carbopol-940 and propylene glycol depending upon their range. Carbopol gels were non-greasy and smooth on application. Evaluation of gel involved (PH, Spreadability, Drug content, Drug diffusion). Evaluation of Final batch of gel involved (PH, Spreadability, drug content, Drug diffusion, Viscosity, Texture analyzer, Muco-adhesive strength, ex-vivo tissue permeation). In optima-zation of Gel, F2 formulation was found better results compared to other formulation.

CONCLUSION: The present study of Quercetin and monoammonium glycyrrhizinate Gel increase patient compliance of gel and also to mask the test of quercetin with the help of sweet and pleasant test of Monoammonium glycyrrhizinate. The combination of Quercetin and MAG would be treating mouth ulcer Disease more effectively due to both drug shows activity and gives more benefits. Advantages of Quercetin and MAG combinational drug therapy doesn’t have any serious side effect after for prolonged use and also maintain good bacterial flora of mouth which is important to minimizing ulcerative condition. Quercetin and MAG also has cardiovascular protection, anticancer, antitumor, anti-ulcer, anti-allergy, anti-viral, anti-inflammatory activity, anti-diabetic, gastroprotective effects, antihypertensive, immune-modulatory and antiinfective activity

ACKNOWLEDGEMENT: I would like to express thanks to the “International Journal of Pharmaceutical Sciences and Research” for supporting us at every point and provide such a huge platform where we can publish our paper.

I would also like to express thanks to Research lab fine chem. Industries, Mumbai for providing me Quercetin and SV Agro Pvt. Ltd. for providing me Monoammonium Glycyrrhizinate for my research work. I would also like to thanks my project guide Dr. Vaibhav R. Vaidya, for helping me at every point of my project work.

CONFLICTS OF INTEREST: The author reports no conflicts of interest.

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    How to cite this article:

    Shivaji SAK and Vaibhav RV: Formulation and development of quercetin and monoammonium glycyrrhizinate loaded gel for the treatment of mouth ulcer disease. Int J Pharm Sci & Res 2021; 12(3): 1789-98. doi: 10.13040/IJPSR.0975-8232.12(3).1789-98.

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