FORMULATION AND DEVELOPMENT OF QUERCETIN AND MONOAMMONIUM GLYCYRRHIZINATE LOADED GEL FOR THE TREATMENT OF MOUTH ULCER DISEASEHTML Full Text
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:
- Avoidance of primary pass metabolism.
- Convenient to use and easy to apply.
- Easily to terminate the medications.
- Drug delivered selectively to a specific site.
- The gastro-intestinal incompatibility will be avoided.
- Provides drug utilization with short biological half-life and narrow therapeutic window.
- It provides effectiveness in low doses and by continuous drug input.
- Avoids fluctuation in drug levels and risks.
- A large area of application compared to other routes.
- Drug delivery at a specific site
Disadvantages of Topical Drug Delivery Systems:
- Possibility of local skin irritation at the site of application.
- Contact dermatitis due to some drugs may occur.
- Some drugs with poor permeability are difficult to penetrate via the skin.
- Drugs with larger particle sizes are difficult to penetrate.
- Possibility of allergenic reactions.
- Drugs with a very small plasma concentration can be used for action
Types of Ulcer:
- Acute Ulcer:
- Traumatic Ulcers
- Necrotizing Sialometaplasia
- Primary Herpetic Gingivostomatitis
- Varicella-Zoster Virus Infection
- Erythema multiforme
- Recurrent aphthous stomatitis [RAS]
- Behçet Disease [BD (Behçet Syndrome)]
- Chronic Ulcer:
- Sustained traumatic ulcers (Decubitus ulcer)
- Squamous cell carcinoma
- Traumatic ulcerative granuloma (EosinopHilic ulcer of the tongue)
- Pemphigus and Pemphigoid:
- Tuberculous ulcers
- SypHilitic ulcers
MATERIALS AND METHODS:
- 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.
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.
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 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
|Factor||Carbopol 940||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
|Ingredients||Different Batches of Formulation (Concentration in Percentage)|
|F 1||F 2||F 3||F 4||F 5||F 6||F 7||F 8||F 9|
|Distill Water||q.s 100 ml||q.s 100 ml||q.s 100 ml||q.s 100 ml||q.s 100 ml||q.s 100 ml||q.s100 ml||q.s 100 ml||q.s100 ml|
Evaluation of Gel: 19-22
- Spreadability test.
- Drug content test.
- 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
- Viscosity Table 13.
- Texture analyzer Table 12, Fig. 6.
- Ex-vivo drug permeation Table 14, 3.
- Mucoadhesive strength determination Table 15.
- Antimicrobial study Table 16, Fig. 7.
- Stability testing Table 17, 18.
- Percentage drug content
Physical Properties of Drug:
TABLE 3: APPEARANCE OF QUERCETIN AND MONOAMMONIUM GLYCYRRHIZINATE
|Appearance||Light yellowish orange color powder with characteristic odor.||Bright yellow-orange powder with characteristic odor||White powder with characteristic odor.||White powder with a pleasant odour.|
Summary of Validation Parameter:
TABLE 4: SUMMARY OF VALIDATION PARAMETER OF QUERCETIN AND MAG IN ETHANOL (AR)
|S. no.||Parameter||Result of Curcumin||Result of MAG|
|1||Absorption maxima (ƛmax)||377nm||248 nm|
|2||Linearity Range (µg/ml)||2-10||10-50|
|3||Correlation Coefficient (R2)||0.996||0.9969|
|4||Standard regression equation||y = 0.0269||y = 0.0118x|
TABLE 5: SUMMARY OF VALIDATION PARAMETER OF QUERCETIN AND MAG IN pH 6.8 PHOSPHATE BUFFER
|S. no.||Parameter||Result of Curcumin||Result of MAG|
|1||Absorption maxima (ƛmax)||372nm||248 nm|
|2||Linearity Range (µg/ml)||2-10||10-50|
|3||Correlation Coefficient (R2)||0.9979||0.9946|
|4||Standard regression equation||y = 0.0577x + 0.0018||y = 0.0129x|
TABLE 6: pH OF DIFFERENT FORMULATION OF GEL
|Formulation No.||1||2||3||Mean PH||S. D|
TABLE 7: SPREADABILITY OF DIFFERENT FORMULATION OF GEL
|Formulation No.||1||2||3||Mean Diameter||S. D|
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
|Formulation No.||1||2||3||Mean % Drug Content||S. D|
TABLE 9: PERCENTAGE DRUG CONTENT OF MAG FOR DIFFERENT FORMULATION OF GEL
|Formulation No.||1||2||3||Mean % Drug Content||S. D|
TABLE 10: % DRUG RELEASE OF QUERCETIN FROM DIFFERENT GEL FORMULATION FOR DIFFERENT TIME INTERVAL
|Time in Minutes||Formulation Number|
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
|Time in Minutes||Formulation Number|
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:
TABLE 12: TEXTURE ANALYZER FOR OPTIMIZED FORMULATION
|Load at Target||55 g|
|Adhesive Force||20 g|
|Stringiness Work done||0.3 mJ|
FIG. 6: TEXTURE ANALYZER OF OPTIMIZED FORMULATION (TIME (S) VS LOAD (G)
TABLE 13: VISCOSITY OF OPTIMIZED FORMU-LATION
|RPM||1||2||3||Mean Viscosity (cP)||S.D|
Ex-vivo Drug Permeation Table 14, Graph 3:
TABLE 14: EX-VIVO PERMEATION OF QUERCETIN AND MAG THROUGH GOAT MEMBRANE OF OPTIMIZED FORMULATION
|Time in Minutes||Cumulative % Drug Permeation Quercetin||Cumulative % Drug Permeation MAG|
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)
|Formulation||1||2||3||Mean Strength||S. D|
TABLE 16: DETERMINATION OF ANTIMICROBIAL ACTIVITY OF OPTIMIZED FORMULATION
|F 2||10 ± 1|
|Marketed||15.667 ± 2.33|
FIG. 7: DETERMINATION OF ANTIMICROBIAL ACTIVITY OF OPTIMIZED FORMULATION
Stability Testing Table 17, 18:
Table 17: pH of Optimized Gel (F2) for Stability Study at 5 ºC and 25 ºC
|Time (Months)||pH at 5 ºC||pH at 25 ºC|
TABLE 18: SPREADABILITY OF OPTIMIZED GEL (F2) FOR STABILITY STUDY AT 5 ºC AND 25 ºC
|Time (Months)||Spread Ability
at 5 ºC
Percentage Drug Content:
TABLE 19: % DRUG CONTENT OF OPTIMIZED GEL (F2) FOR STABILITY STUDY AT 5 ºC AND 25 ºC
|Time (Months)||Drug Content of Quercetin at 5 ºC||Drug Content of Quercetin at 25 ºC||Drug Content MAG at 5 ºC||Drug Content MAG at 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.
- Sivapathasundharam B, Sundararaman P and Kannan K: Oral ulcers - a review. J of Dentis & Oral Disorders 2018.
- Crispian S and Rosemary S: Clinical review on Mouth ulcers and other causes of orofacial soreness and pain. BMJ 2000; 321.
- Tinku G and Mohd M: Ammonium glycyrrhizinate: a comprehensive review of its traditional use, phytochemistry, pharmacology & safety. Ajper 2018; 7(1): 58-65.
- Shaina K, Kumar VS, Neha, Arshvir K and Narinder S: Review on glycyrrhiza glabra (liquorice) and its pharmacological activities. Int J Pharm Drug Anal 2016; 5(4): 234-39.
- Geetha RV and Anita R: In-vitro evaluation of anti bacterial activity of ethanolic root extract of glycyrrhiza glabra on oral microbes. Int J Dru De & Re 2012; 4(4): 161-65.
- Adel AM, Laith AA and Hatem AS: Licorice: A possible anti-inflammatory and anti-ulcer drug. AAPS Pharm Sci Tech 2005; 6(1): 13.
- Yogesh ST, Asha MSA, Shital VK and Avinash HH: Treatment of mouth ulcer by curcumin loaded thermoreversible mucoadhesive gel: a technical note. Int J Pharm Pharm Sci 7(10): 399-02.
- National Center for Biotechnology Information. Pub Chem Database. Quercetin, CID = 5280343, https://pubchem.ncbi.nlm.nih.gov/compound/Quercetin (accessed on Mar. 9; 2020.
- Adler I, Muiño A, Aguas S, Harada L, Diaz M, Lence A, Labbrozzi M, Muiño JM, Elsner B, Avagnina A and Denninghoff V; Helicobacter pylori and oral pathology: Relationship with the gastric infection. World J Gastroenterol 2014; 20(29): 9922-35.
- Ibrahim SA, Hassan MAMM, El-Mahdy and Mohamed AS: Formulation and evaluation of quercetin in certain dermatological preparations. J Drug Del Sc Tech 2007; 17(6): 431-36.
- Suzuki Y, Ishihara M, Tetsuya S and Ito M: Anti-ulcer effects of antioxidants, quercetin, α-tocopherol, nifedipine and tetracycline in rats jpn. J P Harmacol1998; 78: 435-41.
- Ashni V, Sukhdev S, Rupinder K and Upendra KJ: Topical gels as drug delivery systems: a review. Int J Pharm Sci Rev Res 2013; 23(2).
- Shradha M, Gowda DV, Vishal GN and Akhila AR: An overview on topical drug delivery system Updated review. Wed. 08 Jan 2020 in International Journal of Research in Pharmaceutical Sci 2020.
- Alexander V, Anand D, Radhakrishnan A and Subramani P: Overviews of biological importance of quercetin: a bioactive flavonoid. Pharmacogn Rev 2016; 10(20): 84-89.
- Manoj NM, Kailas CM, Balaji VD and Sajid NS: Studies of antibacterial activities of glycyrrhiza glabra root extract. Int. J Pharm Tech Res 2010; 2: 899.
- Rowe RC, Sheskey PJ and Quinn ME: Handbook of Pharmaceutics excipients. Pharmaceutical press. London UK Sixth Edition 2009; 110-113, 592-593, 627-628, 766-769.
- Shaikh S, Shete A and Doijad R: Formulation and evaluation pharmaceutical aqueous gel of powdered guava leaves for mouth ulcer treatment. Pharma Tutor 2018; 6(4); 32-38.
- Divyesh Y, Ghanshyam P, Bhavin B, Sunita C and Upendra P: Formulation and evaluation of gel containg amlexanox for mouth ulcer. IJPRBS 2015; 4(2): 356-64.
- Dantas MJB, Reis SABJ, Damasceno CMD, Rolim LA and Rolim-Neto PJ: Development and evaluation of stability of a gel formulation containing the monoterpene borneol. Hinda Pub Corp Scientif World J 2016; 7394685.
- Kumar RV, Prasad NY, Priyam S, Nidhi M, Suaib L, Harinath D, Koshy MK and Shubhini AS: Development of cellulosic polymer based gel of novel ternary mixture of miconazole nitrate for buccal delivery. © 2013 Elsevier Ltd Carbohydrate Polymers 2014; 103: 126-33.
- Prasad YN, Kumar RV, Nidhi M, Priyam S, Umrao BD, Pal Anirban., Kumar AT and Singh CC: A Novel approach for development and characterization of effective mosquito repellent cream formulation containing citronella oil. Bio Med Research International 2014; 786084.
- Brookfield CT3 Texture Analyzer Operating Instruction Manual No. M/08-371A0708, Brookfield Engineering Laboratories Middleborough Mass USA 2011.
- Himabindu P, Vasudha B and Narender B: Formulation, in-vitro and ex-vivo characterization of mucoadhesive buccal tablets for antihypertensive drug. Asian J Pharm Clin Res2018; 11(8); 402-11.
- Gaikwad V: Formulation and evaluation of in-situ gel of metoprolol tartrate for nasal delivery. J Pharm Res 2010; 3: 788-93.
- Prasad YN, Kumar RV, Nidhi M, Priyam S, Umrao BD, Anirban P, Kumar TA and Singh CC: A novel approach for development and characterization of effective mosquito repellent cream formulation containing citronella oil. BioMed Research International 2014; 786084.
- Narayana CR, Parvathy PD, Jobin J and Veena AS: Formulation and evaluation of mucoadhesive oral gel containing miconazole nitrate for oral candidiasis. Research J Pharm and Tech 2013; 6(11): 1251-57.
- ICH guidelines. Stability testing of new drug substances and products, 27th October 1993.
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.
All © 2013 are reserved by the International Journal of Pharmaceutical Sciences and Research. This Journal licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 3.0 Unported License.
S. A. K. Shivaji * and R. V. Vaibhav
Department of Pharmaceutics, Dr. D. Y. Patil College of Pharmacy, Akurdi, Pune, Maharashtra, India.
23 March 2020
22 June 2020
30 June 2020
01 March 2021