Formulation and Evaluation of Mouth Ulcer Gel Using Basella Alba Extract

Mouth ulcers are localized disruptions in the oral mucosal lining that present as small, painful lesions These lesions usually appear as circular or oval-shaped spots, commonly located on the inner cheeks or lips, and often cause discomfort.^[1] Mouth ulcers are widely prevalent and may develop due to several underlying health conditions or triggering factors, but usually there is no serious underlying cause. Mouth ulcers can arise from several factors, such as inadequate intake of vital nutrients like iron and vitamins (particularly B12 and C), poor oral hygiene, infections, stress, indigestion, mechanical injury, food allergies, hormonal imbalance, skin disease etc. Aphthous ulcers often become more painful during routine activities like chewing, drinking, or brushing.

Factors responsible for the mouth ulcers

Multiple triggers may contribute to the development of mouth ulcers, including SLS-containing oral care products, irritation from sharp teeth or dental appliances, emotional stress, nutritional imbalance, allergies, hormonal fluctuations, genetic factors, and microbial infections.

Mouth ulcer symptoms

Although the symptoms differ based on the underlying cause, common clinical features include

1. Painful sores that may be yellow, white, or red.
2. Sores on the inside of the mouth, such as on your tongue or the insides of your cheeks or lips.
3. Areas of redness surrounding the sores.
4. Pain that worsens when you eat, drink, or talk.
5. Mouth ulcers are not usually contagious unless they’re caused by an infection such as hand, foot, and mouth disease.^[2]

Medicines used to treat mouth ulcers:

• Medicines such as non-prescription analgesics such as paracetamol or ibuprofen may be used to temporarily reduce pain and discomfort.
• Topical anaesthetic gels that contain agents such as lidocaine can numb the area and offer brief relief when applied to the ulcer.
• Antiseptic rinses like chlorhexidine can help limit microbial growth in the mouth and may support faster healing.^[3]

Herbal medicines:

Herbal medicines refer to preparations made from plant materials, including leaves, roots, seeds, or flowers, used for therapeutic purposes.^[4]

Herbal treatments are generally more affordable and are often preferred for minor conditions because they tend to produce fewer side effects and are widely accessible.

Example:

• Aloe vera contains various bioactive compounds and is widely used for its soothing and skin-protective properties.
• Neem is rich in natural antibacterial and anti-inflammatory compounds, making it useful for managing infections and promoting oral health.
• Turmeric contains curcumin, a powerful antioxidant with strong anti-inflammatory activity that supports tissue healing.
• Mint contains menthol and flavonoids that provide a cooling effect and help freshen the mouth while reducing mild irritation.^[5]

The main aim of the study was to formulate and evaluate herbal mouth ulcer gel using Basella alba, because it  is composed of flavonoids and phytochemicals known for their anti-ulcer, anti-inflammatory, and wound-healing actions.

Prevalence of mouth ulcer:

In total,71.1% had experienced oral aphthous ulceration. When both the genders were compared, RAS more commonly affected females (60.33%) than males (39.69). The regression coefficient for gender revealed that females were more prone to RAS than males.705 patients of total 3244 patients presented with recurrent aphthous ulcer at the time of examination. Giving an overall prevalence of 21.7% (table 1). Patients in the third (20.7) and fourth (26.5) decade were most commonly affected.

MATERIALS AND METHODOLOGY

Basella alba

Figure 1: Basella alba

Synonym: Basella rubra Roxb or Basella alba L

Common name: Malabar Spinach

Family: Basellaceae

Chemical constituents: Dried leaves (per 100 gm) contain proteins 20%, fat 3.5%, carbohydrates 54%, fiber 9%, ash 19%.

Uses:

Pharmacological activity:

• Androgenic activity
• Anti-inflammatory activity
• Antiulcer activity
• CNS depressant activity
• Wound healing activity
• Nephroprotective effect
• Antioxidant activity ^[6,7]

The materials used in this study included Carbopol 934 obtained from LOBO Chem, Mumbai; Triethanolamine procured from SDF Chem Ltd; Propylparaben and Methylparaben sourced from LOBO Chem, Mumbai; Sodium saccharin purchased from LOBA Chem Pvt. Ltd; and Glycerin obtained from SDF Chem Ltd.

Preformulation study:

Preformulation testing is essential to support the development of a stable, safe, and efficacious dosage form. It is a stage of development during which the pharmacist characterizes the physico–chemical properties of the drug substances and its interaction with various formulation components.

Goals of preformulation study

To determine the necessary physico-chemical parameter of a new drug substance.

Organoleptic evaluation:

Organoleptic characters such as color, odor, taste should be observed.

pH Determination

Topical cosmetic preparation should be within mouth skin layer pH, i.e., 6 – 7.5.

Measurement of pH (acidity) is performed by calibrating the pH meter.

Solubility

The solubility of herbal extract in various solvents was measured. Solubility was determined by taking 10 mg of drug sample in 10 ml of solvent as water, methanol, ethanol and chloroform.

Compatibility studies

• Compatibility tests between cosmetic ingredients can be performed using Fourier transform infra-red method. FT-IR, we can analyze the correlation between the components used whether they are compatible or not. If the peak of the spectrum from a single ingredient is stable after added to other ingredients are compatible.
• FT-IR is a simple methodology for detection of variations within drug-excipient blends. The disappearance of an absorption peak, a reduction of the peak intensity or the appearance of new peaks are indicative of existence of interaction between the API and the excipient under study.

Method of preparation of gel

The preparation of herbal mouth ulcer gel involves the use of natural ingredients and certain pharmaceutical excipients to create an effective topical medication. In this procedure, Basella alba (Malabar spinach), carbopol 934 (a carbomer), sodium saccharin, methyl paraben, propyl paraben, distilled water and triethanolamine are employed.^[8]

Method of preparation of herbal gel

Step1: Harvesting and drying

• Fresh Basella alba leaves were collected and thoroughly rinsed first with potable water followed by deionized water
• The cleaned leaves were initially shade-dried for 24–48 hours and later subjected to final drying in a hot-air oven.

Step 2: Extraction

• Grind the dried leaf into fine powder.
• Macerate the powder in the ethanol solution (1:2) for 24-48 hours.
• Filter the macerated solution using a fine cloth or filter paper to obtain a clear extract. This extract serves as API.

Step 3: Preparation of gel  

• Combined demineralized water and Carbopol 934 in a beaker on magnetic stirrer and allow the hydration of Carbopol with stirring. 
• Add 5 ml of distilled water with the propyl paraben, methyl paraben and sodium saccharin in another beaker on water bath.
• Then mix the extracts or powdered herbs in varying concentrations.
• All chemicals were combined with Carbopol 934.
• Triethanolamine should be added gradually while stirring to get the required pH.

Step 4: Packaging

• Transfer the finished herbal gel into a clean storage container.
• Store in a cool, dark place away from direct sunlight to maintain efficacy.^[9]

Table 2: Formulation table

Evaluation of gel:

The evaluation of gel formulation including assessments of visual appearance, pH, homogeneity, and spread ability, with spread ability quantified using a standardized formula involving the weight, slide length and separation time.

Visual appearance: The color, clarity, texture, transparency and occurrence of any gritty particles.

Measurement of pH: 1 gm of gel should be taken and distributed in 10 ml of distilled water and keep it aside for 2 hours. The measurement of pH of formulation is carried out in 3 times and the average values are stated.

Homogeneity: Gel formulations is tested for homogeneity by visual appearance after the gels were poured into the container. The gel is examined   for the presence and visual appearance of any aggregate’s masses.

Spread ability Spread ability is expressed in terms of time in seconds taken by 2 slides to slip off from gel that is placed in between the slides under the direction of certain weight. If the time taken for separation of 2 slides is fewer than well the spread ability.

Spread ability is calculated by using the formula:

S = M × L / T

Where, M = weight tied to upper slide

 L = length of glass slides

 T = time taken to separate the slides

Viscosity: The viscosity is analyzed by the brooked fields viscometer with Heli path using spindle number 64 at 10 rpm.

In vitro drug release study:

Franz diffusion cell was used to conduct an in vitro drug release study of an in herbal gel. In the donor compartment, 1 ml of formulation (equal to 1 g of gel) was deposited, and in the receptor compartment, freshly produced phosphate buffer solution (pH 6.8) was poured. The egg membrane was fitted between the chambers. One cell as blank was filled with only filled phosphate buffer solution. The units were then placed on a magnetic stirrer with thermostat. The medium was maintained at a constant temperature of 37 °C ± 0.5. After each 1 h interval, 1 ml of sample was withdrawn and same amount of phosphate buffer solution from blank was transferred into the sample cell for maintaining sink condition. Then, withdrawal amount was diluted to 10 ml in PBS pH 6.8, and concentration was measured using a UV-visible spectrophotometer at 200-400 nm with phosphate buffer solution pH 6.8 as a blank. The calibration curve was plotted and used to determine the percent cumulative drug release. The best fit model was tested for Korsmeyers, Peppas, and Fickinian diffusion mechanism for their kinetics.15,18^[10]

RESULT AND DISSCUSION

Preformulation studies

Any formulation development work has to be preceded by preformulation studies. This preformulation study includes selection of method of preparation, selection of polymers, drug- polymer compatibility study and analytical investigation of drug.

API characterization of the drug

Table 3: Organoleptic properties of Basella alba

Solubility

Solubility studies are carried out in different solvents as per IP and observation were showed.

Table 4: Solubility of Basella alba

Solubility is an important factor focused onto know the solubility of the drug with different solvent which is important criteria for formulating the medication. Basella alba was found to be completely soluble in methanol, soluble in ethanol and in other solvents like water drug was soluble but changes its color and partially soluble in chloroform.

pH studies of Basella alba

Table 5: pH of Basella alba

The pH range of the drug Basella alba is between 5.5 to 6 and the values are in the ranges as shown above.

 Drug excipient compatibility study

The compatibility study between drug and the carriers was carried out using FTIR spectrometer. The peak numbers of the drug exhibiting C-H, C=C, C-H, C-C stretching was observed and are depicted as below.

Figure 8: FT-IR spectrum of Basella alba

6: Interpretation of FT-IR spectral data of Basella alba

Figure 8: FT-IR spectrum of Drug + Polymer

Table 7: Interpretation of FT-IR spectral data of Drug + Polymer

Evaluation of herbal mouth ulcer gel

Physical evaluation

Table 8: Physical evaluation of gel

The physical evaluation of the formulation was carried out to ensure safety efficacy and quality control of the gel. The gel was found to be dark/light green in color and has musky odor and the gel is in semisolid state. The texture of gel was found to be smooth.

Determination of homogeneity

Table 9: Determination of homogeneity

Determination of Homogeneity was performed for ensuring product effectiveness, safety and user satisfaction. The visual appearance of the gel was found to be light green and dark green in colors and semisolid with gel type appearance the touch of the gel was found to be greasy and thick consistency.

Determination of spreadability:

Table 10: Determination of spreadability

Determination of viscosity

Table 11: Determination of Viscosity

Figure 10: Viscosity of gel formulation

The rheological properties of the solution are of importance in viewing of their proposed oral administration. The formulation should have an optimum viscosity that will allow easy swallowing as a liquid, which then undergoes a rapid Sol-gel transition due to ionic interaction. The prepared formulation was evaluated for their rheological property using a Brookfield viscometer. The order of viscosity of all formulations were F2<F3<F1<F4.

Determination of pH

Table 12: Determination of pH of gel

Figure11:  pH of formulation

In-vitro drug release studies

In-vitro diffusion study was carried out for all the prepared formulations from F1 to F4 and the result of diffusion profile for all the formulation were tabulated in the below.

Table 13: In-vitro drug release studies of herbal mouth ulcer gel of F1 formulation

Table 14: In-vitro drug release studies of herbal mouth ulcer gel of F2 formulation

Table 15: In-vitro drug release studies of herbal mouth ulcer gel of F3 formulation.

Table 16: In-vitro drug release studies of herbal mouth ulcer gel of F4 formulation.

CONCLUSION