INTRODUCTION:

Acne is a type of skin infection. It usually happens at the time of puberty, when the sebaceous (oil) glands begin to function. Androgens are produced by the male and female adrenal glands and stimulate the glands. Acne vulgaris is a type of acne that means "common acne."

It is a skin infection caused by sebaceous gland changes. The red colour is caused by skin inflammation caused by infection in the skin. Acne is a general skin condition that is related to pimples and is common among teenagers. Acne affects the majority of people, but it is most common in teenagers who are going through hormonal changes.^1-5

Acne comes in many forms, including acne vulgaris, rosacea, acne cosmos, acne fulminans, and acne mechanique, with symptoms and indications such as erythema, scaling, burning, itching, hyperpigmentation, and pain. Acne can be caused by genetics (people have XYY karyotype), diet (high glycaemic load, insulin resistance, occlusive cosmetics, puberty, etc.), and medicine (some drugs like benzodiazepines, cyclosporine, lithium, ramipril, corticosteroids, isoniazid, and vitamin B).^6-8

Many treatments are designed to prevent new outbreaks and must be applied to the entire affected skin area, not just the affected area. It can take 6 to 8 weeks for any type of treatment to show any benefit, so it's usually worth sticking with one treatment before switching to another.^9-12

The different topical drug groups available such as comedolytics in which Tretinoin, Isotretinoin, Benzoyl peroxide, Azelaic Acid, Benzoyl Peroxide, Tazarotene. Keratolytic include salicylic acid, sulphur and alpha-hydroxy acids are used. Benzoyl peroxide, Benzoyl peroxide, and topical steroids are examples of anti-inflammatory drugs⁵. The Benzoyl peroxide also called as ‘nicotinamide’ its chemical name is Pyridine-3-carboxamide.⁶It showes antiinflamatory as well as sebostatic activity⁷. Silver nanoparticles show strong antibacterial activity, which can completely inhibit the growth of a variety of fungi as well as gram-negative and gram-positive bacteria.^13-16

MATERIAL AND METHOD:

Materials: Benzoyl peroxide was a gift sample from Red Cros Pharmaceuticals (Aurangabad). carbapol, methyl paraben, Triethanolamine, HPMC, Propylene glycol, silver nitrate, potassium dihydrogen orthophosphate was procured from S.D Fine Chem Mumbai.

Methodology: During the procedure, first the cuvette was filled with different concentrations of the stock solution, and then UV scanning of the sample was done, in which Benzoyl peroxide absorption was found to be greatest at 262nm¹⁸.

Fig: 1 Standard calibration curve for Benzoyl peroxide

Preparation of silver nanoparticles:

Put 1 litre of distilled water in a beaker that is covered with aluminium foil. Add 57.6mg of silver nitrate to 100ml of distilled water. Add in 5% Benzoyl peroxide medication. Start heating the solution. Heat the solution to 40-45^◦C Add a thermometer to measure the temperature. When the drug is bound with silver nitrate, in which we see dispersed silver particles, then stop heating. Put it into an ice water bath. Keep it cool and dark in a dark-coloured bottle.^19-23

Preparation of gel containing Benzoyl peroxide with silver nanoparticle:

By using different concentrations of Carbopol, the Benzoyl peroxide-containing antiacne gel was prepared. In all six batches we change the concentration of Carbopol, i.e., in the first batch we take 0.5gm, in the second we take 1 gm, in the third 1.5 and so on. For the preparation of gel, first take a specific quantity of water, then sprinkle the weighed Carbopol. then stirred it well. The stirring was done by using the magnetic stirrer. Then correctly weighed the drug and dispersed it in water in another beaker. The triethanolamine was used for maintaining the pH. It can neutralise the Carbopol mixture. Then it can increase the transparency of the gel, and in that mixture, solvates and other ingredients are added. The uniformity was maintained by using the homogenizer.^24-26

Table 1: Composition of developed formulation

Ingredients	F1	F2	F3	F4	F5	F6
Benzoyl peroxide	5.0	5.0	5.0	5.0	5.0	5.0
Silver Nitrate	0.057	0.057	0.057	0.057	0.057	0.057
Carbopol	0.5	1	1.5	2	2.5	3
HPMC	0.5	0.5	0.5	0.5	0.5	0.5
Propylene Glycol	15.0	15.0	15.0	15.0	15.0	15.0
Methyl paraben	0.1	0.1	0.1	0.1	0.1	0.1
Triethanol amine	Q. S	Q. S	Q. S	Q. S	Q. S	Q. S
Purified water	Q. S	Q. S	Q. S	Q. S	Q. S	Q. S
	Quantity taken Per 100 gm Gel (In gram)

EVALUATION PARAMETERS:

pH: The formulation’s pH can be determined using a digital pH metre. One gramme of gel was mixed in 100mililiter of distilled water and stirred thoroughly for two hours to determine the pH. The pH range of the formulation is five to six, which is considered normal and does not cause skin irritation.²⁷

Spread ability:

The determination of spread ability is very important because it gives us information about the extent to which it spreads when applied to the skin. From the spreading value, we can also determine the bioavailability of gel. For the determination of spread ability, we took two slides in between two slides. We placed gel, and the time is measured when the two slides slip from gel in seconds. Then separating of the two slides was done if it took less time, from which we conclude that the spread ability was improved. When the gel was placed in between the two slides, a 20gram weight²⁸

was put on the upper slide. The time was measured when the upper slide was travelling 6.0 cm and under the influence of weight, time was separated from the lower slide. The average value was calculated by repeating the experiment three times.²⁹

The following formula was used to calculate spread ability:

Spread ability = Wt. tide to upper slide/Time is taken to separate both slides

Viscosity: For the determination of viscosity, we can use the Brookfield viscometer. The measurements were done four times at 2, 2.5rpm, and the readings were multiplied with a factor, and the mean which was obtained was used as the final viscosity in cp. The factor finder of Brookfield was used in the following manner.³⁰

Dial reading × factor = Viscosity in centipois

Antibacterial activity: The modified agar well diffusion method was used for the antibacterial study. In this process, on the nutrient agar plates After 24hours, 0.2ml of staphylococcus aureus broth culture was added. The drying of the plate required one hour. Then in each plate, four equidistance wells are made by a sterile 8mm borer. One gramme of formulation (F1-F6) and commercial Azithromycin gel were used for comparison. A positive control of sliver colloidal with 5% Benzoyl peroxide and a negative control of sodium chloride solution were introduced into the wells at random. The incubation of the plate was done for ‘24 hours at 37^◦C’. After that zone of inhibition was measured in mm. This experiment was repeated three times.³¹

Drug content: A 100mg sample was placed in a 100ml phosphate buffer (pH 5.5) and stirred for two hours on a magnetic stirrer. Then the filtration of the solution was done by using the Whatman filter paper, and one millilitre of filtrate was transferred to a 100-millilitre volumetric flask and diluted to volume. The concentration of the drug in the sample containing the sample was calculated using the slope and intercept obtained from the standard calibration curve of Benzoyl peroxide in distilled water. This required the absorbance value, which is obtained from the U V spectrophotometer at a wave length of 262nm.³²

Extrudability study: When the formulation was filled into the collapsible tubes, The extrudability was measured by placing the weight on it and then the weight in gm was measured to extrude a gel from the tube.³³26.7gm of weight is required to extrude a gel from a tube for 10 sec.

In vitro drug diffusion study: Glass cylinders open on both sides are used as the device. Use adhesive to attach the dialysis membrane that has been dipped in distilled water for 24hours before use to one end of the cylinder. Place the gel containing 10mg of nicotinamide in the cells (donor chamber) and immerse the cells in a beaker containing 100ml of PBS, pH 7.4. Position the entire assembly so that the bottom of the cell-containing gel is just above the surface of the diffusion medium (1-2mm deep) and stir the medium with a magnetic stirrer at 37 °C. Add the same volume of fresh buffer to an aliquot (5 ml). Check the samples using a UV-Vis spectrophotometer set at 262nm.^34-38

RESULT AND DISCUSSION:

Determining the λmax of the Benzoyl peroxide spectrum: Averaging and concentration values, graphing and slope values, calculating correlation and regression, and using these values as computational standards for in vitro diffusion studies are used for standard calibration.

Physicochemical evaluation of gel formulations: From the physicochemical analysis, it was determined that all batches were clear, homogeneous, and watery smooth in texture. The gel formulation had a clear physical appearance and a pH range of 5–6.

Spread ability: The formulation's spread ability ranged from 15.29 to 24.51gm/sec. As the polymer concentration in the formulation increases, the viscosity of the formulation increases and the spread ability decreases. Thus, batch F1 in Table no 2 has a lower concentration of Carbopol polymer, resulting in a lower viscosity and greater spread ability, whereas batch F6 has a higher concentration of Carbopol, resulting in a higher viscosity and less spread ability.

Table no: 2 Spread ability of gel formulations

Formulations	Average spread ability (g.cm/sec)
F1	24.51
F2	23.24
F3	23.27
F4	19.56
F5	16.35
F6	15.298

Drug content: In Table No.3 The gel formulation drug content was to be consistent across all formulations, and the range was 91.26±0.74 to 98.74±0.63%.

Table No.3: Percent drug content of gel formulation

Formulation	Drug content in %
F1	95.58± 0.85
F2	92.45± 0.52
F3	98.74± 0.63
F4	97.25±0.58
F5	91.26±0.74
F6	93.84 ±0.41

Viscosity: Each formulation’s viscosity was determined. The viscosity of the F1 batch in Table no 4 is lower than that of the other formulations. Depending on the concentration of Carbopol and the solvent, the viscosity of the formulation changes, which affects the formulation's release. The viscosity of the formulation increases as the concentration of Carbopol 940 increases, so batch F6 has more viscosity.

Table no. 4: Viscosity of gel formulation

Formulation	Viscosity(cp)
F1	434± 36.56
F2	480± 43.23
F3	491± 34.34
F4	583± 43.56
F5	593 ± 45.32
F6	651± 41.43

Antibacterial activity:

The antibacterial activity zone of inhibition was compared to the standard azithromycin gel which is shown in figure no 3. The zone of inhibition of azithromycin was discovered to be greater than that of all other formulations. In Table no 5 the zone of inhibition was found to be between 12.23 to 21.65mm, which shows that the formula shows good sensitivity against bacteria.

Fig: 2 Antibacterial study of gel formulation

Table no. 5: Viscosity of gel formulation

Formulation	Zone of inhibition (mm)
F1	16.00
F2	19.34
F3	21.65
F4	12.23
F5	17.35
F6	12.67

In vitro drug diffusion study: As Carbopol concentration increases, the percentage of drug released decreases slightly. The drug's release from Carbopol is reduced due to the formation of a tight, hard gel. The viscosity of the system increases as the drug penetrates the hydrogel network, and the drug does not easily come out of the network, so the percentage of drug release decreases as the concentration of Carbopol increases. As a result of the higher viscosity, batch F1 form Table no 6 show that released 80.74 percent, while batch F6 released 71.52 percent. Which also clearly shown in fig no 3

Table no.6 Drug release of gel formulation

Time (HR)	F1	F2	F3	F4	F5	F6
0	0	0	0	0	0	0
1	18.25	16.25	12.58	10.65	9.65	8.47
2	25.25	21.69	19.25	19.02	18.69	18.58
4	29.58	28.67	26.69	24.25	21.52	19.74
6	49.65	47.52	45.32	41.15	39.74	37.63
8	58.62	56.82	54.95	53.21	52.84	51.54
10	64.62	61.28	50.62	48.75	46.24	43.85
12	80.74	78.25	75.63	74.56	73.96	71.52

Fig: 3 Graph showing the release of drug of all formulations

CONCLUSION:

The preparation of silver nanoparticles and incorporation into the Benzoyl peroxide-containing gel were done successfully. Various evaluations, i.e., the physiochemical analysis, spread ability, viscosity, drug content, drug release, and antibacterial study were done. This study demonstrates that the gel has a good texture, is easily spreadable, has high bioavailability, and is effective in treating acne.

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Received on 19.07.2023 Modified on 24.08.2023

Res. J. Pharma. Dosage Forms and Tech.2024; 16(1):1-5.

DOI: 10.52711/0975-4377.2024.00001