Formulation and Evaluation of Anti-acne Herbal Face Wash Using Broccoli Sprouts

The purpose of herbal cosmetics is to improve people's attractiveness. The demand for herbal formulations is very significant worldwide. With synthetic issues, it is more acceptable to think that natural solutions are safer than those with less negative effects. Nearly everyone has had acne vulgaris at some point in their lives, making it an extremely common skin ailment. Although it affects many adults between the ages of 15 and 30, it is most prevalent during adolescence ^[1]. Mild to moderate acne can be treated using an anti-acne face cleanser that has a blend of natural active ingredients. In addition to honey, xanthan gum, rose water, and citric acid, the face wash's three herbal ingredients are broccoli sprouts, flaxseed, and turmeric. The Brassicaceae family includes the broccoli, or Brassica oleracea var. italica. Antioxidants found in broccoli sprouts can help shield the skin from wrinkles and dryness. Additionally, it pulls impurities out of the skin and is used for anti-aging, acne prevention, and anti-inflammatory purposes ^[2] [3]. Sulforaphane, a potent phytochemical found in Brassica vegetables, offers numerous benefits for the skin, including reducing inflammation, soothing irritated skin, and alleviating redness. By preventing bacterial growth and lowering inflammation, it cures acne ^[4]. The Linaceae family includes flaxseed (Linum usitatissimum). Alpha-linolenic acid (omega-3 fatty acid), lignans, and phenolic chemicals are abundant in it. Flaxseed is a possible natural acne treatment because of its significant anti-inflammatory, antioxidant, and antibacterial components. While the antibacterial action can help limit germs that cause acne, the anti-inflammatory effect helps minimize skin redness and swelling. ^[5].The Zingiberaceae family includes turmeric (Curcuma longa). It has antibacterial, anti-inflammatory, antioxidant, and skin-brightening properties ^[6]. The current study's objective is to create and assess a herbal face wash that contains extracts of turmeric, flaxseed, and broccoli sprouts to prevent acne. Following the extraction of active ingredients from turmeric, flaxseed, and broccoli sprouts, an anti-acne herbal face wash was created utilizing these extracts, and the product's efficacy and quality standards were assessed.

Face wash:

Skin washes are another name for facial cleansers. It is a cleanser designed especially to clean the face without making the skin overly dry. It is said to be a good choice for all skin types. Face washes moisturize dry skin while efficiently removing oil and debris. Toxins, extra oil, and pollutants are removed from the skin of the face using both face washes and cleansers. It aids in pore clearance and prevents skin conditions like acne. ^[7]

Advantages of Face wash:

• The skin becomes luminous as a result.
• It maintains healthy, clear skin.
• Excess oil and dead skin cells can clog pores, resulting in acne, black and white heads, and a drained appearance.
• It aids in eliminating dead skin cells.
• It encourages skin renewal and renewal.

Disadvantages of Face wash:

• Dry skin may result from this.

• Allergic reactions or irritation could result from this.
• The skin barrier may be harmed by excessive washing.
• Not appropriate for every kind of skin.

Properties of an ideal face wash:

• It should be stable and aesthetically pleasing.
• It should soften and spread readily after application without creating drag.
• Neither during nor after application, it should leave the skin feeling greasy or oily.
• The cream residue shouldn't get thick when the water evaporates.
• Rather than absorption, its physical activity should concentrate on clearing the pores and flushing the skin.
• After application, the skin should still have a thin coating of emollient. ^[8]

Forms of face wash:

• Cream based face wash
• Gel based face wash
• Liquid based face wash

MATERIALS AND METHODOLOGY

Ingredients: Broccoli sprouts extract, turmeric extract, flaxseed gel, honey, citric acid, glycerin, xanthan gum, methyl paraben, sodium lauryl sulphate, rose water, distilled water.

Collection of plant material:

After being gathered at a nearby market and thoroughly cleaned, broccoli seeds were steeped in water for 6-8 hours. Once the extra water was drained, the seeds were put in a sprouting jar or tray and stored in a warm, dark environment (18–22 °C), where they were rinsed twice a day. 3-8 days after germination, broccoli sprouts were gathered. Before being used, turmeric was purchased at the neighborhood market and examined for cleanliness and purity. After being gathered from the neighborhood market, flaxseed was kept dry in a container.

Preparation of extractions:

1. Extraction of broccoli sprout:

Broccoli is extracted by using solvent extraction method. To get rid of dirt, dust, and microbiological impurities, fresh broccoli sprouts were gathered and thoroughly cleaned with distilled water. The clean sprouts were either dried in an oven at 40 to 45 degrees Celsius or spread out on a tray in a dust-free, shaded location. To stop microbiological growth and maintain the active ingredients, the samples were dried until they were completely dry. A clean grinder or high-speed blender was used to grind the dried sprouts into a fine powder. Prior to extraction, the powdered broccoli sprouts were kept in a dry, airtight container. Solvent extraction was carried out. A popular solvent is ethanol (50% v/v) in water. 1 gram of powdered broccoli sprouts was combined with 15 ml of solvent (1:15 (w/v)) to do the extraction. After gently shaking and stirring the mixture, it was incubated for 72 hours at 40°C. To extract the clear liquid and eliminate any solid residues, the mixture was filtered using muslin cloth or filter paper. ^[9] [10]   

2. Extraction of turmeric:

Soxhlet apparatus was used for turmeric extraction. A thimble-shaped filter paper is filled with turmeric and stored in a glass cylinder. The cylinder had intake tubes and a siphon. At the top of the cylinder was a water condenser. The entire arrangement was placed inside the neck of a solvent-filled round-bottom flask. A water or sand bath was used to heat the flask. Via the inlet tube, the solvent vapors entered the cylinder and traveled upward into the condenser. When the crude organic material comes into touch with the condensed solvent, it dissolves. as soon as the solution reaches the siphon tube's upper end. As a result, the cylinder was kept filled with solvent vapors at all times, and the organic substance that had dissolved returned to the flask. The solution in the flask was distilled to recover the solvent after the heating was finally turned off, leaving the organic substance behind. ^[1] [11]

3. Extraction of flaxseed gel:

A pot was filled with water and flaxseeds. Bring to a boil, then adjust the heat to a simmer. To prevent the mixture from sticking, stir it continuously. It required two to eight minutes for the fluid to become gel-like after stirring. Take off the heat and let it cool a little. Pour the gel into a jar after straining it through cheesecloth or a cloth. Before being used, the mixture was allowed to cool for up to two hours. ^[12]

Figure 1: Preparation of herbal extracts

Formulation of the face wash:

Table 1: Composition of herbal face wash

Procedure:

Overnight, rose water in a beaker was mixed with the necessary amount of xanthan gum. Glycerine, honey, flaxseed gel, and a few drops of citric acid were added to the second beaker and combined. The rose water mixture (xanthan gum + rose water) was filled with the second beaker mixture (flaxseed gel + glycerin + honey + a few drops of citric acid). After that, the two mixes were mixed together. After properly mixing in the extracts of broccoli and turmeric, methyl paraben and sodium lauryl sulfate were added. After that, the mixture was thoroughly combined to produce a thick herbal face cleanser. ^[1]

EVALUATION PARAMETERS

The anti-acne herbal face cleanser was assessed using the following criteria:  ^{[1] [8] [13]}

1. Physical evaluation: Visual assessments were made of physical characteristics like color, consistency, and look.
2. Washability: The mixture was manually applied to the skin and examined under running water.
3. Spread-ability test: This test shows how far the face wash can be applied to the skin with ease. One glass slide was used to hold the prepared formulation, and another glass slide was placed on top of the sample, sandwiched between the two slides. To guarantee that the sample between the two slides was uniformly squeezed with a thin layer, a weight of 100 g was placed on the upper slide, which was 7 cm long.

Spreadability (S)=

weight tide upper slide(W)× Length of glass slide(L)

Time in sec (T)

4. pH measurement: A digital pH meter was used to measure the formulation's pH.
5. Viscosity: A Brookfield viscometer was used to measure the viscosity.
6. Foamability: In the shaking tube method, a 50 mL measuring cylinder was filled with 20 mL of surfactant solution, which was then violently shaken. A frequency of 3 Hz and an amplitude of roughly 5 cm were used to shake the mixture. The maximal foam height was used to calculate the foaming ability.
7. Stability test: Assessing how physical characteristics, like texture and odour, and chemical characteristics, like pH, change over the course of 1 month at room temperature and at elevated temperature(40°C).
8. Antimicrobial activity: Antimicrobial activity is measured by contrasting the growth inhibition of sensitive bacteria produced by a reference substance with that induced by a known concentration of the test material (isolated compound, extract, or synthesized compound).

Procedure:

The agar well diffusion (cup-plate) method was used to assess the sample's antibacterial activity. To create a consistent agar layer, sterile antibiotic test agar was infected with a Staphylococcus aureus suspension and then transferred into Petri dishes. Following solidification, a sterile Cork borer was used to aseptically punch wells with a diameter of 6–8 mm. A base served as a negative control, and the test sample was added to the wells in amounts of 100 µL and 200 µL.

After allowing the sample to diffuse into the agar medium for 15 to 20 minutes at 2 to 8°C, the plates were incubated for 24 to 48 hours at 30 to 35°C. Following incubation, the zones of inhibition that developed around the wells were measured, and the diameter of these zones was used to determine the sample's antibacterial activity.

Anti-acne activity: The anti-acne activity of the formulation was evaluated using agar well diffusion method against Propionibacterium acne. Agar containing antibiotics was poured evenly into petri plates after being inoculated with test organism. When the substance was solidified, wells measuring 6 to 8 mm across were carefully created without any contamination using a sterile Cork borer. The test materials were introduced into the wells at volumes of 100µL and 200µL for this purpose. While the base act our negative control sample. The plates were modified at 2 to 8°C for 15 to 20 minutes so that diffusion could occur and then incubated at 30 to 35°C for 24 to 48 hours. After a period of incubation, the clear areas surrounding the wells were measured to determine the anti-acne activity which was assessed based on the diameter of the incubation zone. 

RESULTS AND DISCUSSION

1. Physical evaluation:  The color of formulation was a foamy yellow-green color. The odour of formulation was a pleasant, acceptable and good. Appearance and consistency was in compliance with the cosmetic’s requirements.

Figure 2: color of formulation

2. Washability:  The formulation was found to be easily washable with water without leaving any oily or sticky residue on the skin. 

Figure 3: Washability test of formulated face wash

3. Spread-ability: The formulation showed good spread-ability, ensuring uniform distribution over the skin with minimal effort.

Figure 4: spread-ability test of formulated face wash

4. pH: The pH of the formulated facewash was found to be 5.13 which is in the range of skin pH. Therefore, the formulation is suitable for skin care.

Figure 5: pH meter showing pH value of formulation

5. Viscosity: The viscosity of facewash was measured using Brookfield viscometer DV-E at room temperature (spindle number - 64, rotation - 4 rpm). The viscosity of face wash was found to be 5400 cP.

Figure 6: Viscosity determination by Brookfield viscometer

6. Foamability: The formulation produced a stable foam that persisted for a sufficient period, indicating satisfactory cleansing properties.

Figure 7: Foamability test of formulation

7. Stability:  No significant changes were observed in the physical appearance like color, odour, texture and chemical parameter like pH of the formulation at both room temperature and 40°C. The formulation remained stable throughout the study period. 
8. Antimicrobial activity: In comparison to normal face wash, studies concluded that sample face wash had good inhibition of microorganism growth against Staphylococcus aureus.

Anti-acne activity:   in this study shows that sample face wash has better anti-acne activity. It has good inhibition of microorganism growth against Propionibacterium acne.

                                 Figure 8: Yeast and Mold                             Figure 9: E.coli negative plate

                  Figure 10:Antibacterial antibiotic sensitivity        Figure 11: Total bacterial count

Table 2: Evaluation table of formulation