Development and analysis of a herbal bath bomb for its antifungal properties

Introduction

AegleMarmelos belongs to the citrus family Rutaceae. ^[1] Its known as a holy tree in Hindu scriptures which is grown near temples and is also an indicator of underground water sources. AegleMarmelos is known as Bael and originates in India ([Figure 1]). Its mentioned in Indian scriptures since 800 BC. The taxonomical classification of A. marmelosis given in [Table 1]. In Hindu culture all parts of this plant were considered medicinal and were used to treat various conditions like asthma, fractures, anemia, swollen joints, wound healing, diabetes, high BP, jaundice, diarrhea, stomachache, cancer, malaria and gastroduodenal disorders ([Figure 2]). ^[2] The bael plant contains furocoumarins such as xanthotoxol and the methyl ester of alloimperatorin, and also flavonoids such as rutin and marmesin, essential oils, and alkaloids such as á-fargarine (=allocryptopine), O-isopentenylhalfordinol, and O-methylhafordinol.] N-[2-hydroxy-2(4-methoxyphenyl) ethyl]-3-phenyl-2-propenamide (N-[2-hydroxy-2(4-ethoxyphenyl) ethyl]-3-phenyl-2-propenamide) is a component that may be isolated from bael leaves. Aeglemarmelosine has been extracted as an orange viscous oil. ^[1], ^[2], ^[3] Bath bombs commonly contain bath salts, which help in muscle relaxation. It also acts as a soft moisturizer and provides nourishment to the skin. The herbal ingredient used in bath bomb has various therapeutic properties such as antifungal, antimicrobial, antiviral, antidiabetic, analgesic, etc.^[3] The antioxidant activity of methanolic and ethanolic extracts of the fruit pulp of the A. marmelos plant was studied in a rat model using the DPPH radical scavenging method and the nitric oxide scavenging method.^[4] The antibacterial activity of petroleum ether, ethanol, and aqueous extract of A. marmelos plant leaves was investigated using the agar well diffusion technique. The extracts were reported to be efficient against E. coli, Streptococcus pneumoniae, Salmonella typhi, Proteus vulgaris, and Klebsiella pneumoniae bacteria.^[5]Antifungal activity was shown in an ethanolic preparation of the root towards A. fumigates and T. mentagrophytes.^[6] The antifungal activity of essential oils extracted from the ethanolic leaf extract of the Bael plant (Aegle marmelos), (Rutaceae Family), was studied against Candida albicans strains.^[7] All A. marmelos leaf extracts were tested for hypoglycemic action in a variety of animal models. At 500 mg/kg body weight, the aqueous and alcoholic extracts of the fruit portion demonstrated glycemic action against rabbits. ^[8] In vitro proliferation of human tumour cell lines such as the leukemic K562, T-lymphoid jurkat, Blymphoid Raji, erythroleukemic HEL, melanoma Colo 38, and breast cancer MCF and MDA-MB-231 cell lines was inhibited by the extract.^[9] The analgesic effect of a methanolic extract of Bael plant leaves was investigated in a mouse model utilising a squirming and tail immersing test at a dose of 200 mg/kg. The plant's analgesic activity was found to be significant. ^[10], ^[11], ^[12]

Kingdom	Plantae
Order	Sapindales
Family	Rutaceae
Subfamily	Aurantioideae
Genus	Aegle Correa
Species	A.Marmelos
Clade	Angiosperms
Division	Magnoliophyta
Common name	Baelpatra, Bael

Table 1 Taxonomical classification of aeglemarmelos

Materials and Methods

Experimental

Aegle Marmelos leaves were taken from the local area of Pimpri, Pune. All the chemical were of AR grade.

All of the leaves of Bael were cleaned and dried with distilled water. Using a mortar and pestle or mixer grinder, fine powder was extracted from the dried leaves. The cold extraction/maceration method was used to extract fine leaf powder using solvents such as ethanol.  50g of powdered sample was soaked in 100ml of each solvent for 4-5 days to allow diverse chemical contents from the leaf to permeate into the solvents, and the filtrate was collected after the filtration procedure. By evaporating extra solvent with the help of rota evaporator equipment, final pure form of extract is obtained. ^[13], ^[14], ^[15], ^[16] The rotovap works by increasing the rate of evaporation of the solvent by reducing the pressure to lower the solvent boiling point, rotating the sample to increase the effective surface area and heating the solution.

Preparation of bath bomb

All the dry ingredient in dist were combined and mixed until uniform. Food Colourant was added in the last step. After the dry ingredient mixed Bael leaves extract ([Figure 2] ) was added and combined well. At the same time, wet ingredients were mixed in separate bowl without adding food colourant. Dry ingredients were then added to wet ingredients and stirred it properly. During mixing the mixture appeared like dry sand. Bath bomb mould was used to give it beautiful shape. Mould was allowed to stand for 20 minutes in refrigerator before removing the opposite side of the mould. After letting the second side to dry, leave the bath bomb to harden up overnight.Prepared formulations were evaluated forvarious parameters like Effervescent time, pH test, Antifungal test.

S.No.	Ingredients	Quantity	Role
F1	F2	F3	F4
1	Sodium bicarbonate	2.55 g	2.50 g	3.20 g	2.85 g	Weak base
2	Citric acid	1.04 g	2.14 g	2.74 g	1.14 g	Weak acid
3	Corn starch	1.14 g	1.14 g	1.98 g	2.14 g	Binder
4	Magnesium sulphate	1.02 g	1.32 g	1.22 g	1.42 g	Muscle relaxant [5]
5	Bael leaves extract	1.00 g	0.99 g	1.00 g	1.00 g	Antifungal & Antimicrobial [17], [18], [19], [20]
6	Turmeric	Q.S	Q.S	Q.S	Q.S	Coloring agent & Antiseptic [13]
7	Honey	Q.S	Q.S	Q.S	Q.S	Moisturizer & Antibacterial [21]
8	Rose water	Q.S	Q.S	Q.S	Q.S	Fragrance & Anti-inflammatory [22]

Table 2 Formulation table for bath bomb

Determination of zone of inhibition

The cup-plate diffusion method is used to compare the zone of inhibition of ethanolic extract of Bael leaves (Aeglemarmelos) to Fluconazole solution in potato dextrose agar media. ^[15], ^[23]

Determination of zone of inhibition

Firstly Candidaalbicans grown on agar media. Plates were allowed to dry before 5 mm wells were made using a sterile cork borer. Plant leaves extract was added using a micropipette into the wells of incubated plates, which were then allowed to stand for 10-15 minutes for extract diffusion before incubation freeze it for 3 to 4 hrs then being incubated at 37° C for 48 hours. Following the incubation period, the plates were evaluated for the presence of a clear zone surrounding the extract-containing wells. Then compare the zone of inhibition of ethanolic extract of Bael leaves to Fluconazole solution in potato dextrose agar media. ^[22], ^[23], ^[24]

Result and Discussion

Effervescent time and pH

The effervescent time and pH of a bath bomb formulation F1 to F4 were tested both in cold and hot water mentioned as below:

Formulations	Hot water	Cold Water	pH
F1	1.21	1.30	6.6
F2	1.25	1.37	6.8
F3	1.18	1.40	6.5
F4	1.29	1.49	6.7

Table 3 Effervescent time and pH

Antifungal activity

The cup-plate diffusion method is used to compare the zone of inhibition of ethanolic extract of Bael leaves (Aeglemarmelos)  to Fluconazole solution in potato dextrose agar media.Based on the results, it was determined that betel leaves had antifungal action. The zone of inhibition of ethanolicBael (Aeglemarmelos) against Candida albicans was nearly identical to the normal (standard) Fluconazole formulation. A bath bomb comprising an ethanolic extract of Bael leaves was effectively made and tested ([Figure 3]).

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