INTRODUCTION:

Fungal infections have increased in recent decades. Fungal infections are a common disease in nursing homes. An increasing number of immunosuppressive diseases and conditions are affecting the epidemiological pattern of mycoses in hospitalized patients. The epidemiology of invasive fungal infections is currently at a critical stage.¹ Fungal infections caused by Candida are more common than E. coli, Pseudomonas, and Aspergillus. and other sp². There are many host factors that predispose patients to fungal infections. These include: mucositis; use of antibiotics; Radiation therapy or certain immunosuppressants. Intensive Care Unit (ICU)³ Candida albicans is the most common species in the genera associated with candidiasis. Infectious diseases range from superficial skin diseases to systemic diseases. C. albicans, C. glabrata and C. krusei are part of the normal human flora and can be isolated from oral, vaginal and other body parts of normal healthy individuals⁴.

Herbal remedies are ancient methods of curing ailments. Since Vedic times, people have used medicinal plant materials to treat ailments or to provide satisfactory remedies for their ailments. The plant is also known to treat infectious and non-infectious skin diseases. The antibacterial effects of some plants are attributed to a number of plant components such as flavonoids, tannins and triterpenes⁵. Current research objectives are also based on the medicinal properties of plants. H. ginger (Zingiber officinale Roscoe) exhibits broad antibacterial effects. Gingerol is the main chemical component of ginger that exhibits antibacterial activity. Ginger extract has been reported to have potent antifungal activity against C. albicans, C. glabrata, and C. glabrata^6,7.The purpose of this study is to produce various cream formulations containing the herbal antifungal ginger extract using the emulsification method.

Also, cream properties such as various organoleptic properties, pH measurement, homogeneity, elimination test, skin irritation test, viscosity, spreadability test, saponification number, acid number, microbiological test to confirm antibacterial properties⁸.

MATERIALS AND METHODS:

1. Materials:

Propylene Glycol, Beeswax, Stearyl Alcohol, Cetyl Alcohol, Triethanolamine, Propylparaben, Methylparaben, Liquid Paraffin, Stearic Acid. Ginger extract is obtained from fresh ginger rhizomes by continuous heat extraction and maceration in the laboratory.

a) Ginger⁹

Synonyms: Adlak (fresh), Shunti (dried).

Biological Source: Ginger consists of the dried rhizome of Zingiber officinale, which belongs to the family Zingiberaceae.

Family: Zingiberaceae.

Chemical Composition: The essential oil is responsible for the fragrance, and the pungent taste of the medicine is due to a yellowish oily substance called gingerol, which is odorless. Volatile oils are composed of sesquiterpene hydrocarbons such as α-zingiberol. α-sesquiterpene alcohol, α-bisabolene, α-farnesene, α-sesquipherandrene. It also contains pungent ingredients such as gingerone and shogaol.

2. Collection of Samples:

Ginger crude drug is purchased from the market.

a) Maceration^10,11

100 g of fresh ginger root was soaked in 500 ml of absolute methanol in a beaker for 72 hours. After 72 hours, the entire mixture was filtered to remove residue.

3. Preparation of Cream Formulation¹²

Table 1: Formulation of two phases

Part A- Oily Phase			Part B-Aqueous Phase
Ingredients	Quantity	Activity	Ingredients	Quantity	Activity
Stearyl alcohol	5%	Emollient	Propylene glycol	5%	Humectants
Cetyl alcohol	6.5%	Binding agent	Triethanolamine	2%	Stabilizer
Mineral oil (Liquid paraffin)	5%	Moisturizer	Methyl paraben	0.01%	Preservative
Stearic acid	2.5%	Emulsifying agent	Propyl paraben	0.04%	Preservative
White Beeswax	1.5%	Thickening agent	Distilled Water	Upto 100%	Solvent base
Ginger Extract	5%	Anti-fungal

a. Oil Phase Preparation¹³

All ingredients were dissolved, including white beeswax, stearic acid, stearyl alcohol, and cetyl alcohol. Liquid paraffin was added to the mixture and dissolved. The temperature was then maintained between 65°C and 70°C.

b. Aqueous Phase Preparation¹³

Water was heated to 65-70°C. All reagents such as propylene glycol, triethanolamine, propylparaben, and methylparaben were added to this pre-weighed aqueous medium. The temperature of the aqueous phase was then maintained at 65-70°C.

c. Cream Formulation Development¹³

The entire oil phase was slowly poured into the water phase at 65-70°C and mixed for 10-15 minutes. When the temperatures of both media were equal, the water phase was slowly added to the oil phase with moderate stirring until the temperature dropped to 40°C. I added ginger extract to this. The O/W emulsion was then cooled to room temperature to obtain a thick cream base.

Preparation of aqueous phase Preparation of oil phase Final formulation

Figure 1: Stages of herbal antifungal cream formulation

RESULTS AND DISCUSSION:

1. Physical Examination (Organoleptic Characteristics):

The prepared herbal antifungal creams were visually examined for color, appearance, odor and consistency.

2. Determination of Ph:

0.50 g of cream was weighed and dissolved in 50 ml of distilled water and the pH was measured with a digital pH meter.

3. Viscosity^13,14,15

The viscosity of the formulated cream was measured using a Brookfield viscometer NDJ-8S using a spindle S 94 at various speeds and shear rates. Measurements were made over a speed range of 0.15, 0.25, 0.35, 0.45 and 0.55 rpm for 60 s between two successive speeds as equilibrium with shear rates ranging from 0.25 s-1 to 1.0 s-1. Viscosity measurements were performed at room temperature.

4. Spredability test^13,14,15,16

The diffusivity properties of formulations were tested using the method developed by Muttimer et al. The developed device has been charged. It consisted of blocks of wood connected at one end by a pulley. I put a rectangular screen on top of this block. An excess amount of cream (approximately 3-4 g) to be tested was placed on this baseplate. An antifungal herbal cream was then placed between this plate and a glass plate of the same dimensions as the solid bottom plate and secured with hooks. A solid load of 1 kg was placed on the slabs for approximately 4-5 minutes to expel any trapped air and create a uniform film of cream between the slabs. Excess cream was scraped off the edges. Next, we added 80 g of resistance to the top plate. Using a string attached to the hook, record the time (in seconds) it takes for the top plate to move 10 cm. A shorter interval indicates better diffusivity. Malleability is measured in units of gm.cm/s. The spreadability of cream can be determined by the following equation:

S = M x L/T

where

L = length moved by glass slide

T = time (seconds)

M = weight in pan

S = ductility

5. Skin Irritation Test⁸

Skin irritation is determined by the fact that herbal antifungal cream formulations do not affect human skin cells and tissues. Using certain creams without testing them can irritate the surface of the skin, causing swelling, redness, and irritation. Therefore, a mark was made on the back of the left hand and a skin irritation test was performed. The cream was applied with a spatula to the marked area and the time was recorded. Irritation, erythema, and edema were checked regularly for up to 24 hours. The applied herbal antifungal cream did not cause any major irritation and was safe to use.

6. Saponification value¹⁸

Take 2 g of material and reflux with 25 ml of 0.5 N alcoholic KOH for 30 minutes. Next, add 0.1 ml of phenolphthalein as an indicator and titrate with 0.5N HCL.

Saponification Number = (b-A)*28.05/W

Where

A = volume of titrant

B = volume of titrant

W = weight of substance in grams

7. Acid value¹⁸

cream Take 10 g dissolved in 50 ml of a precisely weighed mixture of equal amounts of alcohol and solvent ether. The flask is then attached to a condenser and slowly heated to reflux until the sample is completely dissolved. Then add 1 mL of phenolphthalein and titrate with 0.1 N NaOH until a pale pink color appears after shaking for 20 seconds.

Acid Number = n*5.61/W

W = weight of material

N = number of ml required for NaOH

8. Microbiological studies^19,20,21

For antifungal study sample concentration of 5mg and 10 mg stored in a refrigerator till further used. Antifungal activities of the sample were evaluated by means of agar well diffusion assay. The assay was carried out according to the method of (Hufford et al., 1975). Sabouraud dextrose agar (Hi media) was used for the growth of fungus. Media with acidic pH (pH 5.5 to 5.6) containing relatively high concentration of glucose (40%) is prepared by mixing (SDA) Sabouraud dextrose and distilled water and autoclaved at 121oC for 15 minutes. Twenty five ml of molten (450C) SDA medium was aseptically transferred into each 100mm×15mm sterile Petri dish.

For counting of spore (fungi) were suspended in normal saline to make volume up to1ml and then counted with help of heamocytometer (neubar chamber). Once the agar was hardened, 6mm wells were bored using a sterile cork borer. Then 0.1ml (100μl) from each stock solution of the sample having final concentration of 5 mg and 10mg was placed in each the well and the plates were incubated for 72 hour at 29oC. The antifungal activity was measured as the diameter (mm) of clear zone of growth inhibition. (Umadevi et al., 2003)

Antifungal Evaluation

• Ingredients: Herbal antifungal cream.

• Media: Sabouraud Dextrose Agar.

• Sample: Herbal antifungal cream, Candida albicans.

• Standard: Fluconazole.

Candida albicans Showing antifungal activity

Figure 2: Antifungal activity of Herbal antifungal cream on organism

Table 2. Effect of antifungal activity

Organism	Extract	Test	Standard
Candida albicans	Methanol	Susceptible	Susceptible

Table 3. Showing Diameters of Inhibition Zone

Organism	Plant Extract	Zone of inhibition [mm]
		Test Sample (avgas diameter)	Standard
Candida albicans	Methanol	15	22

Herbal antifungal creams exhibit antifungal activity against Candida albicans and can be formulated as antifungal preparations (creams).

OBSERVATION:

Table 4: Compiled evaluations results

Sr. No	Evaluation Parameter	Results
1	Colour	Buff yellow to creamish
2	Appearance	Smooth
3	Odour	Aromatic
4	Consistency	No phase separation
5	Viscosity	65750 cps.
6	Spread ability (gm.cm/sec)	15.26
7	pH	6.7
8	Skin irritancy test	No Irritancy
9	Saponification Value (mg/g)	56.1
10	Acid Value (mg/g)	7.20
11	Microbiological studies [zone of inhibition]	15 mm

The prepared formulations exhibited good spreadability, no evidence of phase separation and good consistency throughout the study period. And the formulation showed no significant difference during the study period.

CONCLUSION:

The use of herbal/bioactive ingredients in creams (cosmetics) influences the biological functions of the skin and provides necessary nutrients for healthy skin against antifungal infections. The prepared formulations exhibited good spreadability, no evidence of phase separation and good consistency throughout the study period. However, there were downsides to stability parameters such as appearance and texture. The odor of the formulation was aromatic.

CONFLICT OF INTEREST:

The authors have no conflicts of interest regarding this investigation.

ACKNOWLEDGMENT:

The authors would like to thank for the successful completion of this research work, I would like to extend my thanks and cordial sense of gratitude to my research guide Mrs. Sujata P. Choudhari (Assistant Professor, Department of Pharmacognosy) for her continuous guidance for the investigation and preparation of this dissertation. Last but not the least, my heartiest thanks to my friend and juniors for helping me directly or indirectly to complete this dissertation work.

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Received on 02.06.2023 Modified on 30.06.2023

Research J. Science and Tech. 2023; 15(3):129-134.

DOI: 10.52711/2349-2988.2023.00021