INTRODUCTION:

Worldwide, The use of hand gel sanitizer and its preparation are widely used because it is easy to dry to be a film which then was easy to wash prevent the infections and to avoid the transmission of harmful germs and prevent the infections^1-2. Isopropyl alcohol based novel hand sanitizer has gained more attention over the last several decades due to its reputable medicinal properties and from different pharmaceutical, health care product has been manufactured. The recent outbreak of the COVID- 19 virus disease in the in Wuhan, China is a good example. To assist microbial load reduction, hand sanitizers can also be used after hand washing. It is therefore very important that more effective and yet affordable hand gel sanitizers are formulated to reduce the infections spread around the global. Hand sanitizers gel are even more important in places where Contaminated hands can serve as vectors for the transmission of microorganisms for outbreaks are spread from the hands of the food handler to others when the food handler contaminates his/her hands and then passes these microorganisms to consumers via hand contact with food or drinks and does not have the luxury of time to wash the hands after attending to each patient ^3-6.

Medicinal properties of bioactives is been studied since long back and various research on it was carried out and from that various pharmaceutical, health care, cosmetic products has been manufactured. This research was aim to formulate the herbal hand sanitizer gel^7-8. Aloe barbadensis miller has been used in as a source of food industries, especially for the preparation of health food drinks and other beverages and finds its application in the pharmaceutical industry is not negligible as far as the manufacturing of topical ointments, gel preparations, and tablets and capsules are concerned⁹. basically, the production process of Aloe products involve crushing, grinding, or pressing of the entire leaf of the Aloe plant to produce an A. vera juice, followed by various steps of filtration and stabilization of the juice^3,10. Aloe barbadensis miller extract is an important source of compounds having anti-microbial, anti-oxidant, anti-fungal, anti-inflammatory and anti-viral properties.

Isopropyl alcohol-based hand sanitizers, if rubbed thoroughly over finger and hand surfaces for a period of 30 seconds, followed by complete air-drying, can effectively reduce populations of bacteria, fungi, and some enveloped viruses^11-12. Hand sanitizers typically come in gel, foam or liquid form. Their use is prescribed when water and soap are not available for hand washing or when repeated hand washing compromises the natural skin barrier (e.g., causing scaling or fissures to develop in the skin). Although the effectiveness of hand sanitizer is variable, it is employed as a simple means of infection control in a wide variety of settings, from day-care centers and schools to hospitals and health care clinics and from supermarkets to cruise ships. Depending on the active ingredient used, hand sanitizers can be classified as one of two types: alcohol-based or alcohol-free^13-15. Alcohol-based products typically contain between 70 and 95 percent alcohol, usually in the form of ethanol, isopropanol, or n-propanol. Isopropyl alcohol-free products are generally based on decontaminate, such as benzalkonium chloride (BAC), or on antimicrobial agents, such as triclosan. The activity of disinfectants and antimicrobial agents is both immediate and persistent. Many hand sanitizers also contain emollients (e.g., glycerin) that soothe the skin, thickening agents, and fragrance^16-17.

S. aureus is a bacterium that often causes infection in humans. A hand sanitizer gel of Aloe vera extract has been developed to prevent the spread of the bacteria on the palms. The gel is easy to apply and is acceptable to people. We Showed different studies and sanitizer formulation were carried out. Sanitizer with complex chemicals as well herbal sanitizer was prepared and their efficacy were investigated against various groups of organisms. Hence in this studies aloe Vera juice is been use their antimicrobial properties were studied and accordingly sanitizers were prepared^18-20.

The main objective of this research work was to formulate and optimize a herbal gel of aloe vera extract containing Carbopol 934 and determine the MIC against S. aureus of ethanol extract of aloe vera. After the MIC was determined, hand sanitizer gel was prepared with two different concentrations of aloe vera extract. The antibacterial effectiveness of the gels was examined on the palms of the respondents, and the number of bacteria was calculated.

MATERIALS AND METHODS:

Materials:

Isopropyl alcohol, hydrogen peroxide and Glycerol were purchased from Himedia Pvt, Ltd. Mumbai, India. Aloe barbadensis miller were collected from shri Rawatpura Sarkar university medicinal garden. Triethanolamine, carbopol934, propylene glycol and all other reagents/chemicals were used as analytical grade. Hand sanitizer Starlon (Marketed Product) Cadila pharmaceuticals, Ahmadabad, Beaker, Filter funnel, Filter paper, foil paper, paper tape, Packaging bottle, digital weighing balance, distilled water.

Methods:

Preparation of aloe vera extract:

Thick succulent leaves of Aloe vera (Aloe barbadensis) plant obtained from Herbal Garden, Shri Rawatpura Sarkar University, Raipur, were used. To obtain Aloe vera extract, the mucilaginous jelly obtained from the centre (the parenchyma) of the plant leaf of Aloe vera, the leaves of Aloe vera were collected, washed with water and a mild chlorine solution and were finally cut transversely into pieces. With a vegetable peeler, the thick epidermis was selectively removed and the inner gel-like pulp in the center of the leaf was separated with a spoon, minced, and homogenized in a mixer. This juice was stored at 4°C for further use^8,21.

Formulation of herbal Hand Sanitizer:

Measure the quantities of Isopropyl Alcohol, Hydrogen Peroxide, and Glycerol in suitable containers. Transfer the Isopropyl Alcohol and Hydrogen Peroxide into a suitable calibrated container and mix gently. Transfer the Glycerol stepwise and quantitatively into the calibrated container. After that mix aloe vera gently after each addition. Rinse the container containing glycerol several times with Water and add the contents to the calibrated container. Add sufficient distilled Water to bring to final volume. Mix well. Transfer the solution into suitable containers. The screw cap is placed on the bottle immediately after mixing to prevent evaporation as shown table 1.^22-23.

Table 1: isopropyl alcohol 70 % containing topical herbal hand sanitizer

S. N.	Ingrient	Quantity	Function
1.	Isopropyl alcohol 96%	824.2ml	Antimicrobial
2.	Hydrogen peroxide 3%	41.7ml	Antibacterial
3.	Glycerol 98%	7.5ml	Hemectant
4.	Aloe-vera	20ml	Antimicrobial
5.	Distilled Water	Up to 10000ml	Vehicle

Preparation of Hand Sanitizer Gel:

Gel base was prepared by varying the proportion of carbomer to triethanolamine in the gel, which also contained propylene glycol. The optimum ratio of carbomer to triethanolamine was found to be 1:4. This ratio was used to formulate the hand sanitizer gel by mixing the components in appropriate proportions, as shown in Table 2. The optimum composition was determined according to a pH that was comparable to skin pH and a fluid consistency suitable for application to the skin²⁴.

Table 2: Formulation of herbal hand sanitizer gel

S. No.	Ingrients	Quantity taken
1.	Carbomer (gm)	0.5
2.	Triethanolamine (gm)	1
3.	Propylene glycol (ml)	15
4.	Hand sanitizer liquid (ml)	20
5.	Clove (ml)	0.25
6.	Distilled water (ml)	100

Characterization of herbal hand sanitizer gel:

pH:

The pH was determined by using digital pH meter (ELICO.LI 610 pH meter) at the room temperature. 1.0 g of hand sanitizer gel was accurately weighed and dispersed in 100ml purified water. The measurements of pH were done in triplicate and average values were calculated.

Viscosity:

The viscosity of hand sanitizer gel was determined by using digital Brookfield viscometer (Model DV-II, USA) equipped with spindle S27. The apparent viscosity was measured at 17 seconds−1 shear rate (50rpm) and room temperature, after a 3min rest time. Measured quantity of herbal hand gel was taken into a beaker and the tip of viscometer was immersed into the hand sanitizer gel and the viscosity was measured in triplicate ²⁵.

Spreadability:

Spreadability measurements of all the hand sanitizer gels were carried out individually in triplicate using a wooden block and glass slide apparatus. 0.5g of sample under study was applied to the lower plate of apparatus to get a uniform layer of the formulation. A weight of 100g was added to the pan, and the time in seconds required for the upper plate to travel across the length of the lower plate was noted. This time was indicative of the spreadability, i.e., the ease with which a formulation may spread on the skin²⁶.

Antimicrobial activity of extract against standard organism:

Antimicrobial activity of leaves extract of aloe-vera juice was carried out by agar well diffusion method against standard organism. After swabbing culture and adding extract the plates were incubated at 37°C±1⁰C for 24hours²⁷.

Phytochemical analysis:

Test for carbohydrates (Fehlings test), amino acid (Ninhydrin Test), phenols and tannins, Flavonoids, Saponins (Foam test), Terpenoids, steroids and sterols (Salkowski’s test) was performed as per given reference ²⁸.

Determination of Minimum Inhibitory Concentration (MIC) of aloe vera extract:

The Minimum Inhibitory Concentration is defined as the lowest concentration that completely inhibits the growth of microorganisms for 24hrs incubation. Determination of minimum inhibitory concentration of extracts was determined by preparing different concentrations of extracts 200μg, 400μg and 800μg were added respectively to the nutrient broth . A 50μl volume of each dilution was added aseptically into the wells of Mueller Hinton agar plates that were already seeded with the standardized inoculums of the test bacteria. All experiments were performed in triplicate. The agar plates were incubated at 37°C for 24 hours. The lowest concentration of extracts showing a clear zone of inhibition was considered as the MIC²⁹.

Radical Scavenging Activity by the DPPH Assay:

DPPH assay was performed to check radical scavenging activity of tulsi leaves extract and aloe-vera juice. Absorbance of all the samples was taken ³⁰. The percentage radical scavenging activity of each extracts was calculated using the following equation

Percentage radical scavenging activity (RSA %) =

(Absorbance of control – Absorbance of sample)

–––––––––––––––––––––––––––––––––––––––× 100%

Absorbance of control

Antibiotic sensitivity using agar diffusion (Kirby- Bauer) method:

Mueller-Hinton agar plates is used for this method. Agar surface of each plate is streaked by a sterile cotton swab with the reference bacterial strain. Then agar plate is punched with a sterile cork borer of 4 mm size and then poured 100μL of each sample (sanitizer) with micropipette in the bore. Plates are allowed to stand for 30 min. Then incubated at 37°C for 24h. Zone of inhibition (mm) of the bacterial strains against herbal sanitizers is measured³¹. The resultant zones of inhibition which serve as an indication of antimicrobial activity were measured.

In Vitro Drug Release Study:

In vitro release studies of drug aloe vera loaded hand sanitizer gel and pure drug suspension and Starlon(Marketed Product) Cadila pharmaceuticals, was carried out for each drug individually using modified Franz diffusion (Mol. Wt. cut of 6,000-8,000, HI Media Ltd, Mumbai, India). Vertical Franz Diffusion cell was designed and validated prior to the release study. The cellophane membrane was mounted on a diffusion cell assembly having diffusion area of 2.5cm. The receptor compartment consisted of a 22.5ml phosphate buffer at pH 5.5 as the receptor fluid agitated at 100rpm, and was maintained at 37±0.5°C throughout the experiments. The prepared formulation was placed in the membrane of donor compartment. An aliquot of 2ml sample was withdrawn at suitable time intervals and replaced immediately with an equal volume of fresh diffusion medium. The % drug release was calculated and graph of % drug release against time was plotted, release studies were performed in triplicate for each formulation ^32-33. Data were expressed as the cumulative amount of Aloe vera permeated through the membrane considering the total amount of drug applied of each formulation.

Stability studies:

Assessment of ability of developed formulation was sealed bottles of freshly prepared hand sanitizer gel were placed in a stability chamber at storing at room temperature and accelerated temperature and humidity (25± 2⁰C, 60±5% RH) and (40±2⁰C, 75±5% RH) for up to six months. After six month storage, the samples were evaluated for organoleptic properties, Ph, viscosity and consistency. [Jangdey et al. 2019].

Statistical Analysis:

Data were presented as mean ± standard deviation and all experiments in this study were repeated at least three times. The results were statistically analyzed by analysis of variance ANOVA test; P values less than 0.05 (P<0.05) were considered as significant.

RESULT AND DISCUSSION:

pH value of hand gel:

The pH of herbal hand gel wash was found 6.2±0.5 for gel which was very near to the neutral pH, thus the formulation can be used without the risk of irritancy to the skin. This also indicated that the selected ingredients of the formulation did not alter the pH of the formulation.

Viscosity determination:

The measured quantity of herbal hand gel was taken into a beaker and the tip of viscometer was immersed into the hand sanitizer gel and the viscosity was measured in triplicate. The viscosity was found 50.02(cps) ±0.03 (cps) pascals.

Spreadability:

The Spreadability of formulations was found to decrease with increasing the concentration of gelling agent. The value of Spreadability for optimized gel was found out to be 9.5±0.12g.cm/sec indicating that the lotion is easily spreadable by small amount of shear. The results indicated that the formulation can be applied easily without being runoff. This assures that the formulation maintain a good wet contact time when applied to the site of application.

Phytochemical analysis:

Presence and absence of phytochemicals in aloe vera juice was as shown in (Table 3).

Table 3: Phytochemicals screening of Aloe vera extract

S. N.	Test	Result
S. N.	Test	Aloe vera juice
1.	Saponins	+
2.	Tannin	+
3.	Steroid	-
4.	Phenol	+
5.	Flavonoids	+
6.	Carbohydrate	-
7.	Amino Acid	-
8.	Terpenoid	+

Where, + = Present, - = Absent

Minimum inhibitory concentration (MIC) determination:

The MIC of ethyl acetate and ethanol was assayed against Staphylococcus aureus. This is because Staphylococcus aureus was the only susceptible microorganism. Staphylococcus aureus showed consistency in being the most susceptible microorganism to the aloe vera extracts even with the different assay methods used i.e. well agar diffusion and paper agar diffusion test. This can be observed in the results reported in table 4. In this studied, Staphylococcus aureus has been reported to be the most susceptible microorganism to aloe vera extract and it have showed in figure 1.^15,26,27.

Figure 1: Inhibition zone diameter of aleo vera extract against four microorganisms using agar diffusion method

Table 4: Antimicrobial and Antifungal activity of extract by agar well diffusion method

Organism	S.aureus	E.coli	C.albican	P.aeruginosa
Organism	Zone of Inhibition (mm)
Aloe vera extract	18	15	-	23
Without extract	10	8	-	-
DMSO	-	-	-	-

Antimicrobial and antifungal of leaf extract of aloe vera:

Antimicrobial and antifungal activity of extract against standard organism was performed by Agar well diffusion method. Aloe vera juice was inhibiting S. aureus, E.coli, P aeruginosa and not to C. albican. Hence extract of aloe vera juice was selected for further study. DMSO was use as Diluent for extract which does not show its antibacterial activity against selected organisms which was showed in (Table 5) and its results were reported in figure 2.

Figure 3. In vitro drug release profile of aloe vera loaded hand sanitizer gel, hand sanitizer liquid and pure drug suspension in skin pH 5.5

Stability studies:

The stability studies were carried out for pH, spreadability, and viscosity according to ICH guidelines. Results of stability studies showed that hand sanitizer gel formulation lost around negligible changes in Ph from 6.2±0.5 to 6.5±0.12 . The stability studies showed that there was negligible decrease in the viscosity from 50.02±0.12cps to 51.01±0.13cps and spreadability was found in slightly changes from 9.5±0.12g.cm/sec to 9.8 ±0.21 during the storage at room temperature and accelerated temperature and humidity in six months. Hence under six month under accelerated temperature, however marginal variation in terms of pH, spreadability and viscosity was observed for hand gel formulation throughout the stability testing period.

DISCUSSION:

The prepared hand gel formulations that were produced were transparent and no sineresis occurred, indicating that the hand gels had good properties and showed significant results. The herbal hand sanitizer gel showed consistent improvement during storage. The pH of herbal hand gel wash was found 6.2±0.5 for gel which was very near to the neutral. The negative control and hand gel met the standards for good gel characteristics by having a pH similar to that of human skin pH so that they would not irritate the skin when used. Increased consistency was due to the increased viscosity of the dosage after storage of the formulation for six months. Formulations are considered to have good stability if they have a viscosity shift of <10%. The change in the viscosity value of the gel preparation did not affect the flow properties of the dosage, which was still plastic and sufficiently dilute properties with the value of the viscosity present. Thus, it could be said that the gel preparations were stable and complied with standards for good hand gel characteristics. The gel formulations did not show color changes, sineresis, or other forms of instability. This indicates that the gel preparations were stable, and degradation of their compounds did not occur due to stress. Herbal hand sanitizer gel against different bacterial isolates, showed that the hand wash prepared with aloe vera extracts had great activity. Statistical analysis findings in table 4 showed that herbal hand wash gel is the broad spectrum antibacterial agent with different response for different bacterial kinds tested. Hand sanitizer gel showed the highest anti-oxidant activity of 42.05%, by DPPH scavenging activity (Table 6). Hence Herbal hand sanitizer formulation is most cost effective hence it can be sold at very low cost as no complex chemicals are use which ultimately increases the cost of sanitizer.

CONCLUSION:

The optimized gel was evaluated for different physicochemical characteristics and antibacterial property. The color, odor, consistency and pH of the hand sanitizer gel were stable and sineresis did not occur after six month storage at room temperature and accelerated temperature and humidity (25±2⁰C, 60±5% RH) and (40±2⁰C, 75±5% RH).The main ingredients of the hand gel formulation are the gelling agent. The concentration of viscosity enhancer or gel former is of immense value as a less concentration will lead to simple solution or lotion with very low consistency, while high concentration may lead to formation of gels with high viscosity leading. Hand sanitizer gel showed the highest anti-oxidant activity of 42.05% and minimum inhibitory concentration of aloe vera extract was 21% extract showed in decreasing the number of bacteria on the palms of hands. The aloe vera loaded hand sanitizer gel was able to improve for retentive sanitizer gel in the entire depth of palms of hands for a sustainable action. Hence, It has concluded that aloe vera extract have showed significant antimicrobial activity and can be use in hospitals for fruitful effective for human’s beings.

ACKNOWLEDGEMENT:

The authors express their gratitude to director, Shri Rawatpura Sarkar University for the moral support thought the writing of research paper.

DECLERATION OF INTEREST:

The authors report has no decleration of interest. The authors alone are responsible for the content and writing of the research paper.

ABBREVIATIONS

· COVID 19: Corona virus disease 2019

· WHO: World Health Organization

· RH: RELATIVE Humidity

· ICH Guideline; International conference of harmonization

· MIC: Minimum inhibitory concentration

· FDA: Food drug administration

· DMSO: dimethyl sulfoxide

· DPPH: 2,2-diphenyl-1-picrylhydrazyl

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Received on 07.12.2025 Revised on 05.02.2026

Accepted on 22.04.2026 Published on 20.05.2026

Available online from May 25, 2026

Research J. Pharmacy and Technology. 2026;19(5):2276-2282.

DOI: 10.52711/0974-360X.2026.00328

This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License. Creative Commons License.

Conc. of Drug (μg/ml)/ *Organisms*	Zone of inhibition (diameter mm)
Conc. of Drug (μg/ml)/ *Organisms*	800	400	200	STD	Control
E. coli	34 ± 1.0	32 ± 1.1	28 ± 1.2	26 ± 1.4	0
S. aureus	30 ± 0.5	28 ± 1.2	28 ± 1.2	21 ± 1.4	0
Salmonella Sp.	26 ± 1.1	18 ± 1.3	19 ± 0.1	28 ± 0.6	0
C. albicans	26 ± 1.6	21 ± 0.4	20 ± 1.0	27 ± 0.5	0

Concentration (µg/ml)	Absorbance at 320 nm
Concentration (µg/ml)	Aloe vera	RSA %
Undiluted	0.75	13.32
1:2	0.66	24.11
1:4	0.60	36.22
1:8	0.52	42.05
Control	0.72	0
Blank	0.0	0

S. No.	Time Interval	% Cumulative drug release from different formulations
S. No.	Time Interval	Pure drug suspension	Hand sanitizer gel	Hand sanitizer liquid
1.	0	0	0	0
2.	2	13.58± 0.65	9.23 ± 1.86	11.65 ± 0.26
3.	4	26.21 ± 1.12	20.71 ± 0.34	23.11 ± 0.45
4.	6	47.3 ± 1.54	28.35 ± 2.07	36.32 ± 0.62
5.	12	66.25 ± 1.28	38.04 ± 1.03	51.45 ± 1.02
6.	24	85.8 ± 0.25	60.25 ± 0.75	70.16 ± 1.34