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American Journal of  
Chemistry and Pharmacy (AJCP)

The Formulation of  Aloe Vera Gel Herbal Soap and Characterized its Physicochemical 
and Antibacterial Activities with Market Available Herbal Soap, Butwal, Nepal

Arjun Bhandari1*, Bipana Sapkota1, Gautam Prasad Chaudhary2

Volume 3 Issue 1, Year 2024
ISSN: 2834-0116 (Online)

DOI: https://doi.org/10.54536/ajcp.v3i1.3410
https://journals.e-palli.com/home/index.php/ajcp

Article Information ABSTRACT

Received: June 30, 2024
Accepted: August 24, 2024
Published: November 21, 2024

The incredible properties of  formulated and commercial Aloe vera herbal soaps have been 
thoroughly investigated revealing their potent phytochemical and antibacterial effects. The 
samples were collected from Rupandehi, Lumbini Province. Aloe vera soap was carefully 
formulated using a cold process. Through disc diffusion, the soap antibacterial effect was 
tested against harmful gram-positive and gram-negative bacteria, including Escherichia coli and 
Staphylococcus aureus. The impressive results showed a maximum zone of  inhibition (ZOI) of  
8 mm and 7 mm for formulated soap, respectively. This was due to the presence of  important 
bioactive compounds such as tannins, saponins, and reducing sugar, which are abundant in 
Aloe vera extract. Interestingly, the ZOI was less noticeable at lower concentrations of  100 
mg/mL, 50 mg/mL, and 25 mg/mL. The formulated soap still exhibited similar properties 
to the commercial soap, with a pH of  9.7, moisture content, and foam stability. Additionally, 
the formulated soap had a mixture content of  4.6%, 0.52% free caustic alkali, and a foam 
stability test measurement of  6.5 cm. These remarkable results highlight the antibacterial 
potential of  Aloe vera-based soap, proving its effectiveness against harmful microorganisms.

Keywords
Antimicrobial Test, Antiseptic 
Soap, Escherichia Coli, 
Phytochemical Screening, 
Qualitative Analysis, 
Staphylococcus Aureus 

1 Department of  Chemistry, Butwal Multiple Campus, Tribhuvan University, Butwal, Nepal
2 Department of  Pharmacy, Crimson College of  Technology, Pokhara University, Butwal, Nepal 
* Corresponding author’s e-mail: bhandariarjun112@gmail.com

INTRODUCTION 
As a cleaning agent, soap is produced as granules, bars, 
flakes, or liquid and is made by the reaction of  potassium 
or sodium salt with different naturally occurring fatty 
acids. Any water-soluble salt of  fatty acids with eight 
or more carbon atoms is also known as soap. Soaps are 
made for many uses, such as cleaning, bathing, taking 
medication, etc. The negative ions on the hydrocarbon 
chain that is connected to the carboxylic group of  the fatty 
acids are what give soap its cleaning properties. Because 
soap’s carboxylic group is more soluble in water than it is 
in oil or grease, soap is mostly utilized in conjugation with 
water for cleaning reasons (Edah et al., 2017). Triglycerides 
are saponified to generate soap; this process reacts with 
a strong alkali, like KOH or NaOH to form glycerol 
and fatty acid salts. The long hydrocarbon chain of  a 
cleaning soap molecule contains an ionic interaction with 
a metal ion, typically potassium or sodium, at one end of  
the chain, where a carboxylic acid group is located. The 
ionic end is soluble in water, but the hydrocarbon end is 
non-polar and extremely soluble in non-polar substances. 

Because of  their capacity to combine water-insoluble 
materials and retain them within the water suspension, 
soaps have the power to clean (Shehu et al., 2020).
The structure of  the soap molecule is represented below: 
(Structure of  soap, 2024).
CH3 - (CH2)16 - COO- Na+

Non-polar hydrocarbon chain Ionic End
(Soluble in non-polar substances) (Soluble in water)
The fatty acids required to produce soap, such as stearic 
acid, myristic acid, palmitic acid, lauric acid, and oleic acid, 
are what give it its lathering and washing qualities. Even in 
seawater, soaps produced with fatty acids with 12 or more 
carbons are highly soluble and readily lather. However, 
because they irritate the skin and have offensive scents, 
fatty acids with 10 or fewer carbons are seldom employed 
in soaps. Because soaps can emulsify or scatter things 
that are insoluble in water and hold them in a suspension 
of  water, they have cleaning properties. This capability 
is demonstrated by the molecular makeup of  soaps. The 
molecules of  soap envelop the oil droplets when they are 
introduced to water containing oil or other substances 

Figure 1: A general scheme of  soap preparation (saponification) process (Besty et al., 2013)



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Am. J. Chem. Pharm. 3(1) 12-19, 2024

that are insoluble in water. The ionic end of  the oil 
permits it to dissolve in water, but the alkyl groups of  the 
soap molecules dissolve it. Consequently, the oil droplets 
should scatter throughout the water and be removed by 
washing (Habib et al., 2016).
The general saponification reaction used in soap 
preparation is shown Figure 1.
Producing natural and handmade soap is a completely 
creative process that calls for a range of  abilities, materials, 
creativity, and careful thought in order to produce soap 
of  the highest caliber. Fatty acids and alkali salts derived 
from plants or vegetables, as well as organic materials 
or natural smells, are characteristics of  herbal soaps. It 
takes the presence of  fatty acids and bases like potassium 
and sodium hydroxides for the hot-and-cold process 
that makes soap (Sindhu et al., 2019).  Natural, organic 
ingredients like herbs and skin-beneficial ingredients are 
used to make herbal soaps. There is very little chance that 
these soaps will cause any harm because they are entirely 
chemical-free. Rather, they contain essential oils like 
saffron, sandalwood, strawberry, and rose water, along 
with natural oils like castor and almond oil, all of  which 
are great for smoothing, brightening, and whitening skin 
(Hrushikesh & Hingane, 2022). Herbal soap preparation 
is a pharmaceutical or medicinal that is generally used to 
treat illnesses and injuries and to boost general health 
using plant components such as leaves, stems, roots, and 
fruits. It has antifungal and antibacterial qualities as well. 
This product can be administered topically and possesses 
antibacterial effects. It is available in many different forms, 
such as ointment, soap, gel, lotion, cream, and solvent 
extract. The qualities of  creams and soaps have been used 
to cure a variety of  skin issues (Dwiyanti et al., 2021).
The evergreen green herb Aloe vera, as Aloe barbadensis, 

is a plant that has fruits that are loaded with seeds, yellow 
tubular blooms, and thick, triangular leaves with sharp 
edges. Each leave is composed of  three layers: Inside is 
a translucent gel that contains 99% water and additional 
ingredients such as vitamins, lipids, sterols, amino acids, 
and glucomannans. Aloe vera leaves are used to create 
a mucilaginous, colourless gel that is frequently used in 
cosmetic and therapeutic applications (Sanchez et al., 
2020). In the past, this medicinal herb was used to cure 
skin ailments like inflammation, burns, and sores. Aloe 
vera has also been shown to have antioxidant, anticancer, 
antidiabetic, and anti-hyperlipidemic properties. Aloe 
vera contains over 75 different compounds, including 
vitamins A, C, E, and B12, enzymes like amylase, catalase, 
and peroxidase, minerals like zinc, copper, selenium, 
and calcium, sugars like monosaccharides like mannose-
6-phosphate and polysaccharides like glucomannans, 
anthraquinones like aloin and emodin, fatty acids like lipol 
and campesterol, hormones like auxins and gibberellins, 
and other substances like salicyclic acid, lignin, and 
saponins (Sopan et al., 2023).
This study was conducted to manufacture the herbal 
soap in assess its biological activity against both gram-
positive and gram-negative bacteria in addition to its 
physicochemical features. 
                        
MATERIALS AND METHODS
The study was conducted in Sainamaina Municipality, 
Rupandehi, Lumbini province, Nepal. According to 
Rupandehi (Wikipedia, 2024), the local government area 
is located between 27º 43’ 12’’ North latitude and 83º 18’ 
36’’ East longitude. The Aloe vera leaves collected from 
home garden. To keep the leaves out of  the sun, they 
were kept in the refrigerator for further use. 

Figure 2: Aloe vera Plant
Source: Fieldwork 2023

Data Analysis and Interpretation
The primary data was collected after the experiments 
and observation in the laboratory work. The data was 

arranged in tables and were evaluated by using appropriate 
statistical methods such as bar diagrams, graphs, and lines 
and other tools. 

Table 1: Analysis and Methods
Parameters Methods Employed
pH pH Measurement



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Am. J. Chem. Pharm. 3(1) 12-19, 2024

Moisture Content Change in initial and final weight
Free Caustic Alkali Titration
Foam Stability Height of  the Foam
Saponification Value Titration 

Chemicals and Standards
Every chemical utilized was of  the best grade available on 
the market. Laminar air flow and Antimicrobial Activity 
Incubator and Autoclave were provided by Indosati 
Scientific Lab Equipment, Delhi, and S.M. Scientific 
Instrument (p) Ltd. 

Extraction of  Aloe Vera Gel
Initially, aloe vera leaves that were just cut were cleaned 
with water to rid of  any dirt or yellow material known 
as aloe latex that leaked out of  the leaves. 400 g of  aloe 
vera leaves were precisely weighed using an automated 
weighing balance following washing. To gently cut away 
the clear gel in the center, the tops of  the leaves and the 
spines at the edges of  each leaf  were removed. To achieve 
the pure gel, the residual gel was spoonfully scooped into 
a mixture and then ground and filtered off. After that, 
the gel was placed in a beaker and kept at 4ºC in the 
refrigerator for further study.  

Formulation of  Aloe Vera Soap 
The formulation of  Aloe vera soap was carried out using 
(Upadhyay et al., 2021). After weighing 185 g of  aloe vera 

gel using an electronic scale, the gel was transferred into 
an ice cube tray to freeze. Subsequently, the beaker was 
filled with the frozen gel of  aloe vera. Then, gradually 
add 65.65 g of  NaOH. The temperature of  the aloe vera 
gel increased when NaOH was added. I had chilled the 
aloe vera gel to stop it from burning for that reason. The 
flakes of  NaOH were dissolved. An R.B. flask was filled 
with 450 g of  mixed oils (350 g olive oil and 100 g of  
coconut oil). Subsequently, the R.B. flask was filled with 
the Aloe vera gel and NaOH combination.
After that, three items were thoroughly combined with 
a shaker machine. Using the glass rod, the mixture and 
the aloe vera gel were rapidly mixed. Aloe vera gel, a 
vegetable oil, and NaOH solution had an exothermic 
reaction. There would be a longer saponification 
procedure. Thus, the mixture was stirred for two to 
three minutes, and then it was allowed to thicken for 
ten minutes. The mixture was thickened after one hour 
of  this operation. To make the mixture thick, it was put 
into the mold and allowed to rest for a few days. A thick 
bar started to form in the mold after one to two days. 
After the prepared soap had been soaked in the filter 
paper, it was ready to use. 

Figure 3: A-Formulated Aloe vera and B-Commercial Herbal Soap

Physiochemical Analysis of  Soap Prepared 
The properties of  formulated and Commercial Soap can 
be characterized in terms of  pH, Moisture Content, Free 
Caustic Alkali, and Foam Stability.

Determination of  pH in Soap
To determine the pH of  the produced soap, the beaker 
holding 20 mL of  distilled water was filled was 2 g of  the 
soaps that had been manufactured. Using the glass rod, 
it was aggressively swirled until the soap was completely 
dissolved. Using a pH meter, the soap solution’s pH was 
determined after 12 hours (Kareru et al., 2010).

Determination of  Moisture Content
The empty clean crucible dishes were dried in the oven at 
a temperature of  105ºC for 30 minutes and cooled for 10 
minutes in a desiccator. 5 g of  the prepared soap sample 
were taken in a dried, and tarred dishes. The sample was 
then dried for two hours at 101ºC to determine the moisture 
content (Benjamin et al., 2022) with some modification. A 
watch glass filled with soap was removed from the oven 
after two hours, and the total weight was determined.
The following formula was used to calculate the moisture 
content;
% Moisture Content= Weight loss/Sample Weight * 100 (1)



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Determination of  Free Caustic Alkali (FCA) 
Dissolved 5 g of  aloe vera soap into 30 mL of  ethanol. 
10 mL of  20% BaCl2 were added along with a few drops 
of  phenolphthalein indicator. Then, titrate the solution 
against 0.05 M H2SO4, by changing the color of  the 
indicator endpoint is recorded (Habib et al., 2016) with 
some modification. 
The free caustic alkalinity was calculated by using the 
following formulae:
FCA = 0.31/W × VA          (2) (Betsy, 2013)
Where, 
VA = Volume of  Titrated Acid
W = Weight of  Aloe vera soap
FCA = Free Caustic Alkali

Foam Stability 
In a 100 mL measuring cylinder, 1 g of  the aloe vera 
soap was weighed and dissolved in 20 mL of  distilled 
water. After giving the mixture a good shake for two to 2 
minutes, it was left to stand for an additional 2 minutes. 
After that, the height of  the foam was measured and 
noted (Kareru et al., 2010).

Determination of  Phytochemicals
Tannins, flavonoids, saponins and reducing sugars were 
determined using qualitative methods described by 
Imohiosen, (2023) with some modifications. 

Test for Tennins
In a beaker, 20 mL of  distilled water and 0.5 g of  Aloe 
vera extract were brought to a boil before being filtered. 
To the filtrate, a few drops of  0.1% ferric chloride were 
applied. A blue-black or brownish-green coloration 
suggested the presence of  tannin. 

Test for Flavonoids
Sulphuric acid (H2SO4) was added to a portion of  the 
plant extract’s aqueous filtrate after 5 mL of  diluted 
ammonia solution was added. The presence of  flavonoids 
was shown by the appearance of  a yellow solution that 
disappears with standing.

Test for Saponins
2.0 g of  plant extract was boiled in a water bath with 20 
mL of  distilled water, then filtered. For a stable, persistent 
fourth, 10 mL of  the filtrate and 5 mL of  distilled water 
were combined and forcefully shaken suggest the saponins. 

Test for Reducing Sugars
1 mL of  Fehling’s solutions A and B was added to 1mL of  
each sample’s aqueous filtrate, which was then heated in 
water. The presence of  non-reducing sugar was indicated 
by the red precipitate. 

Antimicrobial Activity Test 
For the research, gram-positive (S. aureus) and gram-
negative (E. coli) bacteria were obtained from the 

Crimson College of  Technology’s Department of  B.Sc. 
MLT in Butwal, Rupandehi, Nepal. The gram-positive 
and gram-negative commercial standards Gentamycin 
and Ciprofloxacin were utilized. 

Preparation and Dilution of  Soap Samples Extract 
(Chaudhari, 2016)
In a sterile container 1 g of  each soap sample was 
weighed and dissolved in 5 mL of  distilled water. The 
resulting concentration of  formulated and commercial 
soap of  200 mg/mL, 100 mg/mL, 50 mg/mL and 25 
mg/mL were then made serially and utilized for the disc 
preparation. 

Preparation of  Culture Medium (Rahama & Sani, 2020)
Muller Hinton Agar (MHA) media preparation

Activation of  Culture Plates
The previously prepared, frozen at 5ºC media plates were 
sufficiently dried during incubation. In a sterile airflow 
hood, the plates were thereafter allowed to cool. 

Preparation of  Filter Paper Disc
Whatman’s No. 1 filter paper was used to create filter 
paper discs. 5 mm discs were produced in Petri plates and 
autoclaved for 15 minutes at 121ºC to sterilize them. A 
formulated and commercial soap solution of  200 mg/
mL, 100 mg/mL, 50 mg/mL and 25 mg/mL was added 
one by one to each sterile disk. Paper discs soaked in the 
soap solution were allowed to stand for a period of  one 
hour to ensure full saturation of  the soap preparations. 
After being aseptically withdrawn from the soap solution, 
the discs were left to dry in an oven at 25ºC. 

Assay of  Antimicrobial Activity
Overnight cultures were kept ready for anti-microbial 
activity. Assay of  the antimicrobial activity of  soap were 
done by disc diffusion method. 

Disc Diffusion Assay
To find antibacterial assay, the Agar disc diffusion 
method (Chaudhari, 2016; Rahama & Sani, 2020) was 
employed. The surface of  sterile Muller-Hinton agar 
plates was inoculated with a standardized 0.1 mL saline 
suspension of  test organisms. Using sterile forceps, 
aseptically transfer sterile filter paper discs made from 
varying concentration i.e. 200 mg/mL, 100 mg/mL, 50 
mg/mL and 25 mg/mL respectively of  the individual 
formulated and commercial soap samples directly onto 
the plate surface and incubation at 37ºC for 24-48 hours. 

Measurement of  ZOI
The zone of  inhibition (ZOI) was determined by using 
a ruler or Vernier caliper in millimetre (mm.) caused by 
the Aloe vera formulated soap and commercial soap and 
compared with standard antibiotics following a 24 hours’ 
incubation period.



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RESULTS AND DISCUSSION
The physicochemical parameters of  the soap were 
categorized based on multiple factors, including pH, 

moisture content (MC), free caustic alkali (FCA), and 
foam stability of  both prepared and commercial aloe vera 
soap.

Table 2: The Physiochemical Results Obtained from Prepared Soap and Commercial Soap
Characteristics Prepared Soap Commercial Soap
pH 10.75 9.18
Free Caustic Alkali (%) 0.527 0.806
Moisture Content (%) 4.6 8.8
Foam Stability (cm) 6.5 10

Figure 4: The Bar Graph of  Physicochemical Analysis of  Prepared and Commercial Herbal soap 

Physicochemical Properties
The pH values of  the herbal soap that was produced and 
sold were found to be within the permitted range of  9 to 
11, which was in line with earlier research conducted by 
Rama and Sani (2020). Based on the measured pH, the 
soap should be alkaline, less corrosive, safe for the skin, 
and cause fewer skin reactions when used.
Other characteristics that were indicative of  the produced 
and commercial herbal soap were measured, such as 
the moisture content, formability, and percentage of  
free alkali content. The prepared Aloe vera soap had a 
moisture level of  4.6%, according to the analysis’s results, 
while commercial soap had a high moisture content while 
produced soap had a low moisture content. Low moisture 
indicates that the material won’t hydrolyze if  stored for an 
extended amount of  time (Chitkara et al., 2020). 
The aloe vera soap that was prepared had a free caustic 
alkali of  0.57%, whereas commercial soap had a free 
caustic alkali of  8.8%. The prepared soap’s low free 
caustic alkali rating indicates that it is suitable for sensitive 
skin types. The amount of  alkaline-free ingredients in the 
soap that, in excess, might irritate the skin is known as the 
free caustic alkali value. It is a sign that the soap won’t be 
too rough on the skin or fabric. 
According to the trial approach. Homemade aloe vera 
soap had the second-highest foam height at 6.5 cm, after 

commercial soap with a height of  10 cm. For foam height, 
liquid viscosity is the most influential factor. The type of  
oil used to make the soap, particularly palm kernel oil, 
whose main fatty acid is lauric acid and is well-known for 
its high formability, is responsible for the amount of  foam 
height (Rahama & Sani, 2020). The height of  the foam aids 
in removing grime, oil, and bacteria from the skin. 

Phytochemical Screening

Table 3: Results of  Phytochemical Screening
Compounds Results
Tennis +
Flavonoids +
Saponins +
Reducing Sugar +

(+) = Present

Table 3 shows the phytochemical screening result of  
the Aloe vera extract which indicated the presence of  all 
phytochemicals tested. In the previous literature (Kareru 
et al., 2010) almost all the phytochemical screening of  the 
Aloe vera leaves extract. 

Antibacterial Susceptibility Test

Table 4: The Antibacterial activity of  produced and commercial soap of  their ZOI
S Zone of  inhibition against fractions (mm)

Produced Soap (mg/mL) Commercial Soap (mg/mL)
200 100 50 25 200 100 50 25



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E. coli 8 7 8 - 7 - - -
S. aureus 10 9 - 8 - - - -
Ciprofloxacin 
(E. coli) 34 33
S. aureus 20 17
Gentamicin 
(E. coli) 24 25
S. aureus 15 13
Control -

(-) = No zone of  inhibition

Figure 5: The Bar Graph of  Antimicrobial Activity of  Prepared and Commercial herbal soap with E. coli and S. aureus 
bacteria 

Figure 6: Different concentrations of  Solution Prepared by Formulated and Commercial Herbal Soap 



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A = ZOI of  Formulated Soap against S. aureus; B = ZOI 
of  Commercial Soap against S. aureus;
C = ZOI of  Formulated Soap against E. coli; D = ZOI of  
Commercial Soap against E. coli 
The prepared soap and the commercial soap exhibit 
strong inhibition at the highest concentration (200 mg/
mL), which is followed by a steady decline in inhibitory 
character down to the lowest dosage (25 mg/mL) in 
the given anti-bacterial screening results. According 
to Sharma et al. (2022), the ZOI of  gram-positive (B. 
subtiles) and gram-negative (S. typhi) at 400 mg/mL were, 
respectively, 0.82 and 1.62 mm for manufactured herbal 
soap and marketed herbal soap. Higher concentrations of  
ZOI of  produced soap A-500, 250, 125, 62.5, and 32.25 
mg/mL were achieved for S. aureus 20, 16, 12, 9, and 7 as 
well as E. coli 15, 11, 9, 8, and 7 mm, respectively, showing 
anti-bacterial action (Kareru et al., 2010). 
The finding shows that the commercial aloe vera soap had 
a higher concentration of  anti-bacterial activity than the 
produced soap. This is because the commercial aloe vera 
soap had extra ingredients that enhanced its activity, while 
the produced aloe vera only contained olive and coconut 
oil and NaOH. Also by preventing bacterial development, 
this showed that both soaps have anti-bacterial activity. 

CONCLUSION
In addition to its moisturizing and anti-inflammatory 
qualities, Aloe vera seems to enhance the skin’s ability to 
absorb some pharmaceutical molecule. Since, external 
application of  Aloe vera on intact skin is generally 
considered harmless and does not appear to have 
any side effects, the use of  this natural resource as a 
penetration enhancer is intriguing (Sharma et al., 2015). 
This study shows that formulated herbal soap from 
aloe vera leaves can be successful. When compared 
to commercially available herbal soap, the formulated 
soap’s physicochemical qualities show a significant 
improvement. The results that are now available indicate 
that aloe vera extract has antibacterial properties and has 
been successfully employed in the formulated soap. 

Recommendation
To prevent adverse effects on the skin, make sure the right 
amount of  necessary ingredients is used when making 
aloe vera soap. We can create a variety of  herbal soaps 
with easy-to-use methods that don’t include dangerous 
chemicals for everyday usage.

• Before using soap on our bodies, it is important to 
inspect its quality.

• Determining the Minimum Inhibitory Concentration 
(MIC) of  Aloe vera soap.

• Determining the results of  various soap’s Minimum 
Bactericidal Concentration (MBC) may also be conducted.

• Determine the antioxidant character of  herbal soaps. 

Data Availability
The data used to support the findings of  this study are 
included in the article. 

Authors’ Contributions
All author prepared the materials, collected the data, and 
conducted the analysis in addition to contributing to the 
study’s conceptualization and design. Gautam P C has 
specially supported the anti-bacterial analysis procedure. 
Moreover, Arjun B wrote the draft of  the work, offered 
feedback on earlier iterations, and read and approved the 
completed version. 

Acknowledgments
The authors would like to thank Butwal Multiple 
Campus, Tribhuvan University and Crimson College 
of  Technology, Pokhara University for their invaluable 
assistance in testing the study samples and greatly 
enhancing the correctness of  our results.

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Figure 7: Anti-bacterial Activity Shown by Formulated and Commercial Herbal Soap



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