The Mixing of Ethanol Extract of Terminalia catappa L in Transparent Soap Base to maintaining it's the Organoleptic properties, Solid quality and Inhibition of Staphylococcus aureus

 

Munira Munira1*, Ampera Miko2, Muhammad Nasir3, Basri A. Gani4

1Department of Pharmacy, Politeknik Kesehatan Aceh, Aceh Besar, Aceh, Indonesia.

2Department of Nutrition, Politeknik Kesehatan Aceh, Aceh Besar, Aceh, Indonesia.

3Department Biology, Faculty of Mathematics and Natural Sciences, Universitas Syiah Kuala, Darussalam, Banda Aceh,Aceh,  Indoensia.

4Department of Oral Biology, Dentistry Faculty, Universitas Syiah Kuala, Darussalam,

Banda Aceh, Aceh, Indonesia.

*Corresponding Author E-mail: munira.bio@poltekkesaceh.ac.id

 

ABSTRACT:

Staphylococcus aureus (S. aureus) caused skin surface infections. The active ingredients in soap dosage forms of Terminalia catappa L extract have antibacterial properties and retain alkaline properties. The study evaluated Terminalia catappa L extract's quality in a transparent soap formulation in inhibiting S. aureus and maintaining soap organoleptic. Terminalia catappa L was extracted by the maceration method also phytochemical screening. An organoleptic assay examined soap's formulation, and an inhibitory of S. aureus was analyzed by disk method. The ethanol extract of Terminalia catappa L contains alkaloids, saponins, tannins, polyphenols, quinones, flavonoids, and triterpenoids. The transparent soap formulation with the addition of ethanol extract of Terminalia catappa L has good organoleptic properties according to the Indonesian National Standard (SNI) 3532-2016. Also, there is a change in the color of transparent solid soap based on the concentration, at a concentration of 1% (light brown and transparent), 2% (dark brown and transparent), and 3% (dark brown and transparent). Soap with ethanol extract content of Terminalia catappa L has high inhibition of S. aureus at a concentration of 3% (20.09 mm). While the concentration is 1% (13.33mm), 2% (15.58 mm), and soap base (19.36 mm). The transparent solid soap base added with ethanol extract of Terminalia catappa L has good quality, maintains the organoleptic properties of soap, and can inhibit the development of S. aureus.

 

KEYWORDS: Staphylococcus aureus, Transparent soap solid, Terminalia catappa L, Organoleptic.

 

 


INTRODUCTION:

The Staphylococcus aureus (S. aureus) was reported to be the most dominant gram-positive bacteria infecting the skin. The skin has a very high sensitivity to environmental changes. Apart from functioning as a thermoregulator, the skin also provides aesthetic value to the body1. The S. aureus infection can cause dermatitis, impetigo, and cellulitis2. The use of antibacterial soap is intended to prevent bacteria like S. aureus as agents that cause the skin's infection.

 

Vidal (2018) reports that soap is a saponification product made through a triglyceride reaction process with a strong base such as potassium or sodium hydroxide, producing glycerol, salt, and fatty acids3. Transparent solid soap is one of the most frequently used antibacterial soap innovations. Its soap can be used as an emulsion preparation to function as a promising drug agent, including antibacterial4.

 

Triclosan is an active ingredient in antibacterial soaps. Besides, triclosan can cause several health problems, such as endocrine system disorders, cancer, and bacterial resistance5. Using triclosan with levels that are not suitable for a transparent solid soap base can cause skin irritation and reduce UV protection6. Using antibacterial soap with active ingredients derived from nature is an alternative to avoid skin irritation and allergies. One of them is the ketapang leaf (Terminalia catappa L), which is believed to have antibacterial properties while maintaining the organoleptic properties of soap.

 

The Terminalia catappa L was beneficial in diseases of metabolic disorders of joints and muscles, respiratory and digestive tract infections7. Farther, Terminalia catappa L can prevent skin allergies and reduce infection intensity with Staphylococcus aureus and other gram-negative bacteria8. Terminalia catappa L contains flavonoids, alkaloids, tannins, terpenoids, saponins, and phenolics. These chemical compounds' content functions as antioxidants, anti-inflammatory, and anti-pyretic9. The Terminalia catappa L can also prevent oxidation of skin fibroblasts from UV rays' influence10.

 

Munira (2018) proved that red ketapang leaf extract (Terminalia catappa L) has the highest antibacterial activity in inhibiting Staphylococcus aureus11. However, there are few research reports on Terminalia catappa L in antibacterial soap bases. Therefore, this study evaluates Terminalia catappa L's function to maintain organoleptic properties related to its ability to inhibit the growth of S. aureus as a pathogen involved in the pathogenesis of skin infections.

 

MATERIAL AND METHODS:

This study used Terminalia catappa L with 3%, 2%, and 1% as the test material. Meanwhile, S. aureus ATCC 25923 was used as the subject. This research was conducted in vitro to evaluate the quality of the ethanol extract of Terminalia catappa L as anti of S. aureus and maintain the organoleptic soap's quality.

 

Plant material:

The red ketapang leaves (Terminalia catappa L) were obtained from the Aceh Besar area with Latitude coordinates: 5° 35'4.14 "N, Longitude: 95 ° 21'5.34" E. The extraction and storage process at the Pharmacy Laboratory, Health Polytechnic, Banda Aceh, Indonesia. Extract of Terminalia catappa L collected by Munira Munira with voucher number L2231.

 

Plant extract preparation and phytochemical screening:

Plant extract preparation and phytochemical screening were adopted by Yusuf (2021)12. A maceration process carried out 1000grams of Terminalia catappa L by adding 10 L of 96% ethanol, then stirred and let stand for 18 hours. Furthermore, the macerate was separated by filtration. The macerate is then evaporated with a vacuum rotary evaporator until a thick extract is obtained. Moreover, the quote was subjected to phytochemical screening. In the first stage, the alkaloids are examined. Examination of Dragendroff adds a few drops of Dragendroff reagent to the sample extract and positive results containing alkaloids when a red precipitate emerged. The next step is Hager examination by adding a few Hager reagent drops to the sample extract and a positive result containing alkaloids if a yellow precipitate.

 

The flavonoid assay was carried out by adding 0.5mL of concentrated hydrochloric acid and Magnesium metal to 1mL of extract, and the red color showed positive results containing flavonoids. Simultaneously, checking the saponin content by dissolving 50mg of the extract in 20mL of aquadest. The suspension is shaken vertically for 15min. Saponins are positive if it is formed a foam layer. The polyphenol assay added iron (III) chloride solvent to the extract. Positive if a blue-black color solution. Simultaneously, the tannin test adds 1% gelatin to the sample, positive if the white solution is cloudy. Steroid and terpenoid tests were used to add chloroform and concentrated sulfuric acid to the extract, positive result for containing terpenoids if a pink, red or purple color forms. While positive for steroids if a turquoise color. Quinone content examination was adding 5% KOH solution. Positive results if the formation of a yellow to reddish color in the solution.

 

Formulation of the transparent solid soap antibacterial:

The making of transparent antibacterial soap formulation refers to the research results of Febriyenti et al. (2014)13. Table 1 reports the ingredients used to make transparent solid soap. In the first stage, all the ingredients are weighed, and the stearic acid is melted at 60°C, then added with oil (VCO and olive oil) then stirred until homogeneous. Next, 30% NaOH was added at 70°C, then went for 4 min until soap formed. The temperature was lowered to 50°C, and glycerin, TEA, sucrose, coco betaine, and citric acid were mixed stirred for 10 min until homogeneous. Then slowly add 96% ethanol to form a clear solution. Making antibacterial soap is carried out by adding various concentrations of ethanol extract of Terminalia catappa L in a base soap preparation. Then dissolved in the remaining 96% ethanol and stirred at 40°C until homogeneous. Furthermore, the fragrance is added and remixed until homogeneous and put into a transparent soap mould.

 

Evaluation of organoleptic properties of transparent solid soap:

The organoleptic evaluation of antibacterial transparent solid soap was evaluated shape, color, odor, pH, moisture content, and high foam. Checking the soap's pH was started by dissolving it in water and then measuring it with a pH meter (Mettler Toledo, Merck KGaA, Darmstadt, Germany). After that, we checked the moisture content 1 gram of sample in a porcelain dish was heated in an oven at 105ºC for 2 h, then weighed. Then re-heating was carried out until the weight was constant. The subsequent examination is the assessment of high foam. One gram of soap is dissolved in 10mL of aquadest. A total of 5mL of dissolved soap is put into a test tube then homogeneous with vortex for 2 min. The foam formed was then observed. The observations until the fourth week of storage13.

 

Antibacterial assay of transparent solid soap:

Transparent solid soap antibacterial assayed by disc diffusion method adopted from Niyas (2016).14 The S. aureus suspension was calibrated with Mc Farland 0.5 (1.5x108 CFU/mL). In the first stage, the disk paper was adapted in a soap formula solution for 15 min, formula 1 (soap base), formula 2(1%), formula 3(2%), and formula 4(3%). Media Nutrient Agar (Sigma-Aldrich, Merck KGaA, Darmstadt, Germany) is sterilized is poured into a petri dish until it is firm. Furthermore, sterile cotton swabs were inserted into a test tube containing a suspension of S. aureus. Then rubbed it all over the surface to make it medium. Then the disk paper is placed on the surface of the medium based on the concentration formula. Besides, it was incubated at 37°C for 24 h. Calipers (mm) measure the precipitation zone (inhibition zone) is calculated by calipers (mm). The optimum inhibition zone value is measured in horizontal and vertical areas. The average is then taken to obtain an antibacterial image of the soap formula added with the ethanol extract Terminalia catappa L.

 

Statistical Analysis:

One Way ANOVA statistically analyzed a significant difference in the inhibition of S. aureus (p<0.005; 0.000) with a strong correlation (r=1).

 

RESULTS AND DISCUSSION:

Fig 1 shows that the extract of Terminalia catappa L can be homogeneous in a solid transparent soap base formula. The concentration of the test material can affect the color change of the soap base. This property is one of the requirements for making antibacterial soap. The soap preparation added with 1% extract of Terminalia catappa L is light brown and transparent, 2% dark brown, and 3% dark brown, both of which are not transparent. The change in the color of the soap base added with the test material indicated that the homogenization process was going well. Figure 2 and Table 4 show that Terminalia catappa L, which is added in a transparent solid soap base, can inhibit the growth of S. aureus on a substantial scale (1% and 2% concentrations) and is very strong at a concentration of 3%.

 

Fig.1: Profile of transparent solid soap with the addition of ethanol extract of Terminalia catappa L. (A) Soap base, (B) Soap base with 1% extract (C) Soap base with 2% extract, and (D) Base fighting with 3% extract. The concentration of Terminalia catappa L affects the discoloration of soap.

 

Fig.2: The extractive power of Terminalia catappa L concentrations of ethanol against S. aureus. (A) aquadest (control), (B) soap base, (C) 1%, (D) 2%, and (E) 3%. Blue arrows (zone of inhibition), black arrows (S. aureus).

 

Figure 2 explained the transparent solid soap that added Terminalia catappa L extract had excellent qualities in inhibiting the growth of S. aureus. All concentrations of the antibacterial soap solution have excellent sensitivity with a minimum inhibition concentration (MIC) value above 16mm. Therefore, the inhibition of S aureus utilizing a solid transparent soap is determined by the concentration of the essay material used in the soap. Table 1 shows the formula for making solid transparent soap with the addition of extract of Terminalia catappa L. Whereas Table 2 shows the results of the phytochemical screening of Terminalia catappa L. on these results. It indicates that Terminalia catappa L has antioxidant and antibacterial properties.


Table 1: The formula of transparent solid soap from the ethanol extract of Terminalia catappa L

S. No

Material

Composition (% w/v)*

Formula 1

Formula 2

Formula 3

Formula 4

1

Stearic Acid

6.5

6.5

6.5

6.5

2

Virgin coconut oil

15.0

15.0

15.0

15.0

3

Olive oil

6.0

6.0

6.0

6.0

4

NaOH 30%

20.0

20.0

20.0

20.0

5

Ethanol 96%

17.0

17.0

17.0

17.0

6

Glycerin

12.0

12.0

12.0

12.0

7

Sucrose

10.0

10.0

10.0

10.0

8

Triethanolamine (TEA)

2.0

2.0

2.0

2.0

9

Citric acid

4.5

4.5

4.5

4.5

10

Butylated hydroxytoluene (BHT)

0.1

0.1

0.1

0.1

11

Coco betaine

2.0

2.0

2.0

2.0

12

Fragrance

0.5

0.5

0.5

0.5

13

Terminalia catappa L Ethanol extract

0

1(%)

2 (%)

3 (%)

14

Aqua Destillata

100

100

100

100

* w/v = weight/volume

 


Table 2 shows the ethanol extract of Terminalia catappa L containing alkaloids, saponins, tannins, polyphenols, quinones, flavonoids, triterpenoids, and the steroid is undetected. Table 3 explains the organoleptic soap properties. The soap has a solid texture and a citrus aroma. The addition of citrus fragrance causes this citrus scent. When viewed from the color, the soap that contained test material is transparent. Organoleptic properties of water content range from 12.50-13.99%. The water content in this soap preparation follows the Indonesian National Standard (SNI), which is a maximum of 15%. Simultaneously, the high yield of foam in each Terminalia catappa L extract formulation ranged from 4.5cm - 5.5cm. The highest foam is founded in Formula 2 (extract 2%), and the lowest is formula 3 (extract 3%). At the same time, the pH of soap preparations ranges from 8.6-9. The pH meets the SNI between 8-1115.

 

Table 3 shows that soap with the addition of 1% and 3% extract of Terminalia catappa L shows a pH above 9, while soap with 2% Terminalia catappa L has the same pH as a soap base. However, soap with the addition of Terminalia catappa L has a high foam above the soap base value (without Terminalia catappa L extract). At the same time, the texture and aroma did not change. Meanwhile, Table 4 reports the effect of Terminalia catappa L in transparent solid soap base in inhibiting the growth of S. aureus.

 

Table 2: Phytochemical screening of ethanol extract of Terminalia catappa L

Phytochemical Screening

Results

Alkaloids

+

a.     Hager

+

b.     Dragendrof

+

c.     Bouchardat's

+

Saponin

+

Tannin

+

Polyphenol

+

Quinone

+

Steroid

-

Flavonoid

+

Triterpenoids

+

(+) = Contains the tested compound; (-) = Contains no tested compounds

 


 

Table 3: Organoleptic assay performance for transparent soap with the addition of ethanol extract Terminalia catappa L.

Soap formula

Parameter of organoleptic and soap properties

Color

Aroma

Texture

Water Content (%)

Foam Height (cm)

pH

Formula 1 (Soap base)

Transparent

Lemon

Solid

12.50

4.7

8.6

Formula 2 (1%)

Light brown-Transparent

Lemon

Solid

13.35

4.8

9.4

Formula 3 (2%)

Dark brown

Lemon

Solid

13.66

5.5

8.7

Formula 4 (3%)

Dark brown

Lemon

Solid

13.99

4.5

9

 


 

Table 4: Inhibition of S. aureus by transparent solid soap with ethanol extract of Terminalia catappa L.

S. No.

Treatment groups

N

Average

SD

Inhibition Scale

P-Value

1

Aquadest (control)

5

0.00

0.00

No action

 

p<0.05 (0,000)

2

F1 (Soap Base)

5

19.36

0.27

Strong

3

F2 (Soap + 1% Extract)

5

13.33

0.78

Strong

4

F3 (Soap +2% Extract)

5

15.58

0.38

Strong

5

F4 (Soap + 3% Extract)

5

20.09

0.25

Very Strong

Data represent means ± SD (n=5). Table 4 shows the One Way ANOVA statistical analysis showed a significant difference in the inhibition of S. aureus from the three concentrations (p<0.005; 0.000). Based on the average inhibition zone, the soap base with the addition of Terminalia catappa L at a concentration of 3% has strongly inhibitory compared to 1% and 2% concentrations.


 

 

This study explained that Terminalia catappa L has a significant effect on the organoleptic properties of soap and can inhibit the development of S. aureus. These two potentials become references to Terminalia catappa L as active material in antibacterial soap bases. Table 2 reported that Terminalia catappa L has antibacterial and antioxidant properties. Based on phytochemical screening, the extract of Terminalia catappa L contains alkaloid compounds, saponins, tannins, polyphenols, quinones, flavonoids, and triterpenoids. Anand (2015) reports that Terminalia catappa L is a fruit with high nutritional value and medicinal benefits. It contains phenols, flavonoids, and carotenoids anti-inflammatory, antidiabetic, antioxidant, hepatoprotective, and anticancer9.

 

Table 3 reported that Terminalia catappa L could improve soap's organoleptic properties by maintaining alkaline pH, water content, texture, aroma, and high soap foam. This property is needed for soap-based preparations to withstand moisture and temperature and protect the skin from the threat of bacterial infection and other pathogens16. The ability to improve the organoleptic properties of soap by Terminalia catappa L can be possible. Several chemical compounds such as saponins, alkaloids, and phenol flavonoids help reduce water's surface tension17. Surface tension is the force that allows the water lump on the surface to hold its shape from spreading18. Maibaum (2004) reported that this activity is related to the hydrophobic molecule and the solution's ionic activity. This formation is called the micelles process19. Soap in the form of micelles, besides having the ability to clean oily dirt, also has antibacterial properties and ultraviolet protection20. The enzymes in Terminalia catappa L may clean various starch and protein particles that support bacterial development21.

 

Based on the research results that Terminalia catappa L can maintain the soap's alkaline pH, this property is related to safety. It can increase the absorption of the skin so that it allows the skin to avoid irritation. Terminalia catappa L can also maintain the balance of water content in soap to affect the foam produced by soap22. It is due to the soap molecule factor, which has the head bound with water (hydrophilic), and the other end is attached to oil and grease (hydrophobic)23. Besides helping to remove dirt and oil, the active molecules in soap foam prevent bacteria's development on the skin24. Cohen (2017) reports that soap can attract unwanted substances from the skin to enter the water through the hydrophilic head25. Another research finding is that soap changes color after adding Terminalia catappa L with different concentrations. The resulting color is a mixture of oil, NaOH, and Terminalia catappa L extract. According to Wijana (2019), a factor affecting the transparent structure of soap is ethanol, glycerin, and sugar solution26.

 

In Table 4, a transparent solid soap base with the addition of Terminalia catappa L in different concentrations strongly affects the development of S. aureus. It is assumed that several chemical compounds in the essay material have a strong sensitivity to S. aureus. Salares (2018) reports that the methanol extract of ketapang leaves (Terminalia catappa L) has antioxidant and antibacterial effects on the Staphylococcus aureus, Bacillus cereus, Escherichia coli, Salmonella typhi, Serratia marcescens, Zymomonas mobilis27. Meanwhile, Allyn (2018) proved that brown ketapang leaf extract could inhibit the growth of Staphylococcus aureus ATCC 25923 and Pseudomonas aeruginosa ATCC 2785321. Another study has shown that the leaf and flower extracts of Quisqualis indica Linn possess bioactive components for antibacterial activity28. Shahid (2015) said that the antibacterial compounds in soap provide maximum activity in inhibiting bacteria, which is influenced by the hydrophilic-lipophilic properties of soap29. The hydrophilic nature of soap causes the antimicrobial compounds to diffuse in a polar medium. In contrast, soap's lipophilic properties will help penetrate the antibacterial compounds into the bacterial cell membrane30.

 

This research is an effort to develop the phytopharmica value of Terminalia catappa L plants as an additional ingredient in antibacterial soap innovation. Further research is highly recommended to strengthen previous studies' results, including purification and purification of Terminalia catappa L extract. The solid soap base remains transparent even though added with high concentrations of Terminalia catappa L. Besides, an evaluation of the hydrophilic and hydrophobic activity of the soap solution after adding Terminalia catappa L to gram-negative bacteria was related to its virulence and sensitivity to skin allergies.

 

CONCLUSION:

The transparent solid soap base added with ethanol extract of Terminalia catappa L has good quality, maintains the organoleptic properties of soap, and can inhibit the development of S. aureus, especially the strongest at a concentration of 3%.

 

CONFLICT OF INTEREST:

The authors declare no conflict of interest.

 

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Received on 03.04.2021           Modified on 07.11.2021

Accepted on 14.02.2022         © RJPT All right reserved

Research J. Pharm. and Tech. 2022; 15(8):3423-3428.

DOI: 10.52711/0974-360X.2022.00573