Formulation and evaluation of antibacterial Moringa soap prepared by Melt pour and Cold process method

Shashank Nayak N¹, Akanksha², Ashwini S K³, Manoj M J⁴, Srusti B⁵,

Shwetha S⁶, Kamath K⁶, Thimmasetty J⁷

¹Associate Professor, Department of Pharmaceutics, Bapuji Pharmacy College, SS Layout, Shamanur Road, Davanagere, India. Affiliated to Rajiv Gandhi University of Health Sciences, Bengaluru, Karnataka.

²Final year B Pharm Student, Bapuji Pharmacy College, SS Layout, Shamanur Road, Davanagere, India. Affiliated to Rajiv Gandhi University of Health Sciences, Bengaluru, Karnataka.

³Final year B Pharm Student, Bapuji Pharmacy College, SS Layout, Shamanur Road, Davanagere, India. Affiliated to Rajiv Gandhi University of Health Sciences, Bengaluru, Karnataka.

⁴Final year B Pharm Student, Bapuji Pharmacy College, SS Layout, Shamanur Road, Davanagere, India. Affiliated to Rajiv Gandhi University of Health Sciences, Bengaluru, Karnataka.

⁵Final year B Pharm Student, Bapuji Pharmacy College, SS Layout, Shamanur Road, Davanagere, India. Affiliated to Rajiv Gandhi University of Health Sciences, Bengaluru, Karnataka.

⁶Assistant Professor, Department of Pharmaceutics, Bapuji Pharmacy College, SS Layout, Shamanur road, Davanagere, India. Affiliated to Rajiv Gandhi University of Health Sciences, Bengaluru, Karnataka.

⁷Professor and HOD, Department of Pharmaceutics, Bapuji Pharmacy College, SS Layout, Shamanur road, Davanagere, India. Affiliated to Rajiv Gandhi University of Health Sciences, Bengaluru, Karnataka.

*Corresponding Author E-mail: shashanknayak87@gmail.com

ABSTRACT:

In the present investigation, melt-pour and cold-process soaps were made using various Moringa oleifera components, including seed oil, leaf powder, and leaf extracted oil. The produced soaps properties like pH, foaming capacity, foam retention, Total fatty content (TFM), saponification value, and anti-microbial activity, were determined. The 3%, 4%, and 5% moringa seed oil, leaf powder, and leaf extracted oil were used to make the soaps, and the 5% soap made using the melt-pour and cold-process method had the most promising outcomes. Considering these findings, the 5% cold process soaps were made employing the Soapcalc calculator. As advised by the soapcalc calculator, the following ingredients were utilised in this method: moringa powder, moringa seed oil, moringa leaf extract, cocoa butter, coconut oil, castor oil, olive oil, and NaOH lye. Results from the cold process experiment were satisfactory. The foam height was determined to be between 5 and 24 ml, the pH of the prepared soap was between 8.24to10.04, and the zone of inhibition of the prepared soaps for E. coli and Klebsiella was between 14 mm and 27 mm. The TFM of the prepared soap F1 to F9 was 19.6, this indicated that the melt and pour base had lower TFM. Furthermore, formulation F10 to F12 had the TFM in the range of 55.9 to 58.7. The saponification value of F10 to F12 was in the range of 27.73 to 62.74.

KEYWORDS: Soapcalc calculator, Melt pour method (MPM), Cold process method (CPM), TFM, Saponification value, Distilled Water (DW), Antibacterial activity.

INTRODUCTION:

The Moringa plant, which has seeds, pods, and leaves, is a member of the Moringaceae family and is native to Africa, the Indian subcontinent, and South and Southeast Asia. It is grown extensively for myriad benefits of its young pods and leaves. It is regarded as a top notch supplement due to its high protein content. Without that, because of its many beneficial characteristics, it is referred to as the miracle tree. ^1,10

The main chemical constituents of moringa leaves are fibre, fat, protein and minerals like Ca, Mg, P, K, Cu, vitaminA, B,B1. The seeds contain oleic acid, linolic acid, tannins, saponins, terpenoids and lectins. The roots contain alkaloids like morphine and the pods contain lipids, carbohydrates, proteins, and ash. These chemical constituents are found to be useful in the treatment of asthma hyperglycaemia, syphilis, malaria, Crohn’s disease, gout, epilepsy, STDs, ulcer, diarrhoea, and inflammation.²

Soap is a cleansing agent prepared by the action of alkali on the fatty acids. They are produced by MPM and CPM. In MPM, saponified bars are purchased from vendors and will be melted in oven or by double boiler method. When soap is completely melted vegetables or fruit extracts will be added with fragrance, colour or other additives and poured into the mould and allowed to solidify. Whereas, cold process method involves heating of various oils in which lye-water mixture will be added after heating. It will be mixed until it thickens after which fragrance and additives will be added and poured into the mould.³

M. oleifera leaf extracts were used in a study by Yongbing Xu et al. to measure the antielastase as well as anticollagenase activities using in vitro spectrophotometric and fluorometric assays. As potential anti-elastase, anti-collagenase, and antihyaluronidase ligands, phytochemicals 10, 8, and 14 were ruled out. Affinity ultrafiltration, HPLEUV/ESIMS/MS analysis, collagenase, and elastase inhibitory activity were all determined. Molecule docking analysis was also used to provide additional confirmation. These findings imply that the leaves of M. oleifera are a pure source of anti-aging skincare ingredients. ⁴

In order to create a moringa seed oil cream, Sirivan Athikomkulchai et al. researched the advantages of moringa oleifera seed oil. A free radical scavenging assay using 2,2diphenyl1-picrylhydrazyl (DPPH) was used to measure the oil's antioxidant activity. Moringa seed oil was used to create an o/w cream, which was then tested for antioxidant activity. Measurements of the skin's hydration, erythema, melanin levels, and viscoelasticity were made on 32 participants to determine the effectiveness of the moringa seed oil cream. According to the stability study, after 28 days of storage at 4, 30, and 45 C and six heatingcooling cycles, the pH, the viscosity and rheological properties of the cream with moringa seed oil had not been significantly altered. The the moringa seed oil cream showed antioxidant activity in vitro and increased skin hydration in vivo. ^\5

In this current investigation, attempts are made to formulate and evaluate the moringa soaps by melt pour method and cold process method. Dried moringa leaf, moringa seed oil and moringa leaf extract oil are used in the concentration of 3 to 5%. The prepared soaps were subjected to varied evaluation parameters which were reported in the literature.

MATERIALS AND METHODS:

Castor oil, Olive oil, Coconut oil, tea tree oil, Anise oil, Moringa powder, Silicon moulds and other basis needed for making melt and pour soap were purchased from the Akshit sales corporation (Flipkart). All the other ingredients were procured from Rajesh chemicals, Mumbai and Vasa Scientifics, Bangalore.

Melt and Pour method (MPM)³

Preparation of moringa soap using melt and pour base (F1 to F9):

3%, 4% and 5% of moringa powder, moringa seed oil and moringa leaf oil were added in 100 g of Ultra Clear Glycerine Pour and Melt soap Base and melted in a 250 ml beaker on water bath using the double boiler method at a temperature of 100°C.0.5 ml (1.0%) of benzyl alcohol was added to the melted base before solidification. The beaker was removed from the water bath and brought to congealing temperature. Later, five drops of tea tree oil was added. The melted soap base containing the moringa was poured in to the soap moulds lubricated with glycerine, and allowed to solidify for 3 hours at room temp. Finally, the soaps were demoulded and subjected to a varied evaluation parameters like pH, foam forming ability, foam retention, moisture content, antimicrobial activity.

Figure 1: Preparation of Moringa soap by MPM

Figure 2: Preparation of Moringa soap by CPM

Cold Process Method (CPM):

Cold process is the ideal method to prepare soap when we have a series of oils and waxes of varied grades. The properties of the soap can be controlled by preparing it by cold process method. The trial-and-error method of soap preparation can be avoided using soapcalc calculator, the detailed procedure for the use of the calculator is given below.

How to use soap calc calculator

1. Select the type of lye- Click on NaOH for the hard soap preparation by cold process method.

2. Select weight of oils- For ease of use select grams and enter the quantity requires (50 g- Approx 1 soap was obtained).

3. Enter lye concentration

• Enter % (This % can be changed depending on type of oil used)

• Low water- 50% (50% NaOH+50% Water) – Fast drying- Hard soap

• Medium Water- 33% (33% NaOH+ 67 % Water)

• High Water- 25% (25% NaOH+ 75% Water)- Long time to dry- Leads to shrinkage of the soap surface. Hence, approximately 40 % was selected.

• Find out NaOH and Water concentration at the end of the calculation. Check whether the properties like ideal hardness, cleansing, conditioning, bubbly, and creamy properties.

4. Super fat- Ideal 4% of excess oil is added to prevent drying of soap.

5. Enter oils and bases which is preferred to be present in soap.

E.g.- Olive oil- 50%, Coconut oil- 22%, Castor oil- 3%.

6. Click on calculate logo recipe, then click on view and print recipe.

7. Amount of NaOH and Water required for saponification will be displayed on the soapcalc calculator. (Water-10.55 g, NaOH-7.03 g)

8. All the oils are weighed along with 25% of cocoa butter and heated on the water bath. 5 % of Moringa leaf powder, moringa seed oil and moringa leaf extract oil and 2% of Niacinamide was added to the mixture. Prescribed amount of lye is added to above mixture containing oil and wax was homogenized using silverson emulsifier until it obtained pourable consistency. After complete homogenization, the above mixture was transferred to the silicon moulds.

9. Demoulding followed by evaluation of the soaps were performed after 4 weeks as mentioned in F10 to F12.

Figure 3: Soap recipe for moringa soap by Soapcalc calculator

Figure 4: Desired properties of soap before soap preparation by use of Soapcalc calculator

Table 1: Formulation of Moringa soaps by Melt and pour method:

Ingredients	F₁	F₂	F₃	F₄	F₅	F₆	F₇	F₈	F9
Moringa leaf powder	1.5 gm (3%)	2 gm (4%)	2.5 gm (5%)	-	-	-	-	-	-
Moringa seed oil	-	-	-	1.5 ml (3%)	2.0 ml (4%)	2.5 ml (5%)	-	-	-
Moringa leaf extract oil	-	-	-	-	-	-	1.5 ml (3%)	2 ml (4%)	2.5 ml (5%)
Niacinamide (2%)	1 gm	1 gm	1 gm	1 gm	1 gm	1 gm	1 gm	1 gm	1 gm
Tea tree oil	0.3 ml	0.3 ml	0.3 ml	0.3 ml	0.3 ml	0.3 ml	0.3 ml	0.3 ml	0.3 ml
Benzyl alcohol	0.5 ml	0.5 ml	0.5 ml	0.5 ml	0.5 ml	0.5 ml	0.5 ml	0.5 ml	0.5 ml
Ultra clear glycerine soap base	50 gm	50 gm	50 gm	50 gm	50 gm	50 gm	50 gm	50 gm	50 gm

Table 2: The formulation of moringa soaps (55 gms each) by cold process method as suggested by Soapcalc calculator:

Ingredients	F₁₀	F₁₁	F₁₂
Moringa leaf powder	2.5 gm (5%)	-	-
Moringa seed oil	-	2.5 ml (5%)	-
Moringa leaf extract oil	-	-	2.5 (5%)
Niacinamide (2%)	1 gm	1 gm	1gm
Castor oil	1.5 gm	1.5 gm	1.5 gm
Coconut oil	11 gm	11 gm	11 gm
Olive oil	25 gm	25 gm	25 gm
Cocoa butter	12.5 gm	12.5 gm	12.5 gm

Evaluation parameters^{3,7,8,9,11-18}

pH of soap:

10 g of soap were dissolved in 100 ml of DW to create the 10% soap solution. Volume was made up to 100 ml using DW in volumetric flask. Before pH testing was done using pH paper and a follow up pH test was performed using a prepared 10% soap solution using pH electrode (Digital control dynamics pH meter).

Foam forming ability and Foam height:

In a beaker, 1 g of soap was measured and stirred with 50 ml of DW. Additionally, the solution was poured into a graduated 100 ml measuring cylinder and stroked for 25 times. At time zero and ten minutes, the foam height was measured.

Foam retention:

1 g of soap was dissolved in 100 ml of distilled water to make 1% soap solution. Furthermore, 25 ml of the 1% soap solution was transferred into a measuring cylinder of 100 ml. By placing the hands over the aperture at one end of the cylinder, the measuring cylinder was swung ten to twelve times. Throughout the first four minutes, the volume of foam was recorded every minute.

Moisture content determination:

5 g of soap was placed in a dry and moisture free China dish. For around two hours, the China dish was heated on the water bath at 101 °C. The moisture content was found out by the below mentioned equation using the formula shown below.

Difference in weight of the soap

Moisture content = ------------------------------------ × 100

of the soap Initial weight of the soap

Determination of saponification value:

2 g of soap was added to a conical flask having 15 ml of 0.5 M KOH. For one hour, the mixture was heated at 55ºC. In order to transform the pink color into a pale-yellow color, 2 drops of phenolphthalein was added to the above mixture and titrated against 0.5 M HCl.

SV=56.1x (B-S)X N/W

Antimicrobial activity:

The organisms like S. aureus and Klebsiella were used to assess the produced soaps antimicrobial properties using the cup plate method. Using 1% of soap solution, the antibacterial activity of all soap formulations was assessed. The organisms were added to 25 ml of sterile nutrient agar, which was then transferred to plate. Once the agar was solidified, a cavity was created using sterile borer and then filled with 0.1 ml of soap solution. The zones of inhibition were estimated after plates had been incubated for 24 hours at 37ºC.

Determination of TFM⁶

5g of soap was weighed and transferred to 500ml beaker. 100ml distilled water was added and heated till the soap dissolves. 3 drops of methyl orange was added to differentiate the aqueous and fatty layer. Next, 50ml of concentrated H₂SO₄was added with constant stirring (H₂SO₄ plays a pivotal role in the separation of the aqueous and fatty layers). As H₂SO₄is added the fatty layer get separated at the supernatant. After cooling the mixtures were transferred to a separating funnel. The residues left in the beaker was washed with 30 ml of petroleum ether and later transferred to separating funnel. Vigorous shaking was done for 3 minutes to separate the aqueous layer (first). Aqueous layer was transferred to another separating funnel and 30 ml of petroleum ether was added to collect the remaining fatty matter and the process was repeated in triplicate. The fatty layer was then transferred to the first funnel. After the collection of fatty matter in a separating funnel, the funnel was washed with water. To separate the aqueous layer, to remove the excess of water filter the contents into a beaker containing a funnel with filter paper. Sodium sulphate was added to remove the water. Final wash with 30 ml of petroleum ether was done. Fatty matter along with the petroleum ether was heated to evaporate the petroleum ether for 30-40 min. Later the final volume of fatty matter was collected add 5ml of acetone was added. Lastly, the beaker was kept in hot air oven at 90-100º C for 1 hour, after cooling it was transferred to desiccator until a constant weight was reached.

RESULTS AND DISCUSSION:

1) pH determination:

The moringa soaps had a pH which was found to be in between pH 8.0 and pH 9.5. All the formulated soaps had the pH which was closer to the pH as that of commercially available moringa soaps like Naturemix nature moringa. The outcomes showed that none of the prepared soaps pH had an adverse effect on the skin. Table 4 represents the list of the formulated soaps pH values.

2) Foam height/ Foam forming ability:

The ability of all the soaps to create foam without the presence of additional surfactant specifically Sodium Lauryl Sulphate (SLS) as well as other surfactants. When compared to all the formulated soaps F1 had the least ability to create foam and F3 and F10 had the most. The foam height of various prepared soapsis mentioned in Table 4.

3) Foam Retention:

The capacity of the soap to produce long lasting foaming ability is indicated by its foam retention. The higher contact time on the skin surface is achieved by greater foam retention. The foam retention test helps us to evaluate how well soaps can deliver their qualities on surface of skin. Table 4 reports foam retention of the prepared soaps.

4) Moisture content determination:

Moisture content determination is used to find out the amount of moisture present in the soap. Lower moisture content yields higher stability soaps. The moisture content was reduced to the most possible extent with an intention to have better control on deterioration of soaps. The moisture content of prepared soaps ranges from 13.2% to 36.56%. Table 4 reports moisture content of various formulated soaps.

5) Determination of saponification value:

The saponification value is defined as the Quantity of KOH in mg required to completely saponify 1 gof fat or oil.

The saponification value of the prepared soap F10 to F12 are reported in Table 4.

6) Antimicrobial activity test.

The zone of inhibition for various formulated soaps are shown in the figure. For Klebsiella, formulation F3 (5 % moringa powder soap) and F9 (5% moringa leaf extract oil soap) demonstrated higher zones of inhibition. Higher zones of inhibition for S. aureus were seen for formulation F3 (5 % moringa powder soap) and F6 (5% moringa seed oil soap). Table 3 represents zone of inhibition of various prepared soap.

Figure 5: Zone of inhibition of Moringa powder soap (F3)

Table 3: Antimicrobial activity of moringa soaps:

Organism used

F₁

F₂

F₃

F₄

F₅

F₆

F₇

F₈

F₉

F₁₀

F₁₁

F₁₂

Klebsiella

17mm

20mm

23mm

17mm

18mm

22mm

0mm

19mm

23mm

11 mm

12 mm

S. aureus

25mm

26mm

27mm

19mm

21mm

23mm

14mm

16mm

18mm

18 mm

16 mm

Note: 3% moringa leaf oil (F7) did not produce any zone of inhibition for Klebsiella microorganism. Diffusibility is comparatively less and the clear zones are not observed in Klebsiella compared to S. aureus.

Table 4: Evaluation parameters of moringa soaps

Parameters	Melt and pour method									Cold process method
Parameters	F1	F2	F3	F4	F5	F6	F7	F8	F9	F10	F11	F12
pH	8.24	8.52	8.29	9.2	9.5	9.6	8.9	9.0	9.4	10.0	10.04	9.98
Foam height	5 ml	10 ml	15 ml	8 ml	7 ml	8 ml	7 ml	8 ml	9 ml	24 ml	12 ml	14 ml
Foam retention	20 ml	39 ml	30 ml	15 ml	25 ml	32 ml	22 ml	34 ml	35 ml	10 ml	7 ml	18 ml
Moisture content determination	25.8 %	25.0%	31.4%	26.4 %	21.9 %	18.0%	36.56 %	27.5 %	21.4 %	13.2%	15.09%	12.96%
Saponification value	-	-	-	-	-	-	-	-	-	27.73	40.59	62.74
TFM	19.6									57.8	58.7	55.9

7) Total fatty matter content (TFM):

Higher TFM ensures that soaps are least harmful to the skin and do not cause dryness, in bathing bars. The TFM of the prepared soaps are given in Table 4.

CONCLUSION:

As discussed in results and discussion, all the prepared soaps (F1 TO F9) melt and pour technique, (F10 TO F12) cold process method gave prominent results. The physical parameters like appearance, hardness, colour, odour etc. of all the soaps were satisfactory. However, the TFM of the soap prepared by using marketed base and marketed soap were comparatively less (5.36). Literature survey revealed that lower the TFM higher the chances of irritation to the skin. Henceforth this problem was mitigated by preparing soaps by cold process method using soap calc calculator. Although the process of preparation of soap is a tedious process the outcome of TFM is greater than that of cold process method. Finally, it can be concluded that it is advisable for all the cosmetologist to prepare the soap by cold process method in contrast to the use of marketed base in preparation of soap by melt and pour method.

ACKNOWLEDGEMENT:

We the authors thank Bapuji Pharmacy College, Davanagere for providing the necessary chemicals, glassware and equipment’s to perform this research work.

CONFLICT OF INTEREST:

Authors do not have any conflict of interest.

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Received on 10.10.2023 Modified on 03.01.2024

Research J. Topical and Cosmetic Sci. 2024; 15(1):20-26.

DOI: 10.52711/2321-5844.2024.00004