INTRODUCTION:

Microemulsions are a fascinating kind of thermodynamically stable colloidal systems which have received considerable interest in pharmaceutical, cosmetic, agricultural, and industrial application because of their novel physicochemical characteristics. The increasing interest in sustainability and environmentalism has redirected interest to the discovery of bio-based microemulsion systems that use renewable materials instead of petroleum-based materials.^1-3 In this regard, biological surfactants and methyl esters derived in vegetable oil have been put forth as potentials in the preparation of environment-friendly microemulsions. Neem oil which comes as a product of the Azadirachta indica seeds has been long cherished because of its pesticidal, antimicrobial as well as therapeutic qualities. Its methyl ester form has better physicochemical properties such as low viscosity, high miscibility and oxidative stability than the original oil and is a desirable renewable oil phase in microemulsion formulation. In the same way, cardphenol ethoxylate surfactants which are cashew nut shell liquid derivatives are a sustainable alternative to traditional synthetic surfactants with favourable surface-active properties and biodegradability, as well as environmental profile.^4-6

Microemulsions have experienced significant momentum in microemulsions application in agricultural formulations and especially as a delivery system of pesticides and herbicides in recent years. The traditional emulsions commonly use have shortcomings of separation of phases, large droplets, and spreading properties that are not effective at plant surfaces. Microemulsions address these limitations by offering higher wetting, greater spreading, and penetration of active ingredients into the plant cuticles due to the nanoscale size of the droplet.^7-8 The size of the droplet in microemulsions gradually reduces the size of the droplet, which raises the surface area of the platform to absorb the active ingredients and, as a result, uptake of agrochemicals. Moreover, microemulsions have the ability of solubilizing both hydrophilic and lipophilic active compounds in a single formulation thus providing versatility in delivering pesticides. Bio-based components in agricultural microemulsions are in line with the trend of the world to operate on sustainable agriculture and integrated pest management methods, as bio-based components cause a lower impact on the environment and retain crop protection activity.^9-11

Methyl esters of vegetable oil that is commonly referred to as biodiesel when utilized as a fuel source have been applied to other related uses other than energy production. Transesterification of vegetable oils with methanol produces these esters that have desirable characteristics like low viscosity, high solubility of other compounds and biodegradation. Among the various ester forms of neem oil, neem oil methyl ester still has some of the biological activity of the original compound, and it has better handling properties. The chemical stability and solubility of neem oil with surfactant systems are due to its fatty acid composition consisting of oleic acid that is abundant, and moderate levels of palmitic and linoleic acids. Numerous experimental works have already investigated different vegetable oil methyl esters in emulsions, which has proven to be a potential as a renewable alternative to mineral oils and synthetic solvents.^12-14 but the literature still lacks extensive research on microemulsion systems as a substitute of cardphenol ethoxylate surfactants on neem oil methyl esters. Although the use of bio-based microemulsions has been gaining more and more popularity, there are still multiple issues with the development and optimization of bio-based microemulsions. The naturally occurring variability of natural raw materials, possible oxidative instability of unsaturated fatty acid chains, and a lack of systematic research on the impact of surfactant blending and HLB optimization discourage the prevalence of these systems. Moreover, the connection between the parameters of the formulations used (including the surfactant-co-surfactant ratio and HLB value) and the properties of the microemulsions (i.e. droplet size, conductivity, and stability), should be studied in detail to obtain the design principles of bio-based microemulsion systems. The gaps in the knowledge are necessary in the development of commercially viable and environmentally sustainable microemulsion formulations used in agriculture and industrial practice.^15-19

This paper examines the creation and overall description of sustainable microemulsion systems based on the use of cardphenol ethoxylate surfactants and neem oil methyl ester. The experiment includes physicochemical characterization of neem oil methyl ester to assess its use as a renewable oil phase, assessment of cardphenol ethoxylates with various hydrophilic-lipophilic balance (HLB) values as bio-based surfactants and pseudo-ternary phase diagrams to map the microemulsion formation regions. The systems developed are described according to the size of the particles, pH, electrical conductivity, and density, and then the long-term stability is sought in centrifugation and storage analyses. The objectives of this investigation will be to determine the potential of cardphenol ethoxylate-neem oil methyl ester systems to be useful as alternatives in developing stable, bio-based microemulsions, which can be used in the agricultural formulations and sustainable delivery systems.

2. MATERIALS AND METHODS:

2.1. Materials:

Neem oil methyl ester, card phenol ethoxylated 9 (HLB 11.9) and card phenol ethoxylated 15 (HLB 13.9), iso propyl alcohol ethoxylated 2 moles (IPA 2) were gifted by Rossari biotech Ltd., Mumbai, India. All other reagents of analytical grade were purchased through normal suppliers and utilized in their pure state. Otherwise, all materials were of analytical reagent (AR) grade.

2.2. Methods:

2.2.1. Vegetable Oil Methyl Esters Characterization:

The physicochemical properties of the methyl esters of the neem oils were analysed using standard AOCS techniques. Measurement of acid value, saponification value, iodine value, was done to ascertain the purity levels and oxidation resistance of these samples of biodiesel.

2.2.2. Surfactants Characterization:

The physical characteristics such as the appearance, Hydroxyl value, Hydrophilic-Lipophilic balance (HLB) values were evaluated based on the previously prepared protocols of non-ionic surfactant analysis. Measures of surface tension were taken at 25 +- 1 0C using a Du Noy ring tensiometer.

2.2.3 Blend Preparation Method:

The required volume of CP-15 and CP-9 was weighed with a lot of caution following the ratios that are indicated in Table 4. In blends that do not have co-solvent (Blend 1 and Blend 3), the two surfactants were mixed directly and thoroughly with the use of a magnetic stir at room temperature (25 +- 2degC) during 15 minutes as a way of ensuring that the whole mixture homogenized. In the case of blends with a co-solvent, which is isopropyl alcohol (IPA-2), the surfactant mixture was first prepared by mixing CP-9 and CP-15 in the desired ratios, then adding IPA-2 at the required proportions of surfactant to co-solvent. The blends thus formed were stirred continuous in 20 minutes to make all the ingredients uniformly mixed. The weight-average method was used to compute the calculated hydrophilic-lipophilic balance (HLB) of each blend, using the weight-average approach, based on the weight-average of the HLB of CP-9 and CP-15. Each of the prepared blends was kept in closed containers at room temperature and within 24 hours of preparation to preserve physicochemical properties. Homogeneity of each blend was visually checked prior to use and any phase separation in case it occurred was corrected by a mild re-mixing.

2.2.4. Microemulsions Preparation and Characterization:

The neem oil methyl ester was used as the oil phase, an emulsifier to be used as the surfactant, isopropyl ethoxylate (2 mole) as the co-solvent and deionized water as the aqueous phase to compose the microemulsion system. To construct the phase diagram, the surfactant and co-solvent were premixed in different ratios (1:1, 1:1.5 and 1:0.5) to obtain one surfactant system (Smix). At ambient temperature (25 +- 1 ⁰C) pseudo-ternary phase diagrams were developed through the aqueous titration technique in order to determine the range of the microemulsion. Oil phase (neem oil methyl ester) was added in a gradual manner into the surfactant system (Smix) at a weight ratio of 0: 10-10:0 (oil: surfactant). To each oil-to-surfactant ratio, the deionized water was dropwise added to the mixture, and the mixture was gently stirred by magnet at room temperature. The titration was followed until the initial appearance of turbidity which was the transition point between the transparent microemulsion phase and either a turbid emulsion or liquid crystalline phase. Stirring was discontinued at every turbidity level, and the system was left to stabilize between 5 and 10 minutes to ensure that the phases were stable. The actual percentage of the oil, surfactant system and water at the turbidity point were noted. This was done several times when varying ratios of oil to the surfactant were used in order to map the entire phase boundary. The compositions were then estimated in weight percentage, such that the amount of the three components added to 100%. The data of the weight fraction of the titration experiments were plotted on the pseudo-ternary phase diagram, which has three axes, oil, system of surfactants (Smix), and water. The three points on the triangle diagram had 100 percent representation of each component with the opposite side depicting 0 percent of that component. The phase diagram revealed that the turbidity points defined the microemulsion region, which was the region that was thermodynamically stable and optically isotropic, and a part of the phase diagram. Phase diagrams were plotted individually at the various ratios of the surfactant to co-solvent (1: 1, 1:1.5, and 1: 0.5) to analyse how the concentration of the co-solvent changed the region of formation of the microemulsions. The edges of the microemulsion zone were drawn by plucking the turbidity points together and the space between the two points was the representation of those compositions that could form stable microemulsions spontaneously. A representative of the identified microemulsion regions was then selected and subjected to horizons of physicochemical characterization such as particle size analysis, zeta potential measurement, Electrical Conductivity, pH Measurement, appearance and stability during different storing conditions.

3. RESULTS:

3.1 Characterization of Methyl ester

Table -1: - Physicochemical parameters of methyl ester

Sr.no	Parameter	Neem oil methyl ester
1	Physical appearance	Clear Yellow liquid
2	Specific Gravity	0.873
3	Acid Value	0.12
4	Iodine Value	69
5	Saponification value	181
6	Viscosity (mm²/s at 40°C)	4.3

Table -2: - Fatty acid composition of methyl ester

Sr.no	Fatty acid Composition	Neem oil methyl ester
1	Palmitic acid	12.1
2	Stearic acid	4.2
3	Oleic acid	55.78
4	Linoleic acid	12.08

Table -3: - physicochemical parameters of Surfactant

Sr.no	Parameter	Card phenol ethoxylated 9 moles	Card phenol ethoxylated 15 moles
1	Physical appearance	Brown colour liquid	Brown colour liquid
2	pH (5% solution)	6.7	6.9
3	Hydroxyl value	64	35
4	Moisture content	0.21	0.29
5	HLB value	11.9	13.9
6	Surface tension	32.2	33.2

Table -4: - HLB Blend of Surfactant

Blend No.	CP-9 (%)	CP-15 (%)	Co-Solvent	Surfactant to Co- solvent Ratio	Calculated HLB
Blend 1	30	70	No solvent	–	13.3
Blend 2	30	70	IPA-2	1:1	13.3
Blend 3	90	10	No solvent	–	12.1
Blend 4	90	10	IPA-2	1:1	12.1
Blend 5	90	10	IPA-2	1:1.5	12.1
Blend 6	70	30	IPA-2	1:1	12.5
Blend 7	70	30	IPA-2	1:1.5	12.5
Blend 8	70	30	IPA-2	1:0.5	12.5

Figure 1 Pseudo-Ternary Phase Diagram of Blend 1 Figure 2 Pseudo-Ternary Phase Diagram of Blend 2

Figure 3 Pseudo-Ternary Phase Diagram of Blend 3 Figure 4 Pseudo-Ternary Phase Diagram of Blend 4

Figure 5 Pseudo-Ternary Phase Diagram of Blend 5 Figure 6 Pseudo-Ternary Phase Diagram of Blend 6

Figure 7 Pseudo-Ternary Phase Diagram of Blend 7 Figure 8 Pseudo-Ternary Phase Diagram of Blend 8

Table -5: - Microemulsion with different HLB Physical parameter

Sr.no	Formulation	HLB	Type of emulsion	pH	Density	Conductivity µS/cm	Particle size(nm)
1	Blend 1	13.3	W/O	7.10	0.987	100	147
2	Blend 2	13.3	W/O	7.23	0.991	90.3	123
3	Blend 3	12.1	W/O	7.14	0.981	81.4	103
4	Blend 4	12.1	W/O	7.35	0.990	89.4	174
5	Blend 5	12.1	W/O	7.40	0.983	70.3	99
6	Blend 6	12.5	W/O	7.35	0.985	81.9	105
7	Blend 7	12.5	W/O	7.54	0.987	87.4	118
8	Blend 8	12.5	W/O	7.42	0.988	88.1	114
9	Blend 1	13.3	O/W	7.43	1.010	230.4	179
10	Blend 2	13.3	O/W	7.21	1.012	270.9	163
11	Blend 3	12.1	O/W	7.32	1.011	310.5	157
12	Blend 4	12.1	O/W	7.42	1.013	298.6	142
13	Blend 5	12.1	O/W	7.31	1.011	290	121
14	Blend 6	12.5	O/W	7.32	1.014	340.6	163
15	Blend 7	12.5	O/W	7.29	1.011	341.3	174
16	Blend 8	12.5	O/W	7.12	1.012	344.2	144

Table -6: - Microemulsion Centrifuge stability data

Sr.no	Formulation	HLB	Centrifuge stability Before	Centrifuge stability After 30 days
1	Blend 1	13.3	Stable	Stable
2	Blend 2	13.3	Stable	Stable
3	Blend 3	12.1	Stable	Stable
4	Blend 4	12.1	Stable	Stable
5	Blend 5	12.1	Stable	Stable
6	Blend 6	12.5	Stable	Stable
7	Blend 7	12.5	Stable	Stable
8	Blend 8	12.5	Stable	Stable
9	Blend 1	13.3	Stable	Stable
10	Blend 2	13.3	Stable	Stable
11	Blend 3	12.1	Stable	Stable
12	Blend 4	12.1	Stable	Stable
13	Blend 5	12.1	Stable	Stable
14	Blend 6	12.5	Stable	Stable
15	Blend 7	12.5	Stable	Stable
16	Blend 8	12.5	Stable	Stable

4. DISCUSSION:

4.1 The following characterizes the Neem Oil Methyl Ester.

The physicochemical neem oil methyl ester analysis showed characteristics of making microemulsion. Specific gravity of 0.873 is within the normal range of biodiesel and methyl esters and implies that the fuel has good characteristics. A low acid value of 0.12 indicates that the microemulsion system has minimal free fatty acid and this is important in avoiding hydrolysis and allowing the microemulsion system to remain stable over an extended period of time. The iodine number of 69 means moderate unsaturation that is beneficial because it provides a balance between oxidative stability and fluidity. The value of saponification at 181 can be used to confirm the complete saponification of the product and the occurrence of methyl esters and not triglycerides. A review of the fatty acid composition revealed that oleic acid was the dominant one with 55.78 percent, palmitic acid with 12.1 and linoleic acid with 12.08. The high level of oleic acids is what makes the oil fluid and compatible with surfactant systems, which makes it an excellent choice in the form of microemulsion.

4.2 Characterization of Surfactant and Blend selection:

CP-9 and CP-15 surfactants had different HLB values of 11.9 and 13.9, respectively, which gives the flexibility in surfactant system design in various microemulsion types. The value of hydroxyl in CP-9 of 64 and CP-15 of 35 is the extent of ethoxylation and CP-9 contains fewer units of ethylene oxide and is therefore less hydrophilic. Good surface activity is exhibited by the values of surface tension of 32.2 and 33.2 mN/m of CP-9 and CP-15 respectively. The low pH levels (6.7-6.9) of the two surfactants are also beneficial towards the creation of stable systems without the need to add pH modifiers. The blending of these surfactants in diverse proportions gave HLB values of between 12.1 and 13.3 that could be used systematically to explore the impact of HLB on the formation and properties of microemulsions.

4.3 The effect of Co-Solvent on the Formation of Microemulsions was Investigated and the Results Analysed using the Following Method:

The results in comparison between Blend 1 and Blend 2, which contain HLB 13.3, though with a difference in the presence of co-solvent, show that IPA-2 plays a tremendously important role in microemulsion formation. According to the pseudo-ternary phase diagrams, it is evident that the addition of IPA-2 as co-solvent increases the microemulsion zone and thus allows the formation of stable systems easily. This is explained by the fact that the co-solvent is able to lessen interfacial tension, rise surfactant fluidity at oil-water interface and boost solubilization potential of the surfactant system. On the same note, Blends 3, 4 and 5 that have HLB 12.1 indicate that a co-solvent presence has a big effect on the phase behaviour and size of the microemulsion domain. The co-solvent is an intermediate constituent, which separates the oil and aqueous layers, thus enhancing the inflexibility of the interfacial film, and employing stability of the microemulsion.

4.4 Effect of Surfactant to Co-Solvent Ratio:

Combinations of 6, 7 and 8 with same surfactant composition (70% CP-9, 30% CP-15, HLB 12.5) but different surfactant to co-solvent ratios (1:1, 1:1.5 and 1:0.5, respectively) are an insight into the optimum co-solvent concentration. It can be seen that with a higher ratio of the co-solvent (Blend 7, 1:1.5), the microemulsion region grows bigger than under the 1:1 ratio (Blend 6) indicating greater interfacial flexibility and better solubilization. On the other hand, a reduction in the co-solvent ratio (Blend 8, 1:0.5) will cause the size of the microemulsion region to be smaller, which means that the lack of co-solvent will cause the interfacial film to be more rigid and able to accommodate only a limited range of oil-water compositions. These data indicate that there is an ideal concentration of co-solvent that allows maximizing the creation of microemulsion, economic, and practical aspects.

4.5 Physicochemical Properties of W/O and O/W Microemulsions:

The physicochemical characterisation of both W/O and O/W microemulsions indicate the presence of differences in the characterisation. W/O microemulsions had much lower electrical conductivity (70.3-100 uS/cm) than O/W microemulsions (230.4-344.2 uS/cm), which is reasonable because the continuous phase in W/O microemulsions is oil, which is not conductive. The analysis of particle size indicates that the W/O microemulsions tend to have a smaller or similar particle size (99-174 nm) to the O/W systems (121-179 nm), with Blend 5 giving the smallest particle sizes in both groups. The density of the W/O microemulsions (0.981-0.991 g/cm3) is smaller than that of O/W microemulsions (1.010-1.014 g/cm3) as the density of water is higher, being the continuous phase in O/W systems. All the formulations had a near-neutral (7.10-7.54) pH, which is good in agricultural and cosmetic use.

4.6 Optimisation of Formulation Parameters:

Blend 5 (90% CP-9, 10% CP-15 with the same amount of IPA-2 in 1:1 proportion) was the best formulation in terms of its ability to perform well with both W/O (99 nm) and O/W (121 nm) microemulsions, and it was also found to have the smallest particle sizes. This specific system seems to have the HLB value of 12.1 which is the best one to have in terms of giving the right balance between the hydrophilic and the lipophilic character to create tiny and stable droplets. A higher concentration of CP-9 (low HLB surfactant) in Blend 5 might also help the alternative to pack on the oil-water interface and achieve a smaller droplet size. Moreover, the values of conductivity of Blend 5 were moderate (70.3 uS/cm W/O and 290 uS/cm O/W), indicating a clear phase structure. The outstanding centrifuge stability of all formulations beyond 30 days signifies thermodynamic stability and coalescence resistance, which is crucial in the field of practice.

4.7 Stability Considerations:

The centrifuge stability results indicate that the entire sixteen formulations were stable either at the beginning or after 30 days during storage hence showing good long-term stability. This stability is exceptional and can be explained by the combination of the synergistic effect of the co-solvent with the surfactant, which forms a strong interfacial film that stops the coalescence of droplets. The fact that no phase separation or destabilization occurs with time, indicates that such microemulsions are thermodynamically, but not kinetically stable which is a major strength in commercial application. The stability in the presence of various HLB values and surfactant to co-solvent ratios show that cardphenol ethoxylate system is both versatile and robust and can be used to stabilize the system under different compositional conditions.

5. CONCLUSION

The research was very effective in creating and defining stable microemulsions of neem oil methyl ester with the help of cardphenol ethoxylate surfactants of different HLB. The overall study of eight various surfactant mixtures by constructing pseudo-ternary phase diagrams and physicochemical characterization dwells useful information on optimization of the microemulsion formulation. The most significant findings and conclusions are the following: The neem oil methyl ester had good physicochemical characteristics such as low acid value, moderate iodine value, and high content of the oleic acid and therefore formed a good microemulsion formulation. The need to fine-tune the HLB value by mixing led to the use of cardphenol ethoxylate surfactants (with varying degrees of ethoxylation of 9 and 15) to obtain both W/O and O/W microemulsions. The co-solvent (isopropyl alcohol ethoxylated 2 moles) addition as a critical component in expanding the microemulsion region in pseudo-ternary phase diagrams showed that this co-solvent was a crucial component in reducing the interfacial tension, as well as enhanced flexibility of the system. Surfactant to co-solvent ratio was observed to have an effect on the size of the microemulsion region, and in general, large co-solvent ratios favored an increase in the size of the microemulsion region. Of all the formulations that were tested, Blend 5 (90% CP-9, 10% CP-15, and at the O:W ratio of the 1:1 level, HLB 12.1) was the best formulation as it generated the smallest particle sizes in both the W/O (99 nm) and O/W (121 nm) microemulsions. This is an indication that the lower value of HLB with higher content of CP-9 gives better control of droplet size. All formulations were found to be thermodynamically and practically stable (lasting more than 30 days centrifuge) to be used. Unique physicochemical characteristics of W/O and O/W microemulsions, especially conductivity and density, make it possible to select them depending on the peculiarities of application. The pH of all formulations is very close to neutral, which makes it suitable to use in agriculture, cosmetic and pharmaceutical without any need to adjust the pH. In this study, cardphenol ethoxylate-based surfactant systems are shown to have the potential of producing bio-based microemulsions with stable particle sizes and properties. The results give the basis of the future advancement of neem oil methyl ester-based preparations in the delivery of agrochemicals, biopesticide preparations, and sustainable emulsion technology. It may be taken a step ahead in future work, to consider the addition of active ingredients, assess the biological efficacy, and scale-up analysis of commercial production of these promising bio-based microemulsion systems.

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Received on 15.11.2025 Revised on 09.02.2026

Accepted on 11.04.2026 Published on 24.06.2026

Available online from June 30, 2026

International Journal of Technology. 2026; 16(1):35-42.

DOI: 10.52711/2231-3915.2026.00004

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