Optimization of Tween 80 and Lecithin in Diclofenac Sodium Nanoemulsion

Viddy Agustian Rosyidi, Ulfi Mawadatur Rohmah, Lina Winarti, Lusia Oktora Ruma Kumala
Sari, Lidya Ameliana, Kuni Zu’aimah, Eka Deddy Irawan, Dwi Nurahmanto, Budipratiwi
Wisudyaningsih
Faculty of Pharmacy, University of Jember, East Java, Indonesia

Abstract
Diclofenac sodium had some of the deficiencies, the side effects of causing disturbances in the
digestive tract, having low bioavailability and solubility. Development of a system of transdermal
nanoemulsion type oil-in-water (o/w) is able to be used as an alternative to overcome the deficiency
of diclofenac sodium. One important component in nanoemulsion is the surfactant. By optimizing
the number of tween 80 as hydrophilic surfactant and lecithin as lipophilic surfactant, it was
expected to form a good system of diclofenac sodium nanoemulsion. This research was conducted to
determine the effect of the number of tween 80, lecithin, and their interactions on entrapment
efficiency, transmittance, and pH of nanoemulsion system using factorial design method. The
interaction of tween 80 and lecithin gave a significant effect on the decrease of entrapment
efficiency. The amount of lecithin significantly increased transmittance, while the interaction of two
factors significantly reduced transmittance. An increase in the number of tween 80 also increased pH
significantly. The optimum formula composition of diclofenac sodium nanoemulsion covered the
combination of 1.5 mL of tween 80 and 1.5 mL of lecithin with the value prediction of entrapment
efficiency as much as 90.084%, 99.424% of transmittance value and 6.327 as its pH. The optimum
formula of diclofenac sodium nanoemulsion was in pale yellow, and clear appearance, 1.0467 g/mL
of density, 2.6213 mPas of viscosity. O/w-type nanoemulsion had droplet size, polydispersity index,
and zeta potential of 7.7 nm, 0.378 and -2.6 mV.
Keywords: Diclofenac sodium, nanoemulsion, optimization, tween 80, lecithin

1. Introduction
The prevalence of osteoarthritis (OA) joint disease according to Basic Health Research in
2018 was 7.3%. Nonsteroidal Anti Inflammatory Drugs (NSAIDs) is main pharmacotherapy
recommended for OA1. Diclofenac sodium is widely used in oral dosage, but able to provide several
problems if given orally, the side effects causes disturbances in the digestive tract and low
bioavailability, only 50% can reach systemic circulation2,3. The development of transdermal delivery
system can be used as an alternative to overcome the shortcomings of diclofenac sodium. This
research selected nanoemulsion type oil-in-water (o/w) as the transdermal delivery system of
diclofenac sodium. The selection of nanoemulsion systems can be used as an alternative to increase
permeation, delivery system and targeting drugs that are difficult to dissolve like diclofenac sodium4.
One important component in nanoemulsion is surfactant. The function of surfactant is
reducing interfacial tension between water phase and oil phase so that small and stable droplets will
form. Single surfactant is not enough to form stable nanoemulsion5. The combination of hydrophilic
and lipophilic surfactants is recommended to be used as a combination of surfactants. The properties
of different surfactants have synergistic effect to reduce interfacial tension6.
Tween 80 with HLB value of 15 was used as hydrophilic surfactant and lecithin with HLB
value of 4 was taken as a hydrophobic surfactant. The results of Samiun's research (2015) showed
that the combination of tween 80 and lecithin as a surfactant had an important role in determining the
size of the aripripazole nanoemulsion droplet7. The small size of nanoemulsion droplet makes
nanoemulsion more stable8. By optimizing the number of tween 80 and lecithin, it was expected to
form a good nano diculcate sodiumemulsion system.
This research was aimed at finding out the effect of the number of tween 80, lecithin, and
their interactions with entrapment efficiency, transmittance, and pH of the nanoemulsion system. The
factorial design method was used with two factors (the number of tween 80 and lecithin) and two
levels (high and low). The responses observed were transmittance values, entrapment effeciency, and
pH. Analysis of factor and subsequent responses produced the optimum formula.

2. Method
2.1. Tools
The instrumentation were used, such as standard laboratory glass ware, analytical balance
(Adventure Ohaus), hotplate and magnetic stirer (Ika c-mag HS7), centrifuge, pH meter (Denver),
spektrophotometer UV-Vis (Genesys 10S), CAMAG TLC Scanner 3 dan CATS evaluation software,
TLC Silica gel 60 F254 (Merck), (Particel Size Analyzer) HORIBA, picnometer, viscometer
Ostwald.

2.2. Materials
Diclofenac Sodium (Cheng Fong Chemical Co. Ltd., Taiwan), fennel oil (PT. Brataco
Chemica), VCO (Agricultural Product Processing Laboratory of Jember State Polytechnic), olive oil
(Makmur Sejati), tween 80 (Makmur Sejati), lecithin ( PT. Bratachem), span 20 (Makmur Sejati),
span 80 (Makmur Sejati), aquadestilata (Makmur Sejati), technical methanol (True Prosperity),
toluene (J.T. Baker), glacial acetic acid (Merck, USA), ethyl acetate ( Merck, USA).

2.3. Oil Determination

The solubility of diclofenac sodium in various oils (fennel oil, olive oil, virgin coconut oil)
was determined by dissolving 100 mg of diclofenac sodium in 2 ml of each selected oil. The
dissolution process was done by using a magnetic stirrer with 200 rpm for 10 minutes. Determination
of solubility was done qualitatively, which was by looking whether or not there was sediment at the
bottom of the vial. Fennel oil dissolved diclofenac sodium perfectly. Meanwhile, olive oil and VCO,
diclofenac sodium remained in the form of suspension, so that the fennel oil was determined as a
component of diclofenac sodium carrier.

2.4. Surfactant Determination

Tween 80 was chosen as a hydrophilic surfactant with HLB value of 15. The candidates for
hydrophobic surfactants included lecithin (HLB=4), span 80 (HLB=4.3), and span 20 (HLB=8.6).
The determination of surfactant combination was done by titration. Tween 80 as a hydrophilic
surfactant and other hydrophobic surfactants (lecithin, span 80, and span 20) were pipetted and
mixed with volume ratio of 1:1 8. Each mixture of the surfactant was then added with 2 mL of fennel
oil and 3 mL of aquadest and was stirred until it became homogeneous. The selected surfactant
combination was one that reduced water and oil interface stress8.Tween 80 and lecithin were chosen
as a combination of surfactants as they were able to reduce the oil and water interface tension
marked by the formation of clear emulsions. While the combination of tween 80 with span 80 and
span 20 did not form a clear emulsion.
The design of the formula was based on optimization design through factorial design method.
Four formulas were used in the design of the formula with two factors and two levels. Each formula
was replicated three times. The composition of selected nanoemulsion formulations are shown on
Table 1.
Table 1. The formula of Diclofenac Sodium Nanoemulsion
Formula
Composition
1 A B AB
Diclofenac sodium 0,1 g 0,1 g 0,1 g 0,1 g
Fennel oil 2 mL 2 mL 2 mL 2 mL
Tween 80 1,5 mL 3,5 mL 1,5 mL 3,5 mL
Lecithin 0,25mL 0,25 mL 1,5 mL 1,5 mL
Aquadest 3 mL 3 mL 3 mL 3 mL

2.5. The Preparation of Nanoemulsion

The preparation method for diclofenac sodium nanoemulsion used was the method of
spontaneous emulsification. It started by dissolving diclofenac sodium into fennel oil. Tween 80 and
lecithin whose amounts are appropriate to each formula were mixed by using a magnetic stirrer at
250 rpm for 5 minutes. The mixture of tween 80 and lecithin was added to the solution of diclofenac
sodium in oil, stirred with a magnetic stirrer for 5 minutes at 250 rpm. After the mixture became
homogeneous, aquadest was added gradually by using a dropper pipette, stirred with a magnetic
stirrer for 5 minutes at 250 rpm.

2.6. Response Evaluation

Entrapment Efficiency
Entrapment efficiency measurement was done by using TLC densitometry method. The mobile
phase chosen covered toluene: ethyl acetate: glacial acetic acid with the volume ratio of 60:40:1. The
stationary phase used was TLC gel. The analysis was carried out at the wavelength of 282 nm. The
solvent used was methanol9.
Standard solution was made in the concentration series of 50, 100,150, 300, 450 and 500 ppm.
Each standard solution was put in the bottle as much as 2 µL on the TLC silica gel plate. The plot
between the concentration of the standard solution and the area of the densitogram formed a standard
curve to calculate the levels of diclofenac sodium in the sample.
The preparation of sample was begun by adding fennel oil to diclofenac sodium nanoemultion to
make nanoemulsion turbid. The turbid nanoemulsion was then centrifuged for 1 hour at 6,000 rpm.
The oil phase which had been separated was then diluted with methanol and its level was also
determined. Readable level of diclofenac sodium was shown on equation 1 to know the value of
entrapment efficiency. The entrapment efficiency equation of nanoemulsion on Equation 1 is as
follows.
% EE = (A2/A1) x100% ... (1)
A2 referred to readable level of diclofenac sodium and A1 was the amount of diclofenac sodium
included in the formulation10.
Transmittance
Transmittance measurement was carried out by using a UV-Vis spectrophotometer with 650 nm
as its wavelength. The comparison sheet used was aquadest (Hajjar dkk., 2018).
pH measurenment
PH measurement of diclofenac sodium nanoemulsion was carried out by using a digital pH
meter.

2.7. The Determination of Optimum Formula

The optimum formula was determined by using Design Expert 11.0.0 software. The data
from the evaluation of diclofenac sodium nanoemulsion which were used as a response covering
entrapment efficiency, transmittance, and pH in which they were used to find the coefficients of b0,
b1, b2, and b12 and met the equation Y=b0 + b1XA + b1XB + b12XAXB.
The transmittance response was considered important as it was related to the droplet size, the
importance score was given the highest score of +5 (+++++), the goal was set to be maximum. The
optimum formula was expected to have the highest entrapment efficiency so that the goal was set to
be maximum and the importance score was set to +5 (+++++). The optimum nanoemulsion was
expected to have a pH on the pH range of the skin so that the goal was set in range 6,0-7,0 and the
importance score was set to +3 (+++). The software displayed several solutions with the variations in
the composition of the number of tween 80 and lecithin which provided optimum response. The
optimum formula solution for diclofenac sodium nanoemulsion was chosen as it gave the highest
desirability value.

2.8. Optimum Formula Verification

The verification was conducted by using One Sample T-test with a confidence level of 95%.
The optimum formula of diclofenac sodium was evaluated to obtain the transmittance value,
entrapment effeciency, and pH. The results of experiment and prediction were considered to be
significantly different if they provided a significance value of <0.05. On the other hand, it was
considered not significantly different if the significance value was> 0.05.

2.9. Optimum Formula Characterization

Organoleptic Observation
The optimum formula of diclofenac sodium nanoemulsion was observed for its color, smell, and
degree of clarity.
Type Weight Test
The weight of diclofenac sodium nanoemulsion was measured by using a picnometer.
Viscosity Test
The viscosity of diclofenac sodium nanoemulsion was measured by using Ostwald
viscometer ostwald.
Nanoemulsion Type Test
Nanoemulsion type was tested through dye test. Dye test was done by dripping methylene
blue over the nanoemulsion which had been placed on the glass preparation then observed under an
optical microscope. Methylene blue was hydrophilic, so it will be dispersed in the water phase. If
nanoemulsion had the type m/a, then the blue dye diffused evenly at the external phase. On the
contrary, if nanoemulsion was in m/a type, blue dye appeared as droplets in the internal phase11.
Droplet Diameter Test, Polydispersity Index, and Zeta Potential
The average diameter of nanoemulsion droplets was measured by using particle size analyzer
an dynamic light scattering. The 10 mL nanoemulsion was put into the cuvette then placed in the
holder and analyzed by the instrument. The measurement was complete after the monitor showed the
average droplet diameter, polydispersity index, and zeta potential.
3. Results
3.1. Excipient Selection and Diclofenac Sodium Nanoemulsion Formulation
The oil candidates that were used as the oil phase including fennel, olive and VCO oil. The
experimental results showed that diclofenac sodium was only dissolve in fennel oil until a clear oil
solution was formed. While in olive oil and VCO, diclofenac sodium was not be able to dissolve and
settle at the bottom of the vial.
Tween 80 was chosen as a hydrophilic surfactant. Tween 80 was combined with three
hydrophobic surfactant candidates including lecithin, span 80, and span 20. The selected surfactant
combination was the one that was capable of dispersing the oil phase and water phase. The results of
the experiment showed that only a combination of tween 80 and lecithin dispersed fennel oil and
aquadest to form a clear and stable nanoemulsion, so that lecithin was chosen as a hydrophobic
surfactant. The results of the diclofenac sodium nanoemulsion formulation are shown in the
following Figure 1
a b

c d

Figure 1. The Results of making four diclofenac sodium nanoemulsion formula, a) Formula 1;
b) formula A;c) formula B; d) formula AB
3.2. The Result of Response Evaluation
The results of the evaluation of entrapment efficiency, transmittance, and pH responses of the
four formulas of diclofenac sodium nanoemulsion are presented on Table 1.
Table 1. The response evaluation results of four diclofenac sodium nanoemulsion formulas
Response
Formula
Entrapment Efficiency ± SD Transmittance ± SD pH ± SD
1 58,479 ± 6,114 97,022 ± 0,240 6,347 ± 0,070
A 75,098 ± 4,040 99,039 ± 0,417 6,45 ± 0,056
B 90,085 ± 7,287 99,424 ± 0,427 6,327 ± 0,091
AB 59,154 ± 7,804 98,448 ± 0,746 6,583 ± 0,031
3.3. Factorial Response Analysis Design
Entrapment Efficiency
The analysis result of ANOVA on the effect of factor A (number of tween 80) and factor B
(number of lecithin) on entrapment efficiency showed that tween 80, lecithin, and tween 80 and
lecithin interaction gave a significant effect on entrapment efficiency response. The effect of factor A
and factor B on diclofenac sodium nanoemulsion was illustrated in the form of coded factor in which
it produced Equation 4.
Final Equation in Terms of Coded Factors:
Entrapment Efficiency = +70,70 -3,58*A +3,92*B -11,89* AB........(2)
Note: A= Tween 80; B= Lecithin; AB = interaction of two factors
The relationship between Tween 80 and lecithin with the response of entrapment efficiency
was also represented with contour plot. Contour plot of Tween 80 and lecithin combination toward
entrapment efficiency is shown on Figure 2.

Figure 2. Contour plot between Tween 80 and Lecithin relationship toward entrapment efficiency
Transmittance
The statistical result of ANOVA showed that lecithin and the interaction of Tween 80 and
lecithin gave significant effect to the nanoemulsion transmittance of diclofenac sodium. While, the
amount of Tween 80 had no significant effects to the transmittance.
The effect of Tween 80 and lecithin amount and its interaction toward nanoemulsion
transmittance of diclofenac sodium is represented on Equation 3.
Final Equation in Terms of Coded Factors :
Transmittance = + 98,48 + 0,2602* A + 0,453*B - 0,7480*AB…….....................(3)
Note: A= Tween 80; B= Lecithin; AB = the interaction of two factors
The relationship between tween 80 and lecithin with the response of transmittance was also
represented with contour plot. Contour plot of Tween 80 and lecithin combination toward entrapment
efficiency is shown on Figure 3.
 Figure 3. Contour plot of the relationship between tween 80 and lecithin toward the transmittance

pH
The statistical result of ANOVA showed that the factor of tween 80 and interaction of tween
80 and lecithin gave significant effect toward pH of diclofenac sodium nanoemulsion. The analysis
result of the effect of factor A (the amount of tween 80) and factor B (the amount of lecithin) toward
the pH of diclofenac sodium nanoemulsion generated coded factor and actual factor equations as
follows:
Final Equation in Terms of Coded Factors :
pH = +6,43 +0,0900* A +0,0283* B + 0,0383* AB…..............................(4)
Note: A= Tween 80; B=Lecithin; AB = interaction of two factors
The figure of contour plot of the relationship of Tween 80 and lecithin toward pH can be seen
on Figure 4.

Figure 4. Contour plot of the relationship of Tween 80 and Lecithin toward pH

Optimum Formula Determination
The expected optimum formula was the one which had the greatest drug trapping ability
(entrapment efficiency), the smallest droplet size seen from the largest transmittance, and the pH of
the preparation met the criteria for the pH range of the skin. Based on the criteria that had been
established, overlay plot which met the entire criterion was obtained. Based on the result of the
optimization, the chosen optimum formula consisted of the combination of Tween 80 (1,5 mL) and
lecithin (1,5 mL) with desirability value of 0.856. The optimum formula was predicted to have
entrapment efficiency value of 90,085%, transmittance value of 99,424 %, and pH 6,327.

Optimum Formula Verification

Based on the result of One Sample T-test, it showed that the data of entrapment efficiency,
transmittance, and pH responses had value of Sig. (2-tailed) > 0.05, which meant that there was no
significant difference between the result of the experiment and the prediction. The prediction of
factorial deign was the same with the result of the real experiment. The result of verification of
optimum formula of diclofenac sodium nanoemulsion can be seen on Table 3.

Table 2. The verification results of optimum formula of nanoemulsion diclofenac sodium

Response Prediction Result Experiment Result Sig. value

Entrapment Efficiency 58,479 ± 6,114 97,022 ± 0,240 6,347 ± 0,070
Transmittance 75,098 ± 4,040 99,039 ± 0,417 6,45 ± 0,056
pH 90,085 ± 7,287 99,424 ± 0,427 6,327 ± 0,091

Characterization of Optimum Formula

Optimum diclofenac sodium nanoemulsion produced pale yellow color, had distinctive smell
of fennel oil and had clear appearance. The densityof diclofenac sodium nanoemulsion was 1.0467
g/mL, good viscosity characteristics in which the viscosity value was 2.6213 mPas. After being
observed under the microscope, the color of methylene blue was distributed evenly in the water
phase, while the oil droplets looked like clear and shiny granules. It was ascertained that diclofenac
sodium nanoemulsion had type of oil in water (o/w). The measurement results showed that
diclofenac sodium nanoemulsion had a droplet size of 7.7 nm, polidispersity index value was 0.378,
zeta potential value was -2.6 mV. The optimum formula of diclofenac sodium nanoemulsion can be
seen in the Figure 5.

Figure 5. Optimum formula of diclofenac sodium nanoemulsion

4. Discussion
This research used a combination of surfactants covering hydrophilic and hydrophobic
surfactants. The use of a combination of hydrophilic and hydrophobic surfactants improved the
stability of nanoemulsion by reducing interfacial tension and reduced its use in stabilizing
nanoemulsions12. Tween 80 was chosen as a hydrophilic surfactant. Tween 80 was combined with
three hydrophobic surfactant candidates including lecithin, span 80, and span 20. The choice of
surfactant combination was considered important and crucial as it combined the hydrophilic and
hydrophobic surfactants that were appropriate and made it easier to form stable nanoemulsions.
The entrapment efficiency response showed how much diclofenac sodium had been trapped
in nanoemulsion system. From the evaluation result, the four formulas of diclofenac sodium
nanoemulsion successfully trapped >50% of the drugs within the formula. Coded factor equation
showed that Tween 80 gave negative effect toward entrapment efficiency with coefficient value of -
3.58, which meant that by increasing the amount of Tween 80, entrapment efficiency of diclofenac
sodium nanoemulsion was decreased. Whereas, lecithin coefficient value was positive that was
+3.92, which meant that by increasing the amount of lecithin, entrapment efficiency of diclofenac
sodium nanoemulsion was decreased. The coefficient value of interaction between Tween 80 and
lecithin was negative (-11.89). It meant that the interaction between Tween 80 and lecithin decreased
entrapment efficiency.
Surfactant acted as the bridge to disperse oil phase and water phase. Surfactant worked
through being adsorption by the interfacial of oil/water. If the use of surfactant was excessive, the
amount would be too saturated in which caused the oil/water interfacial could not adsorb the whole
amount of surfactant13. Tween 80 had hydrophobic part so it adsorbed diclofenac sodium which was
hydrophobic as well. Tween 80 adsorbed diclofenac sodium to its hydrocarbon chain. Tween 80 that
was not adsorbed formed micelles in water phase by carrying diclofenac sodium partially, as a result
the amount of diclofenac sodium in oil phase decreased along with the increase of formed micelles.
It causes the decrease of entrapment efficiency13.
Lecithin is able to prevent sodium diclofenac partition from oil phase to water phase by
increasing the drug solubility in oil phase and absorbing diclofenac sodium in phospholipid part of
lecithin in oil phase14. Lecithin performance was beneficial because it increased entrapment
efficicency. If the amount of lecithin improved, all droplets of oil phase which absorbed diclofenac
sodium would be covered.
From the Contour plot figure, it can be seen that the combination of high level of Tween 80
and high level of lecithin, and the combination of low level of Tween 80 and low level of lecithin
decreased entrapment efficiency. The high value of entrapment efficiency was obtained by
combining the high level of Tween 80 and low level of lecithin or otherwise.
Transmittance values indicated the clarity of nanoemulsion. Transmittance values that was
close to 100% showed clear, transparent nanoemulsions and droplet sizes estimated in the nanometer
range. Small droplet size was good for drug release as it had a large surface area15. The four
nanoemulsion sodium diclofenac formulas had transmittance values > 97%, so it was estimated that
droplet size was in the nanometer range. Coded factor equation showed that the coefficient value of
Tween 80 and lecithin was positive, in which they were +0,2602 and +0,453. The value of
transmittance would increase if Tween 80 and lecithin amounts also increased. The amount of the
used surfactant determined the size of nanoemulsion droplet. The higher the surfactant concentration
was, the smaller the size of nanoemulsion droplet would be. The result of this research was in line
with the research conducted by Arbain et al. (2018) that the increase of Tween 80 and lecithin
amounts reduced the size of quercetin nanoemulsion droplet16. It happened because with an increase
in the amount of surfactant, the amount of available surfactant to cover the droplet provided more.
Therefore, the sufficient amount of surfactant covered oil droplet to form smaller droplet17.
The interaction value between Tween 80 and lecithin was negative as much as -0.7480, which
meant that the interaction between Tween 80 and lecithin reduced transmittance of diclofenac
sodium nanoemulsion. Droplet size can be attributed to viscosity. Viscocity of nanoemulsion, one of
them, depended on the surfactant18. Tween 80 and lecithin had thick consistency. The increase
amount of Tween 80 and lecithin increased the viscosity of diclofenac sodium nanoemulsion. High
viscosity increased flow resistance and limited the process of droplet collision so it formed bigger
droplet, as the result the value of transmittance decline.
The relationship between the amount of Tween 80, lecithin and its interaction was also
represented in the form of contour plot. The big value of transmittance was obtained by combining
large amount (high level) of Tween 80 and small amount (low level) of lecithin, or otherwise. While,
by combining low level of Tween 80 and lecithin as well as high level of Tween 80 and lecithin, the
value of transmittance decreased.
pH parameters were very important for transdermal preparations because they were related to
skin acceptability. The desired pH value was the one close to the pH of the skin 4-619. The result of
the test showed that four formulas prepared had pH which met the criteria. The coded factor
equations showed that the factor of Tween 80 amount in coded factor equation had the coefficient of
+0,0900, the coded factor of lecithin amount had the coefficient of +0,0283, and the interaction
between Tween 80 and lecithin had coefficient value of +0,0383.
The coded factor equation showed relative impact of incline or decline of each factor and its
interaction toward the response. The increase of Tween 80 and lecithin amounts increased pH.
Tween 80 was alkaline surfactant, so by increasing the amount of its usage, it increased the pH of
nanoemulsion as well. Likewise with lecithin which pH was neutral (tend to be alkaline) that was
7.13. By increasing the amount of Tween 80 and lecithin, it increased pH of nanoemulsion
preparation.
Based on the result of One Sample T-test, it showed that the data of entrapment efficiency,
transmittance, and pH responses had value of Sig. (2-tailed) > 0.05, which meant that there was no
significant difference between the result of the experiment and the prediction. The prediction of
factorial deign was the same with the result of the real experiment. Three models of this equation
was said as valid and gave representation of the effect of tween 80, lecithin, and its interaction
toward the response of entrapment efficiency, transmittance.
Optimum formula diclofenac sodium nanoemulsion had type of oil in water (o/w). This type
was desired type because its usage was more comfortable and easier to rinse after being applied to
the skin. Nanoemulsion o/w type haf lower viscosity than w/o, so that the drug released was expected
to be faster20. The measurement result of polidispersity index value of nanoemulsion diclofenac
sodium was 0.378. The expected participatory index value was <0.2521. However, the polydispersity
index value <0.5 was still said to be monodispers or had a homogeneous and acceptable size
distribution22.
Zeta potential indicated the surface load of nanoemulsion droplet, the desired zeta value was
greater than +30 mV or less than -30 mV21. The zeta potential value of the optimum formula of
diclofenac sodium nanoemulsion was -2.6 mV. This potential zeta value did not meet the desired
zeta potential characteristics. The zeta potential value was usually associated with the stability of
nanoemulsion, but it was not always the main basis for describing the stability of nanoemulsion18.
Diclofenac sodium nanoemulsion with zeta potential value of -2.6 mV was stabilized by steric
obstruction in a non-ionic surfactant polymer chain (tween 80). In the nanoemulsion system, o/w that
contained non-ionic surfactants, the surfactants formed a film layer on the surface of the droplet. The
film layer prevented droplet from being combined in the dispersing medium. The phenomenon of
preventing aggregation due to this film layer was known as steric obstruction23. The zeta potential
value depended on the composition of the constituent and the dispersing medium24. Zeta potential of
diclofenac sodium nanoemulsion was negative and close to 0 (not charged) was possible since the
use of lecithin combination as a hydrophobic surfactant and tween 80 as a hydrophilic surfactant.
Tween 80 was a non-ionic surfactant which has no ion charged, while lecithin was composed of
negative charged phospholipids.
5. Conclusion
The interaction of tween 80 and lecithin had a significant effect on the decrease of
entrapment efficiency. An increase in the amount of lecithin increased transmittance significantly,
while the interaction of two factors decreased transmittance significantly. An increase in the amount
of tween 80 increased pH significantly. The composition of optimum formula on diclofenac sodium
nanoemulsion covered the combination of 1.5 mL of tween 80 and 1.5 mL of lecithin with the
prediction of entrapment efficiency value was 90.084%, transmittance value was 99.424% and pH
was 6.327. The optimum formula of diclofenac sodium nanoemulsion had pale yellow as its color,
distinctive smell of fennel oil, and clear appearance, specific gravity as much as 1.0467 g / mL,
viscosity as much as 2.6213 mPas. Nanoemulsion in o/w type had droplet size, polydispersity index
and zeta potential of 7.7 nm, 0.378 and -2.6 mV.

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