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org/jced Article

Effect of a Nonionic Surfactant on the Pseudoternary Phase Diagram

and Stability of Microemulsion
Chintya Gunarto, Yi-Hsu Ju, Jindrayani Nyoo Putro, Phuong Lan Tran-Nguyen, Felycia Edi Soetaredjo,
Shella Permatasari Santoso, Aning Ayucitra, Artik Elisa Angkawijaya,* and Suryadi Ismadji*
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ABSTRACT: Microemulsion (ME) is a micron-sized droplet that consists of oil and water, with addition of a surfactant and
cosurfactant. Recently, ME is widely used in biomedical application for proper drug delivery in the human body. Castor oil as the oil
phase, Tween 80 or Tween 20 as the surfactant, glycerol or ethanol as the cosurfactant, and DI water as the water phase were used
for ME preparation in this study. The effect of the surfactant-to-cosurfactant ratio on the pseudoternary phase diagram was
investigated. The as-synthesized ME with the composition of 5 wt.% castor oil, 85 wt.% surfactant mixture, and 10 wt.% water was
characterized based on its particle size, polydispersity index, and zeta potential. From that composition, the largest ME was attained
at an Smix 2 weight ratio of tween 80 to ethanol. Astaxanthin as lipophilic drug substance was used as the model drug for the ME
encapsulation study. The thermal and storage analysis test of ME and astaxanthin-loaded ME demonstrated the stability of the as-
synthesized ME and its analogous drug-loaded form.

1. INTRODUCTION of oil spillage from a polluted area to pharmaceutical

Astaxanthin (ASX, C40H52O4, Figure 1) is one of the most application where ME acts as drug carrier.10 Aside from
potent antioxidants, ubiquitously found in red-pigmented promoting absorption of lipophilic drugs, adaptation of ME in
plants, bacteria, marine animals such as shrimp, salmon, and medical usage reportedly may protect the drug against
crab, flamingos, and microalgae.1,2 ASX is classified as a polar oxidation and enzymatic degradation.11,12
xanthophyll that functions as the cells’ and tissues’ protectant Addition of a surfactant and/or cosurfactant in ME
from the damaging effect of free radicals and single oxygen, reportedly can decrease surface tension of the oil−water
thus it is able to inhibit cancers and tumors.3 In addition, it interface, stabilize the connection between lipophilic molecules
also could be used to prevent hypertension and cardiovascular and the aqueous phase, and enhance the solubility of
diseases, as a stabilizer for beverages, and as anti-inflammatory hydrophobic drugs.13,14 Thus, selection of the surfactant and
substance.4−6 Despite its various health benefits, oral cosurfactant is crucial for proper ME formation. The
administration of ASX is impractical due to its eminent hydrophile−lypophile balance (HLB) of different types of
lipophilic nature.7 Thus, it is necessary to combine ASX with surfactants is listed in Table 1. A study by Sharma indicated
lipid for its optimal absorption and functionalization.8
Microemulsion (ME) is a lipid-based formulation that Received: April 16, 2020
combines two immiscible liquids to form thermodynamically Accepted: July 22, 2020
stable droplets with sizes between 100 and 300 nm. The basis
in forming ME is to reduce interfacial free energy to a very low
value, thus formation of ME is spontaneous.9 The application
of ME may vary from the environmental sector by restoration

© XXXX American Chemical Society https://dx.doi.org/10.1021/acs.jced.0c00341

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Figure 1. Molecular structure of (a) astaxanthin, (b) Tween 20, and (c) Tween 80.

Table 1. HLB Value of Different Surfactants is 5000 mg/kg and for Triton X is >1000 mg/kg in rat
(oral).17,18 Therefore, T80 and T20 were selected in this study.
type surfactants HLB ref
For the later applications, oleic acid, salicylic acid, and
anionic sodium dodecyl sulfate 40 43 vegetable oils from soybean, sunflower, peanut, castor, and
sodium stearoyl-2-lactylate 2−3 16 olive were commonly used as the oil phase.19−21 Previous
cationic didecyldimethylammonium chloride n.d.a 44 studies using nonionic surfactants demonstrated the superi-
cetyltrimethylammonium bromide 10 45
ority of ME as a carrier for lipophilic drugs, such as metformin
nonionic T80 15.9 17
hydrochloride, tamoxifen citrate, and probucol. These studies
T20 16.7 17
either used T20 or T80 as the surfactant, with a number of
Triton X 13.4 18
a
cosurfactants such as Cremophor EL, glycerin, Span 80,
n.d. = Not determined. ethanol, and PEG-400. Encapsulation of these lipophilic drugs
was postulated to decelerate the drug release and promote the
drug absorption through an oral route.11 As presented in Table
that the HLB value of 8−18 could synthesize oil in water ME, 2, encapsulation of probucol into MEs improved the
while those ranging from 3 to 6 resulted in water in oil ME.15 bioavailability up to 10.22 folds from the drug solution.22
As a model drug, ASX is lipophilic, which required an oil-in- In this work, the effects of oil, surfactant, and cosurfactant
water ME system for the encapsulation. As listed in Table 1, composition on the formation, size, dispersibility, and stability
several surfactants such as cetyltrimethylammonium bromide, of ME were studied. As the drug carrier matrix, the usage of
T80, T20, and Triton X have an HLB value within this range. harmless initial materials for ME formulation is necessary.
Toxicity of the surfactant was taken into account for the drug Here, castor oil was used as the oil phase since it possesses
delivery purpose. Reportedly, a nonionic surfactant has the anti-inflammatory properties owing to the presence of
least toxicity compared to cationic and anionic surfactants.16 ricinoleic acid as its active fatty acid.23 A study by Putro et
The lethal dose (LD50) of T80 is 22,000 mg/kg while for T20 al. reported the relatively harmless nature of the nonionic

Table 2. Microemulsion Formulated with Nonionic Surfactants for Drug Carrier Application
surfactant cosurfactant oil phase drug bioavailability in MEa ref
b
T80 glycerin, Span 80 sesame oil tamoxifen citrate n.d. 40
T20, Labrafil M 1944 Transcutol P Capmul MCM C8 cilostazol 1.43 46
T80 PEG-400 olive oil famotidine n.d.b 11
T80, Cremophor EL ethanol, Span 80 glyceryl monooleate, linoleic acid glyceride metformin hydrochloride 1.14−1.47 47
T80, Cremophor EL PEG-400 olive oil probucol 2.15−10.22 22

a
Fold change relative to drug without carrier. bn.d. = Not determined.

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surfactant compared to anionic or cationic surfactants.24 Thus, 2.4. Characterization of ME and ME@ASX. The effects
nonionic tween 20 or tween 80 (Figure 1) and ethanol or of various surfactants and cosurfactants on the stability of ME
glycerol were used as the surfactant and cosurfactant, were investigated at three temperatures (277, 298, and 323 K)
respectively, to enhance drug delivery by ME while for 160 days. The stability of ME was examined based on its
maintaining the safeness of the system.25 ASX was used as particle size, polydispersity index (PDI), and zeta potential by
the model drug with the aim to investigate the stability of ME a zeta potential and particle size analyzer (ZetaPALS,
as a drug carrier matrix. Brookhaven Zeta Plus). The analyses were done at 298 K
and a fixed angle of 90°. Zeta potential values were obtained
2. MATERIALS AND METHODS using the Smoluchowski model. ME samples were diluted 50-
2.1. Materials. Astaxanthin (purity >97%) and castor oil fold with DI water for ZetaPALS measurement. Triplicate
were purchased from Sigma-Aldrich (Lanchasire, UK). analyses were carried out for each condition. Phase stability of
Surfactants polyoxyethylene (20) sorbitan monooleate ME with the largest area was also observed by centrifugation at
(Tween 80, T80) and polyoxyethylene (20) sorbitan 3500 ×g for 30 min, % transmittance (%T) measurement by a
monolaurate (Tween 20, T20) were supplied by Wako UV−vis spectrophotometer at 650 nm (UV-2600, Shimadzu,
Chemicals Industry (Osaka, Japan). Their molecular structures Japan), and viscosity determined by a Brookfield DV1
are shown in Figure 1. Ethanol (99.5%) obtained from Echo viscometer.
Chemical (Taiwan) and propane-1,2,3-triol (glycerol) pur- 2.5. Statistics Data Analysis. The data analysis was
chased from JT Baker Chemicals (USA) were used as performed by one-way analysis of variance (ANOVA) and the
cosurfactants. The chemical details are listed in Table 3. Tukey test using Minitab 17. A p-value of <0.05 and different
letters was considered statistically significant. The measure-
Table 3. Chemical Information ments were conducted in triplicate with a confidence interval
(CI) range of 95%.
chemical CAS
chemical name formula number 3. RESULTS AND DISCUSSION
astaxanthin C40H52O4 472-61-7
3.1. Pseudoternary Phase Diagram. The phase diagram
castor oil 8001-79-4
of castor oil, Smix, and DI water with Smix ratios of 2, 1, and 1/2
polyoxyethylene (20) sorbitan monooleate C64H124O26 9005-65-6
was calculated and constructed to obtain the ME region. MEs
polyoxyethylene (20) sorbitan monolaurate C58H114O26 9005-64-5
are classified into four Winsor types: Winsor type I, II, III, and
ethanol CH3CH2OH 64-17-5
IV. This study only focused on ME Winsor type IV, which is a
glycerol C3H8O3 56-81-5
one phase and transparent system.27 As presented in Figure 2,
the black-colored area in the pseudoternary phase diagram
represents the clear, transparent, and one-phase region as ME,
2.2. Construction of the Phase Diagram. The while the white-colored area represents turbid and white-
construction of the pseudoternary phase diagram of ME colored solution. This turbid appearance can be observed
followed the procedure of de Oliveira Neves and colleagues when the weight ratio of oil/Smix was higher than 2, which
with slight modification.26 The surfactant and cosurfactant suggests that the addition of the surfactant and cosurfactant is
were mixed with a fix weight ratio (Smix) of 2:1, 1:1, and 1:2. essential to suppress surface tension between oil and aqueous
Subsequently, mixtures of castor oil and Smix were prepared at phases.
weight ratios of 10:1, 8:1, 6:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:6, 1:8, As presented in Figure 2a−c, with the increasing amount of
1:10, and 1:50 to cover all region in the phase diagram. To ethanol in the system (Smix 2, 1, and 1/2), the ME region of the
obtain a pseudoternary phase diagram of the ME, deionized T20 system decreased from 12.48 to 12.33%. Similarly, in the
(DI) water was added dropwise to castor oil and Smix mixture system containing the T80 surfactant (Figure 2d−f), the ME
and vortexed until homogeneous. From visual observation, the region was reduced from 16.92 to 16.54% with each ethanol
transparent and one-phase liquid is considered as an ME. By addition. A similar trend was reported by Warisnoicharoen et
varying weight ratios, pseudoternary phase diagrams were al., where it was postulated that the addition of a cosurfactant
plotted, and the ME area was calculated using Origin software. may drag the system out of the ME region.28 These results
Calculation of the ME area in the pseudoternary graph was suggest that less ethanol as a cosurfactant may generate a
used to obtain the composition and the weight ratio of the broader ME region. However, excessive reduction of the
components in the mixture. The area of ME was obtained from cosurfactant in the system (Smix ratio 4/1; pseudoternary phase
Origin, and eq 1 was used to obtain the percentage area in the diagram not shown) led to the reduction of the ME region
pseudoternary phase diagram. (11.38 ± 0.01% for T20 and 16.16 ± 0.47% for T80,
respectively). Thus, adequate addition of the cosurfactant is
%area = A /0.5 × 100 (1)
important for the formation of ME. Smix = 2 (Figure 2a,d) gave
where A = ME area from Origin software, and 0.5 = ternary the widest area of the ME region compared to the other
area. studied ratios where T80-containing ME has a greater area
2.3. Preparation of Astaxanthin-Loaded ME. The (16.92 ± 1.69%) compared to the one containing T20 (12.48
optimum weight ratio of the oil, surfactant, and cosurfactant ± 2.35%). The bigger area of the ME region in T80-containing
that generates the largest-sized ME in the pseudoternary phase ME might be attributed to the more hydrophobic nature of
diagram was adopted for ASX-loaded ME (ME@ASX) T80 (HLB 15.9) compared to T20 (HLB 16.7), thus favoring
preparation. Prior to ASX loading, a certain amount of ASX the solubilization of large molecular volume oil.29,30
was solvated in castor oil to produce a 200 μg/mL ME@ASX Besides ethanol, glycerol is another commonly used
solution. Subsequently, this ASX−castor oil solution was mixed cosurfactant due to its cheap and nontoxic nature.31 Since
with 85 wt. % Smix and DI water. the largest ME area for both T20 and T80 was obtained at Smix
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Figure 2. Pseudoternary graph of castor oil with T20 as the surfactant and ethanol as the cosurfactant at an Smix of (a) 2, (b) 1, (c) 1/2; with T80 as
the surfactant and ethanol as the cosurfactant at an Smix of (d) 2, (e) 1, (f) 1/2. The black-colored area refers to the clear, transparent, and one-
phase ME region.

= 2 (surfactant/cosurfactant ratio), substitution of ethanol by castor oil, 85 wt.% Smix (T20/Gly), and 5 wt.% DI water was
glycerol was carried out at the same ratio, and the prepared, and its viscosity was measured. Compared to the one
pseudoternary phase diagrams obtained are presented in synthesized within the ME region (5 wt.% castor oil, 90 wt.%
Figure 3. Addition of glycerol with nonionic surfactants Smix, and 5 wt.% DI water), the viscosity of the LC is
resulted in a significant decrease in the ME area and the significantly higher (430.6 ± 3.39 cP for the LC and 302.15 ±
formation of a unique phase, known as the liquid crystal (LC, 0.35 cP for the ME region). Similarly, the LC formulated with
the gray area), which formed due to the repulsive interaction T80 (5 wt.% castor oil, 90 wt.% Smix, and 5 wt.% DI water) also
between the concentrated surfactant and glycerol. According to showed higher viscosity (581.6 ± 4.8 cP) compared to the one
the study by Li et al., the LC phase could be recognized by in the ME region (551.95 ± 0.64 cP). Since ethanol is a short-
increasing viscosity in the system.32 To confirm the formation chain alcohol, it may prevent the formation of any liquid
of the LC in each system, a mixture composed of 10 wt.% crystal phase due to low interaction force.11 The summary of
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Figure 3. Pseudoternary phase diagram of (a) Tween 20 and (b) Tween 80 with glycerol as the cosurfactant at a weight ratio of 2. The black-
colored area refers to the clear, transparent, and one-phase ME region. The grey-colored area refers to the liquid crystal region.

ME areas generated from different types and weight ratios of capacity of ME, which may later affect the pseudoternary phase
the surfactant and cosurfactant is listed in Tables 4 and 5. diagram structure.34,35 The solubilization capacity of ME at a
Overall, ethanol tends to produce a larger ME area for all Smix salt concentration of 0−0.35 M NaCl was measured according
ratios compared to the system containing glycerol as the to Bera et al.36 The highest solubilization of ME was achieved
cosurfactant. at a salt concentration of 0.14 M, which is desirable since it
mimics the salt concentration in body fluid. To investigate the
Table 4. Selected Microemulsion Formulations (wt.%) and effect of the temperature on the pseudoternary phase diagram,
Characterization of T20 as a Surfactant from Figure 4a a temperature of 310 K was used. This temperature was chosen
to simulate the human body condition for later application. As
parameter 1 2 3a
presented in Figure S2, the area of ME constructed by the
castor oil 0.04 0.10 0.05 same mixture (castor oil, T80, EtOH, and DI water) at 310 K
Smix = 2 0.74 0.80 0.85 was slightly larger (18.49 ± 0.22%) than the one constructed at
DI water 0.22 0.10 0.10 room temperature (16.92 ± 1.69%). This result is in
particle size (nm) 127.60 ± 24.92 163.57 ± 0.90 178.03 ± 9.72 agreement with Bera et al., which reported that a broader
PDI 0.48 ± 0.06 0.22 ± 0.22 0.23 ± 0.04 ME area can be achieved with an increasing temperature and
a
Selected formulation for further study. salinity.37
3.2. Characterization of ME. The larger ME area in the
Table 5. Selected Microemulsion Formulations (wt.%) and pseudoternary phase diagram may facilitate a wider range of
Characterization of T80 as a Surfactant from Figure 4b alternate composition for ME synthesis. Several formulations
parameter 1 2 3a of ME at different ratios of oil, Smix, and water were selected
(Figure 4) and characterized, and the results are shown in
castor oil 0.10 0.11 0.05
Smix = 2 0.70 0.79 0.85
Tables 4 and 5. In all formulations, the formation of ME with a
DI water 0.20 0.10 0.10
droplet size less than 200 nm and a PDI value less than 0.5 was
particle size (nm) 171.7 ± 4.47 108.78 ± 9.22 117.43 ± 0.38
confirmed. It can be observed that the PDI value of T20−ME
PDI 0.25 ± 0.02 0.41 ± 0.11 0.31 ± 0.02 formulation 1 (0.48) is close to the limit of the monodisperse
a system. The same phenomenon can be seen in T80−ME
Selected formulation for further study.
formulation 2 (PDI value = 0.41). Based on this result, ME
with formulation 3, which consists of 5 wt.% castor oil, 85 wt.%
To ensure that similar ME formulation can consistently be Smix, and 10 wt.% water was selected for further investigation.
obtained while using a surfactant produced from different To confirm the presence of the surfactant and cosurfactant in
suppliers, a pseudoternary diagram of a system containing T20 the system, the HPLC chromatogram of ME with formulation
from different brands (obtained from Acros, Belgium) is 3 was plotted (Figure S3).
plotted in Figure S1. The system containing T20 from Acros ME was characterized based on the particle size and
produced 11.91 ± 0.39% area of the ME region (Figure S1), dispersion homogeneity of its micelle. As listed in Table 6, in
which is comparable to the one obtained using T20 from the T20E-1 system composed of T20 as the surfactant and
Wako (12.48 ± 2.35%). A study by Nazar et al. investigated ethanol as the cosurfactant at an Smix ratio of 2 has a particle
the formulation of ME composed of castor oil/T80/ethanol/ size of 178.03 ± 9.72 nm. The particle size in T20E-2 and
phosphate buffer, with T80 obtained from Fluka.33 The ME T20E-3 systems, which consist of T20 and ethanol with Smix
region reported by Nazar et al. is comparable to the one ratios of 1 and 1/2, respectively, are 150.73 ± 2.64 and 213.13
obtained in this study. These results confirm the reliability of ± 0.9 nm. Substitution of glycerol as the cosurfactant greatly
the generated pseudoternary diagrams. affected the particle size of the ME to 280.57 ± 39.83 nm for
Salinity of the aqueous solution and temperature of the the T20G system at an Smix ratio of 2. For the T80-containing
system were postulated to have an effect on the solubilization system, T80E-1, which consists of T80 (surfactant), ethanol
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Figure 4. Pseudoternary phase diagram of (a) T20 or (b) T80 as the surfactant and ethanol as the cosurfactant with an Smix of 2. Yellow
numberings inside the diagrams were the selected compositions for initial characterization.

Table 6. Particle Size and the PDI for Selected Microemulsion at T = 298 K (n = 3, Mean ± SD)
ME at 5 wt.% castor oil, 85 wt.% Smix, and 10 wt.% water
codes cosurfactant Smix ME Area (%) particle size (nm) PDI
Tween 20
T20E-1 ethanol 2 12.48 ± 2.35 178.03 ± 9.72 0.23 ± 0.04
T20E-2 ethanol 1 12.41 ± 0.49 150.73 ± 2.64 0.20 ± 0.04
T20E-3 ethanol 1
2 12.33 ± 0.38 213.13 ± 0.90 0.16 ± 0.02
T20G glycerol 2 3.46 ± 0.21 280.57 ± 39.83 0.15 ± 0.07
Tween 80
T80E-1 ethanol 2 16.92 ± 1.69 117.43 ± 0.38 0.31 ± 0.02
T80E-2 ethanol 1 16.78 ± 0.37 150.50 ± 9.17 0.25 ± 0.06
T80E-3 ethanol 1
2 16.54 ± 1.29 139.17 ± 2.25 0.25 ± 0.01
T80G glycerol 2 5.09 ± 0.06 260.87 ± 2.02 0.21 ± 0.03

(cosurfactant) with an Smix ratio of 2, has a particle size of

117.43 ± 0.38 nm. Similar to the T20-containing system, at the
same Smix, the use of glycerol as the cosurfactant produced
bulkier ME (260.87 ± 2.02 nm). As shown in Table 6, all ME
had an initial size of less than 300 nm, which can be classified
as ME.38
The homogeneity of the system was examined and expressed
as a PDI value. For the T20-containing systems, namely, T20E-
1, T20E-2, T20E-3, and T20G, the PDI values are 0.23 ± 0.04,
0.20 ± 0.04, 0.16 ± 0.02, and 0.15 ± 0.07, respectively.
Similarly, in the T80-containing systems, the PDI value ranges
from 0.21 to 0.31 as listed in Table 6. The PDI values of all
prepared ME are less than 0.5, indicating homogeneous
dispersion and long-term stability of ME.39,40 Besides the PDI,
surface charge of ME is an important property related to
particle repellence in the system to inhibit agglomeration and
prolong its stability.41,42 The surface charges of the MEs
synthesized in various compositions were shown to be negative Figure 5. Zeta potential of freshly selected microemulsions. Different
with magnitude ranging from −11.59 to −8.08 (Figure 5). The letters indicate significant differences among formulations. Multiple
high zeta potential value might be attributed to hydrogen comparisons of means were performed using Tukey’s test at the 0.05
bonding between the hydroxyl group of ricinoleic acid in castor significance level.
oil and the nonionic surfactant.
3.3. Astaxanthin-Loaded ME. Preparation of ME@ASX
was similar to that of ME. Instead of castor oil only, ASX in measurement. From Figure 6a, it can be seen that
castor oil solution was added to the system. The pseudoternary encapsulation of ASX resulted in bigger particle size for T20
phase diagram of ME@ASX (data not shown) for Smix = 2 with or T80 as the surfactant and ethanol as the cosurfactant.
ethanol as the cosurfactant yielded an ME area of 11.54 ± 0.3 Encapsulation of ASX enlarges the particle size of T20E-1 from
and 18.4 ± 0.25% for T20 and T80, respectively. Observation 178.03 to 197 nm and T80E-1 from 117.43 to 198.8 nm.
of ASX loading in the ME was conducted by particle size Additionally, the ME@ASX displayed an orange-red color,
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Figure 6. (a) Particle size comparison of microemulsion loaded with ASX (ME@ASX) and microemulsion without ASX. (b) Physical appearance
and quantitative characterization before and after centrifugation of (i) T80E-1, (ii) T20E-1, and (iii) T80E-1@ASX. Different letters indicate
significant differences among formulations. Multiple comparisons of means were performed using Tukey’s test at the 0.05 significance level.

which demonstrated the successful encapsulation of red- analysis confirms that there is no significant change in the %T
colored ASX into ME (Figure 6b). and viscosity in all systems (including the one after ASX
3.4. Stability Study of ME and ME@ASX. To investigate encapsulation). This suggest that the formulated ME was
the effect of gravitation force on the ME system, MEs with and stable against the centrifugation process, thus centrifugation
without ASX loading were centrifuged at 3500 ×g for 30 min, did not affect the properties of all formulated MEs and ME@
and the results are presented in Figure 6b. After undergoing ASX. No color change, precipitation, nor phase separation was
centrifugation, the transparency and the viscosity of MEs were observed after centrifugation.
quantified. Both T20- and T80-containing MEs show high %T Stability of ME was confirmed through a storage test. After
(> 95), which confirms the transparency of the systems. 160 days of storage at 277, 298, and 323 K, the system
Compared to the one before centrifugation, no significant remained homogeneous with the polydispersity index un-
difference can be observed in the %T of T20E-1, T80E-1, and changed (0.2−0.3, Table 7). MEs with Smix = 2 for T20 and
T80E-1@ASX. Determination of the ME viscosities before and T80 with ethanol were selected for ZetaPALS measurement
after centrifugation further demonstrated the stability of T20E- due to the largest ME region in the pseudoternary phase
1, T80E-1, and T80-1@ASX. The viscosity of all tested diagram. After 160 days, the particle sizes of ME and ME@
systems ranges from 34.96 to 38.67 cP. Statistical significance ASX at 298 K (T80E-1) were maintained at 122.63 and 177.43
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Table 7. Polydispersity Index of Microemulsions with and

without ASX at Different Temperatures for Storage up to
160 Days (n = 3, Mean ± SD)
polydispersity index at T
time storage
(days) samples 298 K 323 K 277 K
0 T20E-1 0.23 ± 0.04
T20E-1@ 0.24 ± 0.05
ASX
T80E-1 0.31 ± 0.02
T80E-1@ 0.25 ± 0.01
ASX
33 T20E-1 0.23 ± 0.04 0.24 ± 0.03 0.24 ± 0.01
T20E-1@ 0.28 ± 0.02 0.29 ± 0.03 0.27 ± 0.01
ASX
T80E-1 0.30 ± 0.02 0.30 ± 0.01 0.27 ± 0.03
T80E-1@ 0.22 ± 0.01 0.24 ± 0.04 0.30 ± 0.02
ASX
160 T20E-1 0.20 ± 0.12 0.24 ± 0.01 0.20 ± 0.05 Figure 8. Stability of T80E-1 at (a) 298, (b) 323, (c) and 277 K
T20E-1@ 0.25 ± 0.03 0.21 ± 0.03 0.26 ± 0.01 during 160 days of storage, all without ASX and loaded with ASX.
ASX Asterisks indicate significance analyzed by Tukey’s test (*, P < 0.05;
T80E-1 0.28 ± 0.01 0.26 ± 0.04 0.28 ± 0,03 **, P < 0.01; ***, P < 0.001; and ****, P < 0.0001)
T80E-1@ 0.29 ± 0.01 0.30 ± 0.01 0.28 ± 0.01
ASX

nm, respectively. No significant difference can be observed in orange and transparent color with an initial particle size slightly
the size of the ME with or without ASX. These suggest that bigger than the parent ME. Within 160 days of storage, both
both ME@ASX and ME without ASX (Figures 7 and 8) were ME and ME@ASX were stable, as indicated by the particle size
and polydispersity index value. From the thermal storage test,
it shows that the temperature did not influence the stability of
the ME. Results of this study suggest the potential of the as-
synthesized ME as a drug carrier matrix. Further studies are
needed for ME@ASX in pharmaceutical application, such as
the release profile and antimicrobial assay.

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sı
The Supporting Information is available free of charge at
https://pubs.acs.org/doi/10.1021/acs.jced.0c00341.
Pseudoternary graph of castor oil with T20 (Acros) as
the surfactant and ethanol as the cosurfactant at an Smix
of 2, pseudoternary graph of castor oil with T80 as the
surfactant and ethanol as the cosurfactant at an Smix of 2
at T = 310 K, and the HPLC chromatogram of ME
(PDF)

Figure 7. Stability of T20E-1 at (a) 298, (b) 323, (c) and 277 K
during 160 days of storage, all without ASX and loaded with ASX.
Asterisks indicate significance analyzed by Tukey’s test (*, P < 0.05;
■ AUTHOR INFORMATION
Corresponding Authors
Artik Elisa Angkawijaya − Graduate Institute of Applied
**, P < 0.01; ***, P < 0.001; and ****, P < 0.0001)
Science, National Taiwan University of Science and Technology,
Taipei 106, Taiwan; orcid.org/0000-0002-4405-5068;
stable. Particle sizes were kept between 96.93 and 204.93 nm
Email: [email protected]
at all three different temperatures studied. The thermal stability
Suryadi Ismadji − Department of Chemical Engineering, Widya
test suggests that the temperature has an insignificant effect on
Mandala Surabaya Catholic University, Surabaya 60114,
ME stability.
Indonesia; Department of Chemical Engineering, National
4. CONCLUSIONS Taiwan University of Science and Technology, Taipei 106,
Taiwan; Email: [email protected]
ME of castor oil, Tween 80, ethanol, and DI water was
successfully synthesized. The largest ME area was obtained Authors
from a combination of Tween 80 and ethanol with a weight Chintya Gunarto − Department of Chemical Engineering,
ratio of 2. ME composition of 5 wt.% castor oil, 85 wt.% National Taiwan University of Science and Technology, Taipei
surfactant mixture, and 10 wt.% water was chosen for further 106, Taiwan; Department of Chemical Engineering, Widya
analysis. Astaxanthin as a model lipophilic drug was Mandala Surabaya Catholic University, Surabaya 60114,
successfully encapsulated in ME. The ME@ASX showed an Indonesia
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J. Chem. Eng. Data XXXX, XXX, XXX−XXX
Journal of Chemical & Engineering Data pubs.acs.org/jced Article

Yi-Hsu Ju − Graduate Institute of Applied Science, Department nanoparticles by nanoprecipitation method. Int. J. Pharm. Technol.
of Chemical Engineering, and Taiwan Building Techonology 2014, 5, 5962−5972.
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