AAPS PharmSciTech ( # 2017)

DOI: 10.1208/s12249-017-0798-x

Research Article

Preparation of Essential Oil-Based Microemulsions for Improving the Solubility,

pH Stability, Photostability, and Skin Permeation of Quercetin

Xia Lv,1 Tiantian Liu,2 Huipeng Ma,3 Yan Tian,2 Lei Li,2 Zhen Li,2 Meng Gao,2
Jianbin Zhang,2,4 and Zeyao Tang2,4

Received 9 March 2017; accepted 30 April 2017

Abstract. Quercetin can bring many benefits to skin based on its various bioactivities.
However, the therapeutic effect of quercetin is limited due to the poor water solubility, pH
instability, light instability, and skin permeation. The aim of the present work was applying
essential oil-based microemulsions to improve the solubility, pH stability, photostability, and
skin permeation of quercetin for topical application. Peppermint oil (PO-ME), clove oil (CO-
ME), and rosemary oil (RMO-ME) were selected as model essential oils. Microemulsions
composed of Cremophor EL/1,2-propanediol/essential oils (47:23:30, w/w) were selected as
model formulations, based on the pseudo-ternary phase diagram and the characterizations. In
the solubility study, the solubility of quercetin was improved dozens of times by
microemulsions. Quercetin was found instable under alkaline condition, with 50% degraded
in the solution of pH 13. However, PO-ME, CO-ME, and RMO-ME could protect quercetin
from the hydroxide ions, with 47, 9, and 12% of quercetin degraded. In the photostability
study, the essential oil-based microemulsions showed the capability of protecting quercetin
from degradation under UV radiation. Where more than 67% of quercetin was degraded in
aqueous solution, while less than 7% of quercetin degraded in microemulsions. At last, the
in vitro skin permeation study showed that the essential oil-based microemulsions could
enhance the permeation capacity of quercetin by 2.5–3 times compared to the aqueous
solution. Hence, the prepared essential oil microemulsions could improve the solubility, pH
stability, photostability, and skin permeation of quercetin, which will be beneficial for its
topical application.
KEY WORDS: essential oil; microemulsions; quercetin; pH stability; photostability.

INTRODUCTION oxidative stress is superior to the defense mechanism of skin,

Skin is the largest organ of human body, playing as a
barrier to protect human body from germs, mechanical
injuries, and exogenous pollutants (1). Skin is continuously
exposed to the environment of ultraviolet irradiation, ionizing
radiation, and toxic chemicals, which can induce oxidative
stress by the increased amounts of reactive oxygen species
(ROS) (2). There are some self-protection mechanisms
against ROS in the skin, such as superoxide dismutase,
metallothionein, and melanin (3). However, once the
Xia Lv and Tiantian Liu contributed equally to this work
1
College of Life Science, Dalian Nationalities University, Dalian,
116600, People’s Republic of China.
2 pathway and the release of several proinflammatory cyto-
College of Pharmacy, Dalian Medical University, Dalian, 116044,
People’s Republic of China.
3
College of Medical Laboratory, Dalian Medical University, Dalian,
116044, People’s Republic of China.
4
To whom correspondence should be addressed. (e–mail:
zhangjb@dmu.edu.cn; tangzeyao@aliyun.com)
skin damage can occur including lipid peroxidation, enzyme
inactivation, and DNA breakage (4). Supporting skin defense
mechanisms by exogenous antioxidants is a promising strat-
egy to avoid skin damage, such as Coenzyme Q10, vitamin C,
β-carotene, and polyphenols (5–7).
Quercetin, a flavonoid compound, is considered as one of
the most powerful natural antioxidants (8). In vitro and
in vivo experiments demonstrate that quercetin can protect
keratinocytes from exogenous oxidizing agents, scavenge free
radicals, prevent endogenous antioxidant depletion, and
inhibit lipid peroxidation upon exposure to UV irradiation
(9,10). Moreover, quercetin has various other biological
activities including anti-inflammatory, antibacterial, antiviral,
and anticancer properties. Quercetin can inhibit NF-κB
kines, making it present broad anti-inflammatory actions (11).
The combined antioxidative/anti-inflammatory actions of
quercetin are also beneficial for wound healing. Furthermore,
quercetin shows anti-aging activity on middle-aged
keratinocyte, rejuvenating activity on terminally senescent

1530-9932/17/0000-0001/0 # 2017 American Association of Pharmaceutical Scientists


cells, as well as whitening effect on skin (12). Hence, it is
obvious that the topical application of quercetin can bring
many benefits to skin. However, the therapeutic effect of
quercetin is limited due to the poor water solubility, low
permeation into skin, and instability under light, pH, and
temperature (13,14). The disadvantages of quercetin can be
overcome by formulation strategies (15). Several transdermal
formulations of quercetin have been developed, including
emulsions, microemulsions, liposomes, vesicles, cyclodextrin,
gels, SLN, LNC, and mesoporous silica (16–21).
Microemulsions are dispersions consisting of oil phase,
water phase, surfactant, and co-surfactant, which are single
optically isotropic and thermodynamically stable liquid solu-
tions with a droplet diameter usually within the range of 10–
100 nm (22,23). In recent years, microemulsions have been
applied to deliver many drugs for topical use, such as
zaltoprofen, zidovudine, sertaconazole, resveratrol, and
curcumin (24–26). They can significantly enhance their
therapeutic effects, due to the advantages such as improving
solubilities of poorly water-soluble drugs, improving the
permeation of drugs across the skin barrier, improving the
stability of unstable drugs (27,28). Moreover, the improve-
ment of solubility, permeation, and stability of drug is highly
related to the components of microemulsions.
Recently, microemulsions containing essential oil for
topical use have gained much attention of researchers
(29,30). Essential oils are natural, complex, multi-component
systems, extracted from different parts of aromatic plants
(31). Essential oils, such as peppermint oil, clove oil, and
rosemary oil, have many biological activities including
antioxidant, anti-inflammatory, anticancer, antibacterial, anti-
fungal, and antiviral activities, leading them widely used in
cosmetics, pharmaceutical, medicinal, and food industries
(32–34). Moreover, essential oils mainly contain terpenes,
which have good penetration enhancing effects on various
drugs with low skin irritation at low concentrations (35). In
theory, quercetin in combination with essential oils can not
only improve its solubility and stability but also improve the
permeation across skin. Moreover, the combination of
quercetin and essential oils might result synergistic effect on
protecting skin.
The objective of the present study was to prepare essential
oil-based microemulsions to improve the solubility and stability
of quercetin. Peppermint oil, clove oil, and rosemary oil were
selected as model essential oils. The peppermint oil, clove oil,
and rosemary oil-based microemulsions (PO-ME, CO-ME, and
RMO-ME) were prepared and evaluated by pseudo-ternary
phase diagram and particle size distribution. The solubilizing
capabilities of three microemulsions on quercetin were subse-
quently studied. Finally, the effect of essential oil-based
microemulsions on the pH stability and photostability of
quercetin was studied.
MATERIALS AND METHODS
Materials
Peppermint oil, clove oil, or rosemary oil were kind gifts
from Hengcheng Natural Flavor oil Co. (Jiangxi, China).
Cremophor EL was purchased from Sigma Chemical Co. (St.
Louis, MO, USA). 1,2-Propanediol was purchased from
Lv et al.
Damao Chemical Reagent Factory (Tianjin, China). Querce-
tin was obtained from Aladdin-reagent Co. (Shanghai,
China). Methanol was HPLC grade and supplied from J&K
Chemical Ltd. (Shanghai, China). All other chemicals and
reagents were of analytical grade.
Pseudo-ternary Phase Diagram
Pseudo-ternary phase diagrams were constructed using a
water titration method at room temperature. Cremophor EL
and 1,2-propanediol were selected as surfactant and co-
surfactant, and they were firstly mixed at the ratio of 2:1 (w/
w) to form the mixed surfactant (Smix). Essential oil
(peppermint oil, clove oil, or rosemary oil) and Smix were
weighted into glass vials with the weight ratios of 1:9, 2:8, 3:7,
4:6, 5:5, 6:4, 7:3, 8:2, and 9:1, where the total weight was 0.5 g.
They were mixed for 2 h on magnetic stirrers, and then the
mixtures were titrated with deionized water in a drop-wise
manner and mixed thoroughly by magnetic stirrers. The
titration of water was stopped until the turbid mixtures
converted into transparent solution. After 30 min equilib-
rium, the samples were clear, transparent, and homogenous,
with light blue opalescence, meaning the form
microemulsions. The amounts of water added in each group
were recorded, and the boundary of microemulsion-forming
domain was determined. Furthermore, the samples were
diluted 20-fold by deionized water and stored 48 h for
observation.
Preparation and Characterization of Microemulsions
Cremophor EL and 1,2-propanediol were firstly mixed at
the weight ratio of 2:1 and stirred for 1 h at room
temperature. Then, Smix and essential oil (peppermint oil,
clove oil, or rosemary oil) were weighted into glass vials with
the weight ratio of 7:3. The glass vials were immediately
sealed with rubber stoppers to avoid volatilization of essential
oils. The mixtures were stirred for 2 h in the dark to prepare
the self-microemulsifying drug delivery systems (SMEDDS).
Microemulsions (PO-ME, CO-ME, and RMO-ME) were
prepared by diluting 1 g SMEDDS formulations with 19 mL
deionized water. For the preparation of quercetin-loaded
microemulsions, quercetin was previously mixed with Smix
and essential oil before adding deionized water.
The particle sizes, polydispersity indexes (PDI), and zeta
potentials of PO-ME, CO-ME, and RMO-ME were mea-
sured by Malvern ZS90 zetasizer (Malvern Instruments Corp,
Malvern, UK). Briefly, 1 mL of microemulsions was put into
the cuvettes for measurement. Light scattering was monitored
at a fixed angle of 90° and maintained the temperature at
25°C. Three parallel samples were measured.
The electrical conductivity of PO-ME, CO-ME, and
RMO-ME was measured using a DDS-307 electric conduc-
tivity meter (Shanghai Precision Scientific Instrument Corp,
Shanghai, China). The electrode was dipped in the
microemulsion samples at the temperature of 25°C, and
electrical conductivity was recorded after equilibrium had
been reached. Three parallel samples were measured.
The refractive index was determined at 25°C using a
2WAJ Abbe refractometer (Shanghai Optical Instrument
Company, Shanghai, China). The transmittance of
Essential Oil-Based Microemulsions of Quercetin
microemulsions was measured at 650 nm using the ultraviolet/
visible spectrophotometer (UV2300, Techcomp, China) keep-
ing distilled water as a blank. Three parallel samples were
measured.
The viscosity of microemulsions was measured by a
cone-plate viscometer (Brook-field, DV-II + Pro, USA). The
electronic cone spindle CPE-40 was applied, and the gap
between the cone and plate was set at 13 μm. The spindle
speed was 3 rpm, and the viscosity was recorded when the
number was stabilized. Three parallel samples were
measured.
Solubility Study
Excess amounts of quercetin were added into test tubes
containing 1 mL of water, essential oil, Cremophor EL, 1,2-
propanediol, Smix, SMEDDS, diluted microemulsions, or
micelles. The test tubes were sealed and shaken for 72 h in
a shaking incubator (HZQ-F, Harbin Donglian Electronic
Technology Development Co., LTD., Harbin, China) at 37°C,
150 rpm. After equipment, the test tubes were centrifuged at
5000g for 30 min, to remove the excess drug. The supernatant
was diluted several times by ethanol, and the concentration of
quercetin was measured using the ultraviolet/visible spectro-
photometer at the wavelength of 370 nm. For each group,
samples were performed in triplicate.
pH Stability Study
4.2 g Smix, 1.8 g essential oil (peppermint oil, clove oil, or
rosemary oil), and 30 mg quercetin were added into glass
vials and sealed by rubber stoppers. They were stirred for 2 h
on a magnetic stirrer in dark, to obtain SMEDDS containing
5 mg/g quercetin. Then, 1 g SMEDDS (or 5 mg quercetin)
was respectively diluted by 19 mL (or 20 mL) solution with
the pH of 1, 4, 7, 10, or 13. The formed microemulsions or
quercetin solutions were stirred for 10 h in the dark, and then
the samples were diluted 10-fold by ethanol to measure the
concentrations of remained resveratrol. Furthermore, the
particle sizes of microemulsions in different pH solutions
were measured. The solutions with different pH were
respectively 0.1 M HCl solution (pH 1), 0.1 mM HCl solution
(pH 4), water (pH 7), 0.1 mM NaOH solution (pH 10), and
0.1 M NaOH solution (pH 13).
Photostability Study
The photodegradation experiments were performed
using a UVB lamp (Philips TL40/12 RS), emitting a
continuous spectrum of 270–400 nm with a peak emission at
313 nm (36). Quercetin-loaded SMEDDS (5 mg/g) were
prepared and diluted 20 times by deionized water. Quercetin
aqueous solution with the concentration of 0.25 mg/mL was
also prepared as control. Quercetin microemulsions and
aqueous solution were kept under the light source with the
distance of 15 cm. At predetermined irradiation time, the
concentration of quercetin was measured by UV analysis. The
photostability was evaluated by the percentage of remained
quercetin at different irradiation times, compared to unex-
posed samples. For each group, samples were performed in
triplicate.
In Vitro Skin Permeation Study
The dorsal skin for permeation study was obtained from
Wistar rats. Subcutaneous fat and excess tissue were carefully
removed, and the integrity was examined. The excised rat
skins were washed by phosphate buffer solution (pH 6.8) and
stored at 4°C before permeation experiments.
The in vitro skin permeation study was carried out by the
Franz diffusion cell apparatus (HC-188, Tianjin Zhengtong
Technology Co., China). The skin was fixed between the
donor and the receptor chamber, with the stratum corneum
side towards to the donor chamber. The receptor chamber
was filed with the receptor phase (saline/ethanol = 8:2) and
kept at 37°C. The skin area contacting with the receptor
phase was 0.636 cm2. Quercetin-loaded SMEDDS (5 mg/g)
were prepared and diluted 20 times by deionized water.
Quercetin aqueous solution with the concentration of
0.25 mg/mL was also prepared as control. A 0.3 mL quercetin
microemulsions or aqueous solution was added on the donor
chamber. After 3, 6, 9, 12, and 24 h, 0.5 mL receptor phase
was taken out and replaced with fresh solution.
The amount of quercetin was analyzed by HPLC (e2695,
Waters Corporation, Milford, USA). The analysis was carried
out on a SinoChrom ODS-BP column (4.6 × 250 mm, 5 μm,
Dalian Elite Analytical Instruments, Dalian, China). The
mixture of methanol/water/phosphoric acid (100:100:1, v/v)
was used as mobile phase. The flow rate was kept at 0.8 mL/
min, and the detection wavelength was set at 370 nm.
RESULTS AND DISCUSSION
Determination of Microemulsion Formulations
Cremophor EL and 1,2-propanediol are commonly used
as the surfactant and co-surfactant of microemulsions. Here,
we used the mixture of Cremophor EL/1,2-propanediol (2:1)
as mixed surfactants and evaluated the feasibility of forming
microemulsions with essential oil (peppermint oil, clove oil,
or rosemary oil) by the pseudo-ternary phase diagrams, the
appearance, and the particle size distribution. In the pseudo-
ternary phase diagrams (see Fig. 1), we found that all the
three types of essential oils could form microemulsions, if the
Smix/oil ratio was high enough. Peppermint oil has the largest
microemulsion area in the pseudo-ternary phase diagrams,
where the Smix/oil ratio should be above 5:5. Similarly,
microemulsions of clove oil and rosemary oil required the
Smix/oil ratio higher than 7:3 and 5:5. At the beginning
titration of water, the mixtures formed sticky, opaque,
gel-like dispersions. As the content of water increased,
they transformed into transparent, homogenous o/w
microemulsions. Moreover, the formed o/w microemulsions
could be diluted by bulk water. Figure 2 shows the visual
appearance of the Smix/essential oil mixtures diluted 20-fold
by water. We found that the mixtures with the Smix/
peppermint oil ratios above 5:5 were clear, transparent, and
homogenous microemulsions with light blue opalescence,
while the other solutions were turbid and heterogeneous.
The mixtures of Smix/clove oil and Smix/rosemary oil could
also form the transparent and homogenous microemulsions at
the ratios above 7:3 and 5:5. Furthermore, we measured the
particle sizes of the essential oil-based microemulsions. As

Fig. 1. Pseudo-ternary phase diagrams of different essential oil-based microemulsions. a PO-ME. b CO-ME. c RO-ME. The
microemulsions regions were marked in gray
seen in Fig. 3, the particle sizes of microemulsions were
gradually increased with the ratio of essential oil/Smix. As
microemulsions could be considered as Bswollen micelles,^
the oil phase incorporated into the hydrophobic core of
micelle leading their particle sized increased. It is worth
noting that the particle sizes of microemulsions were very
small, even less than 30 nm for PO-ME and RMO-ME with
the oil content below 50% and less than 20 nm for CO-ME
with the oil content below 30%. In summary, peppermint oil,
clove oil, and rosemary oil can form o/w microemulsions with
the Smix/essential oil ratio above 5:5, 7:3, and 5:5, and the
particle sizes of microemulsions were smaller than 30 nm.
Characterizations of Essential Oil-Based Microemulsions
In this study, we selected the SMEDDS formulations with
the Smix/essential oil ratio of 7:3 as model for further study,
where the Cremophor EL/1,2-propanediol/essential oil content
was 47:23:30 (w/w). The SMEDDS formulations were diluted 20
times by deionized water, and the characterizations of formed
microemulsions were listed in Table I. The particle sizes,
polydispersity index (PDI), and zeta potentials were measured
by Malvern ZS90 zetasizer. The particle sizes of PO-ME, CO-
ME, and RMO-ME were similar, which were 12.66 ± 0.54,
9.74 ± 0.04, and 11.61 ± 0.08 nm, respectively. The polydispersity
index of CO-ME was smaller than PO-ME and RMO-ME,
indicating that they had better homogeneity. The zeta potentials
Fig. 2. Visual appearance of the Smix/essential oil mixtures diluted 20-
fold by water. From left to right, the Smix/essential oil ratio ranged
from 1:9 to 10:0
Lv et al.
were limited from −5 to 0 mV, because there was no charged
group in the compositions. The small negative charge could be
attributed to the polyoxyethylene group of Cremophor EL. The
electrical conductivities of PO-ME, CO-ME, and RMO-ME
were measured to be 60.0 ± 3.3, 89.4 ± 4.0, and 185.6 ± 5.5 μS/cm,
respectively. Moreover, their viscosities were 3.58 ± 0.36,
3.19 ± 0.16, and 3.37 ± 0.24 cP. The electrical conductivity and
viscosity of microemulsions indicated that they were o/w type
microemulsions. In addition, the refractive indexes of PO-ME,
CO-ME, and RMO-ME were 1.3396, 1.3399, and 1.3392, very
similar to the refractive index of water (1.333). At last, we found
that the transmittance of the diluted microemulsions was more
than 93% at 650 nm.
Solubility
The poor solubility has severely limited the application
of quercetin, leading it essential to improve the solubility of
quercetin by pharmaceutical method (37). The solubilizing
capabilities of microemulsions as well the components are
listed in Table II. The solubility of quercetin in water was just
0.013 mg/mL; however, its solubility was significantly in-
creased in surfactant, co-surfactant, essential oils, SEMDDS,
Fig. 3. Particle sizes of microemulsions with different Smix/essential
oil ratios
Essential Oil-Based Microemulsions of Quercetin

Table I. The components and characterizations of essential oil-based microemulsions

PO-ME CO-ME RMO-ME

Components (w/w) Cremophor EL Cremophor EL Cremophor EL

47%/1,2-propanediol 47%/1,2-propanediol 47%/1,2-propanediol
23%/peppermint oil 30% 23%/clove oil 30% 23%/rosemary oil 30%
Particle size (nm) 12.66 ± 0.54 9.74 ± 0.04 11.61 ± 0.08
PDI 0.265 ± 0.041 0.077 ± 0.008 0.385 ± 0.01
Zeta potential (mV) −4.35 ± 0.25 −0.992 ± 0.05 −4.05 ± 0.14
Electrical conductivity (μS/cm) 60.0 ± 3.3 89.4 ± 4.0 185.6 ± 5.5
Refractive index 1.3396 1.3399 1.3392
Viscosity (cP) 3.58 ± 0.36 3.19 ± 0.16 3.37 ± 0.24
Transmittance (%) 95.38 ± 0.06 99.17 ± 0.06 93.80 ± 0.19

microemulsions, and micelles. Cremophor EL has the best
solubilizing capability among the components, which reached
to 18.17 ± 0.12 mg/mL. The quercetin solubility in essential
oils varied significantly, where peppermint oil showed the best
solubilizing capability of 15.73 ± 0.07 mg/mL. The solubility of
quercetin in clove oil and rosemary oil was 6.84 ± 0.02 and
1.1 ± 0.03 mg/mL. The solubilities of quercetin in Smix and
SMEDDS were between 8 and 21 mg/mL, among which the
solubility in PO-SME was the highest, more than 1600 times
higher than water solubility. Furthermore, we found that the
solubilizing capabilities of Smix and SMEDDS were signifi-
cantly decreased, if they were diluted. After diluted by 100-
fold, the formed micelles and microemulsions have the
solubility of 0.27–1.11 mg/mL, still dozens of times higher
than water solubility.
pH Stability
Quercetin was chemically unstable, and the instability
could be attributed to hydroxyl groups and the instable
pyrone structure of quercetin (38). It is reported that
quercetin was degraded under alkaline conditions (14).
Moreover, the oxidation of quercetin by air oxygen could
take place under moderately basic media (39). Some formu-
lations could protect quercetin from alkaline degradation,
such as lecithin-based nanoparticle, cyclodextrin, and solid
dispersion (40–43). In this work, we studied the ability of
essential oil-based microemulsions protecting quercetin from
degradation at different pH conditions. To evaluate the
stability of microemulsions in different pH solutions, the
particle sizes of PO-ME, CO-ME, and RMO-ME were
measured with the pH ranging from 1 to 13. As seen in
Fig. 4, the particle sizes of three kinds of microemulsions were
between 6 and 12 nm. Despite the particle sizes were slightly
deceased if the pH increased, the changes were negligible.
Hence, the essential oil-based microemulsions were stable in
the wide range pH conditions. The pH stability of quercetin
in aqueous solution and microemulsions is shown in Fig. 5.
Quercetin was stable in aqueous solution and microemulsions
under acidic and neutral conditions; unfortunately, it was
degraded under alkaline conditions. The degradation of
quercetin was limited in aqueous solution of pH 10, with
about 94% of quercetin remained. However, the degraded
quercetin reached to 50% when the pH increased to 13.
Meanwhile, the remained quercetin in PO-ME, CO-ME, and
RMO-ME was respectively 57, 91, and 88% at pH 13. This
indicated that essential oil-based microemulsions could pro-
tect quercetin from alkaline degradation to some extent. The
quercetin protective effect of clove oil and rosemary oil
seemed better than peppermint oil. For drugs in
microemulsions, they can be either located in the hydrophilic
group of surfactant (the shell of microemulsions) or located in
the hydrophobic oil region (the core of microemulsions) (44).
We inferred that quercetin might be located in the core of
microemulsions, where quercetin was separated from the
hydroxide ions. The incorporation of quercetin has many
benefits for protecting it from degradation and prolonging the
shelf-life of quercetin products.

Table II. The solubilities of quercetin in water, micelles, microemulsions, and the components

Solubility (mg/mL) Solubility (mg/mL)

Water 0.013 ± 0.01 PO-ME (100-fold) 0.54 ± 0.01

Cremophor EL 18.17 ± 0.12 Clove oil 6.84 ± 0.02
1,2-Propanediol 3.90 ± 0.03 CO-SME 15.96 ± 0.48
Smix 11.65 ± 0.09 CO-ME (20-fold) 0.91 ± 0.01
Micelles (20-fold) 1.01 ± 0.02 CO-ME (100-fold) 0.78 ± 0.02
Micelles (100-fold) 0.43 ± 0.01 Rosemary oil 1.1 ± 0.03
Peppermint oil 15.73 ± 0.07 RMO-SME 8.27 ± 0.03
PO-SME 20.81 ± 0.02 RMO-ME (20-fold) 0.74 ± 0.01
PO-ME (20-fold) 1.11 ± 0.02 RMO-ME (100-fold) 0.27 ± 0.02

Fig. 4. Particle sizes of microemulsions with the pH ranging from 1 to 13
Photostability
UV exposure to the skin leads the generation of reactive
oxygen species (ROS), which could damage cell membranes
by peroxidation of fatty acids. Quercetin, a powerful antiox-
idant, can scavenge ROS to protect the skin. Moreover,
quercetin could directly absorb UV radiation to prevent the
formation of ROS and the consequent direct DNA damage
(45). However, quercetin is instable under UV irradiation.
Quercetin is prone to be degraded under light exposure,
resulting in the decreased efficacy. The risk of light-induced
degradation is throughout the whole process of preparation,
storage, and application of quercetin formulations. Hence, it
is essential to evaluate photostability of quercetin formula-
tions. In this work, the photostability of quercetin under UV
irradiation was studied. As seen in Fig. 6, the quercetin was
quickly degraded in aqueous solution under UV irradiation.
The remained quercetin in aqueous solution were 70.4, 50.2,
39.7, 36, and 32%, with the irradiation time of 0.5, 1.5, 3.5,
Fig. 5. pH stability of quercetin in aqueous solution and microemulsions
Lv et al.
Fig. 6. Photostability of quercetin in aqueous solution and
microemulsions under UV irradiation
8.5, and 12.5 h. More than two-thirds of quercetin was
degraded after 12.5 h irradiation. The degradation products
of quercetin were complicated; products such as 2,4,6-
trihydroxybenzaldehyde, 2-(3′,4′-dihydroxybenzoyloxy)-4,6-
dihydroxybenzoic acid, and 3,4-dihydroxyphenylethanol had
been reported (46,47). Compared to the solution, quercetin in
essential oil-based microemulsions was very stable. The
percentages of remained quercetin were respectively 93.7,
93.1, and 96.8% in PO-ME, CO-ME, and RMO-ME, much
higher than quercetin aqueous solution. No significant
difference was found among the three microemulsions.
Hence, the incorporation of quercetin into essential oil-
based microemulsions could significantly improve its stability.
The presence of essential oil might play an important role in
avoiding quercetin degradation.
Fig. 7. In vitro permeation profiles of quercetin solution and
microemulsions through rat skin
Essential Oil-Based Microemulsions of Quercetin
In Vitro Skin Permeation
An essential requirement to evaluate the effectiveness of
topical formulations is the permeation capacity through the
skin. In this work, we applied the Franz diffusion cell method
to evaluate the permeation capacity of essential oil-based
microemulsions through rat skin. The area of skin contacting
with the receptor chamber was 0.636 cm2, and the total
volume of the receptor phase was 5.2 mL. SMEDDS
containing 5 mg/g quercetin was diluted 20 times and the
formed microemulsions were added into the donor chamber
with the volume of 0.3 mL, which contained 75 μg quercetin.
For PO-ME, CO-ME, RMO-ME, and the aqueous solution,
the cumulative amount of quercetin permeated through rat
skin is listed in Fig. 7. The amount of permeated quercetin
was gradually increased with the diffusion time. The
cumulative amount of quercetin reached to 2.24 ± 0.35 μg/
cm2 for the aqueous solution, while it reached to 8.54 ± 0.55,
7.98 ± 0.57, and 9.12 ± 0.68 μg/cm2 for PO-ME, CO-ME, and
RMO-ME. Hence, it was obvious that the essential oil-based
microemulsions could significantly improve the skin perme-
ation of quercetin, about 2.5–3 times higher than the aqueous
solution. The permeation enhancement should be attributed
to the combination effect of surfactant and essential oils. The
enhanced skin permeation indicated that the essential oil-
based microemulsions could bring more benefits to the skin,
compared to the aqueous solution.
CONCLUSION
In topical application, both quercetin and essential oils
have many biological activities to protect the skin. In this work,
three kinds of essential oil-based microemulsions containing
peppermint oil (PO-ME), clove oil (CO-ME), or rosemary oil
(RMO-ME) were prepared, for topical delivery of quercetin.
Pseudo-ternary phase diagram demonstrated that essential oils,
Cremophor EL, and 1,2-propanediol could form the
microemulsion region depending on the Smix/oil ratio (at least
5:5). Moreover, the particle sizes of microemulsions were
gradually increased with the decrease of Smix/oil ratio. They
formed microemulsions with similar particle sizes (9–13 nm)
with the Smix/oil ratio of 7:3. Solubility study showed that
SMEDDS and the components can significantly improve the
solubility of quercetin. Despite the solubilizing capability
decreased as SMEDDS diluted, they were still dozens of times
higher than water. In pH stability study, quercetin was found
instable under alkaline condition, especially with the pH of 13.
However, the essential-based microemulsion could protect
quercetin from degradation, and the protective effect of CO-
ME and RMO-ME was better than PO-ME. In the
photostability study, we found that the essential oil-based
microemulsions could protect quercetin from degradation under
UV radiation. Where more than 67% of quercetin was degraded
in aqueous solution, while less than 7% of quercetin degraded in
microemulsions. At last, the in vitro skin permeation was
preformed, founding that the essential oil-based microemulsions
could enhance the permeation capacity of quercetin by 2.5–3
times compared to the aqueous solution. Hence, the prepared
essential oil microemulsions could improve the solubility, pH
stability, photostability, and skin permeation of quercetin, which
will be benefit for its topical application.
ACKNOWLEDGEMENTS
This work was supported by the National Science
Foundation of China (Grant No. 81601825), Program for
Science and Technology Project of Liaoning Province (Grant
No. 201601237), and Educational Committee Foundation of
Liaoning Province (Grant No. L2016026, L2015150).
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