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To cite this article: Hadel A. Abo Enin (2015) Self-nanoemulsifying drug-delivery system
for improved oral bioavailability of rosuvastatin using natural oil antihyperlipdemic, Drug
Development and Industrial Pharmacy, 41:7, 1047-1056
Article views: 75
Download by: [University of Wisconsin - Madison] Date: 14 September 2015, At: 06:41
http://informahealthcare.com/ddi
ISSN: 0363-9045 (print), 1520-5762 (electronic)
RESEARCH ARTICLE
Abstract Keywords
Aim: The aim is improving the antihyperlipidemic activity of Rosuvastatin Calcium (Rs) through Antihyperlipidemia, natural oil, rosuvastatin
improving its solubility using self-nanoemulsifying drug delivery system (SNEDDS) containing calcium, self-nanoemulsifying drug
natural oil full of unsaturated fatty acid and omega 3. delivery system
Methods: A 7 32 full factorial design was adopted for optimization of oil ratio, Surfactant:
Co-surfactant (S:CoS) ratio and oil:S/CoS ratio. Ternary phase diagrams were constructed for History
optimizing the system with drug loading (10 and 20%). The optimized SNEDD systems were
evaluated according to their physical evaluation and drug release. Furthermore, the anti- Received 30 April 2014
hyperlipidemia efficacy was compared with commercially marketed product on rates followed Revised 25 September 2014
by clinical study. Accepted 24 October 2014
Results: The system containing Tween 80:PEG 400 (3:1) and olive oil:garlic oil (1:1) as an oily Published online 18 November 2014
phase has droplet size less than 100 nm, ZP (+23.43 ± 2.58 mV), PDI (50.02) and cloud point
(490 C). In vitro drug release studies showed remarkable enhancement of the Rs release from
Rs-SNEDDS. The antihyperlipidemic effect of Rs-SNEDDS is greater than that of the commercial
tablets and the pure drug on rates and in hyperlipidemic patients.
Conclusion: Rs-SNEDDS is a promising drug delivery system for improving the drug solubility
and antihyperlipidemic effect using natural oils as (olive oil and garlic oil).
alcohols showed detergent activity that can disrupt the cell Screening of oils and surfactants and co-surfactants
membranes. Also, oleic acid in high concentration can cause
To evaluate the oil and surfactant capability to form an emulsion
excessive and continued inflammatory reaction by promoting the
spontaneously, oil was added to surfactant with vigorous vortex in
secretion of pro-inflammatory16,17.
the 1:1 (w/w) ratio. The mixtures were heated at 40–50 C. Fifty
Thus, introducing of Rs in the SNEDDS using natural oil as
milligrams oil–surfactant mixtures were diluted with distilled
olive oil, fish oil, linoelic acid, nigella oil and garlic oil was a
water in a Stoppard conical flask to 50 ml. The number of flask
promising. These oils are rich in unsaturated fatty acid and omega
inversions (time of emulsification) required to yield homogeneous
3. They also have a significant reduction in total cholesterol (TC)
emulsion indicates the ease of emulsification. Emulsions were
averaging 10%18–24. For example, an olive oil intake even with
kept standing for two hours and their percentage transmittance
high fat diet simultaneously decreases the cholesterol and has a
was evaluated at 638.2 nm using distilled water as a blank.
beneficial effect on the progression of hypercholesterolemia25.
Various co-surfactants were screened by mixing with surfactant
Also, garlic oil significantly inhibited the hypercholesterolemia,
in 2:1 (w/w) ratio. Oily phase was added to this mixture in 1:1
decreased tissue cholesterol and minimized the atheromatous
(w/w). The mixture prepared and evaluated as previously
changes in the aorta26.
reported28,30.
The aim of this study is to develop and characterize SNEDDS
formulations containing rosuvastatin calcium using natural oil
Formulation optimization of a blank SNEDDS
full of unsaturated fatty acid and omega 3. An efficient self-
nanoemulsifying system for Rs was selected and optimized using The blank SNEDDS formulations were screened by the factorial
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solubility tests and phase diagram construction. The formulations design using olive oil and garlic oil as an oil phase, T80 as a
were characterized by the evaluation of the self-emulsification surfactant and PEG 400 as a co-surfactant. Oil, surfactants and co-
performance, droplet size analysis and in vitro drug release surfactants were mixed at prefixed ratios in tubes with vigorous
characteristics. Finally, the antihyperlipidemic effect of Rs in vortex. A certain amount of the mixtures was added to distilled
SNEDDS was evaluated on rats and human. water (v/v, 1:100). Four experiments were then performed for
each combination. The solubility test of the samples, emulsifica-
tion rate, turbidity and the mean particle size using a photon
Materials and methods
correlation spectroscope were determined. All measurements
Materials were done in triplicate.
Rs was purchased from Merck (Barcelona, Spain). Gelatin
Construction of ternary phase diagram
capsules, No. 3, were kindly supplied by Arabia gelatinea,
Egypt. All other chemicals, Tween 80 (polyoxyethylene sorbitan The self-emulsifying region of the selected system with different
monooleate), Tween 20 (polyoxyethylene sorbitan monolaurate), drug loading was identified from ternary phase diagrams. The
PEG 200 and PEG 400 (poly ethylene glycol) were obtained from system composed of olive:garlic ratio 1:1 and T80:PEG 400 3:1
Merck, Mumbai. Cremophor El, propylene glycol and oils (Olive was further studied. A series of self-emulsifying systems with
oil, Fish oil, Linoelic acid, Nigella oil and Garlic oil) were varying oil phase and S/CoS concentrations were prepared. Rs
obtained from Sigma (St. Louis, MO), Fine-Chemicals Limited, was added in each system in different concentration (10–20% w/
Mumbai. Other chemicals were in analytical grade. w). The samples were then vortexed and sonicated until oily liquid
mixtures were obtained. Each sample was then added into
Methods distilled water (v/v, 1:100) drop by drop. The self-emulsifying
performance was visually observed of the generated samples.
Determination of rosuvastatin calcium (Rs)
Samples that showed any cracking, drug precipitation or form
Rs was determined by a modified validated high-pressure liquid immediate coalescence of oil droplets when stirring were rejected.
chromatography (HPLC) on an Agilent 1100 HPLC system with Phase diagrams were plotted using the above criteria by Chemix
an ultraviolet detector set at wavelength 242 nm. Separation was software version 3.5131 (Minnesota, MN).
achieved using a mobile phase consist of acetonitrile–water
(40:60, v/v) solution at a flow rate of 1.5 ml/min. Rs was
Characterization and evaluation of SNEDDS
separated by C8 column (HypersilÕ BDS C8 analytical column).
The column was maintained at ambient temperature and injection The mean droplet size and Zeta potential determination
volume was 20 mL. The mobile phase was filtered through
The mean droplet size was measured by Laser Diffractometer
0.45 mm filter before using. The linearity ranged from 0.01 to
(LD) Mastersizer 2000 Version 2.00 (Malvern Instruments,
10 mg/mL with a correlation coefficient 0.9996. Inter-day and
Malvern, UK). The data were presented in terms of median
intra-day precisions were done as per ICH guidelines for 3 days.
(D50) volume and the span, which were calculated by Malvern
All precision were below 2%27.
Software Version 3.0. For measurement, SNEDDS were analyzed
by dispersing it in 100 ml distilled water under slow agitation at
Solubility studies
room temperature 25 C. All systems’ analysis was done in
The solubility of Rs in various buffer media, oils, surfactants (S) triplicate.
and co-surfactants (CoS) was determined as in a previously The zeta potential z of the diluted SNEDDS formulations was
reported method28. An excess amount of Rs was added into each measured using a Malvern zetasizer (Malvern, UK). The
vial containing 5 ml of the selected vehicle. Then, the mixture was SNEDDS were diluted with distilled water (1:2500 v/v) and
heated at 40 C in a water bath to facilitate the solubilization. The mixed for one minute using a magnetic stirrer. Zeta potential of
systems were shaken at 25 C for 48 h. After equilibrium, each SNEDDS was determined in triplicate32.
centrifugation of the mixtures was done at 1000 rpm for 10 min
and followed by filtration through a 0.45 -mm Millipore membrane Cloud-point measurement
filter paper. The supernatant was diluted with methanol. The
dissolved Rs in various vehicles were quantified by a previously The selected formulations were assessed for cloud-point value.
validated HPLC method29. The formulations were diluted with distilled water (1:100) and
DOI: 10.3109/03639045.2014.983113 Nanoemulsifying system of natural antihyperlipdemic oil 1049
placed in a water bath with gradual increase in temperature. related diseases recorded in pharmacological method (Third
The drop in the sample percentage transmittance from zero point edition)37.
(the cloud point) was measured spectrophotometrically at Eighteen days after the start of the study (after insuring
638.2 nm33,34. of NAFLD case), two positive control groups were used [Group 1
(S/CoS) and oil (olive oil:garlic oil, 1:1) in Group 2]. Group 3
Physical robustness to dilution received Rs with the dose 10 mg/kg/day. The commercial market
tablets were tested in Group 4 (10 mg/kg/day). The selected
Robustness of the selected formulations to dilution was assessed
formula was tested in Groups 5. Each animal was received a drug
in the absence and the presence of Rs (10 or 20%). The formulae
equivalence to 10 mg/kg/day. Animals received corresponding
were exposed to different dilution folds (100, 500 and 1000) using
treatments every day for another 24 days with continuous feeding
different media as distilled water, 0.1 N HCl and phosphate buffer
with high fat diet. All rats were fasted for two hours every day
pH 6.8. Percentage transmittance was then measured spectro-
before the therapeutic agent administration.
photometrically at 650 nm. The diluted nanoemulsions were
stored for six hours and monitored for any physical changes
Assessment of total cholesterol, triglyceride, lipoprotein and liver
(such as phase separation or precipitation)33,35.
marker enzyme in blood
Transmission electron microscopy Samples of blood 1.5–2 mL in volume were collected from the
retro-orbital venous plexus once every 6 days throughout the
Transmission electron microscopy (TEM) was employed as a
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Results and discussions other hand, linoleic acid showed poor emulsification efficiency
with T80 as it requires a higher number of flask inversions
Solubility study (screening of oil)
(Table 2).
The SNEDDs mainly consists of oil, surfactant, co-surfactant and It has been reported that the well-formulated SNEDDS is
the drug. All components should produce a clear monophonic dispersed within seconds under gentle stirring conditions which
solution when introduced in an aqueous phase at ambient finally depends on the emulsification ability of the used
temperature to allow the presentation of the drug in solution39. surfactant42. The lower emulsification capacity of T20 than T80
Solubility studies were aimed at identifying a suitable oily phase, was due to its HLB value (16.7). It has a more hydrophilic nature;
surfactant and co-surfactants for the Rs-SNEDDS develop. It also therefore, it has the minimum oil solubilization capacity. The
used to determine the most suitable media for in vitro release solubilization capacity of oils increases as the hydrophobic chain
study. Identifying the suitable oil having the maximal solubilizing length of surfactants increase (more hydrophobic)43. In addition,
potential for the drug under investigation is very important to T80 has a biological activity and, less affected by pH change in
achieve optimum drug loading40. The solubility of Rs in various gastrointestinal drug to facilitate the release profile of Rs44. The
oily phases and 10% (w/w) surfactant or co-surfactant solutions is aforementioned results suggested that, the use of garlic oil and
presented in Table 1. olive oil as an oily phase with T80 as a surfactant is a promised for
From the various oily phases that were screened, linoleic this study.
acid improves the maximum solubilizing potential. The solubil-
ity of Rs in it was about (25.026 ± 1.23 mg/mL). The solubility Preliminary screening of co-surfactants
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led to the system is less stable due to its intrinsic high aqueous will lead to decrease the nanoemulsion area in the phase diagrams
solubility which lead to an increase in droplet size47,48. Hence, (Figure 1a and b). The aim of optimization of the pharmaceutical
the selected surfactant to co-surfactant ratio would be 3:1. The preparation was the determination of the best required criteria.
saturated solubility of the drug also increases with increase the The highest drug solubility and the highest oil phase concentra-
T80 ratio which related to the solubility test results. Also, increase tion are required, in addition, the most stable and the smallest
the olive oil ratio than garlic oil required increase of the T80 ratio. particle size. Based on the results shown in Figure 1, S1, S2, S3
This may be due to an olive oil which is composed of oleic acid and S4 were chosen for further studies (Table 4). They have a
(monounsaturated fatty acid) has up to the 18-carbon (medium large amount of oil ratio, the smallest particle size and the most
chain carbon length)49. It was reported that the medium chain stable self-nanoemulsifying systems.
carbon length with a high HLB value produces more stable
SNEDDS (small particle size and low turbidity)50. Therefore, In vitro characterization of optimized SNEDDS
using S:CoS (3:1) ratio, the oil ratio 1:1 is the most promising.
Droplet size and zeta potential determination
Another reason for selecting this combination of S/CoS was
the presence of a lower amount of PEG 400. Mei et al.51 reported Droplet size distribution is a critical factor to evaluate a self-
that the possibility of instability of SNEDDS formulations which nanoemulsion system. The particle size decrease gives the
are intended to be filled in hard gelatin capsule shells when they meaning of enhancing the drug bioavailability. The mean
contain PEG 400 at high concentrations. The selected system was globule size of the selected Rs-SNEDDS formulations was
stable even with increasing oil ratio and produce high drug shown in Table 4. It is clear all formulae had an acceptable
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solubility with smaller particle size. nanoparticle size ranged from 50 to 100 nm. This nanoparticle
size indicated the ability of the present formula to offers
Construction of ternary phase diagram large interfacing surface area needed for drug absorption52.
Smaller values of the polydispersity index (PI50.2; within
The pseudo ternary phase diagram of RS-SNEDDS in Figure 1
the acceptable limits) were used as a measure of homogeneity
indicates the nanoemulsion region. Increasing the drug loading
(Table 4). Formula S4 had the lowest mean particle size
(68 ± 4.11 nm; Figure 2a). The results showed a significant
difference between the different formulae.
Table 3. The factorial 7 32 design for formulation of the different The optimized Rs-SNEDDSs showed a significant different
SNEDDS systems were prepared using mixtures of garlic oil and olive oil zeta potential’s value as shown in Table 4. The zeta value and the
as an oily phase in different ratio.
positive charge of increases as the concentration of the drug
increase. This may be due to increasing the Ca ion concentration
Oil mixture (olive oil:garlic oil)
(rosuvastatin calcium). It was reported that the emulsion stability
Oil ratio:S/CoS mix is directly related to the magnitude of its’ surface charge53. In
in the system A (1:1) B (1:2) C (2:1) S*:CoS** general, an increase of electrostatic repulsive forces between
Low (10:90) 10A 10B 10C 1:1 nanoemulsion droplets prevents the droplets’ coalescence. On the
1:2 other hand, a decrease of electrostatic repulsive forces will cause
1:3 phase separation. This indicated that the formulation would
Medium (30:70) 30A 30B 30C 2:1 remain physically stable in storage. Also, the formulation’s zeta
2:3 potential positive charge undergoes an electrostatic interaction
High (50:50) 50A 50B 50C 3:1
3:2 with the mucosal surface of the intestine (mucus layer in the
lumen carries a negative charge) which would lead to enhance the
*S: Tween 80. **CoS: PEG 400 oral bioavailability of the drug54.
Figure 1. The pseudo ternary phase diagram of nanoemulsion region(Rs-SNEDDS). (a) 10% Rs drug loading. (b) 20% Rs drug loading.
1052 H. A. A. Enin Drug Dev Ind Pharm, 2015; 41(7): 1047–1056
Table 4. The composition of the chosen formulae and their evaluation results.
Formulae S1 S2 S3 S4
Composition (W/W) 10% oil 10% oil 20% oil 20% oil
87.6% S/CoS 86.6% S/CoS 77.6% S/CoS 76.6% S/CoS
1.4% water 1.4% water 1.4% water 1.4% water
10 mg drug 20 mg drug 10 mg drug 20 mg drug
Emulsification rate 3 3 3 3
*Solubility% 50.21 ± 2.01 49.71 ± 3.12 53.65 ± 4.08 59.87 ± 3.54
*Mean particle size (nm) 73.68 ± 1.02 75.70 ± 2.01 69.1 ± 1.80 68 ± 4.11
*Polydispersity index (PI) 0.099 ± 0.014 0.1175 ± 0.024 0.1325 ± 0.03 0.2004 ± 0.02
Turbidity after dilution with water (1:100) (NTU) 4.65 11.465 3.58 1.23
*Zeta potential (mV) +15.65 ± 1.71 +16.43 ± 1.76 +20.65 ± 1.51 +23.43 ± 2.58
Mean Globule size (nm)
Physical robustness
*Water (100) 234 ± 23.81 103 ± 2.41 209 ± 6.98 87 ± 1.11
*Water (500) 571 ± 11.87 105 ± 1.44 611 ± 10.45 90 ± 0.99
*Water (1000) 843 ± 45.88 103 ± 1.45 798 ± 5.31 88 ± 1.02
*0.1 N HCl (1000) 914 ± 23.65 103 ± 3.81 960 ± 19.7 88 ± 2.43
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*Phosphate buffer pH 6.8 (1000) 886 ± 33.13 103 ± 8.36 988 ± 21.11 89 ± 1.23
Cloud-point ( C) 60.98 77.85 93 93.8
% Transmittance 93.14–28.14 91.34–23.64 90.67–44.23 –
Cloud-point measurement
The cloud point is the temperature above which the formulation
clarity turns into cloudiness. At the cloud point, dehydration of
polyethylene oxide moiety of the non-ionic surfactant, phase
separation and a drop in sample percentage transmittance would
occur. Therefore, the drug solubility and the formulation stability
will decline. Therefore, the cloud point of the formulation should
be higher enough than 37 C (body temperature)35. Different
factors affect on the cloud point value such as drug hydropho-
bicity and mixing ratio of oils, surfactant and co-surfactant used55.
In our study, all formulations were stable as the formed clouds
were reversible after a few minutes. S2 and S4 exhibited the
highest cloudiness point at 93 and 93.8 C, respectively (Table 4).
There was no significant difference between both results. The
%transmittance dropped from 90.67% to 44.23% in S2. Hence, in
S4, the cloudiness did not persist enough time to be measured.
The results confirmed that the Rs-SNEDDS formula (S4) is stable
regarding the separation at gastrointestinal tract temperature.
Figure 3. The in vitro release study of the prepared formula S4 (Rs-SNEDDs equivalent to 10 mg rosuvastatin calcium) was compared in this study
with respect to the plain drug and the commercial product (10 mg rosuvastatin calcium) in 0.1 N HCl media and in phosphate buffer pH 6.8.
by a thick monolayer. This result was explained by the previously and alanine aminotransaminase (ALT) blood levels were
reported that reducing the interfacing energy, and barrier coales- evaluated on days 0, 18 and 42 (data not shown). No change or
cence forming result from the formation of monolayer around the even increases in AST or ALT levels were observed, which
emulsion droplets56. indicating no liver damage will occur during the research period.
The decrease in the raised levels of TG, TC, LDL-C and HDL-
In vitro dissolution studies C were illustrated in Figure 4. TC, TG, HDL-C and LDL-C levels
in all groups (G2–G5) were significantly lower than in group B
The in vitro release study of the prepared formula S4 was
after treatment (except group 1). Group 1 shows no significant
compared in this study with respect to the plain drug and the
difference than the GpB. The control and all other groups showed
commercial product (Figure 3). The Rs-SENDDs have a higher
similar AST and ALT levels, indicating the normal liver function
dissolution rate than the plain drug and the marketed product in
(data not shown).
0.1 N HCl media and in phosphate buffer pH 6.8. After 10 min,
At the end of the experiment, TG, TC, LDL-C and HDL-C
more than 85% of the drug was released from the prepared
lowering effects were the greatest in Group 5 (Rs-SNEDDS
formula while the plain drug released 530% in SGF and 40% in
group), even after one week treatment. The TG, TC, LDL-C and
SIF after 60 min. The statistical study of the dissolution result of
HDL-C levels in Group 5 were dropped to 188.0 ± 50.93,
Rs-SENDDS (S4) did not show any significant difference between
87.94 ± 40.14, 89.70 ± 96.70 and 82.04 ± 10.14 mg/dL, respect-
SGF and SIF (p ¼ 0.324). By that comparison, the Rs-SNEDDS
ively, which was significantly lower than that of model group
completely dissolved in 10 min without pH influence. That can be
(Group A; p50.001). The rats given market product, oil or even
explained by the drug in both media was not only in free form, but
the drug showed higher results than those gives Rs-SNEDDS at
also entrapped in nanoemulsion particles (micelles form) which
the end of the study (Groups 2, 3 and 4; p50.05).
helps in increasing the rate of the drug release through two
The results of the statistical analysis revealed that the
reasons. First, the very small particle size (nanoparticle) helps to
formulation had a significant effect on all the tested parameters
disperse rapidly in aqueous media. The other is the drug particle
at p50.05 (F5, 17; n ¼ 11.424, 6.469, 2.002 and 43.890 for TC,
dispersal in the interface formed by the surfactant and co-
TG, LDL-C and HDL-C, respectively). On the other hand, for all
surfactant.
the tested parameters, there was no significant difference between
the subjects (rats).
Antihyperlipidemic effect of the selected SNEDDS
Multiple comparisons using the Tukey’s HSD test revealed that
The antihyperlipidemic effect of the selected formula was TG, TC, LDL-C and HDL-C results in all groups extremely
represented in Figure 4. The TG and TC contents in Group B differed significantly from each other with the lowest value
were higher than those of Group A on day 6. The TG and TC observed for Rs-SNEDDS followed by the market product, drug
contents of Group B were increased to the highest level on day 24, and finally the oil.
359.9 ± 61.90 mg/dL and 300.258 ± 38.79 mg/dL, respectively, In our previous studies, Rs-SNEDDS showed more pro-
almost three times than the control group (72.97 ± 3.12 and nounced effects on TG and TC regulation. The significant
64.63 ± 5.21) for TG and TC, respectively. decrease in the TG, TC, LDL-C and HDL-C in the Group 5 than
Also, LDL-C and HDL-C contents in Group B were higher Group 3 or Group 4 (drug and market product, respectively) was
than in Group A from the day 6 and through the experiment. At for many reasons. The first was poor oral absorption of the drug
the end of the study, LDL-C and HDL-C were 260.97 ± 70.74 and and the market product (low solubility and large particle size)
200.74 ± 67.28, respectively. Aspartate aminotransaminase (AST) compared to Rs-SNEDDs. The other is using an oil mixture (olive
1054 H. A. A. Enin Drug Dev Ind Pharm, 2015; 41(7): 1047–1056
oil and garlic) in Rs-SNEDDS. These oils were rich in unsaturated Clinical study
fatty acid and omega 3 which have a main effect on the TG and
In Table 5, the mean cholesterol levels, LDL levels and the TG
TC7,8. Also, allicin and diallyl sulfide in the garlic oil have a
levels decreased while the mean HDL levels also increased after
reducing effect on LDL and HDL-C levels than pure drug can be
treatment of the patients by Rs-SNEDDS and the market product.
attributed to their antioxidant effect24,57.
DOI: 10.3109/03639045.2014.983113 Nanoemulsifying system of natural antihyperlipdemic oil 1055
Table 5. The antihyperlipidemic effect after oral administration of drug 3. Ridker PM, Danielson E, Gotto AM, et al. Rosuvastatin to prevent
in suspension and Rs-SNEDDS (S4) to hyperlipidemic patient vascular events in men and women with elevated C-reaction protein.
(Mean ± SD, n ¼ 30). N Engl J Med 2008;359:2195–207.
4. Michael S. Chemical pharmacokinetic and pharmacodynamic
Mean difference properties of statins: an update. Fundam Clin Pharmacol 2004;19:
117–25.
Variable RS-SNEDDS Commercial tabletsÕ p Value 5. Shah SR, Parikh RH, Chavda JR, Sheth NR. Self-nanoemulsifying
drug delivery system of glimepiride: design, development, and
TC 16.53 ± 1.56 5.31 ± 1.11 50.01
optimization. PDA J Pharm Sci Technol 2013;67:201–13.
TG 39.98 ± 5.46 19.89 ± 2.87 50.01
6. Porter CJ, Trevaskis NL, Charman WN. Lipids and lipid-based
LDL 2.64 ± 0.98 1.056 ± 0.57 50.01
formulations: optimizing the oral delivery of lipophilic drugs. Nat
HDL 9.08 ± 1.08 3.56 ± 1.98 50.01
Rev Drug Discov 2007;6:231–48.
LDL/HDL ratio 2.45 ± 2.01 2.81 ± 2.14 50.01
7. Iosio T, Voinovich D, Grassi M, et al. Bi-layered self-emulsifying
pellets prepared by co-extrusion and spheronization: influence of
formulation variables and preliminary study on the in vivo absorp-
tion. Eur J Pharm Biopharm 2008;69:686–97.
A statistical significant difference between these two groups was 8. Myoung KB, Jong HL, Young HC, et al. Self-microemulsifying
also found for mean change p50.01. drug-delivery system for improved oral bioavailability of pranlukast
In contrast to the above findings, the decrease in the same hemihydrate: preparation and evaluation. Int J Nanomedicine 2013;
parameters was highly significant (p50.01) when using Rs- 8:167–76.
9. Jyothi BJ, Sreelakshmi K. Design and evaluation of self-nanoemul-
Downloaded by [University of Wisconsin - Madison] at 06:41 14 September 2015
25. Jain RC, Konar DB. Effect of garlic oil in experimental cholesterol 42. Jaydeep P, Garala K, Anjali P, et al. Design and development of a
atherosclerosis. Atherosclerosis 1978;29:125–9. self-nanoemulsifying drug delivery system for telmisartan for oral
26. Saez Lancellotti TE1, Boarelli PV, Romero AA, et al. Semen quality drug delivery. Int J Pharm Investig 2011;1:112–18.
and sperm function loss by hypercholesterolemic diet was recovered 43. Mehta SK, Kaur G, Bhasin KK. Tween-embedded microemulsions–
by addition of olive oil to diet in rabbit. PLoS One 2013;8:e52386. physicochemical and spectroscopic analysis for antitubercular drugs.
27. Kaila HO, Ambasana MA, Thakkar RS, et al. A new improved RP- AAPS PharmSciTech 2010;11:143–53.
HPLC method for assay of rosuvastatin calcium in tablets. Indian J 44. Xianyi S, Juan W, Yanzuo C, Xiaoling F. Self-microemulsifying
Pharm 2010;72:592–8. drug-delivery system for improved oral bioavailability of probucol:
28. Borhade V, Pathak S, Sharma S, Patravale V. Clotrimazole preparation and evaluation. Int J Nanomedicine 2012;7:705–12.
nanoemulsion for malaria chemotherapy. Part II: stability assess- 45. Chen ML. Lipid excipients and delivery systems for pharmaceutical
ment, in vivo pharmacodynamic evaluations and toxicological development: a regulatory perspective. Adv Drug Deliv Rev 2008;
studies. Int J Pharm 2012;431:149–60. 60:768–77.
29. Persson LC, Porter CJ, Charman WN, Bergström CA. 46. Borhade V, Pathak S, Sharma S, Patravale V. Clotrimazole
Computational prediction of drug solubility in lipid based formu- nanoemulsion for malaria chemotherapy. Part I: preformulation
lation excipients. Pharm Res 2013;30:3225–37. studies, formulation design and physicochemical evaluation. Int J
30. Sara Z, Jingyuan W, Raid GA. Formulation and physicochemical Pharm 2012;431:138–48.
characterization of Imwitor 308 based, self microemulsifying drug 47. Lu Y, Wu K, Li L, et al. Characterization and evaluation of an
delivery system. Chem Pharm Bull 2010;58:1332–8. oral microemulsion containing the antitumor diterpenoid com-
31. Jinjie Z, Qiang P, Sanjun S, et al. Preparation, characterization, and pound ent-11alpha-hydroxy-15-oxo-kaur-16-en-19-oic-acid. Int J
in vivo evaluation of a self-nanoemulsifying drug delivery system Nanomedicine 2013;8:1879–86.
(SNEDDS) loaded with morin-phospholipid complex. Int J
Downloaded by [University of Wisconsin - Madison] at 06:41 14 September 2015