Content uploaded by Somnath Vibhute
Author content
All content in this area was uploaded by Somnath Vibhute on Jul 21, 2015
Content may be subject to copyright.
Available online at www.scholarsresearchlibrary.com
Scholars Research Library
Der Pharmacia Lettre, 2015, 7 (3):7-16
(http://scholarsresearchlibrary.com/archive.html)
ISSN 0975-5071
USA CODEN: DPLEB4
7
Scholar Research Library
Design and characterization of Moringa oleifera seed oil impregnated anti-
inflammatory topical micro-dispersion
Somnath Vibhute*, Veena Kasture, Sanjay Kasture, Prakash Kendre, Shruti Rupnar
and Vishal Pande
Sanjivani College of Pharmaceutical Education and Research, Kopargaon, Dist: Ahmednagar (M. S.), India
______________________________________________________________________________________________
ABSTRACT
Rheumatoid arthritis is one of the most serious non-fatal conditions with significant prevalence around the globe.
Objective: The objective of present work was to design and characterize Moringa oleifera seed oil impregnated
anti-inflammatory topical micro-dispersion. The ternary phase diagrams for microemulsion regions were
constructed using Moringa oleifera seed oil as oil phase, Tween 80 and Span 80 as surfactants. The microemulsion
was characterized for its percentage transmittance, refractive index, pH, globule size, zeta potential, viscosity, in-
vitro drug release, skin irritancy and anti-inflammatory activity. The batch F3 with 25.30% of oil, 59.02 % of S
mix
and 15.68% of water; was found to be more stable over other batches. The percentage transmittance and refractive
index (RI) of formulation was found to be 99.60 and 1.469 respectively; whereas pH and average globule size was
7.1 and 167nm respectively. zeta potential and viscosity of optimized microemulsion was found to be -35.8 and 88.7
cps respectively. The permeability of Moringa oleifera seed oil microemulsion was 76.86% as compared to 54.06%
of pure oil. The prepared microemulsion was found to be non-irritant and showed significant (p<0.05) anti-
inflammatory effect. The Moringa oleifera seed oil can be used to develop stable micro-dispersion system with better
skin permeation and promising anti-inflammatory activity without any skin irritation.
Keywords: Rheumatoid arthritis, Moringa oleifera seed oil, micro-dispersion, ternary phase diagram, anti-
inflammatory activity.
_____________________________________________________________________________________________
INTRODUCTION
Inflammatory conditions are the common afflictions irrespective of ages. Rheumatoid arthritis (RA) is a systemic
auto-immune disease beginning in the small joints of the hands and the feet, spreading later to the larger joints
involving severe inflammation. The inflamed joint lining or synovium extends and then erodes the articular cartilage
and bone, causing joint deformity and progressive physical disability [1]. It is 31
st
leading cause of disabilities;
Years Lived with Disability (YLD) at global level, accounting for 0.8% of total global YLDs [2]. Numbers of
NSAIDs are commonly used to treat rheumatic conditions and the most widely cited side-effect of NSAIDs
includes, gastrointestinal ulcer, accompanied by anemia due to the bleeding. Hence it makes it desperate to look for
other ways for the management of rheumatism. In order to avoid the gastric irritation, minimize the systemic toxicity
and achieve a better therapeutic effect, one promising method is to administer the drug via skin [3]. Green
alternatives could be the best answer to this quest as anti-inflammatory herbal preparations cause much less adverse
effects. Moringa oleifera Lam widely distributed in tropical and sub-tropical regions and has nutritional and
therapeutic values. Moringa oleifera is traditional medicine used in rheumatoid arthritis[4]. Moringa is rich dietary
source of Omega 3 poly unsaturated fatty acids (PUFA) [5]. A number of studies showed that Omega 3 fatty acid
helps to reduce inflammation and pain related to rheumatoid arthritis[6].
The concept of micro-dispersions(microemulsions) was first introduced by Hoar and Schulman during 1940s. It is
defined as a system of water, oil and amphiphile which is an optically isotropic and thermodynamically stable liquid
Somnath Vibhute et al Der Pharmacia Lettre, 2015, 7 (3):7-16
______________________________________________________________________________
8
Scholar Research Library
micro-dispersion with 10-100nm droplet size. Microemulsion is composed of oil, water, surfactants and co-
surfactants[7].Microemulsions offer several advantages such as transparency, low viscosity, enhanced drug
solubility, good thermodynamic stability, long shelf life, ease of manufacturing and enhancing effect on transdermal
delivery of drug compared to conventional formulations [8].Recently, more attention has been focused on
microemulsions for transdermal delivery of drugs. This is due to their ability to incorporate large amount of drug,
microemulsion components combined synergistically to increase drug flux and enhanced rate of permeation [9].The
use of novel approaches for herbal drugs is need of hour for their overall better performance. In this study, we tried
to develop a new microemulsion formulation using Moringa oleifera seed oil for topical application which may lead
to an improvement permeation and better therapeutic effect. The anti-inflammatory activity was evaluated using
vascular permeability test and carrageenan induced paw edema in rats.
MATERIALS AND METHODS
Materials
Moringa oleifera seeds were procured from local market. Span 80, Tween 80 and other materials used were of
analytical grade and purchased from SD Fine Chemicals, Pvt. Ltd, Mumbai.
Methods
Collection and authentication
Fresh Moringa oleifera seeds were authenticated by Prof. (Dr.) Sandanshiv, Dept. of Botany, S. S. G. M. College,
Kopargaon Dist: Ahmednagar (M.S.)India.
Extraction of oil
Moringa oleifera seeds were air dried for a week and placed in hot air oven at 40°C (Lab Star, Mumbai), ground and
sieved through #40 and #60 to geta coarse powder. Extraction of seeds was carried out by Soxhlet extraction using
petroleum ether (60-80
O
C) as solvent [10].
Preliminary physicochemical study[11]
1. Organoleptic properties
Moringa oleifera seed oil was analyzed for organoleptic properties like color, odor, and appearance.
2. Saponification value
About 2g of oil and 25ml of alcoholic potassium hydroxide was taken in flask. The resultant mixture was heated for
1h; to which 1mlof 1% phenolphthalein was added and titrated with 0.5N HCl.
3. Iodine value
About 2g of oil and 10ml of carbontetrachloride were taken in flask and20ml of Wij’s solution (1.5% iodine
monochloride in 98% acetic acid) was added and kept in dark for 30min. Fifteen ml of potassium iodide and 100ml
of water added in above solution. The resultant solution wastitrated with 0.1M sodium thiosulphate solution using
starch as an indicator.
4. Peroxide value
About 1g of oil,20ml acetic acid-chloroform (2:3), 1g potassium iodide was taken in test tube and boiled. This
mixture was transferred to 20mlof 5% potassium iodide solution. The resultant solution was titrated with 0.1N
sodium thiosulphate solution using starch as an indicator.
5. Identification of oil actives
Moringa oleifera seed oil was run over TLC plate using silica gel-G as stationary phase. After the sample has been
applied on the plate, solvent mixture (known as the mobile phase) is drawn up the plate via capillary action. The
qualitative evaluation of the plate was done by determining the migrating behavior of the separated substances given
in the form of Rf value[12].
Analytical methods
1. UV spectroscopy
The ultra-violet absorption spectrum of Moringa oleifera seed oil in n-hexane was obtained using UV-Visible
spectrophotometer (Shimadzu 1650) in the range of 400-200 nm[13].
Somnath Vibhute et al Der Pharmacia Lettre, 2015, 7 (3):7-16
______________________________________________________________________________
9
Scholar Research Library
1. Infrared spectroscopy
Oil-excipients interaction study
FT-IR spectroscopy used to determine the molecular interaction between oil and excipients. FT-IR measurement of
oil was taken at ambient temperature. The blend of Moringa oleifera seed oil and other excipients (Tween 80 &
Span - 80) used for final formulation were analyzed by FT-IR spectrophotometer (FT-IR 8400-S; Shimadzu, Japan).
Drop of oil and blend placed on the thin polymeric film were scanned over a wave number range of 4000 to 400 cm-
1
in FT-IR instrument and spectral analysis was done[14].
Formulation of Moringa seed oil microemulsion
1. Construction of ternary phase diagram
The ternary phase diagrams were constructed using water titration method[15]to determine the microemulsion
region and to detect the possibility of making microemulsions with different possible compositions of oil,
surfactant/s and water. Various ratios (1:9, 2:8, 3:7… to 9:1) of Tween 80 and Span 80 mixed with oil phase. These
mixtures were titrated with double distilled water by drop wise addition using micropipette and stirred on magnetic
stirrer at room temperature. After each addition, the system was examined for the appearance and flow properties.
The end point of the titration was the point where the solution becomes cloudy or turbid. The quantity of the
aqueous phase required to make the mixture turbid was noted. The microemulsion region was identified as
transparent and isotropic mixture. All these value substituted in CHEMIX School Software and ternary phase
diagram was constructed by plotting amount of oil, surfactant, and water phase combination.
2. Formulation of microemulsion
Appropriate quantities of Tween 80, Span 80 and Moringa oleifera seed oil were weighed mixed in glass beaker to
make transparent mixture and vortexed for 15min. The appropriate amount of double distilled water was added with
help of micropipette followed by stirring on magnetic stirrer at room temperature to obtain microemulsion system.
While formulating the microemulsion the surfactant blend of 83% Span 80 and 17% of Tween 80 (S
mix
) was taken
by considering the required HLB for the preparation. Different compositions as shown in Table 1;were used to
develop microemulsion formulation using Moringa oleifera seed oil.
Table 1: Formulation of microemulsion using Moringa oleifera seed oil
Sr. No.
Batch No.
Oil (%)
S
mix
(%)
Water (%)
1
F1
8.57
77.12
14.31
2
F2
17.16
68.66
14.16
3
F3
25.30
59.02
15.68
4
F4
33.81
50.71
15.46
5
F5
44.17
44.17
11.66
6
F6
57.58
38.38
4.03
7
F7
65.54
28.08
6.37
8
F8
73.87
18.47
7.66
9
F9
82.72
9.19
8.09
Characterization of prepared herbal microemulsion
1. Thermodynamic stability
To overcome the problem of metastable formulation, thermodynamic stability tests were performed in following
manner.
a. Heating-Cooling Cycle: Six cycles were carried out between temperature 4
o
C and 45
o
C for 48h and the
formulations were observed for phase separation. Formulations that did not show any phase separation were taken
for centrifugation test[16].
b. Centrifugation Test: Formulations were centrifuged at 3000 rpm for 30 min. The formulations with no phase
separation were used in a Freeze–Thaw Cycle Stress Test[17].
c. Freeze–Thaw Cycle Stress Test: The formulations were subjected to a total of three complete freeze–thaw
cycles, each cycle consisting of 24 h at 25°C followed by 24 h at −5°C[18].
The formulations that survived thermodynamic stability tests as described earlier were selected for the further
characterization.
2. Globule size andZeta Potential
Average globule size, Zeta Potential and Polydispersity Index of microemulsion were determined by using dynamic
light scattering Malvern Zetasizer Nano-ZS90 (Malvern Instruments, UK). Sample was prepared by dissolving 1g of
sample in 10ml of dispersion medium i.e. distilled water in glass test tube. The sample was loaded into transparent
cuvette having volume of 1cm
3
in the thermostatic chamber at 25
o
C. Laser light scattering was monitored at fixed
angle 90
0
.
Somnath Vibhute et al Der Pharmacia Lettre, 2015, 7 (3):7-16
______________________________________________________________________________
10
Scholar Research Library
3. Transparency and Refractive Index
Transparency and homogeneity of microemulsion was determined by measuring the percentage transmittance
at650nm by UV spectrophotometer against water as reference. The Refractive Index of pure oil and microemulsion
was determined using an Abbe’s refractometer at 25±0.5°C. One to two drops of sample applied and reading was
recorded.
4. Determination of viscosity
The viscosity of optimized microemulsion was determined using Brookfield viscometer (LVII, Brookfield Inc.,
USA) using spindle no.4 with shear rate 100 rev/ min. The measurement was done at room temperature[19].
5. pH measurement
The pH of optimized formulation was determined by pH meter (PICO+ Lab India); which was calibrated by using
phosphate buffer solutions i.e. pH 4, 7 and 9 (triple point calibration). The electrode was inserted into the sample 10
min prior to study at room temperature[20].
6. In-vitro permeation study
In-vitro drug release study of microemulsion containing Moringa oleifera seed oil was carried out using modified
Franz diffusion cell with cellulose acetate membrane having 0.45µ pore size and 1mm thickness. The cellulose
acetate membrane was hydrated for 24h prior to study. The membrane was placed between donor and receptor
compartment. The receptor compartment was filled with methanolic phosphate buffer (pH 6.8) and maintained
temperature at 37 ± 0.5°C using a circulating water bath. The receptor cell was stirred with a small magnetic bar at a
500 rpm. The donor compartment was filled with accurately measured 1g optimized microemulsion and pure oil
separately. The amount of drug released from microemulsion and pure oil was analyzed using UV
spectrophotometer (Shimadzu 1650, Japan) at 240 nm[21].
Anti-inflammatory activity
1. Vascular permeability test
Vascular permeability test [22]was performed to find out anti-inflammatory effect of prepared microemulsion
formulation. The Institutional Animal Ethics Committee had approved the experimental protocols for animal study
(IAEC No. 1093/A/07/CPCSEA) and care of animals was taken according to CPCSEA guidelines. Wistar albino rats
of either sex weighing 180–200g were used for this study. The animals were grouped in polyacrylic cages and
maintained under standard laboratory conditions (temperature 25 ± 2 °C) and relative humidity (50±5%) with dark
and light cycle (14/10 h). They were allowed free access to standard dry pellet diet and water ad libitum. The rats
were acclimatized to laboratory condition for 10 days before commencement of experiment. The five groups of
animal were categorized as follows
Group 1: Vehicle (Inflammation control)
Group 2: Piroxicam gel 0.5% topically
Group 3: Moringa oleifera seed oil 0.5% topically
Group 4: Microemulsion 0.5% topically
Group 5: Microemulsion 1.62%, topically
Animals were anaesthetized using 0.1ml of 0.1% chloral hydrate solution given orally. The effect on blood vessels
was observed under motic microscope at 10min time interval for 1h.
2. Carrageenan induced paw edema in rats
Carrageenan induced paw edema method [23] was performed to confirm anti-inflammatory effect of prepared
microemulsion formulation. The Institutional Animal Ethics Committee had approved the experimental protocols
((IAEC No. 1093/A/07/CPCSEA) and care of animals was taken according to CPCSEA guidelines. Anti-
inflammatory activity of prepared formulation was evaluated and compared with marketed preparation. Healthy
Wistar albino rats weighing 180-200g were used in this experiment and animals were starved overnight. Four groups
consisting of five animals were employed in the present study.
Group 1: Vehicle
Group 2: Piroxicam gel 0.5% topically
Group 3: Moringa oleifera seed oil 0.5%topically
Group 4: Microemulsion 1.62% topically
The animals were housed in cage maintain temperature at 25±1°C and 60±5% relative humidity. 0.1ml of 1% w/v
solution of carrageenan in normal saline solution was injected into sub planer tissue of left hind paw of each rat. The
Somnath Vibhute et al Der Pharmacia Lettre, 2015, 7 (3):7-16
______________________________________________________________________________
11
Scholar Research Library
paw is marked with ink at particular level and immersed in mercury up to this mark. Microemulsion, pure oil and
standard formulation were applied topically to the paw of the animals and spreaded gently; before and after 1, 2, and
3 h the injection of carrageenan into left hind of paw. Measurement of foot volume was performed using digital
Plethysmometer.
The % inhibition of edema was calculated as:
% inhibition of edema = (C
t
– C
0
)/C
0
×100
Where,
C
t
= Paw thickness after injection of carrageenan
C
0
= Paw thickness before injection of carrageenan
Skin irritation study
Three healthy albino rabbits of around 18 months age and weighing 1.5-2kg were selected for this study. Dorsal
surface of rabbit was cleaned and hairs were removed by shaving of three different region of dorsal surface prior to
study. Shaved skin was cleaned with rectified spirit. Pure oil, microemulsion were applied on shaved surface area.
The animals were observed for erythema, edema, and inflammation for 72h after application[24].
Accelerated stability study
The optimized microemulsion was stored in closed glass vials at 40°C and 75% RH for three months in stability
chamber. The physical parameters were determined at the end of study and compared with initial results. The
microemulsion was subjected to centrifugation at 3,500 rpm for 30 min at different time intervals to observe phase
separation[25].
Statistical analysis
All data are reported as Mean ±SEM and groups were compared using ANOVA with p < 0.05 considered
statistically significant.
RESULTS AND DISCUSSION
Physicochemical properties of Moringa oleifera seed oil
Table 2: Physicochemical characteristics of Moringa seed oil
Characteristics Observation
% yield of oil in petroleum ether 24%
Color Pale yellow
Odor Characteristic
Refractive index 1.458
Saponification value 195.1mg KOH/g
Iodine value 86.7 gI
2
/100g
Peroxide value 7.52 mEq/Kg
The color and odor of Moringa oleifera seed oil found to be pale yellow and characteristic respectively. The
Refractive Index of M. oleifera seed oil (1.458) was in close agreement with values reported for conventional oils
from soybean (1.466- 1.470) and palm kernel (1.449- 1.451). The high refractive index of this oil seems to confirm
the high number of carbon atoms in their fatty acids [26]. Refractive index also increases as the double bond
increases [27]. The saponification value indicates average molecular weight of fatty acid contents as glyceride in oil.
Generally higher number of saponification value of oil used in manufacturing of soap. Iodine value is reflection of
unsaturated degree of fats and oil and therefore, the high value; indicate high number of unsaturated double bonds.
Peroxide value is an index of rancidity, thus low peroxide value indicates resistance of the oil to peroxidation during
storage. The peroxide value of Moringa oleifera seed oil is low (7.52 mEq/Kg) compared to the maximum
acceptable value of 10 meq KOH/g set by the Codex Alimentarius Commission for groundnut seed oils [28]. The oil
is thus stable and would not easily go rancid.
Analytical methods
UV- spectroscopy
As shown in Fig 1 Moringa oleifera seed oil containing oleic acid exhibited absorption maxima at 240 nm; whereas
palmitic acid and stearic acid showed absorbance at 281 nm and 411 nm respectively in n-Hexane.
Somnath Vibhute et al Der Pharmacia Lettre, 2015, 7 (3):7-16
______________________________________________________________________________
12
Scholar Research Library
Fig 1: UV spectra of M. oleifera seed oil in n-Hexane
Infrared spectroscopy
Fig 2: IR Spectrum of Moringa oleifera seed oil
IR spectral analysis of Moringa oleifera seed oil has shown peak 3200-2800cm
-1
C-H alkane stretch, 1450-1375cm
-
1
CH
3
bending, 1600-1900cm
-1
C-C alkane, 1000-650cm
-1
alkene out of plane bend, 1850-1650cm
-1
C=O carbonyl
stretch.
Fig 3: Ternary phase diagram for Moringa seed oil microemulsion
Construction of ternary phase diagram and microemulsion formulation
Ternary phase diagram constructed using various ratio of Moringa seed oil, Tween 80, Span 80 and water.
Microemulsion existence regions were identified from ternary phase diagram. There was no phase inversion from
water-in-oil to oil-in-water microemulsion observed. Ternary phase diagram with 83% Span 80 and 17% Tween 80
Somnath Vibhute et al Der Pharmacia Lettre, 2015, 7 (3):7-16
______________________________________________________________________________
13
Scholar Research Library
(S
mix
), oil and water with different concentrations is depicted in Fig 3. Batch F3 with maximumwater and more
stability for a week after formulation was selected for further characterization. It was observed that when S
mix
ratio
becomes more lipophilic, the microemulsion zone moved to the lipophilic side of the diagram, vise-versa.
Thermodynamic stability of microemulsions
Visual examination showed that all microemulsion systems were stable after being subjected to heating-
cooling,centrifugation and freeze–thaw cycles.
Globule size and polydispersity indexand zeta potential analysis
Globule size and polydispersity index are important physical characteristics of microemulsion; as they can affect the
stability of microemulsion. Globule size of microemulsion was observed between 10-200 nm ranges. This micro size
is useful for topical drug delivery; as it promotes the skin permeation. The mean diameter of globule denoted by Z-
Average (d. nm) was found to be 167nm. The globule size distribution graph was found to be bell shaped with even
distribution range (Fig 4). The low polydispersity index indicates higher uniformity of particle size in the
formulation. Polydispersity index of Moringa microemulsion of was found to be 0.3; indicating higher uniformity of
globule size in formulation. Zeta potential of optimized microemulsion was found to be in the range of -35.8
indicating no agglomeration and stability of the microemulsion[29].
Fig4: Globule size and size distribution of optimized microemulsion
Transparency
Microemulsion was found to be transparent and homogeneous phase system. The percentage transmittance of
formulation was found to be 99.60. The Refractive Index (RI) of M. oleifera seed oil microemulsion was found to be
1.469 suggesting no change RI value of Moringa seed oil (1.458) after formulating into microemulsion system.
Viscosity
Viscosity is important parameter for topical delivery. Higher viscosity is preferred as it increases residence time but
permeation rate also decreases with increase in viscosity and hence formulation should have moderate viscosity. The
viscosity of optimized microemulsion was found to be 88.7 cps.
pH measurement
The pH deviations may cause irritation to the patient; hence it should be within range of skin pH. The pH of
formulation was found to be 7.1;that is suitable for topical application.
In-vitro permeation study
The drug release study of Moringa oleifera seed oil through cellulose acetate membrane was observed using Franz
diffusion cell. Microemulsion showed higher permeation rate than pure oil. Various factors affecting on
percutaneous penetration study such as oily phase nature, globule size, and composition of microemulsion. Small
globule size of droplets can interact with number of vesicles on fixed area of stratum corneum and increasing
efficacy of percutaneous uptake. The thermodynamic activity in formulation is a significant driving force for the
release and penetration of drug into skin. The drug get released from inner phase to outer phase and then further into
skin[21]. Tween 80 not only acts as surfactant but also act as penetration enhancer[30]. The percentage release of
drug from pure oil and microemulsion was found to be 54.06 and 76.86 respectively.
Somnath Vibhute et al Der Pharmacia Lettre, 2015, 7 (3):7-16
______________________________________________________________________________
14
Scholar Research Library
.
Fig 5: In vitro drug release from Moringa oil and Moringa microemulsion Anti-inflammatory activity
1. Vascular permeability study
Inhibitory effect of Moringa oleifera seed oil, microemulsion, and standard formulation on blood vessels of rat was
observed. The vascular permeability indicates acute phase of inflammation where there is increased vascular
permeability and migration of leukocytes in to the inflamed area occurs. Constriction of blood vessels indicates
reduction in vascular permeability[31]. The microemulsion preparation caused constriction of blood vessels
indicating reduction in vascular permeability and thus anti-inflammatory potential. The results in Fig 6; indicate
anti-inflammatory potential of Moringa seed oil microemulsion.
.
Fig 6: Inhibitory activity of pure oil and microemulsion
Fig7: % inhibition in carrageenan induced paw edema in rats
2. Carrageenan induced paw edema
Carrageenan injected in sub-planter region of hind paw induced inflammation is time dependent. Carrageenan
induced paw edema is standard experimental model for acute inflammation and widely used as a model for the
evaluation of anti-inflammatory activity of drugs. Carrageenan-induced oedema is a biphasic event, with early
hyperemia due to the release of histamine and serotonin and the delayed oedema due to the release of bradykinin and
0
20
40
60
80
100
01234
% drug release
Time (h)
Moringa oil
Moringa
microemulsion
0
20
40
60
80
100
10 20 30 40 50 60
% of inhibition
Time (Min)
Standared 0.5%
Pure oil 0.5%
Microemulsion 0.5%
Microemulsion
1.62%
0
2
4
6
8
10
0123
% inhibition
Time (h)
Vehicle
Standard
Pure oil
Microemulsion
Somnath Vibhute et al Der Pharmacia Lettre, 2015, 7 (3):7-16
______________________________________________________________________________
15
Scholar Research Library
prostaglandin[24].Standard formulation (Piroxicam 0.5%), pure oil, microemulsion decreased inflammation about
29.96%, 35.16% and 46.84% respectively. The microemulsion containing Moringa oleifera seed oil was found to be
superior to standard formulation. Treatment with pure oil and microemulsion significantly inhibited paw edema. The
effect of Moringa seed oil and Moringa microemulsion was compared with standard formulation (Fig 7).
Skin irritation study
Skin irritation study did not show erythema, edema and inflammation on rabbit skin after 72h. Skin irritation study
revealed that optimized formulation wasnon-irritant and non-sensitizing to skin.
Accelerated stability study
Optimized microemulsion was found to be stable over period of three months as there was no significance change in
parameters of microemulsion found (Table 6). There was no phase separation even after centrifuged test.
Table 3: Observations of stability study
Sr. N
o
.
Parameter
0 month
1 month
2 months
3 months
1
Appearance
Clear
Clear
Clear
Clear
2
pH
7.1
7.0
6.9
6.8
4
Viscosity
(cps)
88.7
88.0
87.8
87.2
5
% Transparency
99.60
99.40
98.50
98.
45
CONCLUSION
Topical microemulsion of Moringa oleifera seed oil was successfully developed using widely accepted safe
excipients and reproducible methodology. This microemulsion system has shown acceptable parameters with better
skin permeation as compared to pure oil. Microemulsion shown promising anti-inflammatory activity up to 3h. This
microemulsion proved to be non-irritant to skin and stable over study period.
Acknowledgement
Authors are grateful to All India Council for Technical Education, New Delhi, for providing fund for this study
under Research Promotion Scheme.
REFERENCES
[1] AM Abdel-Nasser; JJ Rasker; HA Valkenburg. Seminars in arthritis and rheumatism, 1997, 27(2), 123-140.
[2] World Health Organization, World Health Report. Reducing Risks, Promoting Healthy Life. Geneva: World
Health Organisation, 2002.
[3] SC McNeill; RO Potts; ML Francoeur. Pharm Res, 1992, 9, 1422-1427.
[4] JF Morton. Econ Bot, 1991, 45, 318–333.
[5] S Lalas; J Tsaknis. Journal of Food Composition and Analysis, 2002, 15, 65-77.
[6] P Delaveau.PlantesMédicinalesetPhytothérapie, 1980, 14(10), 29-33.
[7] TP Hoar; JH Schulman. Nature 1943, 152, 102-103.
[8] J Lawrence; G Rees. Adv Drug Deliv Rev, 2000, 45, 89-121.
[9] S Prajapati; K Shailendra; S Ajit; S Ankush. World J of Pharm and Pharm Sci, 2013, 2, 5935-5947.
[10] EG Bligh; WJ Dyer. Can J BiochemPhysiol, 1959, 37, 911-917.
[11] Association of Official Analytical Chemists (AOAC). Official Methods of Analysis, 15
th
ed., Washington DC,
1990, pp. 955-972.
[12] J Mendham; RC Denney; JD Barnes; MJK Thomas. In Vogel’s Textbook of Quantitative Chemical Analysis,
Pearson Education Pvt. Ltd., London, 2002; pp256-257.
[13] S Khokhra. Int J of Drug Dev and Res, 2010, 3, 191-196.
[14] M Farnaz; H Davoud; V Hadi; R Mohammad; S Siahi; SM Javid et al. Eur Journal of Pharm Biopharm,2009,
73, 404–413.
[15] PK Sharma; M Bajpai. Journal of Pharmacy Research, 2011, 4(7), 2216-2220.
[16] SK Jha; R Karki; DP Venkatesh;AGeethalakshami. International Journal of Drug Development & Research,
2011, 3(4), 336-343.
[17] NG Tiwari;AN Bajaj. Indian J Pharm Sci, 2007, 69, 731-737.
[18] B Brime; MA Moreno; GFrutos; MA Ballesteros; P Frutos. J Pharm Sci, 2002, 91(4), 1178–85.
[19] K Purushothamrao; K Khaliq; P Sagare; SK Patil; SS Kharat; K Alpana. Int J Pharm Sci Tech 2010, 4, 32-41.
[20] K.Rupali; RSA Mulik; L Badgujar; AR Paradkar; KR Mahadik; SL Bodhankar;S Sharma. International Journal
of Drug Formulation & Research,2010, 1(1), 359-386.
[21] J Siepman;AAinaoui; JM Vertgnaud; R Bodmeier. J Pharm Sci,1998, 87, 827-832.
Somnath Vibhute et al Der Pharmacia Lettre, 2015, 7 (3):7-16
______________________________________________________________________________
16
Scholar Research Library
[22] NK Jain; JL Vegad; AK Katiyar; RP Awadhiya. Avian Pathol, 1995, 24(4), 723-729.
[23] NMK Ito; AM Noronha; GM Boham. Research in Veterinary Science, 1989, 46, 192-197.
[24] CW Park; JY Kim; YS Rhee; TO Oh; JM Ha; NY Choi; SC Chi et el. Arch Pharm Res, 2012, 35(11), 1935-43.
[25] ICH Q1A (R2),Guidance for Industry: Stability Testing of New Drug Substances and Products, International
Conference on Harmonization, Geneva, 2003, 1-14.
[26] OS Falade; SA Adekunle; MA Aderogba;OS Atanda; C Harwood; SR Adewusi. J Sci. Food Agric, 2008, 88,
263-268.
[27] CO Eromosele; NH Pascal. J Bioresoures Tech, 2003, 86, 203-205.
[28] OJ Abayeh; EA Aina; CO Okounghae. J of Pure and Applied Sci,1998, 1(1), 17-23.
[29] MR Patel; RB Patel; JR Parikh; AB Solanki; GB Patel. APPS PharmSciTech, 2009, 10, 917-923.
[30] N Akhtar; MU Rehman; HMS Khan; F Rasool; T Saeed;Murtaza G. Tropical Journal of Pharmaceutical
Research,2011,10(3), 281-288.
[31] AOJ Ojewole. J Ethnopharmacol, 2003, 85, 217-220.
