Indo American Journal of Pharmaceutical Research, 2017 ISSN NO: 2231-6876

FORMULATION AND EVALUATION OF TOPICAL DOSAGE FORM CONTAINING

MICROSPHERES FOR MODEL ANTI INFLAMMATORY DRUG
Atefeh Shabani*, Narmada G.Y
Visveswarapura Institute of Pharmaceutical Sciences, Bangalore -560070, Karnataka, India.
ARTICLE INFO ABSTRACT
Article history The purpose of this study was to formulate a sustained release topical dosage form containing
Received 27/11/2016 microspheres for Ketorolac Tromethamine (KT). Oral consumption of KT has significant
Available online gastric irritation, hence tried to prepare topical formulation to prevent possible side effects of
09/02/2017 oral consumption. The drug-excipients compatibility studies were carried out by FT-IR and
DSC studies. Based on the results, excipients used were found to be compatible with KT. The
Keywords formulations were prepared by using different concentration of polymer and plasticizer along
Anti Inflammatory, with other excipients, and using 0.008% benzalkonium chloride as a preservative. The
Ketorolac Tromethamine, concentration of polymers was in the concentration range of 1.0%-2.5%, where plasticizer
Solvent Evaporation, concentration range was 15-25%. The formulation F8 resulted in drug content 99%±0.36,
Adverse Effect, spreadability 9±0.50gm-cm/sec, viscosity of 13420cps and maximum in-vitro diffusion of
Non-Narcotic. 47.37±0.39% over a period of 5 hrs which was selected as a best batch. The microspheres
prepared by solvent evaporation method were evaluated and M3 batch was selected. M3
formulation of microspheres was incorporated into gel formulations F5 to F8. F11 selected as
an optimized gel formulation containing KT microspheres which diffused 43.97±0.23% after
5 hrs. The selected optimized formulation F11 was subjected to stability studies as ICH
guidelines and were found to be stable. Hence the formulation F11 will eliminate the GIT side
effect, enhance the percutaneous absorption and exhibit sustained release anti-inflammatory
activity.

Corresponding author
Atefeh Shabani
Department of Pharmaceutics,
Visveswarapura Institute of Pharmaceutical Sciences,
Bengaluru-560070,
Karnataka, India.
atefeshabani@gmail.com
+91-9036643670

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Please cite this article in press as Atefeh Shabani et al. Formulation and Evaluation of Topical Dosage form Containing
Microspheres for Model Anti Inflammatory Drug. Indo American Journal of Pharmaceutical Research.2017:7(01).
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Research, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
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Vol 7, Issue 01, 2017. Atefeh Shabani et al. ISSN NO: 2231-6876

INTRODUCTION
Ketorolac tromethamine is a potent non narcotic analgesic with moderate anti-inflammatory activity. It has been investigated
extensively for use in post-operative analgesia both as a sole agent and supplement opioid analgesics and excellent applicability in the
emergency treatment of post operative cancer pain and in the treatment of migraine pain [2]. The biological half-life of KT is 4–6 h.
Therefore frequent dosing is necessary to sustain the action of drug to alleviate pain in post operative patients with a possibility of
patient non compliance. When administered as the conventional formulation, it causes gastro intestinal complications including
irritation, ulcer, bleeding and perforation [3-5]. Therefore, recent focus of the researchers has been to deliver such potential NSAIDs in
a controlled manner by using a dosage form that will minimize its release in stomach and thereby overcomes its chronic adverse effect
[6]
.
Due to GIT upset which happens on oral consumption of KT, this research tried to provide a topical dosage form of KT to
prevent GIT problems associated with the same drug. Recently several advancements have been made which have resulted in
development of new techniques of drug delivery. These techniques are capable of controlling the rate or drug delivery, sustaining the
duration of therapeutic activity and targeting the delivery of the drug to the tissue [1]. The main aim of the research work is to develop
a multiparticulate drug delivery system for topical dosage form, using model anti-inflammatory drug (Ketorolac tromethamine), in
view to maintain sustained release action and reduce dosing frequency.
Several approaches have been tried to develop non-oral formulations in addition to injections. Among the non-invasive
routes, topical administration has a promising potential as a viable alternative for systemic medication of drugs which shows merits
such as non-invasive, easy, local effect, and high level of patient satisfaction [7].
Topical preparations circumvent GI irritation, prevent the metabolism of drug in the liver and increase the bioavailability of
the drugs and provide its action directly at the site of action. Gel base formulations for dermatological use have several favorable
properties such as greaseless, easily spreadable, easily removable, thixotropic, water soluble or miscible, non- staining and emollient
[8,9]
.
Microspheres carrier systems made from the naturally occurring biodegradable polymers have attracted considerable
attention for several years in sustained drug delivery. Recently, dosage forms that can precisely control the release rates and target
drugs to a specific body site have made an enormous impact in the formulation and development of novel drug delivery systems.
Microspheres form an important part of such novel drug delivery systems [10]. They act as reservoir releasing an active ingredient over
an extended period of time maintaining effective drug concentration in the skin and at the same time, reducing undesired side effects
[11]
.
Hence an attempt was made to formulate Ketorolac tromethamine as gel incorporated in microspheres in order to eliminate
the GIT side effect, to enhance the percutaneous absorption and to achieve sustained release anti-inflammatory activity.

MATERIALS AND METHODS

Materials
Ketorolac Tromethamine drug (molecular weight 376.403 gm/mol) received as a gift sample from Mylan Laboratories,
Bengaluru (India). Carbopol 934P, ethyl cellulose, poly vinyl alcohol(PVA), dichloro methan, Benzalkonium chloride, sodium
hydroxide flakes, were received from Central drug house (Pvt) Ltd., New Delhi (India). Propylene glycol, polyethylene glycol 6000,
methanol, ethanol, glycerin, menthe oil, potassium Dihydrogen orthophosphate, were received from Industrial Estate, Mumbai (India)
and Triethanolamine was received from Merck Limited, Mumbai (India). All other chemicals and reagent used in this study were of
analytical grade.

Methods
Pre-formulation studies
Preparation of calibration curve
A standard curve was prepared in the concentration range of 0-18 µ/ml with phosphate buffer pH 7.4. The absorbance of
resulting solutions was measured at 324 nm and recorded. Concentrations versus absorbance values were plotted.

Drug-excipients Compatibility
The compatibility studies were carried out at room temperature by Fourier transform infrared (FTIR) spectroscopy to
determine the interaction of drug with other excipients used in the formulation. The IR spectra of drug alone and physical mixtures of
the drug with proposed excipients (ethyl cellulose) in the ratio of 1:1 were prepared. 1 part of this mixture was triturated with 100
parts of KBR (400–4000−1) with a scanning speed of 2 mm/sec with normal slit and compressed to form pellet which was then
analysed by FTIR (FTIR, 8400S, Shimadzu, Germany).

Differential Scanning Calorimetric (DSC)[12]

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Samples were weighed and sealed in 40 ml aluminum crucibles with a pierced aluminum lid. The analyses were performed
under nitrogen (nitrogen flow rate 50ml/min) in order to prevent oxidative effect at the standard heating rate of 10C/min over a
temperature range of 100 -270 , under static air, using a Mettler-Toledo STAR system.
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Vol 7, Issue 01, 2017. Atefeh Shabani et al. ISSN NO: 2231-6876

Formulation of Ketorolac Tromethamine gel[13-16]

The formulations were prepared by using different concentration of polymer and plasticizer along with other excipients, and
using 0.008% benzalkonium chloride as a preservative. The concentration of polymers was in the concentration range of 1.0%-2.5%,
where plasticizer concentration range was 15-25%. The accurately weighed quantity of polymer was mixed in three by fourth quantity
of distilled water. The resultant solution was viscous. The accurately weighed quantity of drug was dissolved in ethanol in separate
beaker. Then the drug solution was mixed with polymer solution. Then the remaining excipients are mixed to the viscous solution.
The above solution was thoroughly mixed by stirring for 30minutes. The prepared gel was evaluated for various parameters.

Table No. 1: Formulation of Ketorolac Tromethamine gels: F1-F8.

Ingredients F1 F2 F3 F4 F5 F6 F7 F8
Ketorolac Tromethamine (gm) 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1
Carbopol 934P (gm) 1 1.5 2 2.5 2 2 1.5 1.5
Propylene Glycol (ml) 20 20 20 20 15 25 15 25
Polyethylene Glycol (ml) 5 5 5 5 5 5 5 5
Glycerine (ml) 10 10 10 10 10 10 10 10
Ethanol (ml) 10 10 10 10 10 10 10 10
Mentha Oil (ml) 1 1 1 1 1 1 1 1
Triethanolamine (ml) 1 1 1 1 1 1 1 1
Benzalkonium Chloride (ml) 0.008 0.008 0.008 0.008 0.008 0.008 0.008 0.008
Distilled water (ml) q.s to 100 q.s to 100 q.s to 100 q.s to 100 q.s to 100 q.s to 100 q.s to 100 q.s to 100

Preparation of sustained release Ketorolac Tromethamine microspheres [17-21]

Microspheres were prepared by solvent evaporation method. Drug and polymer (ethyl cellulose) were dissolved in 10 ml of
Dichloromethane and methanol mixture which are present in the proportion of 8:2 as shown in Table No. 2. Then the slurry was
slowly introduced into 50ml of 0.5% w/v of polyvinyl alcohol while being stirred at 900 rpm by a mechanical stirrer, equipped with a
three bladed propeller at room temperature. The solution was stirred for 2 hrs to allow the solvent to evaporate completely and the
microspheres were collected by filtration. The microspheres were filtered by using Whattman filter paper. The collected microspheres
were dried for 1hr at room temperature and subsequently stored in desiccators over fused calcium chloride.

Table No. 2: Formulation of Ketorolac Tromethamine M1-M3.

Ingredients M1 M2 M3
KT (gm) 0.1 0.1 0.1
Ethyl cellulose (gm) 0.20 0.30 0.40
Dicholoromethan (ml) 8 8 8
Methanol (ml) 2 2 2
0.5% W/V Polyvinyl Alcohol (ml) 50ml 50ml 50ml

Preparation of sustained release Ketorolac Tromethamine gels containing microspheres

Best formulation of prepared gels and microspheres were selected to prepare Ketorolac Tromethamine gel containing
microspheres in different ratios. The formulated microspheres were uniformly dispersed in the gel base by mechanical stirring for 30
minutes to get microspheres loaded gel. The prepared gels were packed in wide mouth glass jar covered with screw capped plastic lid,
covering the mouth with an aluminum foil and were kept in dark and cool place.

Table No. 3: Formulation of Ketorolac Tromethamine gel containing microspheres (F9-F12).

Ingredients F9 F10 F11 F12

(gm or ml)
Prepared Microspheres of Ketorolac Tromethamine (1:4) 0.1 0.1 0.1 0.1
Carbopol 934P 1.5 2 2 1.5
Propylene glycol 20 20 15 15
Polyethylene glycol 5 5 5 5
Glycerin 10 10 10 10
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Ethanol 10 10 10 10
Mint oil 1 1 1 1
Triethanolamine 0.5 0.5 0.5 0.5
Benzalkonium chloride 0.008 0.008 0.008 0.008
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Distilled water q.s to 100 q.s to 100 q.s to 100 q.s to 100

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Evaluation of gel:
Appearance of gel
The prepared gels were inspected visually for clarity, color and presence of any particle. The test is important regarding
patient compliance.

pH
Measurement of pH for all the formulations were done by dissolving one gram of formulation in 100ml distilled water for
2hrs and pH was measured by digital pH meter.

Drug content
One gram of gel was taken and dissolved completely in phosphate buffer of 7.4 pH. Made up the volume to 50ml by
phosphate buffer and withdrawn 1, 2 and 3ml of the stock solution and made up the volume to 10ml with the phosphate buffer of same
pH. Then it was analyzed by validated UV spectrophotometric method at max of 324nm.

Spreadability22
Spreadability of the formulations was determined by an apparatus suggested by Multimer, which was fabricated itself in
laboratory and used for slide fixed on wooded block and upper slide with one end tied to glass slide and other end tide to weight pan.
Excess of gel (2gm) was placed between two glass slides and then 100gm weight was placed on slides for 5mins to compress the
sample to a uniform thickness. Weight in the increasing order i.e. 20gm, 40gm, 80gm was added to pan. The time (sec) required to
separate the two slides was taken as a measure of spreadability. It was calculated using the below formula:

S=L
Where,
M = Weight tied to the upper slide.
L = Length of glass slide moved.
T = Time taken (sec).
Shorter time interval to cover the distance of 6.5cm indicates better spreadability.

Extrudability
For a good gel formulation, it should extrude easily from the selected container. This is also type of measurement of viscosity
of formulation. In this test, sample is extruded from the tube by the visual procedure. A closed collapsible tube containing gel was
passed firmly at crimped end. When the cap was removed, the gel extrudes until pressure was dissipates. The result for each
formulation were recorded as extrusion pressure.

Viscosity

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Figure No. 1: Measurement of viscosity by Brookfield Viscometer.

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The viscosity of the gels prepared was determined using Brookfield Viscometer model. The gel sample is filled in the
sample holder and a particular spindle (No.64) inversed into the sample. The spindle is attached to the viscometer and it is
allowed to rotate at particular speed (50rpm). The viscosity of the formulation is measured after 2mins.

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Preservation Efficacy Test

Weighed 3.7gm of nutrient agar was dissolved in 100ml distilled water, sterilized the media and pour 20ml into each petri-
dish which were previously sterilized. The petri-dishes containing nutrient agar media were refrigerated for 2hours for solidification.
Using a sterile swab, a suspension of the pure culture (S. aureus) is spread evenly over the face of sterile solid agar medium. A hole is
bored in the center of the solid agar medium and 1gm of gel was added. Incubated at 30 -35 and the product testing was done after
24hrs, 48hrs and 72 hrs. The zone of growth of microorganism is measured and noted.

In-Vitro diffusion study

The apparatus used is Franz diffusion cell that is consists of donor and receive compartment. One gm gel equivalent to
100mg of KT, spread uniformly on the surface of cellophane membrane (previously soaked in phosphate buffer pH of 7.4 for
overnight) and was fixed to the end of donor compartment such that the preparation occupied inner circumference of the tube. The
whole assembly was fixed in such a way that the low end of the tube containing gel was just touched the surface of diffusion media
i.e.50ml phosphate buffer of pH of 7.4, and maintained at 37 2⁰C. A quantity of 2ml of samples was withdrawn from receptor fluid
at the time intervals of 30, 60, 90, 120, 150, 180, 210, 240, 270, 300 min. The released drug was estimated by using Shimadzu UV –
Visible spectrophotometer at 324 nm.

Evaluation of microspheres
Particle size analysis of microspheres23
The particle size of the microspheres was determined by using optical microscopy method. Approximately 100 microspheres
were counted for particle size analysis by using calibrated optical microscopy.

Scanning Electron Microscopy24

The shape and surface morphology of the dried microsphere was examined by scanning electron microscopy. Prior to
examination, samples were gold coated under vacuum using 10.0-KV accelerating voltage, a 30mm working distance, 50 m objective
aperture and a probe current of 6 10-11 amps. The object was exposed to two different magnifications to show the nature of the
surface of the microspheres. The samples after 10hrs dissolution was also magnified at two different magnifications to show the
formation of pores and the nature of release of drug from the matrix.

In-vitro drug release of Microspheres25

Microspheres equivalent to 100mg of KT were subjected for dissolution. Dissolution tests were carried out using USP Type-I
rotating basket (40 mesh). The stirring rate was 50 rpm. Dissolution medium was 900ml Phosphate buffer 7.4 pH and the temperature
was maintained at 37 . Aliquots of 1ml were withdrawn at predetermined intervals for 6hrs with pipette, filtered and volume
was made to 10ml using Phosphate buffer 7.4 pH. The collected samples were analysed spectrophotometrically at 324nm.

Evaluation of gel loaded microspheres

In-vitro diffusion studies26
The apparatus used is Franz diffusion cell that is consists of donor and receive compartment. One gm gel equivalent to
100mg of KT, spread uniformly on the surface of cellophane membrane (previously soaked in phosphate buffer pH of 7.4 for
overnight) and was fixed to the end of donor compartment such that the preparation occupied inner circumference of the tube. The
whole assembly was fixed in such a way that the low end of the tube containing gel was just touched the surface of diffusion media
i.e.50ml phosphate buffer of pH of 7.4, and maintained at 37 2⁰C. A quantity of 2ml of samples were withdrawn from receptor fluid
at the time intervals of 30, 60, 90, 120, 150, 180, 210, 240, 270, 300 min. The released drug was estimated by using Shimadzu UV –
Visible spectrophotometer at 324 nm.

Stability studies27
Stability studies were carried out at 0 , 25 /60% RH and 40 /75% RH for the optimized formulation for one month. The
selected formulation was kept in suitable container and tightly closed. They were then stored at 0 , 25 /60% RH and 40 /75% RH
for 1 month and evaluated for drug content.

RESULTS AND DISCUSSION

Pre-formulation studies
A standard curve ranging from 0-18 µ/ml with phosphate buffer pH 7.4 at 324 nm was found to be linear with y=0.0549x and
R2=0.9994 as shown in Figure No. 2.
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Figure 2: Standard calibration curve of Ketorolac Tromethamine.

Drug-excipients Compatibility
The FT-IR peaks of pure Ketorolac Tromethamine is presented in the Figure No. 3 showed characteristics of –OH stretch at
3354.68cm-1, aromatic asymmetric and symmetric groups at 2926.32cm-1 and 2872.10cm-1, C-OH and C-OH stretch at 1057.03cm-1
and 1197.83cm-1 respectively. The mixture of Ketorolac tromethamine with carbopol 934P is shown in Figure No. 4 and mixture of
Ketorolac Tromethamine with ethyl cellulose is shown in Figure No. 5. The observed characteristic peaks are in limit and thus chosen
excipients for formulation were found to be compatible and have no chemical interaction with the Ketorolac Tromethamine.
Therefore, the FTIR studies ruled out the possibility of any drug polymer interaction and hence contribution of drugs along with
excipients can be formulated safely.

Figure No. 3: FT-IR spectra of Ketorolac Tromethamine.

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Figure No. 4: FT-IR spectra of Ketorolac Tromethamine + carbopol 934P.

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Figure No. 5: FR-IR spectra of Ketorolac Tromethamine + ethyl cellulose.

Differential Scanning Calorimetric (DSC)

Differential Scanning colorimetry was conducted on Ketorolac Tromethamine pure drug and in combination with carbopol
934P and ethyl cellulose. The thermogram of pure drug Ketorolac Tromethamine with carbopol 934P exhibited an endothermic peak
at 146.97⁰C (Figure No. 6) corresponding to its melting point, where as in case of Ketorolac Tromethamine with ethyl cellulose
exhibited an endothermic peak at 143.70⁰C (Figure No. 7). The combined mixture of pure drug Ketorolac Tromethamine with both
carbopol 934P and ethyl cellulose exhibited endothermic peak at 163.62⁰C (Figure No. 8) corresponding to its melting point. Based on
results of compatibility studies it may be concluded that the excipients used were found to be compatible with Ketorolac
Tromethamine.

Figure No. 6: DSC thermogram of Ketorolac Tromethamine + carbopol 934P.

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Figure No. 7: DSC thermogram of Ketorolac Tromethamine + ethyl cellulose.

Figure No. 8: DSC thermogram of Ketorolac Tromethamine + carbopol 934P + ethyl cellulose.

EVALUATION OF KETOROLAC TROMETHAMINE GEL

Visual examination revealed that all the systems were transparent. The addition of liquid excipients improved the consistency
of gels. The results for evaluation of gel parameters such as Viscosity, pH, drug content uniformity, spreadability, extrudability and
viscosity are given in Table No. 4. The pH values of all developed formulae was in range 6-7 which is considered acceptable to avoid
the risk of irritation upon application to the skin. The values of spreadability indicated that the polymer used gave gels spread by small
amount of shear. The formulation F8 resulted in drug content 99%±0.36, spreadability 9±0.50gm-cm/sec along with viscosity of
13420cps was selected as a best batch.

Table No. 4: Drug content uniformity of Ketorolac Tromethamine gel:

Formulation Drug content (%) pH Spreadability Extrudability Viscosity

code (gm-cm/sec) (CPS)
F1 95%±0.60 6.4±0.52 9±0.26 Good 12550±5.56
F2 76%±0.36 6.6±0.20 11±0.36 Better 14378±8.54
F3 82%±0.45 6.9±0.26 8±0.50 Poor 15895±3.60
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F4 107%±0.10 7.4±0.26 9±0.43 Poor 17210±11.13

F5 103%±0.10 7.2±0.10 8±0.36 Good 13412±11.13
F6 102%±0.26 7.4±0.45 10±0.30 Better 14951±1.0
F7 98%±0.60 7.1±0.36 10±0.10 Better 12980±8.0
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F8 99%±0.36 7.0±0.36 9±0.50 Good 13450±6.24

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Preservation efficacy test for Ketorolac Tromethamine gel

The Zone of Inhibition of Ketorolac Tromethamine gel containing 0.005%, 0.008%, 0.01% and 0.02% of Benzalkonium
chloride was calculated as shown in Table No. 5 and Figure No. 9. Since the ZOI was maximum in 0.008% of BKC, this
concentration was selected for the further formulations.

Table No. 5: Preservative efficiency by Zone of Inhibition Method.

Time (Hour) Concentration of BKC (%) Zone of Inhibition (cm)
0.005 1.1
0.008 1.4
24
0.01 1.2
0.02 1.3
0.005 1.1
0.008 1.5
48
0.01 1.3
0.02 1.3
0.005 1.1
0.008 1.7
72
0.01 1.3
0.02 1.3

Figure No. 9: Effect of preservative efficiency after 72 hrs.

In-vitro diffusion study of gel

The results of in-vitro diffusion studies showed the cumulative drug release of the prepared gel for the formulations F1 to F8
as shown in Table No. 6 and Figure No. 10. The percentage cumulative drug release of F1 to F8 was 36.44±0.23, 43.86±0.12,
47.14±0.54, 37.70±0.68, 41.2±32, 39.50±0.51, 43.40±0.50 and 47.37±0.39% respectively. Among the eight batches, F8 showed the
maximum release of 47.37±0.39% over a period of 5 hrs.

Table No. 6: In-vitro diffusion study of gel (F1 to F8).

Time Percentage cumulative release

(min) F1 F2 F3 F4 F5 F6 F7 F8
30 9.47±0.23 9.21±0.36 10.36±0.38 4.67±0.31 8.82±0.63 6.60±0.32 8.25±0.52 8.08±0.28
60 12.42±0.30 12.21±0.28 11.30±0.51 5.32±0.63 11.56±0.35 6.80±0.64 9.02±0.41 10.4±0.46
90 15.4±0.10 14.36±0.61 15.29±0.90 6.65±0.84 14.34±0.43 10.00±0.52 13.59±0.62 13.13±0.95
120 20.56±0.51 17.71±0.19 18.53±0.47 9.15±0.73 18.17±0.96 13.51±0.75 17.68±0.74 16.32±0.62
150 17.97±0.62 17.05±0.28 21.80±0.80 12.89±0.91 14.74±0.73 15.93±0.35 22.81±0.38 20.98±0.38
180 19.07±0.38 21.50±0.34 26.97±0.82 16.14±0.64 20.54±0.83 19.38±0.42 23.00±0.47 23.31±0.47
210 25.89±0.90 29.15±0.72 29.26±0.31 21.30±0.51 25.92±0.41 23.85±0.61 32.00±0.58 24.30±0.68
240 30.69±0.27 34.94±0.24 31.58±0.61 26.63±0.73 31.45±0.29 30.93±0.83 32.52±0.68 29.93±0.41
270 34.56±0.51 40.90±0.28 39.63±0.31 33.17±0.34 35.25±0.84 37.38±0.34 39.61±0.79 37.20±0.97
300 36.44±0.23 43.86±0.12 47.14±0.54 37.70±0.68 41.2±0.30 39.50±0.51 43.40±0.50 47.37±0.39
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Figure No. 10: In-vitro diffusion study of gel (F1 to F8)

EVALUATION OF KETOROLAC TROMETHAMINE MICROSPHERES

Particle size analysis
The resulting microspheres formulated by solvent evaporation method were found to be spherical and free flowing in nature.
The mean particle size of microspheres ranged from 96.18±0.04µm to97.33±0.07µm as shown in Table No 7.

Table No. 7: Particle size of microspheres:

Formulation code Mean particle size ( m)

M1 97.33±0.07
M2 94.84±0.03
M3 96.18±0.04

Scanning Electron Microscopy

SEM analysis of Ketorolac Tromethamine microspheres revealed that all microspheres prepared were spherical in shape and
is having porous outer skins as shown in Figure No. 11.

Figure No. 11: SEM of Ketorolac Tromethamine microspheres.

Drug entrapment efficiency

The drug entrapment efficacy of all the formulation was in the range of 81.0 to 95.9%. The drug entrapment efficacy of
microspheres was noted to increases with increase in concentration of polymers. Amongst formulations (M1, M2 and M3) M3 have
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shown good entrapment efficiency as shown in Table No. 8.

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Table No. 8: Drug entrapment efficiency of microspheres:

Formulation code Ratio of Ketorolac Tromethamine :Polymer Drug content (%)

M1 1:2 81.0%±0.79
M2 1:3 94.0%±1.30
M3 1:4 95.9%±0.79

In-vitro drug release of Microspheres

In-vitro dissolution profile showed the cumulative drug release of all the three batches M1, M2 and M3 as shown in Table
No. 9 and Figure No. 11. The percentage cumulative drug release of M1, M2 and M3 was found to be 28.26±0.27, 35.29±0.16 and
36.66±0.24 respectively. Among the three batches, M3 showed the maximum release of 36.66%±0.24 over a period of 5 hrs.

Table No. 9: In-vitro dissolution study of microspheres (M1 to M3).

Time % cumulative release

(min) M1 M2 M3
0 10.13±0.23 10.84±0.35 11.25±0.52
60 12.56±0.96 12.55±0.67 14.88±0.24
90 14.40±0.64 15.28±0.86 16.60±0.38
120 16.24±0.52 18.72±0.95 20.41±0.75
150 19.51±0.84 21.10±0.24 23.80±0.25
180 20.70±0.54 24.67±0.38 25.78±0.19
210 21.21±0.34 27.19±0.46 28.54±0.64
240 23.35±0.24 29.22±0.29 32.45±0.38
270 25.99±0.57 32.39±0.34 34.52±0.29
300 28.26±0.27 35.29±0.16 36.66±0.24

Figure No. 11: In-vitro dissolution study of microspheres (M1 to M3).

PREPARATION OF KETOROLAC TROMETHAMINE GEL CONTAINING MICROSPHERES

The formulation F5 to F8 was selected as the best formulation of gel containing Ketorolac Tromethamine. The formulation
M3 was selected as the best formulation of microspheres containing Ketorolac Tromethamine. M3 formulation of microspheres was
incorporated into gel formulations F5 to F8. The resultant gel containing microspheres of formulations F9 to F12 were transparent.

EVALUATION OF KETOROLAC TROMETHAMINE GEL CONTAINING MICROSPHERES

Visual examination revealed that all the systems were transparent. The results of pH, drug content, pH, spreadability,
extrudability and viscosity are shown in Table No. 10. The pH values of all developed formulae were in range 6.9 – 7.2, which is
considered acceptable to avoid the risk of irritation upon application to the skin. The values of spreadability indicated that the polymer
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used gave gels spread by small amount of shear. The formulation F11 resulted in drug content 100±0.52%, spreadability 9±0.50gm-
cm/sec along with viscosity of 16610±7.81cps was selected as a best batch.
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Vol 7, Issue 01, 2017. Atefeh Shabani et al. ISSN NO: 2231-6876

Table No. 10: Drug content uniformity in Ketorolac Tromethamine gel containing microspheres:

Formulation pH Drug content (%) Spreadability Extrudability Viscosity

(gm-cm/sec) (CPS)
F9 6.9±0.36 97%±0.45 10±0.62 Better 16245±5.56
F10 7.2±0.10 80%±1.22 10±0.50 Good 15310±2.64
F11 6.9±0.30 100%±0.52 9±0.50 Good 16610±7.81
F12 7.1±0.10 98%±0.17 8±0.52 Good 16120±3.0

In-Vitro diffusion studies of Ketorolac Tromethamine gel containing microspheres:

The in-vitro diffusion profile of the designed formulations, F9-F12 was studied by using Franz diffusion cell under specified
conditions. The formulations were subjected to diffusion in the media of phosphate buffer of pH 7.4 for 5hrs. In first 30 minutes of
diffusion studies, all the formulations exhibited less than 10% of cumulative drug release. But in further case of diffusion studies,
variations in drug release profile are observed and it was found to be depended upon the amount of excipients used (polymer
plasticizer ratio of microspheres). At the end of diffusion studies for 5hrs none of the formulations of gel containing microspheres
showed drug release more than 45% and F11 showed 43.97±0.23% (Table No. 11). After thorough comparison of evaluation
parameters, F11 was found to show sustained release of drug and fulfilled needed criteria of topical gel containing microspheres.

Table No. 11: In-vitro diffusion study of microspheres (F9 to F12).

Time % cumulative release

(min) F9 F10 F11 F12
0 6.18±0.23 6.8±0.56 7.14±0.63 7.65±0.52
60 9.52±0.35 10.9±0.27 8.95±0.57 9.48±0.17
90 11.13±0.68 14.1±0.39 10.61±0.85 11.18±0.28
120 13.75±0.29 16.57±0.42 13.67±0.54 12.70±0.24
150 13.71±0.64 15.87±0.85 17.71±0.29 15.81±0.78
180 16.08±0.41 20.00±0.96 22.32±0.46 19.85±0.49
210 22.06±0.27 23.00±0.75 22.00±0.57 25.48±0.54
240 27.09±0.57 29.37±0.54 27.83±0.38 29.77±0.38
270 33.13±0.48 35.32±0.34 32.79±0.63 34.47±0.64
300 40.51±0.23 40.15±0.31 43.97±0.23 37.53±0.52

Figure No. 12: Comparison of In-vitro Diffusion Profiles F9-F12.

Stability studies:
The evaluation parameters showed satisfactory results and by comparison with previous formulation, it showed no
degradation formulation. Thus it can be concluded that stability studies on F11 formulation was within the specification and stable
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(Table No. 12).

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Vol 7, Issue 01, 2017. Atefeh Shabani et al. ISSN NO: 2231-6876

Table No. 12: Stability Studies of Formulation F11.

Parameters F11
pH 6.9±0.1
Viscosity 16610 cps±5.29
Drug content 98.86%±0.07
In-vitro Diffusion studies 38.22%±0.09

CONCLUSION
From the result obtained formulation F1-F8, we can conclude that carbopol 934P 2% and 1.5% and propylene glycol
concentration of 15% and 25% (F5-F8) were found to be more acceptable for the development of topical gel containing Ketorolac
Tromethamine microspheres and among the KT gel formulations F1-F8, F8 showed better sustained release profile of 47.37±0.39 for
5 hrs along with satisfactory pH, drug content, spreadability and viscosity compared to other formulations. Microspheres of 3 different
concentrations of drug and polymer were prepared with 1:2(M1), 1:3(M2) and 1:4(M3) ratios of KT and ethyl cellulose. Microspheres
ratio of 1:4 (M3) showed better drug entrapment of 95.9% and release profile of 36.66% over 5hrs in comparison to 1:2 and 1:3. SEM
of optimized M3 formulation were discrete, spherical with nearly smooth surface. The best formulation of microspheres M3 was
incorporated into selected gel formulations F9-F12. The formulations F9-F12 were further evaluated for drug content, pH,
spreadability, extrudability, viscosity and in-vitro diffusion studies to select the best sustained release formulation. After thorough
evaluation of F9-F12 formulation, F11 showed the satisfactory sustained release profile of 43.97±0.23% after 5 hrs with suitable drug
content, pH, spreadability and viscosity. From the stability studies result, it was concluded that the F11 formulation containing
microspheres was successfully formulated and evaluated to sustain the release of KT over a period of time.
Further research can be done regarding its in-vivo result. This research can be considered as one illustration of topical
NSAIDs, which combined with new drug delivery system (microparticulate). Therefore, it can be applicable for some other NSAIDs
as well.

ACKNOWLEDGEMENT
I immensely thank the principal of Visveswarapura Institute of Pharmaceutical Sciences and my guide, Dr. G.Y. Narmada for
providing facilities and help me out during this research.

Conflict of interest:
The authors declare that there are no conflict of interest.

LIST OF ABBREVIATIONS:

ABBREVIATIONS FULLFORMS
ABS Absorbance
amps Ampere second
BP British pharmacopeia
CAP Cellulose acetate phthalate
CDR Cumulative drug release
CLA Cumulative loss amount
Cm Centimeter
cm2 Centimeter square
Degree Celsius
Cmc carboxymethylcellulose
COX Cyclooxygenase
Conc. Concentration
Cps Centi poise
DEE Drug entrapment efficiency
DSC Differential scanning colorimeter
EC Ethylcellulose
FT-IR Fourier transform infrared spectroscopy
gm Gram
HPC Hydroxypropylcellulose
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HPLC High pressure liquid chromatography

HPMC hydroxypropylmethylcellulose
HPTLC High pressure thin layer chromatography
hrs Hour
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ICH International council of harmonisation

JP Japan pharmacopeia
KT Ketorolac tromethamine

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KV Kilovolt
mg milligram
Mps Mean particle size
µg/ml Microgram per milliliter
µm Micrometer
ml Milliliter
mm Millimeter
Min Minute
mol Mole
MP Melting point
N Normality
NaoH Sodium hydroxide
Na-cmc Sodium carboxymethylcellulose
NDDS Novel drug delivery system
nm Nanometer
NSAID Non-steroidal anti-inflammatory drug
PVA Poly vinyl alcohol
RH Relative humidity
rpm Rotation per minute
Sec Seconds
SEM Scanned electron microscopy
s.aureus Staphylococcus aureus
TRA Trans-retinoic acid
USPNF United states pharmacopeia and national formulary
USP United states pharmacopeia
UV Ultraviolet
w/w Weight by weight
w/0/0 Water in oil,oil in oil
XRD X-ray diffraction
pH Power of hydrogen ion concentration
% Percentage
λmax Absorption maxima
α Alpha
β Beta
γ Gamma

REFERENCES
1. Chien YW. Rate-controlled drug delivery systems. Indian J Pharm Sci. April 1988:63-88.
2. Andrade JR, Maslanka M, Maneatis T, Bynum L, Burchomore M. The use of Ketorolac tromethamine in the management of post
operative pain. Orthopedics. 1994; 17: 157–166.
3. Shankar C, Mishra M. Development and in-vitro evaluation of gelatin A microspheres of Ketorolac tromethamine for intranasal
administration. Acta Pharm. 2003; 53: 101–110.
4. Shyamala B, Sanmathi BS. Poly(lactic acid) microspheres of Ketorolac tromethamine for parenteral controlled drug delivery
system. Indian J Pharm Sci. 2001; 63: 538–540.
5. Shyamala B, Sanmathi BS. Influence of manufacturing parameters and the release profile of Ketorolac tromethamine from poly
(Lactide-co-glycolide) microspheres. Indian Drugs. 2001; 38: 383–385.
6. Abitha M H, Flowerlet M. Formulation and Evaluation of Nanoparticles as Sustained Release Topical Formulation Containing
Non‐Steroidal Anti ‐Inflammatory Drug. World J. Clin. Pharmacol. Microbiol.Toxicol. Vol 1 [3] September 2015. 35‐42.
7. Parthiban KG, Manivannan R, Kumar BS, Ahasan MB. Formulation and evaluation of ketorolac ocular pH-triggered in-situ gel.
IJDDR. 2010;2(2):379-387.
8. Velissaratou AS, Papaioannou G. In vitro release of chlorpheniramine maleate from oinment bases. Int J Pharm 1989; 52: 83-6.
9. Klich CM. Jels and Jellies. In: Swarbrick J, Boylan JC, eds. Encyclopedia of PharmaceuticalTechnology. New York, NY: Marcel
Dekker Inc;1992;6:415-39.
10. K.V. Ranga Rao, K.P. Devi (1988). Swelling controlled release systems: recent developments and application. Int J Pharm. 48, 1-
7569

16.
11. Basarkar G, Shirsath G, Patil S. Development of microspheres containing Diclofenacdiethylamine as sustained release topical
formulation. Bull Pharm Res 2013;3:14-22.
12. Akash MSH, Iqbal F, Raza M, Rehman K, Ahmed S, Shahzad Y, et al. Characterization of ethyl cellulose and hydroxypropyl
methylcellulose microspheres for controlled release of flurbiprofen. J Pharm Drug Deliv Res. 2013;2(1):1-10.
Page

13. Bhagat S, Keny RV, Bobde S. Formulation development and evaluation of ketorolac tromethamine gel for topical application in
pain management. Int J Pharm Phytopharmcol Res. 2014 Jun;4(1):70-75.

www.iajpr.com
Vol 7, Issue 01, 2017. Atefeh Shabani et al. ISSN NO: 2231-6876

14. Saroha K, Aggarwal A, Nanda S. Comparative evaluation of semisolid bases for transdermal use of ketorolac tromethamine.
IJPBR. 2014;2(2):1-6.
15. Fasale PD, Wadkar KA, Mohite SK. Development and evaluation of topical gel containing ketorolac tromethamine. Int J Uni
Pharm Bio Sci. 2013 Aug;2(4):258-266.
16. Hosny KM, Tayeb MM, Fallatah OM, Mahmoud AA, Mandoura MS, Al-sawahlin MM, et al. Preparation and evaluation of
ketorolac tromethamine hydrogel. Int J Pharm Sci Rev Res. 2013 Jun;20(2):269-274.
17. Yadav N, Mohite DD, Pawar KR, Pawar UR, Bhise SB, Sande TA, et al. Synthesis and characterization of sustained release
atenolol microspheres by solvent evaporation technique. J Pharm Sci Tech. 2011;3(2):559-562.
18. Chella N, Yada KK, Vempati R. Preparation and evaluation of ethyl cellulose microspheres containing diclofenac sodium by
novel w/o/o emulsion method. J Pharm Sci Res. 2010;2(12):884-888.
19. Ganesh S, Kumar SD, Kumar SB, Abhilash R, Bharadwaj SP, Mohammed I, et al. Controlled release formulation and evaluation
of idarubicin microsphere using biodegradable hydrophilic and hydrophobic polymer mixtures. Asi J Pharm Cli Res.
2010;3(3):179-182.
20. Avachat MA, Bornare NP, Dash RR. Effect of process parameters Sustained release microspheres of ropinirole hydrochloride.
Acta Pharm. 2011 Jul;61:363-376.
21. Dhoka VM, Pande VA, Vaidya DP, Arora A. In vitro and in vivo evaluation of ethyl cellulose based floating microspheres of
cefpodoxime proxetil. Int J Pharm Biomed Res. 2010;1(4):122-128.
22. Jyothi D, Koland M, Priya S. Investigation of anti-inflammatory activity of ointments containing fenugreek extract. AJ Pharm Cli
Res. 2014;7(2):66-69.
23. Yadav N, Mohite DD, Pawar KR, Pawar UR, Bhise SB, Sande TA, et al. Synthesis and characterization of sustained release
atenolol microspheres by solvent evaporation technique. J Pharm Sci Tech. 2011;3(2):559-562.
24. Chella N, Yada KK, Vempati R. Preparation and evaluation of ethyl cellulose microspheres containing diclofenac sodium by
novel w/o/o emulsion method. J Pharm Sci Res. 2010;2(12):884-888.
25. Yang JH, Yun MY, Kim DK, Kim MJ, Kim Y, Kim TY, et al. Preparation and evaluation of ketorolac tromethamine gel
containing genipin for periodontal diseases. Arch Pharm Res. 2007;30(7):871-875.
26. Zia H, Nasseri AA, Aboofazeli R, Needham TE. Lecithin- stabilized microemulsion- based organogels for topical application of
ketorolac tromethamine. In vitro release study. Irani J Pharm Re. 2003;2:117-123.
27. Basarkar GD, Shirsath GN, Patil S.B. Development of microspheres containing diclofenac diethylamine as sustained release
topical formulation. Bull Pharm Res. 2013;3(1):14-22.

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