http://informahealthcare.com/drd ISSN: 1071-7544 (print), 1521-0464 (electronic) Drug Deliv, Early Online: 1–7 ! 2013 Informa Healthcare USA, Inc. DOI: 10.3109/10717544.2013.859186

ORIGINAL ARTICLE

Formulation development, in vitro and in vivo evaluation of microemulsion-based gel loaded with ketoprofen Kishor V. Nikumbh, Shailesh G. Sevankar, and Moreshwar P. Patil

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Department of Pharmaceutics, MET’s Institute of Pharmacy, Bhujbal Knowledge City, Adgaon, Nashik, India

Abstract

Keywords

Background: Anti-inflammatory agents are widely used to relieve inflammation caused by various factors. Aim: This study was initiated with the intention to deliver low aqueous soluble ketoprofen to enhance its solubility by developing microemulsion system as a template and then incorporating it into gel phase. Materials and methods: Initially ketoprofen was solubilized into microemulsion preparation made up of clove oil, Tween 20 and propylene glycol as oil phase, surfactant and co-surfactant respectively, then it was incorporated into different concentration of gelling phase using gelling agents namely Carbopol 940, Carbopol 934 and hydroxypropyl methyl cellulose K4M (HPMC K4M). Formulated emulgels were evaluated for their physical appearance, pH, rheological properties, globule size, extrudability, drug content, spreadability, bioadhesion strength, in vitro and ex vivo drug release, skin irritation test and anti-inflammatory activity. Results: Microemulsion had shown globule size 396 nm, pH 6–6.7, viscosity 29.4 cps and zeta potential 12 mV indicating good stability. Formulated emulgels showed good physical appearance, skin acceptable pH 6–6.9, non-Newtonian shear thinning system, drug content 99.28  0.16%, bioadhesion strength 48.4 gram force, globule size 473 nm, spreadability 22.96 gm.cm/s, good extrudability, in vitro release, ex vivo release did not showed any irritation reaction and possess a good anti-inflammatory activity. Conclusions: Selected batch showed enhanced drug release (92.42  4.66%) as compared to marketed gel (65.94  3.30). Similarly ex vivo release of formulation showed 72.22% release through mice skin compared with marketed gel. Formulations followed Korsmeyer–Peppas diffusion kinetic model. It was observed from the results that the formulated emulgel can provide promising delivery of ketoprofen.

Anti-inflammatory activity, emulgel, ex vivo release, globule size, zeta potential

Introduction Inflammation is an important nonspecific local protective reaction to the tissue injury, caused by pathogens or wound. Acute inflammation is the immediate response of the body to injury or cell death (Willey et al., 2008). In last few decades, research demonstrated that inflammation is regulated by many pro and anti-inflammatory chemical mediators like histamine, prostaglandins (PEG2 and prostacyclins), leukotrien B4, serotonin, bradykinin, cytokines (interleukin [IL]-1, IL-6, IL-11 and tumor necrosis factor-a), reactive oxygen species, growth factors and lysosomal enzymes of neutrophils (Khatib et al., 2010). Inflammation is the result of concerted participation of a large number of vasoactive, chemoactive and proliferative factors at different stages (Tripathi, 2003). Ketoprofen is chemically a propionic acid derivative, non steroidal anti-inflammatory drug (NSAID). Its Address for correspondence: Kishor V. Nikumbh, Department of Pharmaceutics, MET’s Institute of Pharmacy, Bhujbal Knowledge City, Adgaon, Nashik 422 003, Maharashtra, India. Tel: +919850630624. Email: [email protected]

History Received 5 September 2013 Revised 22 October 2013 Accepted 22 October 2013

anti-inflammatory action is due to inhibitory effect on cyclooxygenase-2, an enzyme involved in prostaglandin synthesis that is responsible for inflammation. Emulgels are emulsions or microemulsions, either of the water-in-oil type or oil-in-water type, which are gelled by using suitable gelling agents. Emulsified gel is stable and better vehicle for delivery of hydrophobic or poorly water soluble drugs. They have a high patient compliance since they possess the advantages of both emulsions and gels. Direct (oil-in-water) systems are used to entrap lipophilic drugs, whereas hydrophilic drugs are entrapped in the reverse (water-in-oil) system (Jain et al., 2010; Joshi et al., 2011; Singla et al., 2012). Emulgels for dermatological use have several favorable properties such as being thixotropic, greaseless, easily spreadable, easily removable, emollient, long shelf life, biofriendly, transparent and pleasing appearance (Magdy, 2004). Microemulsion is defined as a dispersion consisting of oil, surfactant, cosurfactants and aqueous phase, which is a single optically isotropic and thermodynamically stable liquid solution usually within the range of 10–100 nm. Microemulsions

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have several advantages such as enhanced drug solubility, good thermodynamic stability, ease of manufacturing and enhancement effect on transdermal ability over conventional formulations. Recently, increasing attention has focused on microemulsions for transdermal delivery of hydrophobic drugs (Chen et al., 2004; Hyun et al., 2012; Sahle et al., 2012). Carbopols are high molecular weight polymers of acrylic acid cross-linked with allyl ethers of pentaerythritol. The molecular weight of carbopol resins is theoretically estimated at 7  105 to 4  109. Carbopol disperse in water to form acidic colloidal solutions of low viscosity, these solutions when neutralized produce highly viscous gel (Bugay & Findlay, 1999; Rowe et al., 2009). Ketoprofen possesses poor water solubility and high hydrophobicity (Gordon et al., 2006), it also causes gastric irritation when taken orally, hence creates limitation in formulating as oral dosage forms (Renceber et al., 2009). This study was aimed to develop and evaluate emulgel formulation containing ketoprofen, as the market survey indicated the absence of ketoprofen emulgel to be applied directly on the affected area with the objective of releasing drug locally and more effectively. The study was initiated using microemulsion as a template consisting of clove oil as oil phase, Tween 20 as surfactant and propylene glycol as a cosurfactant, which helps in solubilization of hydrophobic ketoprofen in its fine globule droplets, it was then incorporated into separately prepared gel phase using Carbopol 940, Carbopol 934 and HPMC K4M to get homogeneous and thickened emulgel; this could result in low concentration of drug in emulgel as compared to gel. To achieve these objectives, the emulgel was evaluated for the influence of pH, rheological properties, bioadhesion strength, spreadability, in vitro drug release, globule size, ex vivo release, zeta potential, extrudability and drug content. The anti-inflammatory activity of selected ketoprofen containing formulation using carrageenan-induced paw edema had been evaluated and compared with commercial gel formulation (Fastum gelÕ ).

Materials and methods

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surfactants, Tween 20, Tween 80 and cosurfactants like Capmul MCM L8, propylene glycol and Captex. An excess amount of ketoprofen was added to 3 ml of selected oils, surfactants and cosurfactants separately in 10 ml capacity stopper vials. Then the mixture was vortexed using a cyclomixer (Remi motor, Mumbai, India) for 10 min in order to facilitate proper mixing of drug with the vehicles and then stirred for 48 h at 40  0.5  C. Furthermore, the mixtures were kept for 24 h at room temperature to reach equilibrium. The equilibrated samples were centrifuged at 3000 rpm for 15 min followed by filtration through a 0.45 mm membrane filter. The filtrates were diluted with methanol subsequently quantified by ultraviolet (UV)-spectrophotometer at 260 nm. Pseudo ternary phase diagrams Ketoprofen showed maximum solubility in clove oil as compared to other oils; hence, it was selected for further studies. Tween 20, as a surfactant, and propylene glycol, as cosurfactants, showed better solubility for ketoprofen and good emulsifying properties with clove oil. Pseudo ternary phase diagrams were constructed using water titration method. Surfactant and cosurfactant (Smix) were mixed in different weight ratios (1:1, 1:2, 1:3, 2:1 and 3:1). Oil and Smix mixture were mixed thoroughly in different weight ratios (1:9, 2:8, 3:7, 4:6, 5:5, 6:4, 7:3, 8:2 and 9:1). Distilled water was added drop wise to the different mixtures of oil/Smix until cloudy dispersion was obtained. Pseudo ternary plots were constructed using Chemix School Software, trial version 3.6 (Oslo, Norway), and microemulsions were prepared based on ternary phase diagram. Formulation of microemulsion From all five-phase diagrams, the ratio of 2:1 S/Cos concentration showed higher self microemulsifying region (Figure 1), hence selected for formulation of microemulsion. Right part from boundary line in phase diagram shows us the region in which self microemulsifying region exists.

Materials Ketoprofen was kindly gifted by Cipla Ltd., (Daman, India). Carbopol 940 was purchased from Loba chemicals (Mumbai, India). Carbopol 934 was supplied from Research Lab Fine Chemicals (Mumbai, India). HPMC K4M was purchased from Dow chemicals (Mumbai, India). Tween 20, clove oil, triethanolamine (TEA), methylparaben, propylparaben, sodium hydroxide, potassium dihydrogen phosphate and methanol were supplied from Thomas Baker (Mumbai, India). All the chemicals used during study were of analytical reagent grade and used further without dilutions. Albino mice were obtained from Haffkine Institute (Mumbai, India). Solubility studies Screening of oils, surfactants and cosurfactants for microemulsion To find out suitable oil, surfactant and cosurfactants phase in microemulsion, the solubility of ketoprofen was screened in various oils, almond oil, clove oil, liquid paraffin, oleic acid

Figure 1. Pseudo ternary phase diagram of microemulsion (2:1).

Microemulsion-based ketoprofen gel

DOI: 10.3109/10717544.2013.859186

A larger microemulsion region is responsible for the higher microemulsifying potential of the combination. Thus, it is helpful in finding regions having better ability at lower proportion of cosurfactants and having higher drug loading potential.

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Preparation of emulgel For the preparation of emulgel, initially microemulsion was prepared. The oil phase was prepared by dissolving methyl and propylparabens in propylene glycol and it was added into clove oil. Ketoprofen was added into oil phase. Aqueous phase was prepared by incorporating Tween 20 into distilled water, and then both the phases were mixed using constant stirring to get transparent microemulsion. For the preparation of gelling phase, Carbopol 940 and Carbopol 934 were dispersed in distilled water with continuous stirring to get homogeneous dispersion, while HPMC K4M was separately dispersed in hot distilled water (75  C) and the stirred continuously until room temperature was reached. The pH was adjusted in between 6 and 6.5 using TEA. The gelling phase was slowly mixed with microemulsion in 1:1 ratio with constant and uniform stirring to get milky white homogeneous emulgel. The composition of different formulations is given in Table 1. Evaluation of microemulsion Measurement of globule size and zeta potential The globule size and zeta potential were measured using Zetasizer Nano – ZS (Malvern instruments, Worcestershire, UK). The measurement was performed at 25  C. A sample, 1 ml, was diluted using double distilled water (Yang et al., 2006; Graf et al., 2009).

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pH determination The pH of all formulations was determined by using calibrated pH meter MK VI (Systronic, Mumbai, India). A dispersion, 10%, was made up with distilled water and measurements were carried out at ambient temperature (Bhanu et al., 2011). Rheological study Viscosity was determined at 37  1  C using Brookfield digital viscometer-LV DV E (Brookfield, Massachusetts, USA) with spindle 96 at different rpm (Mostafa et al., 2011). Spreadability Spreadability was determined by apparatus suggested by Mutimer. Apparatus consist of a wooden block, which was attached to a pulley at one end. Spreadability was measured on the basis of ‘‘Slip and Drag’’ characteristics of emulgel. A ground glass slide (7.5  2.5 cm) was fixed on the wooden box. Emulgel formulation under the study, 1 gm, was placed on this ground slide. The formulation was then sandwiched between ground slide and upper glass slide having same dimensions as that of ground slide and it was provided with the hook. Weight of 50 gm was allowed to rest on upper slide for 2 min to expel air and to provide uniform film of formulation. Known weight was placed in the pan attached to the pulley with the help of hook. The time required to cover distance of 7.5 cm was recorded. A shorter interval indicates better spreadability (Joshi et al., 2012). It is calculated using the formula: S ¼ M  L=T where M is the weight tied to upper slide, L is length of glass slide and T is time taken to separate the slide.

Phase separation

Globule size measurement

Microemulsions were subjected to centrifugations at 10 000 rpm for a period of 30 min and observed for any phase separation (Patravale & Bachhav, 2009).

Globule size of emulgel formulations was determined by using Zetasizer Nano – ZS (Malvern instruments). Sample preparations were made by dissolving 1 gm of formulation in double distilled water (Khunt et al., 2012).

Evaluation of prepared emulgel Physical appearance

Drug content determination

The prepared ketoprofen emulgel formulations were inspected visually for their color, homogeneity and consistency (Thakur et al., 2012).

Ketoprofen content in emulgel was measured by dissolving known quantity of jellified emulsion in solvent (methanol) by sonication. Absorbance was measured after suitable dilution

Table 1. Composition of emulgel formulations. Component (% w/w)

EG 1

EG 2

EG 3

EG 4

EG 5

EG 6

EG 7

EG 8

Ketoprofen Carbopol 940 Carbopol 934 HPMC K4M Clove oil Tween 20 Propylene glycol Methylparaben Propylparaben Triethanolamine Water

2.0 1.5 – – 10 27 13 0.03 0.01 q.s. q.s.

2.0 2.0 – – 10 27 13 0.03 0.01 q.s. q.s.

2.0 – 1.5 – 10 27 13 0.03 0.01 q.s. q.s.

2.0 – 2.0 – 10 27 13 0.03 0.01 q.s. q.s.

2.0 1.0 – 0.5 10 27 13 0.03 0.01 q.s. q.s.

2.0 1.0 – 1.0 10 27 13 0.03 0.01 q.s. q.s.

2.0 – 1.0 0.5 10 27 13 0.03 0.01 q.s. q.s.

2.0 – 1.0 1.0 10 27 13 0.03 0.01 q.s. q.s.

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at 255 nm in UV-spectrophotometer and % drug content was calculated.

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In vitro drug release This study was carried out using the modified diffusion cell. Samples, each 1 gm of the different formulations, were spread on egg membrane previously soaked overnight in the diffusion medium. The loaded membrane was firmly stretched over the edge of glass tube of 3.10 cm diameter. The tube was then immersed in the receptor medium, which contained 200 ml of the diffusion medium (phosphate buffer, pH 7.4) previously warmed and maintained at 37  1  C and stirred at 100 rpm using magnetic stirrer. Aliquots of 10 ml were withdrawn from the receptor medium at different time intervals. Withdrawn samples were replaced by equal volume of fresh medium. The samples were analyzed at 260 nm against blank using UV-spectrophotometer. Experiments were carried out in triplicates.

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In vivo anti-inflammatory activity The study was conducted in accordance with the approval of the Animal Ethical Committee, MET’S Institute of Pharmacy, CPCSEA Reg. no-1344/ac/10/CPCSEA/IAEC. Edema was induced on the left hind paw of the mice by subplanter injection of 1% w/v carrageenan. Emulgel formulation EG 3 and standard (Fastum gelÕ ) were applied 30 min before carrageenan injection. The paw volume was measured at intervals of 30, 60, 90, 120 and 180 min by vernier caliper.  Group 1 (control group): carrageenan (1% w/v) was injected in the plantar surface of mice.  Group 2 (standard group): topical marketed fastum gel þ carrageenan (1% w/v).  Group 3 (test group): emulgel formulation EG 3 þ carrageenan (1% w/v). Each group contained five mice. The % inhibition of paw edema in drug treated group was compared with carrageenan control group and calculated using the formula: %inhibition ¼ Vc  Vt =Vc  100

Bioadhesion study The fresh mice skin was cut into pieces (2.5 cm2) and washed with 0.1 N NaOH. Two pieces of mice skin were tied to the two glass slides separately; from that, one slide was fixed on the wooden platform and another piece was tied with the balance on right-hand side. The right and left pans were balanced by adding extra weight on the left-hand side. About 1 gm of emulgel was placed between these two slides containing hairless mice skin, and extra weight from left pan was removed to sandwich the two pieces of skin and applied pressure to remove entrapped air. The balance was kept in this position for 5 min, then water was added slowly to the left hand pan until the both skins were separated. The weight (gram force) of water required to detach emulgel from the skin surfaces was noted as bioadhesive strength. The bioadhesive strength is calculated using formula (Khullar et al., 2011):  Bioadhesive strength ¼ Weight requiredðin gramsÞ=Area cm2 Ex vivo diffusion study This study was performed by using the freshly shaven mice skin as a diffusion membrane. Skin was soaked into diffusion medium (phosphate buffer, pH 7.4) overnight and then it was stretched to the diffusion tube. Emulgel formulation, 1 gm was placed on the membrane and dipped it into receptor medium and maintained the temperature at 37  1  C, aliquots of 10 ml were withdrawn at different time intervals, and same volume of buffer was added to maintain sink conditions. The release profile data of prepared emulgel formulation was compared with the marketed gel (Fastum gelÕ ). Skin irritation study A set of five mice was used in the study. The emulgel formulation (EG 3) was applied on the properly shaven skin of mice. Undesirable skin changes, i.e. change in color and changes in skin morphology, were observed for a period of 24 h (Khullar et al., 2012).

where Vc is the inflammatory increase in paw volume control group and Vt the inflammatory increase in paw volume in test group (Khullar et al., 2012).

Results and discussion Evaluation of microemulsion Measurement of globule size and zeta potential The microemulsion had the less globule size as compared to the coarse emulsion due to presence of co-surfactant, which reduces the interfacial tension to ultra low value. Globule size of microemulsion was found to be 396 nm (Table 2), which did not show a conclusive pattern to correlate with formulation components. The small globule size of microemulsion was due to large percent of Smix. Similarly, zeta potential was observed to be 12 mV due to the presence of non-ionic surfactant, which provides stable microemulsion due to the neutral charge present at the diffusive boundary. Phase separation Emulsion is thermodynamically unstable system, which may separate when subjected to physical stresses like centrifugation. Though microemulsions are homogeneous single phase system, they were subjected to centrifugation to confirm the absence of phase separation. Microemulsion did not show any sign of phase separation when subjected to centrifugation, which confirms physical stability of microemulsion. Evaluation of prepared emulgels Physical appearance Emulgel formulations were milky white creamy preparations with a smooth homogeneous texture and glossy appearance, which is presented in Table 3. Table 2. Globule size and zeta potential of microemulsion. Formulation ME 1

Globule size (nm)

Zeta potential (–mV)

396

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Microemulsion-based ketoprofen gel

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DOI: 10.3109/10717544.2013.859186

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pH determination

Globule size measurement

The pH of the topical formulations should be compatible with skin pH. A change in the pH may cause irritation or disruption of skin. The pH of all formulations was modified with the help of TEA and when measured, it was found between 6 and 6.8, which is acceptable for skin preparations.

Globule size of all formulations is given in Table 4. There is slight increase in globule size of all emulgel formulations due to addition of gelling agent into it, as it causes entrapment of oil globule in its network thus, responsible for slight increase in interfacial tension between oil water phase.

Rheological study

Drug content determination

Rheological behavior of the emulgel formulations exhibited non-Newtonian shear thinning pseudo plastic type of flow, i.e. decreases in viscosity at increasing shear rates. As the shear stress is increased (Figure 2), the disarranged molecules of the gelling material are caused to align their long axes in the direction of flow. Such orientation reduces the internal resistance of the material and decreases viscosity. An increase in the concentration of Carbopol 940 and Carbopol 934 (1– 2%) were expected to show increase in viscosity. Emulsifier and clove oil concentrations could be contributing to the rheological properties of the formulations.

The drug content of all formulations was found to be in the range of 98–101%, which is considered as normal according to official monograph (Indian Pharmacopoeia, 2007).

Spreadability Spreadability is the term expressed to denote the extent of area to which the gel readily spreads on application to the skin (Figure 3). One of the essential criterias for an emulgel is that it should have good spreadability. It depends upon the type and concentrations of polymers used in the formulation. More viscous formulation would have poor spreadability. The spreadability values of all formulations were given in Table 4. The formulation EG 3 showed more spreading coefficient, i.e. 22.96, as compared to other formulations, this is because formulation contained optimum concentration of Carbopol 934, i.e. 1.5%.

In vitro drug release In vitro release profiles of ketoprofen from various emulgel formulations are represented in Figure 4. Microemulsion system contains oil globules with entrapped drug molecules which provides enhanced solubility to drug molecule, thus, increases the release of ketoprofen from emulgel than gel. From the data obtained, it was observed that most of emulgel formulations gave better release as compared to marketed gel formulation at the end of 8 h. The higher drug release was observed with formulations EG 3 and EG 7, i.e. 99.42% and 88.92%, respectively. The release profile up to 8 h of all formulation is given in Table 4. The release data were analyzed according to various kinetic models. Most of the studied formulations followed Korsmeyer–Peppas kinetic model. Bioadhesion study The bioadhesion strength of emulgel formulation is given in Table 5.

Table 3. Physical appearance of emulgel formulations. Formulation EG EG EG EG EG EG EG EG

1 2 3 4 5 6 7 8

Color Milky Milky Milky Milky Milky Milky Milky Milky

white white white white white white white white

Homogeneity

Consistency

Homogeneous Homogeneous Homogeneous Homogeneous Homogeneous Homogeneous Homogeneous Homogeneous

Excellent Excellent Excellent Excellent Excellent Excellent Excellent Excellent

Figure 3. Spreadability of all formulations..

Table 4. Data showing spreadability, globule size and percent drug release.

Figure 2. Rheological behavior of emulgel formulations.

Formulations

Spreadability (gm.cm/s)

Globule size (nm)

Percent cumulative drug release (mean  SD)

EG 1 EG 2 EG 3 EG 4 EG 5 EG 6 EG 7 EG 8 Marketed gel

14.67 16.07 22.96 12.05 13.5 17.76 19.85 16.07 14.06

481.3 526.5 473 419.1 448.5 494.7 468.2 516 –

61.91  1.25 59.48  0.85 92.42  4.66 64.43  0.14 87.34  0.38 67.47  0.91 88.92  2.50 77.05  2.70 65.94  3.30

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Ex vivo diffusion study The ex vivo diffusion study was performed using fresh mice skin. Study was performed using marketed gel and formulated emulgel (EG 3) (Figure 5). The marketed gel showed 54.44% release of ketoprofen, whereas formulated emulgel showed 72.22% at the end of 8 h (Table 6). Both of these figures indicated that formulated emulgel gave higher flux and permeation as compared to standard marketed gel preparation.

The p value is 50.0001, which is considered as extremely significant by applying analysis of variance (ANOVA) followed by paired ‘‘t’’ test. Skin irritation study This study was performed on mice. After application of emulgel and observed for 24 h, formulation did not indicate any evidences of skin irritation such as redness of skin or any change in morphology of skin. Thus, it may be concluded that formulation does not have skin irritation potential and hence, safe for topical application.

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In vivo anti-inflammatory activity The anti-inflammatory activity of the formulation EG 3 (test) was compared with marketed ketoprofen gel (fastum gel) i.e., standard group. The % inhibitions of standard and test group were 65.92% and 51.08%, which indicated that formulated emulgel was more effective than marketed gel (2.5%) in its anti-inflammatory activity even at low concentration, i.e. 2% (Table 7). The p value is 50.0001 is considered as extremely significant by applying ANOVA test followed by Dunnett’s test.

Figure 4. In vitro release profile of all formulations.

Conclusion Table 5. Bioadhesion strength. Formulation

Bioadhesion strength (gram force)

EG 3

48.4

Amongst all formulations, emulgel prepared with oil (10%), S/Cos (40%) and Carbopol 934 (1.5%) was better with respect to overall formulation qualities. Developed microemulsion system provides solubilization of hydrophobic drug, thus impart availability of ketoprofen in formulation, whereas globule size and zeta potential was 396 nm and 12 mV, respectively, indicating the stability and proper formulation of microemulsion. The prepared emulgel can be considered as cost effective formulation because of reduction of topical dose of ketoprofen in formulation. The highest release was showed Table 6. Comparative ex vivo release profile. Percent cumulative drug release

Time (h)

Figure 5. Comparative ex vivo release profile of formulated emulgel and marketed gel.

1 2 3 4 5 6 7 8

EG 3

Marketed gel

9.43 11.08 18.81 28.01 36.38 53.63 63.06 72.22

8.34 9.32 13.63 19.26 24.09 29.98 46 54.44

Table 7. Percent inhibition of paw edema. Paw volume (mm) (mean  SEM)

Group Control Standard Test

Percent inhibition

0 min

30 min

60 min

120 min

180 min

2.43  0.13 17 2.58  0.11 86 2.2  0.06 48

4.26  0.26 48 4.24  0.24 47 4.02  0.18 14

4.57  0.29 73 3.8  0.20 07 3.62  0.23 66

4.7  0.29 71 3.33  0.15 49 3.32  0.20 33

4.82  0.31 92 3.52  0.27 76 3.48  0.22 90

0 65.92 51.08

DOI: 10.3109/10717544.2013.859186

by EG 3 batch (92.42  4.66%), which was increased and prolonged when compared with marketed gel. In addition, their permeation and appearance was found to be more acceptable. All formulations exhibited non-Newtonian pseudo plastic behavior, i.e. it was shear thinning system. Formulation showed no skin irritation potential as confirmed by skin irritation study. Thus, the results of this research study clearly indicated a promising potential of the ketoprofen emulgel as an alternative to the conventional dosage forms.

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Acknowledgements The authors are thankful to Cipla Ltd., Daman, for generously providing gift sample of ketoprofen. They also thank MET’s Institute of Pharmacy, Bhujbal Knowledge City, Adgaon, Nashik, for providing necessary facilities.

Declaration of interest The authors report no conflicts of interest. The authors alone are responsible for the content and writing of this article.

References Bhanu VP, Shanmugam V, Lakshmi PK. (2011). Development and optimization of novel diclofenac sodium emulgel for topical drug delivery. Int J Comprehen Pharm 9:1–4. Bugay DE, Findlay WP. (1999). Pharmaceutical excipients. Vol 94. New York: Marcel Dekker, 116–7. Chen H, Chang X, Weng T, et al. (2004). A study of microemulsion system for transdermal delivery of triptiolide. J Control Rel 98: 427–36. Gordon LA, Jennifer JS, Nehal AK, et al. (2006). Solubalization and dissolution of insoluble weak acid, ketoprofen: effects of pH combined with surfactants. Eur J Pharm Sci 29:306–14. Graf A, Rades T, Hook SM. (2009). Oral insulin delivery using nanoparticles based on microemulsions with different structure-types: optimization and in-vivo evaluation. Eur J Pharm Sci 37:53–61. Hyun JC, Wan SK, Ubovan T, et al. (2012). Development of udenefil loaded microemulsions for intranasal delivery: in vitro and in vivo evaluations. Int J Pharm 423:153–60. Indian pharmacopoeia Vol II. (2007). Government of India, Controller of Publications, Ghaziabad: Ministry of health and family welfare, 1259.

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Jain A, Gautam S, Gupta Y, et al. (2010). Development and characterization of ketoconazole emulgel for topical drug delivery. Pelagia Res Lib 1:221–31. Joshi B, Singh G, Rana AC, et al. (2011). Emulgel: a comprehensive review on the recent advances in topical drug delivery. Int Res J Pharm 2:66–70. Joshi B, Singh G, Rana AC, Saini S. (2012). Development and characterization of clarithromycin emulgel for topical delivery. Int J Drug Dev Res 4:310–23. Khatib NA, Pawase K, Patil PA. (2010). Evaluation of anti-inflammatory activity of Garcinica Indica fruit rind extracts in wistar rats. Int J Res Ayu Pharm 1:66–70. Khullar R, Kumar D, Seth N, Saini S. (2012). Formulation and evaluation of mefenamic acid emulgel for topical drug delivery. Saudi Pharm J 20:63–7. Khullar R, Rana AC, Seth N, Saini S. (2011). Emulgels: a surrogate approach for topically used hydrophobic drugs. Int J Pharm Bio Sci 1: 117–28. Khunt MD, Mishra AD, Shah DR. (2012). Formulation design and development of piroxicam emulgel. Int J Pharm Tech Res 4:1332–44. Magdy IM. (2004). Optimization of chlorphenesin emulgel formulation. The AAPS J 6:1–7. Mostafa S, Hady S, Hammad M, Mortada N. (2011). Optimized formulation for topical administration of clotrimazole using pemulen polymeric emulsifier. Drug Dev Ind Pharm 27:1083–97. Patravale VB, Bachhav YG. (2009). Microemulsion based vaginal gel of flucanazole: formulation, in-vitro and in-vivo evaluation. Int J Pharm 365:175–9. Renceber S, Karavana S, Ozyazici M. (2009). Bioavailability file: ketoprofen. J Pharm Sci 34:203–16. Rowe CR, Sheskey PJ, Quinn M. (2009). Handbook of pharmaceutical excipients. 6th ed. New York: Pharmaceutical Press, 108–9. Sahle FF, Metz H, Wohlrab J, Neubert R. (2012). Polyglycerol fatty acid ester surfactant-based microemulsions for targeted delivery of ceramide AP into the stratum corneum: formulation, characterization, in vitro release and penetration investigation. Eur J Pharm Biopharm 3:139–50. Singla V, Saini S, Joshi B, Rana AC. (2012). Emulgel: a new platform for topical drug delivery. Int J Pharm Bio Sci 3:485–95. Thakur V, Prashar B, Arora S. (2012). Formulation and in-vitro evaluation of gel for topical delivery of antifungal agent flucanazole using different penetration enhancers. Drug Invent Today 4:414–19. Tripathi KD. (2003). Essentials of medical pharmacology. 5th ed. New Delhi: Jaypee Brothers Medical Publishers Pvt. Ltd, 168. Willey MJ, Sherwood ML, Woolverton JC. (2008). Prescott, Harley and Klein’s Micribiology. 7th ed. New York: McGraw-Hill, pp. 756–9. Yang X, Chen H, Chang X, et al. (2006). Microemulsion-based hydrogel formulation of ibuprofen for topical delivery. Int J Pharm 315:52–8.

Formulation development, in vitro and in vivo evaluation of microemulsion-based gel loaded with ketoprofen.

Anti-inflammatory agents are widely used to relieve inflammation caused by various factors...
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