Innovative Vesicular Drug Delivery System of Ibuprofen with Unsaturated Fatty Acids: Formulation and Evaluation
T.G. Rukari*, P.A. Gavali, V.J. Patil, S.S. Gaonakr, K.S. Rathod, Y.M. Patil,
Y.R. Dhongade, V.A. Jagtap
Yashwantrao Bhonsale College of Pharmacy, Sawantwadi.
*Corresponding Author E-mail: tushar.rukari@gmail.com
ABSTRACT:
Inflammation is a complex physiological response involved in numerous pathological conditions, requiring effective treatment. Non-steroidal anti-inflammatory drugs (NSAIDs) have long been utilized to alleviate inflammation associated symptoms. Conventional topical formulations of NSAID suffer from limitations such as poor skin penetration, low drug retention, and inadequate therapeutic efficacy. To overcome this limitations, novel vesicular drug delivery systems, particularly transferosomes, have emerged as promising alternatives. Transferosomes, a specialized type of deformable liposomes, offers unique advantage for targeted drug delivery through the skin. They can deform and squeeze through narrow intercellular spaces, enhancing drug permeation into underlying tissues. This facilitates deepest penetration of NSAIDs into inflamed regions, maximizing therapeutic effects while minimizing systemic exposure and adverse effects. Transferosomes, formed via thin film hydration method with oleic acid, cholesterol, surfactant, and organic solvent. They have a particle size of 297nm and a mean zeta potential of -24.2mV. Entrapment efficiency is 83% and demonstrates effective drug retention. The optimized batch is chosen on particle size and entrapment efficiency, and these incorporated into carbopol 934 gel and evaluated for appearance, grittiness, texture pH, spreadability, extrudability, drug content and in-vitro drug release. This comprehension evaluation ensures formulation efficiency. Overall, the development and evaluation of novel vesicular drug delivery systems, particularly transferosomes, represent significant advancement in drug delivery technology. By utilizing their unique properties, these systems hold promise for improving therapeutic efficacy and enabling sustain release in the treatment of inflammatory conditions.
KEYWORDS: Oleic Acid, Transferosomes, Anti-inflammatory, In-vitro drug release etc.
INTRODUCTION:
Inflammation is a biological response involving blood cells and fluid release in damaged tissues often linked to conditions like cardiovascular disease and arthritis1,2.
Non-steroidal anti-inflammatory drugs inhibit COX enzymes to reduce prostaglandin production, alleviating pain and inflammation without affecting consciousness3. Among other NSAID Ibuprofen has lower risk of GI ulcers according to the epidemiological research, notwithstanding concern4. Its racemic form can harm the gastrointestinal tract and heart, particularly with frequent dosing due to its short half-life. Topical formulations have been developed to improve bioavailability and reduce systemic side effects associated with its lipophilicity and poor water solubility5.
Topical drug delivery systems like gels, ointments and creams offer advantages such as by passing first pass metabolism, improving patient compliance, and allowing for easy dose adjustments. However, they often face challenges including poor skin penetration and low bioavailability6. To address this obstacle, vesicular drug delivery system such as liposomes, niosomes, transferosomes have been developed to enhance drug delivery through the skin by improving stratum corneum penetration7. Transferosomes stand out due to their flexible structure, which aids in overcoming skin barrier limitations compared to more rigid nano-vesicular systems like liposomes and niosome.
Transferosomes are advanced vesicular carriers designed for transdermal drug delivery of both hydrophilic and hydrophobic molecules as shown in Figure 1. Transferosomes are complex aggregates of highly deformable and stress responsive vehicle. The term transferosomes was introduced in 1991 by Gregor Cevc. Structural composition of transferosomes includes phospholipids, cholesterol which form as lipid bilayer and surfactant such as span and tween. The role of surfactant is that of edge activator which decreases rigidity of lipid bilayer and increases flexibility of transferosomes8. In this study transferosomes were prepared using unsaturated fatty acid like oleic acid which further improves their flexibility and penetration into the skin9.
MATERIAL AND METHODS:
Material:
Oleic acid, Cholesterol, Span 80, Tween 80, Methanol, Chloroform, Ibuprofen, Phosphate Buffer (pH 7.4) were used of analytical grades and issued from central chemical store of the institute.
Determination of ʎ max:
The dilution was obtained to the concentration of 1000 µg/ml for the Ibuprofen and the solution was scanned in the UV range 200-400 nm in 10 mm cell against solvent blank Phosphate buffer (pH 7.4). The study of spectrum revealed that Ibuprofen shows well- defined at ʎ max 262nm. This wavelength was selected for the development of a standard calibration curve.
Calibration Curve of Ibuprofen:
For determination of the calibration curve of ibuprofen 100 mg drug dissolved in 50 ml of Phosphate buffer (pH 7.4) then sonicate for 10 min. to get standard stock solution. The final volume of the stock solution makes up to 100ml with the same solvent to get stock solution obtaining 1000 µg/ml. 1ml of above stock solution was pipette out in another volumetric flask and diluted with Phosphate buffer (pH 7.4) up to 100 ml to get second stock solution obtaining 10 µg/ml. Further, 1 ml to 10 ml of second stock solutions diluted up to 10 ml to get concentration 1 µg/ml to 10 µg/ml. These prepared dilutions were analyzed by UV- Visible Spectrophotometer at 262 nm10.
Preparation of Transferosomes:
In this research, transferosomes are prepared by using thin film hydration technique. Ibuprofen, oleic acid, and cholesterol were used to prepare transferosomes as per the quantity mentioned in Table 1. In a 20 ml mixture of two organic solvents (methanol: chloroform 1:1) dissolved oleic acid, cholesterol, and ibuprofen. This solution is placed in clean, dry bottom flask. The organic solvent was evaporated at reduced pressure to create lipid layer on the flask wall. A phosphate buffer solution (pH 7.4) was hydrated by rotating for 1h at room temperature at 60 rpm. Using a sonicator multilaminar lipid vesicles (MLV) are sonicated for 10 min11.
Table 1: Formulation of Transferosomes
|
Sr. No. |
Chemicals |
TFS1 |
TFS2 |
TFS3 |
|
1. |
Oleic acid (mg) |
100 |
200 |
300 |
|
2. |
Cholesterol (mg) |
25 |
50 |
75 |
|
3. |
Methanol (ml) |
10 |
10 |
10 |
|
4. |
Chloroform (ml) |
10 |
10 |
10 |
|
5. |
Span 80 (ml) |
10 |
10 |
10 |
|
6. |
Tween 80 (ml) |
10 |
10 |
10 |
|
7. |
Ibuprofen (mg) |
100 |
100 |
100 |
|
8. |
Phosphate buffer (pH 7.4) (mg) |
10 |
10 |
10 |
Preparation of Transferosomes Entrapped Ibuprofen Gel:
Transferosomes loaded gel is prepared as per Table 2. The transferosomal suspension were transferred into the beaker, which was placed on the magnetic stirrer. The rpm was set between 200-400. 0.1 N NaOH was added to adjust the pH 6-7. Then add carbopol 934 and HPMC K4M. Stirred continued until a gel like consistency formed. Then propylene glycol, and glycerin were added. Then add triethanolamine to adjust the pH. Stirred for 5-10 minutes until a gel with desired consistency was achieved.
Table 2 Formula of Transferosomes Entrapped Ibuprofen Gel
|
Sr. No. |
Ingredients |
TFS-G1 |
Uses |
|
1. |
Transferosomal Suspension (ml) |
10 |
Anti-inflammatory |
|
2. |
Carbopol 934 (gm) |
0.25 |
Gelling Agent |
|
3. |
HPMC K4M (gm) |
0.05 |
Co-Polymer |
|
4. |
Triethanolamine (ml) |
0.5 |
pH-adjuster |
|
5. |
Propylene Glycol (ml) |
1 |
Viscosity modifier |
|
6. |
Glycerin (ml) |
1 |
Humectant |
Evaluation of Transferosomes:
1. Particle size:
Particle size can be determined using dynamic light scattering system by Anton Parr.
2. Zeta potential:
The significance of zeta potential is that it’s value can be related to the stability of colloidal dispersion. The zeta potential indicates the degree of repulsion between adjacent, similarly charged particles in dispersion. Zeta potential can be determined by using dynamic light scattering system by Anton Parr.
3. Entrapment Efficiency:
In a slightly modified method, 1ml of transferosomal suspension was diluted to 10ml with ethanol and then filtered using Whatman filter paper with a pore size of 2.5µm. The absorbance of the filtrate was measured at 262 nm. The entrapment efficiency was calculated from the following equation12:
%Entrapment Efficiency= Amount of drug entrapped/ Total Drug× 100
Evaluation of Transferosomes Entrapped Ibuprofen Gel:
1. Physical appearance: The formulation of ibuprofen gel was developed, and its physical appearance was visually evaluated.
2. Grittiness: The existence of any particles in the produced gels was also assessed. Gel smears were made on glass slides, and they were examined under a microscope to check for any particles or grittiness11.
3. Texture analysis: Consistency, elasticity and adhesiveness of gel can be determined using compression-relaxation-traction (CRT) test by Tex’an touch 50 N.
4. Extrudability: The gel formulation was filled in standard capped collapsible aluminum tubes and sealed by crimping to the end. The weights of the tube were recorded. The tube was placed between two glass slides and were clamped. 500 g wight was placed over the slides and then, the cap was removed. The amount of extruded gel was calculated (>90% extrudability: excellent, >80% extrudability: Good, and >70% extrudability: Fair)13.
Extrudability= (Weight of gel extruded/ Weight of formulation) × 100
4. Spreadability: One of the criteria for gel meet ideal quality is that it should possess good spreadability. Spreadability of gel measured by compressing the gel into the two slides. To compress the samples to consistent thickness an excess of gel (1 gm) was sandwiched between two slides and about 100gm of weight was applied to the slides for 60 seconds. After the 60 seconds the diameter of circles formed from the spreaded gel were measured. The reading was put into the formula.
S= M×L/T
S: Spreadability, M: Mass, L: Diameter, T: Time
5. pH Measurements:
1g gel was accurately weighed and dispersed in 100ml purified water. The pH of the dispersion was measured using digital pH meter, which was calibrated before use with standard buffer solution at 4.0, 7.0, and 9.0. The measurements of pH were done.
6. Viscosity Measurements:
The measurement of viscosity of the prepared gel was done using Brookfield digital Viscometer. The viscosity was measured using spindle no. 64 at 100 rpm and 25 °C. Before measurements deaeration of gel was done and gel was filled in appropriate wide mouth container. Samples of the gels were allowed to settle over 30 minutes at the assay temperature (25 ± 10 °C) before the measurement.
7. Skin Irritation Test: Make an area 1 sq.cm on left -hand side dorsal surface. The gel was applied to the specified area and time was noted. Irritancy, erythema, edema was checked if any, for regular interval up to 24 h & reported.
8. Drug Content: 1 gm of the prepared gel was mixed with 10 ml of ethanol and sonicated for 5 minutes. The prepared solutions were filtered using Whatman filter paper with pore size of 2.5 µm. Subsequently, the absorbance was measured at 262 nm, and the drug content was calculated by linear regression analysis of the calibration curve14.
9. In-vitro Drug Release:
i. Preparation of Egg Membrane: A small hole was made at the bottom of raw egg and content was removed. The eggshell was wash with dil. HCl (150 ml conc. HCl + 200 ml water) Calcium carbonate of eggshell leaving the membrane. Further wash membrane with water and used for study15.
ii. In-vitro Drug Release: The Ibuprofen drug diffusion from Phosphate buffer across egg membrane was performed using the validated Franz cells and equipment. A clean, dried receptor cell was filled with Phosphate buffer and allowed to equilibrate at 37 °C in the heated magnetic stirrer for 2 h. The egg membrane placed into the Phosphate buffer before placed into the Franz diffusion cell. After that egg membrane mounted between matched donor and receptor compartment. 1 g of Ibuprofen gel was placed on the membrane surface in the donor compartment and an adequate quantity of Phosphate buffer is added to make the gel thin. The receptor compartment stirred at 300 rpm. Using a syringe, the sample volume of 1 ml was extracted. The Phosphate buffer of the same volume was reintroduced into the receptor. The sample volumes were assayed under UV-Vis spectroscopy at 262 nm. Air bubbles formed below the membrane were removed by carefully tilting the Franz diffusion cell for the air bubbles to escape via the sampling arm. Intervals between sampling varied from 15 minutes to 1 h. The comparative study between transferosomal gel and Ibuprofen suspension was performed. The Ibuprofen suspension was prepared by dissolving Ibuprofen in water that contained 1% w/v of Tween 80 and Span 80 as dispersing agents, followed by stirring for 3 h16.
RESULT:
Calibration curve:
The ʎ max of Ibuprofen in Phosphate buffer (pH 7.4) was found to be 262nm and all obtained values are given in the table 3. It obeys Beer lambert’s law. Regression analysis revealed a linear relationship between absorbance and concentration, with regression coefficient value is 0.9948.
Table 3: Calibration Curve of Ibuprofen
|
Sr. No. |
Concentration (µg/ml) |
Absorbance |
|
1. |
1 |
0.0171 |
|
2. |
2 |
0.0185 |
|
3. |
3 |
0.0201 |
|
4. |
4 |
0.021 |
|
5. |
5 |
0.0219 |
|
6. |
6 |
0.0229 |
|
7. |
7 |
0.0243 |
|
8. |
8 |
0.0253 |
|
9. |
9 |
0.0264 |
|
10. |
10 |
0.0271 |
Figure 1. Calibration curve of Ibuprofen
Evaluation of Transferosomes:
1. Particle size:
Particle size is determined by particle size analyzer using DLS technique. The particle size of formula TFS1 had smallest size of 297nm while the formula TFSF3 had largest size of 7115nm. The particle size of optimized transferosomes formulation as presented in figure 2.
2. Zeta Potential:
Light Scattering method using Zeta sizer was utilized to determine the zeta potential by using Anton paar instrument. The zeta potential values of formulation were detected in range of -48mV to -24.2mV. The zeta potential of optimized formulation is shown in figure 2.
3. % Entrapped Efficiency:
The entrapped efficiency percent of transferosomes was detected to be in range of 73.96% to 83%. The formula TFS1 significantly showed the highest percent of entrapped efficiency approaching 83%. The formula TFS1 showed the smallest particle size (297nm) and good percent entrapped efficiency (83%). Accordingly, the formula TFS1 was incorporated to formulate gel. Evaluation parameters are summarize in Table 4.
Table 4: Characterization of Transferosomes
|
Formulation |
Particle Size |
Zeta Potential |
%Entrapped Efficiency |
|
TFS1 |
297nm |
-24.2mV |
83% |
|
TFS2 |
3688nm |
-48.0mV |
79.37% |
|
TFS3 |
7115nm |
-32.2mV |
73.96% |
Figure 2. Particle size analysis and zeta potential of optimized batch
Evaluation of Transferosomes Entrapped Ibuprofen Gel
1. Appearance:
The appearance of formulation was found to be gel like without any lump.
2. Grittiness:
The prepared formulation is free from any particulate matter, showing no grittiness.
3. Texture analysis:
The curve identifies three phases based on force over time. Key parameters include F max at 373.41 g, representing the products consistency under defined compression conditions. % Relaxation at 56.33 % indicates the product’s elasticity inversely. F min, measured at 138.18 g, reflects the traction or adhesion force when probe is removed from the sample. Therefore, gel shows favorable characteristics, in terms of consistency, elasticity and adhesiveness.
Figure 3. Texture analysis of gel
4. Extrudability:
The extrudability of gel formulation was found to be 92.5%. Thus, extrudability of formulation is excellent.
5. Spreadability:
The spreadability of formulation is 6.4 cm. Thus, spreadability was found to be satisfactory.
6. pH Measurement:
The prepared formulation was evaluated for its physiochemical properties, and it was found that the pH of formulated Ibuprofen gel was found to be 6.15.
7. Viscosity measurement:
The viscosity of formulation was found to be 5892 Cp.
8. Skin Irritation Test:
To study any irritant effect patches of formulation were applied on skin and noted for 24 hours under normal condition. After 24 hours no itching, redness, edema, erythema was found. This depicts no irritancy of formulation. Thus, prepared formulations can be applied on skin.
9. Drug Content:
The drug content of transferosomal gel was found to be 63mg, which represent good content uniformity.
10. In-vitro Drug Release:
Drug diffusion studied were done on Franz diffusion cell apparatus with help of egg membrane and by maintaining proper pH with buffer. Drug diffused through egg membrane was checked with help of UV analysis. % Drug release from 1gm of gel was found to be 90.51%. % Drug release from Ibuprofen suspension was found to be 87.67%.
Table 5 In-vitro Drug Release
|
Time (h) |
% Drug Release of Transferosomal Gel |
% Drug Release of Ibuprofen Suspension |
|
0.25 |
37.16712 |
19.38381 |
|
0.5 |
48.72627 |
26.14147 |
|
0.75 |
49.08194 |
28.27546 |
|
1 |
49.9711 |
77.00173 |
|
2 |
52.1051 |
81.62539 |
|
3 |
57.79576 |
88.56089 |
|
4 |
57.97359 |
87.6712 |
|
5 |
66.50958 |
- |
|
6 |
81.0919 |
- |
|
7 |
82.87023 |
- |
|
8 |
88.56089 |
- |
|
9 |
90.51705 |
- |
Figure 4. In-vitro drug release
DISCUSSION:
The project originated from discussion surrounding the insufficient topical anti-inflammatory activity NSAID drugs. To address this concern, we opted for novel drug delivery system utilizing Ibuprofen as the anti-inflammatory agent. Following this decision, we proceeded to formulate transferosomes batches. Three formulations varied in Oleic acid, Cholesterol, Span80, Tween80 showing ranges from 297 nm to 7115 nm and entrapment efficiency 73.96-83% for Ibuprofen. From this evaluation the formulation showing the smallest particle size (297nm) and highest entrapment efficiency (83%) is selected for formulating a gel. Gel formulations were developed with varying quantities of Carbopol 934, from which an optimum batch was selected for further evaluation. The evaluation of gel formulations encompassed several parameters including appearance, grittiness, texture analysis, extrudability, pH, viscosity, irritancy, spreadability, drug content, and percentage drug release. Upon completion of these assessment, a comparative analysis was conducted between the gel formulation and the transferosomal suspension. Notably, the gel dosage from exhibited sustained release characteristics, a favorable trait for prolonged therapeutic activity. From the comprehensive evaluation and comparison, it was evident that the transferosomal gel formulation outperformed the conventional drug delivery system. This suggests that oleic acid enriched transferosomal gel as a promising approach for enhancing the therapeutic efficacy of ibuprofen in topical application.
CONCLUSION:
The study successfully developed and evaluated an innovative transferosomal drug delivery system containing oleic acid for Ibuprofen aimed at improving its topical anti-inflammatory effects. The optimized batch was selected based on particle size, and entrapment efficiency. The optimized formulation F1 showed entrapment efficiency EE (83%), and small particle size (297nm). The optimized formulation of transferosomes was further formulated to gel with Carbopol 934 along with HPMC k4M, Propylene glycol, Glycerin and Triethanolamine. The transferosomal gel shows whitish color, favorable consistency, elasticity, and adhesiveness. The spreadability is 6.4 cm, and pH of 6.15. The actual drug content of transferosomal gel was 62mg, which represent good content uniformity. The viscosity of Ibuprofen transferosomal gel is found to 5892 cp. The Ibuprofen transferosomal gel released 90% of the drug, indicating efficient drug release. These optimized transferosomal formulation demonstrate promising potential for targeted and sustained delivery of nonsteroidal anti-inflammatory drug (NSAISs). Upon comparative analysis with transferosomal suspension, the gel formulation demonstrated sustained release characteristics, including its potential for prolonged therapeutic activity. Overall, the transferosomal gel formulation exhibited superior performance compared to convential drug delivery system, highlighting its promise for enhancing the therapeutic efficacy of NSAID drug in topical application.
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Received on 02.09.2024 Revised on 26.10.2024 Accepted on 30.11.2024 Published on 18.12.2024 Available online on December 21, 2024 Asian J. Pharm. Tech. 2024; 14(4):319-324. DOI: 10.52711/2231-5713.2024.00052 ©Asian Pharma Press All Right Reserved
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