Formulation and Evalutaion of Ethosomal Transdermal Patch of Antidiabetic Drug
Purvi Yadav1, Neha Thakur1, Anju Mishra2, Geetanjali Sahu2, Anish Chandy3
1Students, School of Pharmacy, Chouksey Engineering College, Bilaspur (CG) India.
2Assistant Professor, School of Pharmacy, Chouksey Engineering College, Bilaspur (CG), India.
3Associate Professor, School of Pharmacy, Chouksey Engineering College, Bilaspur (CG), India.
*Corresponding Author E-mail: anishpharma@gmail.com
ABSTRACT:
Objective: The primary goal is to prepare appropriate transdermal patches of the antidiabetic medication glimepiride using varying ratios of polymers (hydroxyl propyl methyl cellulose K-100) in accordance with patient needs (patient compliance) and for superior drug release compared to alternative formulations. These formulations use ethanol and chloroform (1:1) as a solvent system and 0.2 ml of glycerol as a plasticizer. The in-vitro skin penetration experiments were conducted using a Franz diffusion cell and phosphate buffer pH 7.4. Method: A magnetic stirrer was used to prepare the transdermal patches utilizing the solvent casting process. Results: Beer's law was adhered to. between 2 and 12 μg/ml in phosphate buffer at a pH of 7.4. Linear regression equation of pure drug was established as Y=0.058x + 0.1331 and the R 2 value 0.999. Zero order release kinetics implies that the drug release of formulation GTP7 is better than that of the other formulation. Conclusion: Formulation GTP7presented a statistically insignificant change of the percent drug content and permeation profile after 90 days of studies.
KEYWORDS: Glimepiride, HPMC K-100, Franz diffusion cell, T2DM.
Diabetes mellitus (DM) is a set of metabolic diseases having different root causes but sharing a symptom (hyperglycemia) in common. Diabetes mellitus presents a significant clinical and economic burden on the society1. India is leading in the number of cases of diabetes in the world (32 million which is expected to reach 78 million in 2030 based on WHO estimates).
The number of people affected by diabetes will exceed 350 million by 2025 which is according to the estimates of World Health Organization, more than 75% of these will be in the developing world2.
Glimepiride is a third-generation sulfonylurea medication that is employed in the treatment of diabetes. This drug is commonly used on patients with type 2 diabetes. Nonetheless, its insufficient solubility and, low bioavailability Glimepiride would be preferable drug to be used in the formulation of transdermal patches. The most competent drug application method employed in case of potent drug is the transdermal drug delivery. It has been in the past years that renewed interest has been generated in the development of transdermal drug delivery systems of existing drug molecules. An advance in the form of the development of a transdermal delivery system of the already existing drug molecules enhances the performance of the drug in terms of safety, efficacy, and therapeutic benefit in addition to patient compliance3.
To have convenient, safe, and successful anti-diabetic activities a transdermal delivery method of glimepiride should be experimented. In the case of transdermal patches preparation, we used polymers called HPMC in varying concentrations in the form of nine formulations. The patches were tested in regards to examination of its physical characteristics, as well as in-vitro dissolution properties4.
A gift sample of Pure Glimepiride was collected from Dey, Medical private Ltd Kolkata. Propylene glycol, phosphatides, cholesterol, chloroform, HPMC, and glycerine were taken out of the local market. The reagents that were used in this research were of analytical grade.
Capillary tube method was used to determine melting point of Glimepiride (5mg approx.).
Standard Calibration Curve of Glimepiride and Lambda max was prepared with the help of the UV Visible Spectrophotometer, in a concentration between 2 and 12 mg/mL.
Solubility The experiment was conducted through the Saturated solution of drug in different solvents by shake flask method.
The cholesterol, ethanol (X1) and phospholipids (X2) were placed in a glass container at room temperature with the known quantity of propylene glycol. The mixture was incinerated to 30C in the rain of water. Distilled water was added to the fine stream with the help of an attractive stirrer in a shunt vessel rotating at 1500 rpm and constant flow rate of 1ml/min and thoroughly mixed. The temperature was maintained at 30 degrees Celsius in the experiment. The mixture was stirred for another 5minutes, and the temperature maintained at 30oC. Then the mixture was kept at 4 degrees Celsius. Moreover, the sonication of ethosomes was done at 4oC in 3 cycles of 5 minutes with a 5 minutes gap between each cycle5.
Zetatrac measured the zeta capacity and the molecular size of the prepared vesicles using dynamic light dissipating technique6.
Each formulation was placed in an Eppendorf tube (n =3) and centrifuged at 16000rpm and 40C in 30 minutes. (Sigma Laboratory axis, 3K30, Ostrode, Germany). The concentration of free drug of glimepiride (288nm) was calculated by isolating and sifting the supernatant.
The scanning electron microscopy (SEM) was used to examine the morphology of the chosen ethosomal formulation.
Solvents cast technique was used to prepare Ethosomal Transdermal patches in varying concentrations of HPMC K 100. A solvent system is the mixture of ethanol and chloroform (1:1). a proper amount of HPMC K100M in a solvent system and a homogenous solution was prepared through a permanent stir over a magnetic stir. The drug loaded ethosomal suspension and plasticizer (glycerol 0.2ml) were then added to the system. The overall formulation was transferred to pre lubricated Petri disc. It is stored inside hot air oven at 350C degree in order to remove all the additional solvent within 40 minutes. Then the Petri plates are transferred to the room temperature and stored there 24hrs. 24hour patches are cut into the appropriate size to be evaluated further7.
Table 1: Preparation of Ethosomal patch
|
Batch |
Ethosomal suspension |
Ethanol(ml) |
Chloroform (ml) |
HPMC K100M (mg) |
|
GTP1 |
10ml |
2.5ml |
2.5ml |
50 |
|
GTP2 |
10ml |
2.5ml |
2.5ml |
55 |
|
GTP3 |
10ml |
2.5ml |
2.5ml |
60 |
|
GTP4 |
10ml |
2.5ml |
2.5ml |
65 |
|
GTP5 |
10ml |
2.5ml |
2.5ml |
75 |
|
GTP6 |
10ml |
2.5ml |
2.5ml |
80 |
|
GTP7 |
10ml |
2.5ml |
2.5ml |
85 |
|
GTP8 |
10ml |
2.5ml |
2.5ml |
90 |
|
GTP9 |
10ml |
2.5ml |
2.5ml |
95 |
The transdermal patches formulated were tested with regard to their Physical appearance.
In the case of drug-loaded patches, the screw gauge micrometer was used to measure the thickness at three points. The average, the standard deviation and the mean of every drug-loaded patch were calculated through averaging the three measurements obtained8.
A digital scale was applied to make sure that nothing weighed more than the other. In all the formulations, we weight all the patches and measured them three times. The data was then used to calculate the mean and the standard deviation.
This will be an experiment on the effectiveness of the plasticizer and the patch. A folding endurance test was conducted in order to establish the durability of the materials manufactured using different polymers. It is the folding motion that is required to tear a patch of polymer. Fold endurance testing of the film was conducted manually by way of folding a 2 cm x 2 cm strip of the film in the same position until it broke. To measure the value of endurance of folding, we used how many times we could fold the patch at the same location before it was able to crack. Three samples of each type were taken in order to evaluate.
The stability and integrity of the films in very humid conditions was tested by a percent moisture absorption test. The 3.14 cm2 films that were individually made and weighed, were put in a desiccator containing 100 ml of a saturated solution of potassium chloride and left to be exposed to 75% relative humidity at room temperature. The films were weighted at 24, 48 and 72 hours during this time.
The following formula was used to calculate the moisture content retention:
Percentage Moisture Content = [(Initial weight - Final weight) / Final weight] x 100
The test was conducted so that films would not deteriorate when in air. The transdermal films were kept in a desiccator containing fused anhydrous calcium chloride at room temperature. The films at this period were weighed at 24, 48 and 72hours’ periods.
Percentage moisture content calculated using the following formula:
% Moisture Content = [(Initial weight – Final weight) / Final weight] x 100
The percentage content of drugs (UV method- stock-solutions of Glimepiride) was estimated simultaneously. The stock solutions were further diluted between 2-12ug/ml of drug. The double beam UV spectrophotometer was used to measure the wavelength with maximum absorbance and absorptivity of was. The maxima of absorbance of Glimepiride are 288 nm. The values of absorptivity of Glimepiride are(ay1) at 288 nm= 0.215 and (ay2) at 226nm= 0.039.
Franz diffusion cell was used to measure drug release of a synthetic membrane combination patch (9, 10). A Franz diffusion cell has two components: the donor and the receptor. Receptor compartment has a volume of 5-12ml and an effective surface area of 0.5-1.5 cm2. Diffusion buffer is stirred by using magnetic beads at a constant rate of 50 revolutions per minute. The temperature is regulated by pumping bottle-expressed water through water coat which encircles the receptor compartment after the time interval of 1, 2, 3, 4, 5, 6, 7, 8, 12, 14, 24hours. Every time 1 ml of test was taken off the receptor with 6.8 pH an equivalent volume of new receptor support (6.8pH) solution was added to make the receptor medium remain in its sink condition. UV spectroscopy at 288 nm is concurrently done in order to establish the composition of a drug.
The obtained sample of the glimepiride was white in colour and was crystalline in nature and the melting points of glimepiride were measured using a melting point instrument and were found to be 208.45 degrees Celsius. This was within the standard range of the melting point of Glimepiride that is 207-209ºC.53
The UV spectrum presents absorption peaks at 288 nm and this is consistent with the earlier reported ones5.
The calibration curve was found to be linear at a range of concentration of 2.0 -12.0ug/mL, which was in accord with the law of Beer and Lambert. The correlation co-efficient of the curve that was drawn using linear regression technique was 0.999 (table 2 and figure 1). The data of the linear regression of the calibration plot is showing that there is good linear relationship between the absorbance and the concentration over a broad range. The correlation coefficient was representative of high significance. The low value of intercept of the ordinate indicated that the calibration plot was not out of line (6).
Table 2: Standard curve of Glimepiride in 0.1N HCI at 288 nm
|
S. No. |
Concentration(µg/ml) |
Absorbance |
|
1 |
2 |
0.248 |
|
2 |
4 |
0.366 |
|
3 |
6 |
0.487 |
|
4 |
8 |
0.589 |
|
5 |
10 |
0.711 |
|
6 |
12 |
0.832 |
R2= 0.999 Y=0.058x + 0.1331
Figure 1: Calibration curve of Glimepiride in 0.1N HCI at 288 nm
The solubility of glimepiride is very low (5.01mg/ml) in water, decreasing with an increase in the pH of the water and the solubility level almost matches DMSO level (28.32mg/ml) but is even lower in phosphate buffer at pH 6.8 (28.14mg/ml).
Table 3 gives sizes of ethosomal vesicles. The change in the ethanol and cholesterol levels led to the development of nine different formulations. Glimepiride ethosomal formulations increased in size as the content of ethanol increased. The reason is that the concentration of the ethanol increases the elasticity of the vesicular structures hence causing the size of the vesicles to increase. The size of the vesicles was found between 234+-11.03nm and 824+-11.45nm. The capacity of formulation it is the measure of stability -17.32+-0.86mV to-23.11+-1.03. PDI is a measure of mono dispersity of ethosomal formulation.
Table 3: Zeta potential, Vesicle size and PDI of Glimepiride
|
Formulation |
Zeta potential (-mV) |
PDI |
Vesicle size(nm) |
|
GE1 |
-20.12±1.12 |
0.11±0.02 |
768±10.14 |
|
GE2 |
-23.11±1.03 |
0.22±0.01 |
400±11.33 |
|
GE3 |
-18.27±0.98 |
0.42±0.12 |
234±11.03 |
|
GE4 |
-21.14±0.87 |
0.34±0.01 |
300±12.09 |
|
GE5 |
-19.24±0.76 |
0.305±0.03 |
824±11.45 |
|
GE6 |
-17.32±0.86 |
0.457±0.02 |
628±13.29 |
|
GE7 |
-19.24±1.25 |
0.213±0.01 |
639±11.23 |
|
GE8 |
-21.23±1.24 |
0.256±0.04 |
521±10.05 |
|
GE9 |
-18.21±0.83 |
0.366±0.02 |
826±11.45 |
All Nine formulation containing Glimepiride were tested on EE%. The concentration of ethanol and cholesterol increased EE%s. The entrapment efficiency has a range between 60.83+-2.15%to96.78+-1.17% as indicated in table 4 below.
Table 4: Entrapment Efficiency of Glimepiride
|
Formulation |
EE% |
|
GE1 |
80.11±2.14 |
|
GE2 |
85.76±1.34 |
|
GE3 |
60.83±2.15 |
|
GE4 |
72.47±2.17 |
|
GE5 |
89.11±1.29 |
|
GE6 |
92.03±2.03 |
|
GE7 |
93.47±2.14 |
|
GE8 |
96.78±1.17 |
|
GE9 |
83.11±1.29 |
NOTE: According to the mentioned parameters (maximum EE percentage and minimum vesicle size), the optimum formulation was selected. The best formulation was developed using 5 ml of ethanol and 25 mg of cholesterol as the formulation factors. The ethosomal morphological study was considered to be carried out using the selected formulation.
Morphology of Optimized Glimepiride Ethosome formulation:
The surface morphology of an optimised Glimepiride ethosomes was analysed using SEM. The spiracular features were demonstrated in ethosomes. The development of ethosomal lipids formulations was shown to have a specific multilamellar vesicular structure (Fig. 2).
Figure 2: SEM Image of optimized Glimepiride ethosomal formulation
Nine transdermal patches (GTP1 to GTP9) were made with optimized ethosomal suspension of Glimepiride and tested in terms of thickness, folding strength, moisture contentment and moisture uptake percentage. All the patches were transparent, a little yellowish in colour, and smooth.
The thickness of GTP1-9 patch has varying levels between 0.4108+-0.69mm and 0.5169+-0.11mm. The tabulated results can be seen in table 5 below.
Patches when tested were found to be stable after exposure to the strains in skin folding that are normally present. The endurance to folding of GTP 1 -GTP 9 formulations is 198+ -1.11-287 + -1.07 in their transdermal patches, respectively. Folding using HPMC K100M formulation has produced more successful cases without fractures as compared to any other formulation. The results can be observed in table 5, below.
The transdermal patch took up 4.17+-0.44 percent to 12.67+-0.53 percent moisture across the entire range of formulations, F1-F9. The results can be observed in table 5, below.
The moisture content of the transdermal patches of GTP 1- GTP 9 formulations is 8.42%+/-0.62-19.65%+/-0.51. The results are compiled in Table 5 presented below.
Table 5: Evaluation of Transdermal Patch of Glimepiride
|
Formulation |
Thickness (mm) |
Folding endurance |
Moisture uptake (%) |
Moisture content (%) |
Uniformity Weight (mg) |
|
GTP 1 |
0.5169±0.11 |
298±1.19 |
11.73±0.49 |
5.66±0.34 |
300±1.11 |
|
GTP 2 |
0.4210±0.22 |
218±1.09 |
19.65±0.51 |
4.87±0.44 |
299±1.16 |
|
GTP 3 |
0.5470±0.31 |
262±1.10 |
9.42±0.62 |
12.67±0.53 |
301±1.17 |
|
GTP 4 |
0.5496±0.62 |
287±1.07 |
8.42±0.62 |
5.16±0.34 |
298±1.16 |
|
GTP 5 |
0.4460±0.42 |
232±1.18 |
10.87±0.71 |
4.17±0.44 |
299±1.14 |
|
GTP 6 |
0.4517±0.51 |
211±1.09 |
16.44±0.81 |
11.67±0.53 |
301±1.16 |
|
GTP 7 |
0.4108±0.69 |
203±1.19 |
9.42±0.62 |
5.66±0.34 |
300±1.10 |
|
GTP 8 |
0.4437±0.62 |
229±1.09 |
10.07±0.71 |
4.87±0.44 |
299±1.19 |
|
GTP 9 |
0.4903±0.78 |
198±1.11 |
15.44±0.81 |
12.67±0.53 |
301±1.18 |
The values of Standard deviation of the weight of the manufactured transdermal patches were low at all formulations. Data enlisted in table 5, under.
Between 87+-0.21percent and 99+-0.61percent of the drug was contained in transdermal patches known as GTP1 through GTP 9. The results have been compiled in table 6 together with figure 3.
Table 6: Drug content Percentage of Glimepiride
|
Formulations |
Percentage of drug content |
|
GTP 1 |
98±0.12 |
|
GTP 2 |
93±0.13 |
|
GTP 3 |
95±0.22 |
|
GTP 4 |
90±0.32 |
|
GTP 5 |
89±0.43 |
|
GTP 6 |
93±0.31 |
|
GTP 7 |
91±0.52 |
|
GTP 8 |
99±0.61 |
|
GTP 9 |
87±0.68 |
All the nine formulations (GTP 1 to GTP 9) containing HPMC-K100M showed complete drug release after 24 hours. The release of glimepiride varies between 74.77 to 99.13 in 24hr patch. The highest release of glimepiride 99.13% was found in 24hr with formulation GTP 7 which contained 85 mg of HPMC-K100M. All the results are tabulated in Table 7. It has been represented in figure 4 below.
Figure 3: Drug content Percentage of Glimepiride TD patch formulations
Figure 4: Cumulative Drug release % of Glimepiride from TD patch formulations
Table 7: Cumulative Drug release % of Glimepiride from TD patch formulations
|
Time (Hrs.) |
GTP 1 |
GTP 2 |
GTP 3 |
GTP 4 |
GTP 5 |
GTP 6 |
GTP 7 |
GTP 8 |
GTP 9 |
|
2 |
15.33±1.12 |
14.3±1.10 |
15.99±1.11 |
16.01±1.15 |
16.98±1.19 |
18.11±1.20 |
19.11±1.25 |
19.69±1.26 |
16.11±1.15 |
|
4 |
42.12±1.15 |
39.32±1.12 |
47.11±1.19 |
45.77±1.12 |
46.02±1.22 |
39.65±1.19 |
49.65±1.28 |
45.65±1.20 |
35.02±1.21 |
|
6 |
58.16±1.19 |
55.12±1.14 |
51.13±1.12 |
49.32±1.15 |
62.16±1.25 |
51.12±1.25 |
68.02±1.26 |
64.22±1.25 |
59.16±1.29 |
|
8 |
68.76±1.20 |
69.32±1.21 |
62.32±1.25 |
69.98±1.20 |
67.76±1.28 |
79.79±1.22 |
80.33±1.30 |
76.03±1.16 |
72.76±1.35 |
|
12 |
71.11±1.21 |
73.31±1.22 |
70.32±1.16 |
76.01±1.23 |
86.11±1.30 |
88.12±1.28 |
89.12±1.29 |
86.12±1.32 |
82.11±1.33 |
|
24 |
78.21±1.25 |
74.77±1.29 |
82.19±1.22 |
88.12±1.26 |
93.21±1.28 |
91.12±1.30 |
99.13±1.14 |
97.32±1.33 |
81.21±1.36 |
Thus, in pre-formulation study the drug was tested regarding melting point. The regression R2 was 0.999 and the standard curve of drugs was developed in 7.4 pH. The size of the vesicles was observed at 321+-10.23 nm to 573+-13.09nm. Entrapment efficiency is found to be 68.69+-1.15% to 96.02+-2.54. The resultant better formulation had an EE percentage of 91.22 and a size of the vesicles of 460 nm, which was very close to their expected value. The optimised formulation was found to be 0.77 percent error of EE percent, 0.21 percent error of Vesicle size with a predicted value of EE percent and Vesicle size of 91.33 percent and 461nm, respectively. Cold method was used to prepare ethosomes of Glimepiride. The size of the vesicle, which is affected by Glimepiride, was measured between 234+-11.03nm and 824+-11.45nm. The efficacy of Glimepiride ranges between 60.83+-1.17% to 96.78+-2.15%.
The newer developed formulation possessed EE percentage of 87.24 percent and vesicle size of 402nm which was in excellent agreement with their projected values. Improved formulation error percentages were 0.77EE percent and 0.21 Vesicle size with EE percent and vesicle size values of 87.92percent and 403nm respectively. On the tune of a perfect ethosomal suspension of Glimepiride, nine transdermal patches (GTP1 to GTP9) were produced and tested on parameters such as thickness, folding strength, moisture contentment and moisture uptake percentage. In-vitro drug release study is assessed on all formulation between GTP1 and GTP 9.
The permeability values that ranged between 345μg/cm2 and 979μg/cm 2 indicated that the concentration of cumulative drug permeation declined with the increasing concentration of HPMC K100 M polymer.
The method of preparation of glimepiride transdermal patches that has been revealed in this research is simple. The physicochemical characteristics of all formulations including the thickness and weight change, release information, which revealed that the polymer type and the weight concentration of the polymer influenced the drug release of the patch formulation. The in vitro penetration of penetration enhancers like tween-80 on drug absorption has been investigated. In diabetic therapy, Glimepiride transdermal patches could be useful moderately in the preparation of the extended release matrix transdermal patch formulations as transdermal delivery of the drug over prolonged periods. The selected formulation GTP7 was able to satisfy all the pharmaceutical characteristics of transdermal films and seems promising. Formulation GTP7 released close to 99.13+-1.14 percent medication superior drug release in comparison to other type formulation. It may be advantageous as it may offer the benefit of longer release of drugs, decreased administration frequency, increased bioavailability and consequently that may aid in enhancing patient compliance. Other antidiabetic molecules can as well be subjected to this technology to ensure that the control of the disease is better.
1. Nagarathna R, Madhava M, Patil SS, Singh A, Perumal K, Ningombam G, Nagendra AH. Cost of management of diabetes mellitus: a pan India study. Annals of Neurosciences. 2020 Jul; 27(3-4): 190-2.
2. Standl E, Khunti K, Hansen TB, Schnell O. The global epidemics of diabetes in the 21st century: Current situation and perspectives. European Journal of Preventive Cardiology. 2019 Dec; 26(2_suppl):7-14.
3. Chauhan L, Vashisht S. Formulation and evaluation of novel herbal antidiabetic transdermal patch. Inno Pharm Pharmacother. 2018; 6(4).
4. Ahad A, Al-Saleh AA, Akhtar N, Al-Mohizea AM, Al-Jenoobi FI. Transdermal delivery of antidiabetic drugs: formulation and delivery strategies. Drug Discovery Today. 2015 Oct 1; 20(10):1217-27.
5. Kinani AA, Taghi HS. Formulation and characterization of orodispersible tablet of glimepiride. Journal of Advanced Pharmaceutical Technology & Research. 2022 Oct 1; 13(4):252-60.
6. Induri M, Raju M B, Prasad Y R, Reddy K P. Development and validation of a spectrophotometric method for quantification and dissolution studies of glimepiride in tablets. Journal of Chemistry. 2012; 9(2):993-8.
7. Shabnam H, Singh DJ, DN P, Anchal P, Sahil K. Formulation and Characterization of Transdermal Patch of Candesartan Celexitil. Journal of Drug Delivery & Therapeutics. 2019 May 2; 9.
8. Singh A, Bali A. Formulation and characterization of transdermal patches for controlled delivery of duloxetine hydrochloride. Journal of Analytical Science and Technology. 2016 Nov 28; 7(1):25.
9. Jain S, Gupta S, Sahu N. Formulation and evaluation of TDDS of Indian medicinal plant. Int J Drug Discov Herb Res. 2021; 11:1003-10.
10. Torlak C, Güleli M, Kızılok Ş, Kartop RA, Çalışkan C, KurşunBaysak F. Comparison of permeability of topical Cream drug through polymer synthetic membranes of different structures using Franz Cell diffusion test. Journal of Dispersion Science and Technology. 2024 Aug 9:1-9.
|
Received on 14.10.2025 Revised on 13.11.2025 Accepted on 08.12.2025 Published on 20.01.2026 Available online from January 27, 2026 Asian J. Pharm. Tech. 2026; 16(1):19-24. DOI: 10.52711/2231-5713.2026.00004 ©Asian Pharma Press All Right Reserved
|
|
|
This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License. Creative Commons License. |
|