Study on Effective Entrapment of Drug Serratiopeptidase in Transfersomes and Assessing its Transdermal delivery K.Santhosh, T.Shankar, N.Balchandar Department of Physics and Nanotechnology, SRM University, kattankulathur, chennai-603203
Abstract Drug delivery is a process of administering a pharmaceutical compound to achieve therapeutic effect in humans or animals. Transdermal drug delivery is one of the efficient ways of drug administration. Transfersomes are artificial nano vesicles designed like a cell vesicle suitable for control and targeted drug delivery. Standard protein estimation by /RZU\¶V method and transfersome synthesis was done. Anti inflammatory drug called Serratio peptidase used in the treatment of swelling and bronchitis which is taken orally was delivered transdermally by a new protocol. Entrapment efficiency of conventional transdermal drug delivery vesicles, initially found to be 20% was optimized to increase the entrapment efficiency. We were able to achieve a higher efficiency of about 78%. The drug was tested on JRDW¶V skin and characterization was done using UV Spectrophotometer and in scanning electron microscopy. The methods and techniques adopted to increase the efficiency are discussed in detail.
Methods
Transfersomes under microscope Image 2 shows transfersomes entrapment efficiency was found to be 78%.
USING SN15 Using SN15 we got only 25% entrapment efficiency of drug in transfersomes. Because this tablet is a complex of serratiopeptidase and nimusulide.
USING TRIOSERA Triosera (new tablet-pure serratiopeptidase) was used.Vesicles were viewed under light microscope. Clear vesicles were found Triosera was dissolved in NaOH (isotonic solution).Protein estimation was done by /RZU\¶V method and OD value was taken at 660nm and is tabulated below: Concentration(mg/ml)
OD at 660nm
0.2
0.188
0.4
0.316
Introduction
Transdermal Drug delivery is proving to be superior to the conventional oral delivery owing to its distinct virtues. Transfersome is an ultra deformable vesicle, elastic in nature which can squeeze itself through a pore. Transfersomes are applied in a non-occluded method to the skin. Transfersomes are made up of a phospholipids component along with a surfactant mixture.These drugs find place in different places in the elastic vesicle before they get delivered beneath the skin. It was also observed that transdermal drug delivery is proving to be superior to conventional oral delivery of drugs owing to its distinct virtues.
Table 2 concentration of known
Test for transdermal efficiency SEM Characterization Figure 1 VKRZV WKH VDPSOH¶V 6(0 LPDJHV LQ WKH UDQJH RI QDQRVFDOH ZKLFK ZDV done by us and characterized by scanning electron microscopy.
Materials used for synthesis 1. Two tablets-SN15 and Triosera. 2. BSA stock solution (1mg/ml).
Sample S1 S2 S3 S4 S5 Table 3 shows Absorbance of unknown
3. Analytical reagents 4. Folin - ciocalteau solution (1:1 diluted in water). 5. Soya lecithin. 6. Tween 80. 7. SDC (sodium deoxy cholate). 8. Chloroform.
Solubility test
9. Cholesterol. 10. Diethyl ether. 11. Goat skin. 12. Phosphate Buffer: 0.1M of Potassium Dihydrogen Phosphate and 0.1 M of
The tablet SN15 which contains 15mg-serratio peptidase and 100mg-nimesulide was dissolved in neutral, acidic(2.52) and alkaline pH(10.1) and was found to be water soluble. Then the dissolved solution was centrifuged and Standard Protein estimation was done by /RZU\¶V method .
Dipotassium Hydrogen Phosphate.
Transfersomes under microscope Image 1 shows the transferosomes obtained at first and the entrapment efficiency was found to be 25%.
Concentration(mg/ml) 0.1
OD at 660nm 0.93
0.2
0.184
0.3
0.231
0.4
0.366
0.5
0.359
Table 1 Absorbance value of known taken at 660nm
Transfersome preparation In 2 different round bottom flasks the below mentioned formulations were taken and was kept in the shaker overnight: METHOD 1 (i) Soya lecithin (10mg) + Tween80 (10microlitre) + Chloroform. (ii)Soya lecithin (10mg) + SDC + Cholesterol + Chloroform. METHOD 2 (iii) 5mg cholesterol + 10mg SDC + 10mg lecithin dissolved in 5ml diethyl ether. Poster template by ResearchPosters.co.za
Transdermal delivery efficiency was checked using goat skin. Efficiency was found using a system called ³(;9,92´. The equipment was filled with buffer of following composition: Phosphate buffer 0.1M potassium dihydrogen phosphate and 0.1M dipotassium hydrogen phosphate. A small part of the cleaned goat skin was placed at the brim of the apparatus such that on stirring the buffer touches the skin. The processes were started .For every 2 hours 2ml of buffer was removed and was replaced by new buffer.2ml of buffer removed is collected in an eppendorf tubes. Nearly 5 samples were collected and estimation was done by /RZU\¶V method and OD values were taken at 660nm and are tabulated below: OD at 660nm 1.3 0.32 0.482 0.845 0.829
Entrapment Efficiency was calculated by: (Absorbance of unknown) / (Absorbance of Standard) * Concentration of Standard (mg/ml) The entrapment efficiency was calculated as 78 %.
Results and Conclusion Transfersome synthesis was done with various formulations and vesicles were observed. Better results (rigid vesicles) were obtained when cholesterol was added along with BSA. Exvivo was done with goat skin and transdermal efficiency was found to be 78%. Topical applications of enzymes offer potential advantages of delivering the enzyme directly to the site of action. Transdermal delivery of SRP would be useful to treat local inflammations and may prove to be more effective compared to topical NSAIDs(Non-steroidal anti-inflammatory drugs)
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