Chitosan Microspheres Lab Report

Preparation of chitosan microspheres49
In this method, the polymer chitosan was initially dissolved 1% acetic acid, by magnetic stirring for about 2-3 hrs, to the prepared chitosan solution, diltiazem hydrochloride was added, stirred for 1hr, the resulted polymer solution was added to liquid paraffin containing span 60 as surfactant. The resultant emulsion was stirred for one hr further cross linking agent gluteraldehyde was added stirred over night for rigidisation of coating. Microsphers were filtered, washed with petroleum ether, and air dried.

Formulation code Diltiazem hydrochloride
(mg) Chitosan Gluteraldehyde (ml) Surfactant Conc.
FCD1 90 90 3 1
FCD2 90 180 3 2
FCD3 90 240 3 3
FCD4 90 360 3 4
FCD5 90 450 3 5
FCD6 90 90 3 5

Initially eye piece micrometer was calibrated by using stage micrometer. A drop of suspension of microspheres in liquid paraffin was taken on glass slide, which was observed under 45 X. Sizeof 100 particles were measured.
Drug content and drug loading Microsphers equivalent to 100 mg of DLTZH/ND was taken, crushed and dissolved in methanol, made up to 100 ml with methanol in a volumetric flask. This solution was diluted suitably to Beers’ range of the drug. Absorbance of the solution was measured at 238 nm. Drug content was calculated by the formula absorbance/ slope* dilution factor.
Drug loading
Microsphers equivalent to 100 mg of DLTZH/ND taken and washed with 3 x 10 ml of methanol, which removes the free unloaded drug. Filtrate was diluted suitably to Beer’s concentration range and free drug concentration was determined spectrophotometrically at 238 nm and 236 nm for DLTZH and ND respectively. Further microspheres were crushed, dissolved in methanol and made up to 100 ml, which was further diluted suitably to Beer’s rangeand drug concentration was determined. Percentage drug loading was obtained from the formula=Total drug content-free drug content/ total drug content *

The fresh intestine was collected, which was cut through lumen. The microsphers equivalent to 100 mg of the drug was taken on mucosal side of the intestine, was placed b/n donor and receptor compartment; in such a way that mucosal side is facing towards donor compartment. One ml of phosphate buffer pH7.4 was added to donor compartment, the receptor compartment is filled with 25 ml of phosphate buffer pH7.4. The rate of drug permeation was studied by using collection of receptor fluid at regular time intervals, and analysed for drug