IN VITRO EVALUTION OF IN SITU GEL

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K. JAYA RAJ KUMAR DEPT.PHARMACEUTICS RAO’S COLLEGE OF PHARMACY. NELLORE.(AP) INDIA Email:jayaraj2775@gmail.com IN VITRO EVALUATION OF IN SITU GEL

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Candidiasis is an opportunistic infectious condition caused by ubiquitous, saprophytic fungi of the genus Candida, which includes eight species of fungi, the most common of which is Candida albicans. Candidiasis is usually limited to the skin and mucous membranes, which includes oropharyngeal (affecting the oral cavity and pharynx). Factors that alter the oral mucosal environment are xerostomia, antibiotic therapy, poor oral or denture hygiene, malnutrition/gastrointestinal malabsorption, Iron, folic acid, or vitamin deficiencies, acidic saliva/carbohydrate-rich diets, heavy smoking, and oral epithelial dysphasia. INTRODUCTION

Evaluation of buccal gel:Preliminary evaluation of gel (Gelation temperature): : 

Evaluation of buccal gel:Preliminary evaluation of gel (Gelation temperature): The gelation temperature was determined by heating the solution (1-20c) min in a test tube with gentle stirring until gel was formed. The gel was said to have formed when there was no flow after container was overturned. The gelling capacity of the formed gel was determined visual inspection and the different grades were allotted as per the gel integrity, weight and rate of formation of gel with respect to time. Gelling Capacity:

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A sample of 50 gm of gel was placed in a 100 ml graduated cylinder and gelled in a thermostat at 370c. The apparatus for measuring gel strength (weigh or apparatus as shown in figure 1, weighing 27 gm) was allowed to penetrate in gel. The gels strength, which means the viscosity of the gels at physiological temperature, was determined by the time (seconds), the apparatus took to sink 5 cm down through the prepared gel. Measurement of Gel Strength: Assemble

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Determination of pH The pH of the gel was determined using a calibrated pH meter. The readings were taken for average of 3 samples. Determination of gel strength The method by which the properties of polymericsystem may be conveniently determined is texture profile analysis. A TA-XT2 Texture analyzer The experiment was done by placing the gels in standard beaker below the probe. In this an analytical probe is then immersed into the sample. The Texture Analyzer was set to the ‘gelling strength test’ mode or compression mode with a test-speed of 1.0 mm/s. An acquisition rate of 50 points per seconds and a trigger force of 5 g were selected. An aluminum probe of 7.6 cm diameter was used for all the samples. The study was carried out at room temperature[9]. The force required to penetrate the gel was measured as gel strength in terms of g.

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The mucoadhesive force of all the optimized batches was determined as follows, a section of mucosa was cut from the chicken cheek portion and instantly fixed with mucosal side out onto each glass vial using rubber band. The vials with chicken cheek mucosa were stored at 370C for 5 minute then next vial with a section of chicken cheek mucosa was connected to the balance in inverted position while first vial was placed on a height adjustable pan. In situ gel was added onto the buccal mucosa of first vial. Then the height of second vial was so adjusted that the mucosal surfaces of both vials come in intimate contact. Two minutes time of contact was given. Then weight was kept rising in the pan until vials get detached. bioadhesive force the minimum weight required to detach two vials. The buccal mucosa was changed for each measurement. Determination of mucoadhesive Force Detachment stress (dynes/cm2) = mg/A Where m is the weight added to the balance in grams; g is the acceleration due to gravity taken as 980 cm/s2; and A is the area of tissue exposed. All the above mentioned experiments were carried out in triplicates. Assemble

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The bioadhesive potential of the gel was evaluated. Briefly, an agar plate (1%, w/w) was prepared in pH --- buffer. Test sample, 50mg was placed at the center of plate. After 5min, the agar plate was attached to a USP disintegration test apparatus as shown in Fig. 12 and moved up and down in pH ---buffer at 37±1 0 C. The sample on the plate was immersed into the solution at the lowest point and was out of the solution at the highest point. The residence time of the test samples on the plate was noted visually.

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The rheological studies were carried out using Brookfield programmable DV II+ Model pro (USA). The gel under study was placed in a small sample holder with a facility of water circulation. Water was circulated in the jacket with the help of Water immersion pump. Initially ice-cold water was circulated and then hot water to raise the temperature gradually. The temperature sensing probe was lowered in the gel was recorded. Spindle number SS64 was lowered vertically in it. The spindle was rotated at varying speed. The rheological studies were carried out using Brookfield programmable DVII+ Model pro II type (USA). The viscosity of in situ gel and the solution were determined at different angular velocities(0,10,20,30,40….to 100 rpm) average of two reading were used to calculate the viscosity. Evaluation was conducted in triplicate. Viscosity Studies:

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For the determination of spreadability, excess of sample was applied in between two glass slides and was compressed to uniform thickness by placing 1000 g weight for 5 min. weight (50 g) was added to the pan. The time in which the upper glass slide moves over to the lower plate was taken as measure of spreadability (S) S= ML/T Where, M = weight tide to upper slide. L = length moved on the glass slide. T = time taken. Spreadability:

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Buccal cavity of Isolation of chicken cheek mucosa from the anterior healthy chicken was obtained from the local slaughter house. It was cleaned and the mucosa was re-moved from the anterior buccal cavity. The mucosa was stored in normal saline with few drops of gentamycin sulphate injection, to avoid bacterial growth. After the removals of blood from the mucosal surface it become ready for use Content Uniformity

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Assembly of diffusion cell for in-vitro diffusion studies the oral diffusion cell was designed as per the dimension given. The diffusion cells were placed on the magnetic stirrers. The outlet of the reservoir maintained at 37±0.50 C and was connected to water jacket of diffusion cell using rubber latex tubes. The receptor co presentment was filled with fluid. Then the prepared chicken cheek mucosa was mounted on the cell carefully so as to avoid the entrapment of air bubble under the mucosa. Intimate contact of mucosa was ensured with receptor fluid by placing it tightly with clamp. The speed of the sitting was kept content throughout the experiment .With the help of micropipette 1ml of sample was withdrawn at a time intervals of 30 min. from sampling port of receptor compartment and same volume was the replaced with receptor fluid solution in order to maintain sink condition. The samples were appropropriately diluted and the absorbance was measured at --- nm using Schimadzu 1700UV-VIS spectrophotometer. Diffusion Medium: Assemble diffusion cell

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