The cylinder plate assay of drug potency is based on measurement of the diameter of zone of inhibition of microbial growth surrounding cylinders (cups), containing various dilutions of test compounds.A sterile borer was used to prepare four cups of 8 mm diameter in the agar medium spread with micro-organism and 0.1 ml of inoculums was spread on the agar plate by spread plate technique.
Accurately measured (0.05m1) solution of each concentration and reference standards were added to the cups with a micropipette. All the plates were kept in a refrigerator at 2°C to 8°C for effective diffusion of test compounds and standards for 2 hours. Later, they were incubated at 37 °C for 24 hours. The presence of definite zone of inhibition of any size around the cup indicated antibacterial activity. The solvent control (DMSO) was run simultaneously, which was used as vehicle. The diameter of the zone of inhibition was measured and recorded.Anti fungal activity: Test samples The antifungal activity of EEAN was performed using agar cup plate method.
The ethanolic extract of Allmania nodiflora was used in four dose levels of 50mg/ml, 100mg/ml, 150mg/ml, and 200mg/ml.Procedure:Anti fungal activity was performed by using agar cup plate method. The nutrient PDA medium (Hi media) was prepared and inoculated with 0.5 ml of aqueous suspension of the above mentioned test organism , which were prepared from 48 hours culture, are thus transferred in to sterile petri dishes. The medium in the plates were allowed to set at room temperature for 1e 0 min. 5 cups of 5mm diameter were made in each plate at the equal distances. Stock solutions of the tests were prepared in concentrations 50 mg/ml, 100mg/ml, 150 mg/ml, 200mg/ml of each stock concentration were placed in cups by means of sterile pipettes.
In each plate one cup was used for control and standard Fluconazole (0.1 mg/ml) is used as reference standard. The petridishes thus prepared were incubated for 48 hours at room temperature and were later examined by measuring the zone of inhibition and the results were tabulated.