Specification

Description: Yellow colored viscous liquid. Identification: Positive pH: (3---------4.5)

Label Claim: Nystatin Assay: Limit: Nystatin 100,000 IU/ mL

(95----120 %)

TESTING METHOD FOR DAVISTIN DROPS Assay for Nystatin: Microbiological Assay: (Method A) Carry out the microbiological assay of antibiotic (diffusion method). Protect the solution from the light through out the assay. Dissolve the substance to be examined and nystatin CRS separately in dimethylformamide R and dilute with a mixture of 5 volumes of dimethylformamide R and 95 volume of buffer solution pH 6.0. (Diffusion Method) Liquefy a medium suitable for the conditions of the assay and inoculate temperature, for example 48C to 50C for the vegetative forms with the known quantity of a suspension of microorganisms sensitive to the antibiotic to be examined. Immediately pour into Petri dishes or large rectangular dishes a quantity of the inoculated medium to form a uniform layer 2mm to 5mm thick. Store the dishes so that no appreciable growth or death of the micro-organisms occurs before the dishes are used and so that the surface of the medium is dry at the time of use. Using the buffer solution. Apply the solution to the surface of the medium for example in sterile cylinder of porcelain or other suitable material or in cavities prepare in the agar. The same volume of solution must be added to the cylinder or cavity. Alternatively use sterile absorbent paper discs of suitable quality; impregnate the discs of the solution of reference substance or the solution of the antibiotic to be examined and place on the surface of the agar. Incubate at suitable temperature for about 18 hours periods of the diffusion prior to incubation usually 1 h to 4 h, at room temperature or about 4C, as appropriate, may be use to minimize the effect of the variation in time between the application of the solution and to improve the regression slope.

Measure the diameter with precision of at least 0.1mm or the area of the circular inhibition zones with the corresponding precision and calculate the potency using appropriate statistical methods. Assay for Nystatin: (Method B alternative method) Standard Preparation: Take Reference std Nystatin 200,000 IU (36.88 mg) in 100 ml of volumetric flask add 5 ml of glacial acetic acid shake mechanically for 15 min to dissolve, make the volume up to the mark with methanol. Take 5 ml from this solution into 100 ml volumetric flask and make the volume with methanol up to the mark. Sample Preparation: Accurately measure 2ml of syrup equivalent in 200,000 IU (36.88 mg) of Nystatin in 100 ml of volumetric flask add 5 ml glacial acetic acid of shake mechanically for 15 min to dissolve, make the volume up to the mark with methanol. Take 5 ml from this solution into 100 ml volumetric flask and make the volume with methanol up to the mark. Procedure: Take the absorbance for both sample and standard at UV rang max 319 nm. Methanol as a blank. Calculation: %age= As x Wr x 100 = C Ar x Ws Nystatin/ml = 100,000 x C = D IU/mL 100

Where; Ar= absorbance of reference As= absorbance of sample Wr= weight of reference Ws= weight of sample