After the addition of oxidant the contents color had slowly changed to dark green color indicating the polymerization of aniline to polyaniline. The final contents have been stirred for 10 min and kept in refrigerator at
0 °C for 24 h. After that the contents were filtered by washing with deionized water for several times till all unreacted surfactant is washed. Finally washed with methanol to terminate polymerization. The dark green colored precipitate was dried overnight at 100 °C.Similarly pure PANi is also prepared without adding fluconazole. Antifungal activity for PANi and PANi combined with fluconazole nanofibres was performed by agar diffusion method BMN673 in Sabouraud agar. Sabouraud agar was prepared as per the manufacturer protocol. The agar medium was sterilized in aquilots of 15 ml at a pressure of 15 lbs for 15 min. This agar medium was transferred into sterilized petri dishes in a laminar air flow unit and allowed to solidify. After solidification of the media, a 24 h culture of each organism was standardized to 0.5. McFarland standard was cultivated as lawn culture by spreading the organism on the agar media using sterile cotton swab. Cup plate method was used to test BIBW2992 nmr the antifungal activity by using sterile bore with the diameter of 9 mm. Four different concentrations were prepared such as 10 μg/ml, 5 μg/ml, 2.5 μg/ml and 1.25 μg/ml of PANi and PANi doped fluconazole in dimethylsulfoxide
solution. To this media, 100 μl of respective dilution were added using micropipette and incubated for 2 days at 37 °C in the incubation
chamber. Average zone diameters were measured after repeating the experiment for three times. The prepared PANI combined with fluconazole nanofibers were studied by SEM The morphological structure of the synthesized PANI doped fluconazole nanofibers was identified by scanning electron microscope (SEM). A fixed from working distance of 5 mm and a voltage of 5–25 kV were used. Normally, sample preparation for the SEM measurement will be carried out inside the glove box by covering the sample holder with parafilm for minimal exposure to oxygen while transferring it to the secondary emission chamber. First of all, we investigated the influence of the parameters such like ratio of oxidant to monomer, the concentration of the surfactant, aging temperature and time and reaction temperature on the fiber formation of PANI doped fluconazole to discover the optimal conditions for the formation of PANI doped fluconazole nanofiber structure. It was found that the reaction temperature and to some extent aging temperature and time strongly affect the microstructure and the formation probability of PANI doped fluconazole nanofibers. In all the cases we have obtained nanofiber like structures but with different lengths and diameter. The SEM image of PANI doped nanofibers which shown in Fig. 1 which indicates the nanofiber diameter about 10 nm.