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Ultra fine fibers were electrospun from regenerated silk fibroin/formic acid solution. Effect of some process parameters on the morphology, diameter and variation in fiber diameter of electrospun fibers were experimentally investigated. Scanning electron microscope was used for the measurement of fiber diameter. Fibers with diameter ranging from 80 to 210 nm were collected depending on the solution concentration and the applied voltages. Response surface methodology (RSM) was used to obtain a quantitative relationship between selected electrospinning parameters and the average fiber diameters and its distribution. It was shown that concentration of silk fibroin solution had a significant effect on the fiber diameter and the standard deviation of the fiber diameter. Applied voltage had no significant effect on the fiber diameter and its standard deviation.