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In this paper the design, implementation and performance analysis of four wave mixing (FWM) in optical communication system for different number of input channels is presented using various values of channel spacing. Here, all the input channels have been spaced evenly at various values like 6.25 GHz, 12.5 GHz, 25 GHz, 40 GHz, 50 GHz with the different number of channels at the input i.e. with 2, 4, 6, 8, 12 input channels. The simulation results reveal that the four wave mixing is minimum when the channel spacing is maximum i.e. 50 GHz with minimum number of channels i.e. 2 input channels. It is observed that on increasing the channel spacing, the interference between the input frequencies decreases and hence the four wave mixing also decreases. Also, on increasing the number of input channels/users, the interference between the input frequencies increases and thus, the four wave mixing also increases.