Inertial separation, is one of the promising techniques, separates the particles based on size and without any external force. The 3D simulation along with the particle tracing makes them computationally expensive. To reduce the computation cost, the present study is done using 2D simulations. Parametric investigations are done to get a primary idea of successful focusing of particles. The present study explores the effect of number of turns and Reynolds number on the focusing of 5 and 10 μm microparticles in serpentine microchannel. The study has been carried out for Reynolds number 20, 30, 40, 50, 60 and 80 and number turns of 16, 18, 20, 22 and 24 for the serpentine channel. The number of turns and Reynolds number has significant effect on focusing behaviour of 5 μm particles whereas no significant effect is seen for 10 μm particles. Promising result is seen for Reynolds number 40–60 for 5 μm particles for all turns where as promising results are seen at Reynolds number 20–50 for 10 μm particles irrespective of number of turns. The computational time is significantly reduced for 2D simulation making primary parametric investigation fast and cost-effective compare to 3D simulations.