This study presents a voltage loss breakdown method based on in-situ experimental data to systematically analyze the different overpotentials of a polymer electrolyte membrane fuel cell. This study includes a systematic breakdown of the anodic overpotentials via the use of a reference electrode system. This work demonstrates the de-convolution of the individual overpotentials for both anode and cathode side, including the distinction between mass-transport overpotentials in cathode porous transport layer (PTL) and electrode, based on in-situ polarization tests under different operating conditions. This method is used to study the relationship between mass-transport losses inside the cathode catalyst layer (CL) and the PTL for both a single layer and two-layer PTL configuration. We conclude that the micro-porous layer (MPL) significantly improves the water removal within the cell, especially inside the cathode electrode, and therefore the mass transport within the cathode CL. This study supports the theory that the MPL on the cathode leads to an increase in water permeation from cathode to anode due to its function as a capillary barrier. This is reflected in increased anodic mass-transport overpotential, decreased ohmic losses and decreased cathode mass-transport losses, especially in the cathode electrode. © 2015 Elsevier B.V. All rights reserved.