Lithium-ion (Li-ion) batteries are of great interest to the automotive industry due to their higher power and energy density, higher cell voltage, longer cycle life and lower self-discharge compared to other battery chemistries. Electrochemical impedance spectroscopy is a powerful tool employed to investigate the fundamental electrochemical reactions within a Li-ion battery cell, which relates to state of charge, internal temperature and state of health. Its effectiveness has established it as a core method to study electrochemical behaviour of batteries in both off-line and on-line applications. In this work it is shown that in addition to state of charge, internal temperature and state of health, the time period between the removal of an electrical load and the impedance measurement affects the results. The study of five commercially available cells of varying capacities and electrode chemistries show that, regardless of cell type, maximum impedance change takes place within the first 4 h of the relaxation period. The root cause of this impedance change has been discussed from an electrochemical perspective.