The possibility of using different times between laser pulses (Δt) in a PIV (Particle Image Velocimetry) measurement of the same real flow field for error assessment has already been proposed by the authors in a recent paper Nogueira et al. (Meas Sci Technol 20, 2009). It is a simple procedure that is available with the usual PIV setup. In that work, peak locking was considered basically as a bias error. Later measurements indicated that, using appropriate processing algorithms, this error is not the main peak-locking effect. Scenarios with the rms (root mean square) error due to peak locking as the most relevant contribution are more common than initially expected and require a differentiated approach. This issue is relevant due to the impact of the rms error in the evaluation of flow quantities like turbulent kinetic energy. The first part of this work is centred on showing that peak-locking error in PIV is not always a measurement bias towards the closest pixel integer displacement. Insight in the subject indicates that this is the case only for algorithm-induced peak locking. The peak locking coming out of image acquisition limitations (i.e. resolution) is not ‘a priory’ biased. It is a random error with a peculiar probability density function. Discussion on the subject is offered, and a particular approach to use a simple multiple Δt strategy to asses this error is proposed. The results reveal that in real images where amplitude of the peak-locking bias error is assessed to be as small as 0.02 pixels, rms errors can be in the order of 0.1 pixels. As PIV approaches maturity, providing a quantitative confidence interval by estimating measurement error seems essential. The method developed is robust enough to quantify these values in the presence of turbulence with rms up to ~0.6 pixels. This proposal constitutes a relevant step forward from the traditional histogram-based considerations that only reveal whether strong peak-locking error is present or not, without any information on its magnitude or whether its origin is bias or rms.