Description: Off-resonant spinlock (SL) enables an NMR imaging technique that can detect dilute metabolites similar to chemical exchange saturation transfer. However, in clinical MR scanners, RF pulse widths are restricted due to recommended specific absorption rate limits. Therefore, trains of short RF pulses that provide effective saturation during the required irradiation period are commonly employed. Quantitative evaluation of spectra obtained by pulsed saturation schemes is harder to achieve, since the theory of continuous wave saturation cannot be applied directly. In this paper we demonstrate the general feasibility of quantifying proton exchange rates from data obtained in pulsed SL experiments on a clinical 3 T MR scanner. We also propose a theoretical treatment of pulsed SL in the presence of chemical exchange using an interleaved saturation–relaxation approach. We show that modeling magnetization transfer during the pauses between the RF pulses is crucial, especially in the case of exchange rates that are small with respect to the delay times. The dynamics is still governed by a monoexponential decay towards steady state, for which we give the effective rate constant. The derived analytical model agrees well with the full numerical simulation of the Bloch–McConnell equations for a broad range of values of the system parameters. Copyright © 2014 John Wiley & Sons, Ltd. Saturation of a two-pool-CEST system by pulsed spinlock can be modeled by an interleaved saturation–relaxation (ISAR) approach. Taking into account the chemical exchange during the pause is crucial, and leads to an analytical solution that agrees well with the full numerical simulation of the Bloch–McConnell equations for a broad range of values of the system parameters. This analytical solution allows the quantitative evaluation of Z -spectra obtained from creatine model solutions on a clinical 3 T scanner.