In-line valves are devices typically used for isolation or flow regulation in pipe systems, playing a key role in the operational management of transmission mains (TM). However, there is no fast and expeditious procedure available for checking the efficacy of the sealing mechanism, and its ability to prevent leakage, unwanted flow or partial blockages, which is a crucial action for any maintenance operation. Due to the different values of the conveyed discharge, the diameter changes along the TM at a series junctions which therefore makes diameter changes a very common singularity. This paper has two aims. The first one is to evaluate the feasibility of Inverse Transient Analysis (ITA) for checking the sealing of in-line valves. In particular, the primary objective of the numerical model is to identify the distinctive features of the measured pressure signals that correspond to the status of an in-line valve, discerning whether it is fully sealed or partially closed. The second objective is to use Direct Analysis (DA) of the pressure signals to appropriately capture the transient response of the series junctions. To address these issues, safe transients have been generated in a real TM by means of a Portable Pressure Wave Maker (PPWM) device, refined at the Water Engineering Laboratory (WEL) of the University of Perugia, Italy. The results of the field tests and numerical model point out that the positive pressure wave reflected by the in-line valve is smaller than the one expected if it were perfectly sealed. Moreover, the transient response of the series junction has been properly captured by the DA of the pressure signal. Accordingly, the proposed procedures have been demonstrated to be suitable tools for the management of long transmission pipelines.