A transient inline spiking system is a promising tool for evaluating the performance of a virus filter in continuous operation. For better implementation of the system, we performed a systematic analysis to understand the residence time distribution (RTD) of inert tracers in the system. We aimed to understand the RTD of a salt spike, not retained onto or within the membrane pore, to focus on its mixing and spreading within the processing units. A concentrated NaCl solution was spiked into a feed stream as the spiking duration (tspike) was varied from 1 to 40 min. A static mixer was employed to mix the salt spike with the feed stream, which then passed through a single-layered nylon membrane inserted in a filter holder. The RTD curve was obtained by measuring the conductivity of the collected samples. An analytical model, the PFR-2CSTR model, was employed to predict the outlet concentration from the system. The slope and peak of the RTD curves were well-aligned with the experimental findings when τPFR = 4.3 min, τCSTR1 = 4.1 min, and τCSTR2 = 1.0 min. CFD simulations were performed to describe the flow and transport of the inert tracers through the static mixer and the membrane filter. The RTD curve spanned more than 30 min, much longer than tspike, since solutes were dispersed within processing units. The flow characteristics in each processing unit correlated with the RTD curves. Our detailed analysis of the transient inline spiking system would be helpful for implementing this protocol in continuous bioprocessing.