Abstract Funding Acknowledgements Type of funding sources: Foundation. Main funding source(s): Professor Nanthakumar is a recipient of the Mid-career Investigator Award from the Heart & Stroke Foundation of Ontario. Background Ablation targets in the era of early ventricular tachycardia (VT) ablation will likely be driven by substrate-based physiological target approaches rather than activation mapping. Decrement evoked potentials (DeEP) have been co-localized to the diastolic VT circuit and are used as a method to perform VT ablation when VT cannot or should not be induced.1 Traditionally these potentials are evoked by a single extrastimulus. The incremental utility of a second extrastimulus in detecting these potentials has not been evaluated. Purpose We studied the utility of the second extra stimulus (S3) in detecting substrate ablation over the first extra stimulus (S2) following a drivetrain (S1) of 8 beats at 600ms. Methods DeEP was measured as the time difference between the pacing stimulus and the nearfield local activation delay on the bipolar electrogram during the S1 drivetrain and after the S2 or S3 respectively. Decrement of ≥10ms was considered a positive DeEP site. A total of 157 sites from 6 patients with VT were analysed. The incremental value of S3-induced DeEP was compared to the S2-induced DeEP. Results The mean decrement was significantly greater on the S3 compared to the S2 (41.8±35.6ms for S3 vs 25.8±20.4ms for S2, P <0.0001). The mean incremental value of the S3 was 15.9ms (95% CI, 11.5-20.3ms). From the 157 intracardiac electrograms studied, 79.1% had a positive DeEP with S2 alone. At the same electrode locations, an additional S3 evoked DeEP in 82.2% of sites. Conclusion When physiological substrate is present on the first extra stimulus, the second extra stimulus magnifies the identification of the target. However, we found in our study that the second extrastimulus uncovered a hidden arrhythmogenic substrate in only 3.1% additional mapped sites. Consequently, the utility of the second extrastimulus in identification of hidden substrate needs to be balanced with the risk of induction of potentially unstable rhythms.