ABSTRACT: BACKGROUND:Smartphone apps for heart rate variability (HRV) measurement have been extensively developed in the last decade. However, ultra-short-term HRV recordings taken by wearable devices have not been examined. OBJECTIVE:The aims of this study were the following: (1) to compare the validity and reliability of ultra-short-term and short-term HRV time-domain and frequency-domain variables in a novel smartphone app, Pulse Express Pro (PEP), and (2) to determine the agreement of HRV assessments between an electrocardiogram (ECG) and PEP. METHODS:In total, 60 healthy adults were recruited to participate in this study (mean age 22.3 years [SD 3.0 years], mean height 168.4 cm [SD 8.0 cm], mean body weight 64.2 kg [SD 11.5 kg]). A 5-minute resting HRV measurement was recorded via ECG and PEP in a sitting position. Standard deviation of normal R-R interval (SDNN), root mean square of successive R-R interval (RMSSD), proportion of NN50 divided by the total number of RR intervals (pNN50), normalized very-low-frequency power (nVLF), normalized low-frequency power (nLF), and normalized high-frequency power (nHF) were analyzed within 9 time segments of HRV recordings: 0-1 minute, 1-2 minutes, 2-3 minutes, 3-4 minutes, 4-5 minutes, 0-2 minutes, 0-3 minutes, 0-4 minutes, and 0-5 minutes (standard). Standardized differences (ES), intraclass correlation coefficients (ICC), and the Spearman product-moment correlation were used to compare the validity and reliability of each time segment to the standard measurement (0-5 minutes). Limits of agreement were assessed by using Bland-Altman plot analysis. RESULTS:Compared to standard measures in both ECG and PEP, pNN50, SDNN, and RMSSD variables showed trivial ES (<0.2) and very large to nearly perfect ICC and Spearman correlation coefficient values in all time segments (>0.8). The nVLF, nLF, and nHF demonstrated a variation of ES (from trivial to small effects, 0.01-0.40), ICC (from moderate to nearly perfect, 0.39-0.96), and Spearman correlation coefficient values (from moderate to nearly perfect, 0.40-0.96). Furthermore, the Bland-Altman plots showed relatively narrow values of mean difference between the ECG and PEP after consecutive 1-minute recordings for SDNN, RMSSD, and pNN50. Acceptable limits of agreement were found after consecutive 3-minute recordings for nLF and nHF. CONCLUSIONS:Using the PEP app to facilitate a 1-minute ultra-short-term recording is suggested for time-domain HRV indices (SDNN, RMSSD, and pNN50) to interpret autonomic functions during stabilization. When using frequency-domain HRV indices (nLF and nHF) via the PEP app, a recording of at least 3 minutes is needed for accurate measurement.