ObjectivesHeart rate variability (HRV) has gained recognition as a noninvasive marker of autonomic activity. HRV is considered a promising tool in various clinical scenarios. The optimal electrocardiogram (ECG) sampling frequency required to ensure sufficient precision of R–R intervals for HRV analysis has not yet been determined. Here, we aimed to determine the acceptable ECG sampling frequency range by analyzing ECG signals from patients who visited an emergency department with the chief complaint of acute intoxication or overdose.MethodsThe study included 83 adult patients who visited an emergency department with the chief complaint of acute poisoning. The original 1,000-Hz ECG signals were down-sampled to 500-, 250-, 100-, and 50-Hz sampling frequencies with linear interpolation. R–R interval data were analyzed for time-domain, frequency-domain, and nonlinear HRV parameters. Parameters derived from the data on down-sampled frequencies were compared with those derived from the data on 1,000-Hz signals, and Lin's concordance correlation coefficients were calculated.ResultsDown-sampling to 500 or 250 Hz resulted in excellent concordance. Signals down-sampled to 100 Hz produced acceptable results for time-domain analysis and Poincaré plots, but not for frequency-domain analysis. Down-sampling to 50 Hz proved to be unacceptable for both time- and frequency-domain analyses. At 50 Hz, the root-mean-squared successive differences and the power of high frequency tended to have high values and random errors.ConclusionsA 250-Hz sampling frequency would be acceptable for HRV analysis. When frequency-domain analysis is not required, a 100-Hz sampling frequency would also be acceptable.