QT interval on the surface electrocardiogram (ECG) reflects the time for repolarization of myocardium and prolongation of QTc is strongly associated with sudden cardiac death. Studies using novel techniques on beat-to-beat QT interval variability (QTV) have shown that it is influenced by the autonomic nervous system and is a predictor of sudden cardiac death. In this study, we examined the awake and sleep changes in QTV in 39 normal adults (mean age, 35 years) and 10 children (mean age, 11 years) using 24-hour ECG records. We obtained eight 5-minute segments of ECG sampled at 1000 Hz from the 24-hour records. Our results show that there is a diurnal variation of QTvm, detrended QT interval variance corrected for mean QT, and QTvi, an index of QTvm divided by heart rate variability corrected for mean heart rate. There was a significant increase in mean QT during sleep, whereas there was a significant decrease in QTvm and QTvi. QTvi significantly increased during the early morning hours. There were significant but modest correlations between the average 24-hour and awake QTvi and age (p < 0.01). There were also decreased low-frequency and high-frequency powers of QT during sleep. Coherence between heart rate and QT interval fluctuations in the range of 0-0.5 Hz, and especially in the high-frequency range (0.15-0.5 Hz), was significantly lower in adults than in children (p < 0.0002). These findings demonstrate diurnal fluctuations in ventricular repolarization lability. We speculate that these effects may relate to changes in cardiac autonomic function and may contribute to the well-known diurnal variation in the incidence of ventricular arrhythmias.