STUDY OBJECTIVES: The association between daytime/nighttime heart rate variability (HRV) and the apnea-hypopnea index (AHI) remains unclear. We sought a relationship between AHI and the daytime-to-nighttime HRV ratio as measured by 24-hour Holter monitoring in patients with obstructive sleep apnea. METHODS: We prospectively enrolled 66 patients who visited our sleep clinic complaining of habitual snoring or sleep apnea. All underwent 24-hour Holter monitoring (to measure HRV) combined with full-night polysomnography. Sixty-two met our enrollment criteria. We evaluated the associations between HRV frequency domains and the polysomnography indices. We also considered medical histories and anthropometric data. RESULTS: The nighttime very-low-frequency (VLF), low-frequency (LF), and high-frequency HRVs were significantly higher than the daytime values. On correlation analysis, the day/night VLF (r = .550, P < .001), LF (r = .556, P < .001), and high-frequency (r = .303, P = .017) HRVs were significantly related to the AHI. Of the day/night HRV ratios, the VLF (P for trend = .003) and LF (P for trend = .013) ratios decreased significantly by obstructive sleep apnea severity. Multivariable analysis showed that the day/night VLF (beta = 16.387, P < .001) and day/night LF (beta = 25.248, P < .001) were independently (and significantly) associated with the AHI. CONCLUSIONS: Twenty-four-hour Holter monitoring may usefully predict AHI. The day/night VLF and day/night LF ratios tended to decrease by obstructive sleep apnea severity and were independently associated with the AHI. CITATION: Nam E-C, Chun KJ, Won JY, Kim J-W, Lee WH. The differences between daytime and nighttime heart rate variability may usefully predict the apnea-hypopnea index in patients with obstructive sleep apnea. J Clin Sleep Med. 2022;18(6):1557-1563.