We assessed the test validity of the Apple Watch series 6 measure of heart rate variability (HRV) by comparing it with the reference measure assessed via a Biopac 3-lead electrocardiogram (ECG). We recruited 78 healthy adults (aged 20–75 years). HRV was measured using an in-lab protocol while resting, talking, watching a movie, before walking, and after walking. We conducted a synchronized countdown for each condition to guarantee that the recordings would be aligned between the two devices by using event markers in the Biopac at the exact time that the Apple Watch Breathe app began and ended. We assessed test validity using the Bland–Altman method, and both precision and accuracy were estimated using Lin’s concordance correlation coefficient. The highest level of agreement and concordance between devices occurred during rest. We observed near-perfect agreement for R-R intervals and beats per minute (BPM) measures, with mean absolute percentage errors (MAPE) of 1.15% during resting conditions. We observed moderate levels of agreement and concordance for N-N intervals at rest with a MAPE of 31.31% during resting conditions. The Apple Watch provides a high level of validity for measuring R-R intervals and BPM in healthy adults. Further research is needed to determine if HRV measures with the Apple Watch offer a significant opportunity for the surveillance of CVD risk.