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A common clinical practice during single-point Doppler analysis is to measure the centerline maximum velocity and to recover the time-averaged flow rate by exploiting an assumption on the shape of velocity profile (a priori formula), either a parabolic or a flat one. In a previous study, we proposed a new formula valid for the peak instant linking the maximum velocity and the flow rate by including a well-established dimensionless fluid-dynamics parameter (the Womersley number), in order to account for the hemodynamics conditions (Womersley number-based formula). Several in silico tests confirmed the reliability of the new formula. Nevertheless, an in vivo confirmation is missing limiting the clinical applicability of the formula. An experimental in vivo protocol using cine phase-contrast MRI (2-D PCMRI) technique has been designed and applied to ten healthy young volunteers in three different arterial districts: the abdominal aorta, the common carotid artery, and the brachial artery. Each PCMRI dataset has been used twice: 1) to compute the value of the blood flow rate used as a gold standard and 2) to estimate the flow rate by measuring directly the maximum velocity and the diameter (i.e., emulating the intravascular Doppler data acquisition) and by applying to these data the a priori and the Womersley number-based formulae. All the in vivo results have confirmed that the Womersley number-based formula provides better estimates of the flow rate at the peak instant with respect to the a priori formula. More precisely, mean performances of the Womersley number-based formula are about three times better than the a priori results in the abdominal aorta, five times better in the common carotid artery, and two times better in the brachial artery.