ObjectivesTo develop a robust algorithm for estimating ultrasonic axial transmission velocity from neonatal tibial bone, and to investigate the relationships between ultrasound velocity and neonatal anthropometric measurements as well as clinical biochemical markers of skeletal health.MethodsThis study presents an unsupervised learning approach for the automatic detection of first arrival time and estimation of ultrasonic velocity from axial transmission waveforms, which potentially indicates bone quality. The proposed method combines the ReliefF algorithm and fuzzy C‐means clustering. It was first validated using an in vitro dataset measured from a Sawbones phantom. It was subsequently applied on in vivo signals collected from 40 infants, comprising 21 males and 19 females. The extracted neonatal ultrasonic velocity was subjected to statistical analysis to explore correlations with the infants' anthropometric features and biochemical indicators.ResultsThe results of in vivo data analysis revealed significant correlations between the extracted ultrasonic velocity and the neonatal anthropometric measurements and biochemical markers. The velocity of first arrival signals showed good associations with body weight (ρ = 0.583, P value <.001), body length (ρ = 0.583, P value <.001), and gestational age (ρ = 0.557, P value <.001).ConclusionThese findings suggest that fuzzy C‐means clustering is highly effective in extracting ultrasonic propagating velocity in bone and reliably applicable in in vivo measurement. This work is a preliminary study that holds promise in advancing the development of a standardized ultrasonic tool for assessing neonatal bone health. Such advancements are crucial in the accurate diagnosis of bone growth disorders.