Objective To investigate the application of ultrasound‑guided posterior tibial arterial catheterization in neonatal anesthesia. Methods A total of 120 neonates [American Society of Anesthesiologists (ASA) grades Ⅱ or Ⅲ] who were scheduled for surgery from January 2019 to February 2020 were enrolled. During the surgery, invasive arterial pressure was monitored, while arterial blood gas was analyzed. According to the random number table method, the neonates were divided into three groups (n=40): a posterior tibial artery group (group J), a radial artery group (group R) and a dorsalis pedis artery group (group Z). Arterial catheterization was performed using ultrasound‑guided out‑of‑plane procedures. The primary outcome was the first‑attempt success rate of puncture. Meanwhile, the time of catheterization, a total success rate within 10 min, arterial diameter and depth were determined. Results Compared with group Z, groups J and R presented an increased first‑attempt success rate of puncture (P<0.05) and an decreased time of catheterization (P<0.05), with an increased arterial diameter (P<0.05). The arterial depth in group J was larger than those in groups R and Z (P<0.05). Both groups R and J presented a less cumulative success rate than group Z within 10 min (P<0.05). There was no statistical difference in other indicators among those groups (P>0.05). Conclusions Ultrasound‑guided posterior tibial arterial catheterization has a first‑attempt success rate of puncture and catheterization time which is similar to the radial artery, and superior to the dorsalis pedis artery. It can be an alternative for catheterization of the radial artery during neonatal anesthesia.