Objective: To investigate the application value of contrast-enhanced ultrasound in liver mass biopsy. Methods: A retrospective analysis was performed on 190 patients (190 tumors) who underwent liver mass biopsy in Nantong Cancer Hospital from January 2019 to December 2021, including 118 cases of conventional ultrasound-guided biopsy (conventional ultrasound group) and 72 cases of contrast-enhanced ultrasound-guided biopsy (contrast-enhanced ultrasound group). The visualization of puncture needles, the display of necrotic area and adjacent blood vessels, the number of punctures, and diagnosis rate were compared between the 2 groups. Results: The display rates of necrosis and adjacent large vessels in contrast-enhanced ultrasound group were 33.33% and 40.28%, respectively, which were significantly higher than 12.71% and 23.73% in the conventional ultrasound (P<0.05). In the contrast-enhanced ultrasound group, 93.06% of patients obtained satisfactory specimens when punctured less than or equal to 2 times, which was significantly higher than 75.42% in the conventional ultrasound group, and the difference was statistically significant (P<0.05). The diagnostic rate of CEUS puncture biopsy was 95.83%, which was significantly higher than that of conventional ultrasound (89.83%), and the difference was statistically significant (P<0.05). Conclusion: Compared with conventional ultrasound, contrast-enhanced ultrasound-guided liver mass puncture can better display necrotic areas and adjacent blood vessels, which helps to reduce the number of punctures and improve the diagnostic rate of puncture.