Objective <p>The objective of this study is to estimate the maternal knowledge and practice levels regarding neonatal umbilical cord care and their associated factors in different provinces of Pakistan.</p> Methods <p>A descriptive cross-sectional study design was used to collect data from the respondents. Multistage sampling techniques were carried out for this study. Semi-structured questionnaire was designed for collection of data after acquiring ethical approval and informed consent. Descriptive statistics were carried out by taking frequencies and percentages of the collected data and associated factors were analyzed by applying chi-square test at the p value of &lt; 0.05.</p> Results <p>245 (65.3%) of the respondents had poor knowledge levels about umbilical cord care and 220 (58.7%) respondents practiced poor cord care. More than half of the respondents had poor knowledge. 285 (76%) about the usage of chlorhexidine (CHX) on the cord for seven consecutive days. Some conventional and unhygienic practices were also recorded in this study. More than half of the mothers used cord clamps for tying the cord 245 (65.3%) followed by tailor’s threads 77 (20.5%). The associated factors including parity, occupation and income were not showing statistically significant results at the p value of &lt; 0.05 except for place of delivery (*p = 0.01), (*p = 0.005) and educational status (*p = 0.04), (*p = 0.03), which are showing statistically significant results.</p> Conclusion <p>Paucity in both knowledge and practices of umbilical cord care were observed among mothers attending postnatal care in the hospitals. Programs need to be initiated for the improvement of knowledge of UCC and discouragement of unhygienic practices.</p> Graphical Abstract <p></p>