Trace element distribution in the human body varies across regions and countries due to their different living environment and lifestyle. Thus, it is of great significance to investigate the reference level of trace element in a specific population. Wuhan is the largest metropolitan area in central China with highly developed heavy industries. This study aimed at determining the reference urinary distribution in general populations of Wuhan for nine trace elements (Cr, Mn, Cu, As, Se, Cd, Hg, Tl, Pb), and analyzed their associations with age, sex, and the kidney function. In total, 226 healthy adults not exposed to these trace elements were recruited, and the first-morning urine specimens were analyzed by using ICP-MS-based method. Our results showed higher urinary levels for As and Cd in Wuhan population when compared with other countries, while other element levels were almost equivalent. Sex difference existed for urinary Cu, Mn, As, Tl, and Pb. And urinary Cd, Tl, and Pb levels were associated with the glomerular filtration rate. Almost all these urinary elements showed significant inter-correlations, especially for Cu but except for Mn. This study provides systematic information regarding urinary trace element levels in residents of Wuhan in central China, and shall be of importance for future environmental and occupational biomonitoring.