The herbicide-tolerant soybean ZH10-6 was developed by modifying the Zhonghuang 10 (ZH10) variety with the G2-EPSPS and GAT genes, conferring resistance to glyphosate. This study aimed to assess the potential health effects of ZH10-6 in Sprague-Dawley rats through a 90-day subchronic toxicity test. Seven groups of rats (n = 10/sex/group) were fed a commercial AIN93G diet or diets containing 7.5%, 15%, or 30% ZH10-6 or ZH10 soybeans. General behavior, body weight, and food consumption were monitored weekly. At the end of the study, clinical pathology, including hematology, serum chemistry, urinalysis, and histopathology, were conducted. Throughout the study, all rats remained healthy and showed no abnormal clinical signs. Although some coagulation and serum biochemistry parameters showed statistical differences between groups, all values fell within the historical control ranges and were considered normal biological variability rather than treatment-related effects. The results indicate that ZH10-6 soybean consumption did not cause any adverse health effects in rats. These findings suggest that ZH10-6 is as safe as its nontransgenic parental variety, ZH10, with no evidence of toxicity after 90 days of exposure.