This study aimed to examine the effect of nitrogen (N) application rate and time on yield, grain filling, starch metabolizing enzymes, and hormones of maize based on a long-term field experiment initiated in 2012. The total N fertilizer dose [(0 (N0), 100 (N1), 200 (N2), and 300 (N3) kg N ha–1] was split into two (T1, one-third at sowing and two-thirds at the six-leaf stage) or three (T2, one-third each at sowing, six-leaf, and eleven-leaf stage) times application. The results showed that the highest yield was obtained under N3T2, N2T1, and N3T2 in 2018, 2019, and 2020, which was 222.49, 185.31, and 194.00% than that of N0 in each year, respectively. N2 and N3 significantly increased the yield through enhancing ears ha–1, grains per plant, and 100-grain weight; however, N2 and N3 did not show a significant difference in yield and above-yield components. In addition, N application time did not significantly change yield under the same N rate. N0 limited the activities of starch metabolizing enzymes, resulting in insufficient accumulation of sucrose and starch. The contents of indole-3-acetic acid, cytokinin, abscisic acid, and gibberellin were decreased under N0 during grain filling. The average grain-filling rate and maximum grain-filling rate (Gmax) and grain weight increment achieving Gmax increased under N2 and N3, and the grain-filling parameters were positively correlated with 100-grain weight. In conclusion, 200 kg N ha–1 with one-third application at sowing and two-thirds application at the six-leaf stage is a suitable N supply way to improve starch metabolizing enzymes, regulate hormone content, and enhance grain-filling rates, and thus increasing the maize yield in the semiarid Loess Plateau of China.