A pot experiment was conducted in the net house of the Department of Agricultural Chemistry, Bangladesh Agricultural University, Mymensingh, to examine the effect of different levels of Zn and B on the major biochemical and nutritional quality of tomato fruits. The experiment was laid out in a completely randomized design (CRD) with 4 replications along with two treatment factors viz., (i) four levels of Zn- like control, Zn @ 4.0 kg ha-1, Zn @ 6.0 kg ha-1 and Zn @ 8.0 kg ha-1; and (ii) three levels of B- like control, B @ 2.0 kg ha-1 and B @ 3.0 kg ha-1. Among the biochemical parameters- lycopene, total acidity, and vitamin C contents in tomato fruits ranged from 3.24-3.76 mg 100 g-1, 0.26-0.36%, and 21.76-26.40 mg 100 g-1 samples, respectively. The study results revealed that the highest amounts of lycopene and vitamin C were recorded from B @ 2.0 kg ha-1. Combined Zn and B applications showed a highly significant effect on total acidity, lycopene, and vitamin C contents of tomato fruits. Similarly, the application of Zn and B alone or in combination significantly affected the major nutrient contents of tomato fruits. The highest amounts of Ca, Mg, Na, K, and P were obtained from the application of Zn @ 4.0 kg ha-1, while the maximum amounts of Fe and Zn were recorded from Zn @ 8.0 kg ha-1 treatment. However, the application of B alone significantly reduced the contents of Ca, P, S, Fe, and Zn in tomato fruits while the contents of Mg, K, and Na remained almost unchanged. Finally, the study results concluded that the combined application of Zn and B (@ 4.0 and 2.0 kg ha-1, respectively) had a significant effect on major nutrients and biochemical characters of tomato fruits.