This study aimed to evaluate in ovo feeding of vitamins E, B1, and B2 on hatchability characteristics, and blood parameters of hatched chicks. Total of 1140 eggs (Cobb 500, 63.8 g/egg) were divided into five groups (228 in each). Experimental groups included negative control, positive control (0.2 ml sterile saline), eggs injected with Vit. E (1000 mg/100 µl sterile oil solvent), Vit. B1 (8.92 mg/100 µl sterile saline), or Vit. B2 (4.30 mg/100 µl sterile saline). Materials were injected into embryo on 17-day incubation. All eggs were subjected to the same incubation temperatures, relative humidity, and CO₂ concentration. After hatching, blood samples were collected from 6 hatched chicks in each group for analysis. Results showed no significant effect for vitamins on hatchability rates and hatched chick quality. Hatched chick weight was higher (P<0.05) with all treatments than in positive control. Vitamins E and B2 decreased (P<0.05) late embryonic mortality. Vit. B1 increased glucose and LDL. Vit. E decreased (P<0.05) HDL and triglycerides. Total protein was increased (P<0.05) by all vitamins injected. ALP activity was increased only by Vit. B1. Plasma T₃ concentration was increased by vitamins E and B1, but did not differ significantly from that in positive control and Vit. B2 groups. The highest corticosterone level was in the negative control, being lower with all materials injected, particularly with Vit. B1, reflecting positive impacts on eliminating hatching stress. Although hatchability rates were not affected by material injected, all vitamins had beneficial effects on hatched chick weight and health performance.