This study investigated the effects of Fe3O4 nanoparticles released from synthesized Thiourea catalyst to Chlorella vulgaris as an essential primary producer in aquatic systems. A range of Fe3O4 concentrations (0, 10, 100, 250, 500, 750, and 1000 mg L-1) was applied for the exposure test. Biological parameters of C. vulgaris, including cell density, cell viability, and pigment content were assessed. Bioconcentration factor and bioaccumulation were evaluated for contaminated microalgae. Non-carcinogenic risks were then assessed using target hazard quotient (THQ) for potential human consumptions. Findings showed that C. vulgaris cell numbers increased from 0 to 500 mg L-1 of Fe3O4. Chlorophyll a represented a time-dependent response, and greatest values were detected in 250 and 500 mg L-1 Fe3O4 at 4.2 and 4 mg/g, respectively. Chlorophyll b content showed a time-related manner in exposure to Fe3O4 with the highest values recorded at 250 mg L-1 after 96 h. Moreover, bioaccumulation displayed a dose-dependent response as bioaccumulated iron was in the largest amount at 15000 µg/g dw in 1000 mg L-1, whereas the lowest one was in the control group at 1700 µg/g dw. The bioconcentration factor showed a concentration-relevant decrease in all iron treatments and 10 mg L-1 of Fe3O4 represented the greatest BCF at 327.3611. Non-carcinogenic risks illustrated negligible hazard (THQ < 1) in a dose-response pattern and the largest EDI and THQ were calculated in 1000 mg L-1 at 7.4332E-07 (mg kg-1 day-1) and 1.06189E-09, respectively. In essence, iron is an essential trace element for biological aspects in aquatic systems, but in exceeding concentrations could impose toxicity effects in C. vulgaris populations.