The present study evaluates the teratogenic and toxicological effects of citric acid (E330), a food additive, on Xenopus laevis embryos. The embryos were exposed to a range of E330 concentrations, from stage 8 to 11 of development, for 96 h under static renewal test conditions. The median lethal concentrations, no-observed adverse effect concentration (NOAEC), lowest observed adverse effect concentration (LOAEC), and minimum concentration to inhibit growth (MCIG) values were calculated. The lethal concentration (LC) values LC10, LC20, LC30, LC40, and LC50 determined for E330 exposure were 0.0113, 0.0117, 0.0119, 0.0122, and 0.0124 g/L, respectively. NOAEC and LOAEC values were calculated as 0.001 and 0.01 g/L. Since the effective concentration (EC50) value could not be determined, the E330 teratogenic index (TI), which is LC50/EC50, was not calculated. MCIG was calculated to be 0.010 g/L. The anomaly rate in Xenopus embryos treated with E330 was quite low. Therefore, the endpoint for the Xenopus embryos was whether they were alive or dead at the end of the study. It can be concluded from these observations that using unlimited amounts of E330 may result in preterm birth or abortion in humans, and E330 usage must be reevaluated and, potentially, limited. Moreover, our results confirm that the Frog Embryo Teratogenesis Assay Xenopus (FETAX) can be a useful pretest for integrated biological hazard assessment.