The usage of coloring agent and preservative including sunset yellow, tartrazine and sodium benzoate in orange juice is forbidden in Iran because of bad effects on public health. The present study included 30 samples of orange juice purchased from local market of Tehran in Iran for simultaneous identification and quantification of sunset yellow, tartrazine and sodium benzoate through high performance liquid chromatography (HPLC) with UV detection and ACE C18, 5µm (4.6 × 250 mm) column. The additives were eluted with a mixture of methanol and ammonium acetate as mobile phase at flow rate of 1.0 ml/min and monitored at 230 nm. Exposure assessment (estimated daily intakes) of consumers calculated through official reports on annual orange juice production in Iran. Among the samples analyzed, at least one additive detected in all orange juices. The amount of sunset yellow, tartrazine and sodium benzoate were ranging from 5.84 ± 0.05 to 23.12 ± 0.19; 0.70 ± 0.01 to 2.32 ± 0.01 and 12.23 ± 1.50 to 56.80 ± 2.13 µg/ml, respectively. Satisfactory repeatability with relative standard deviation < 2.03, excellent sensitivity (Limit of detection: 0.20, 0.25, 0.22 µg/ml; Limit of quantification: 0.70, 0.90, 0.80 µg/ml) with recovery more than 96.3% were obtained. Estimated daily intake of sunset yellow, tartrazine and sodium benzoate were 0.85, 0.96 and 1.11mg/kg body weight through orang juice, respectively which was lower than acceptable daily intakes in society of Iran. We concluded that the level of additives found in orange juice does not affect adversely on consumer’s health.