Simple simultaneous alkaline extraction (saponification) coupled with HPLC was validated and applied for the analysis of ten vitamers of A (retinol and β-carotene) and E (α-, β-, γ-, and δ-tocopherols (T) and corresponding tocotrienols (T3)) in seafood. Limits of detection for retinol, β-carotene and α-T were 0.380 μg/100 g, 0.602 μg/100 g and 0.0002 mg/100 g, respectively. Analysis precision was excellent for all vitamers (RSD: repeatability <2.4%, reproducibility <4.5%). Recoveries of vitamers ranged from 92% to 110%, showing good accuracy. Total vitamin A and E activities varied among seafood types, with ranges of 0.00-311.77 μg retinol activity equivalent/100 g and 0.01-0.62 mg α-tocopherol equivalent/100 g, respectively. β-Carotene was found in some seafoods such as shellfish (ark shell, blue mussel and spiny top shell) and crustacean (gazami crab) samples as well as marine algae due to the presence of intestinal organ containing algae in edible parts. The highest β-carotene content (689.84±29.86 μg/100 g) was observed in seasoned oyster whereas the retinol content was highest in blackthroat seaperch (311.77±7.99 μg/100 g). α-T was found in all samples and ranged from 0.07 to 4.72 mg/100 g. This study provides reliable vitamin A and E profiles for domestic seafood in Korea.