In this study, biotin (vitamin B7) contents of frequently consumed foods in Korea were determined by usingimmunoaffinity column in conjunction with high-performance liquid chromatography (HPLC). The biotin contents of 24foods of plant origin and 27 foods of animal origin were selected. The highest biotin contents in frequently consumed foodsof plant origin were found in red beans (Huinguseul; 11.475 μg/100 g). On the other hand, biotin was not detected in anyvarieties of sorghum. For frequently consumed foods of animal origin, salted pollack roe (7.486 μg/100 g) showed thehighest biotin content. However, beef and fish contained less biotin. All biotin analyses were conducted under analyticalquality control. The limits of detection and limits of quantification of biotin were 0.007 and 0.023 μg/100 g, respectively, andthe accuracy/recovery percentage was 95.35-105.02%. The precision values were 4.041% (repeatability) and 3.835%(reproducibility). Taken together, our data provide reliable data on the biotin contents of frequently consumed foods inKorea.