Biotin is contained in a wide variety of foods. However, the 5th Revised Japanese Standard Food Table does not include the biotin content of foods, nor does it consider the loss of biotin resulting from food processing and cooking, or the bioavailability of biotin. Therefore, we determined the biotin contents of 330 typical foods available in Japan, and compared them with those of foods available in various other countries. The content of biotin found in peanuts, red peppers, liver (pork and chicken), egg yolk, instant coffee (powder), yeast (dried) and royal jelly exceeded 50 μg/100 g. With regard to various food groups, nuts, mushrooms and meats (including liver) contained, on average, a large amount of biotin, whereas fruits and oils contained only a small amount. Although the biotin content differed according to the method of analysis and the form of the foods, there were no marked differences in the biotin content of foods between Japan and other countries.