Chinese Star anise, Illicium verum Hook, is a well known spice in many cultures and has also been used to treat infant colic. Recent publications report that Chinese Star anise might be adulterated with the toxic Japanese Star anise, Illicium anisatum L. We have developed a molecular method that helps with the detection of I. anisatum as adulterant of I. verum. We PCR-amplified the internal transcribed spacer (ITS) region and analyzed it with the endonucleases PSTI and BFMI. Based on fragment length polymorphisms (RFLP), we were able to detect and distinguish between I. anisatum and other Illicium species in powdered samples.