This document specifies a high-performance liquid chromatography (HPLC) method for determining the 16-O-methylcafestol content in green and roasted coffee. The method described is suitable with a reasonable precision for a content of 40 mg to 1 600 mg of 16-O-methylcafestol per kg of green and roasted coffee respectively. The collaborative study has shown that concentrations also between 20 to 40 mg/kg can be successfully analysed depending on the laboratory equipment.