A method for analysis of oat kernel saponins is presented. The saponins were extracted from defatted oatmeal with methanol for 24 h in a Soxhlet apparatus. Separation of the saponins was accomplished by high-performance liquid chromatography (HPLC) using an octyl-silica column and gradient elution with acetonitrile in water. Two main peaks, avenacoside A and B, were detected. The identification was achieved by comparing retention times and photodiode array UV spectra with those of an avenacoside A and B standard preparation from oat leaves. The identity of the avenacoside A and B peaks was further supported by incubation of oatmeal suspensions in water, after which the retention times were shifted to those obtained with the corresponding desglucoavenacoside standard. The saponin content in oatmeal containing a mixture of Swedish commercial oat varieties was 0.040% (dry matter basis).