Phytoestrogens are nonsteroidal isoflavones present for example in soy foodstuffs. They have many health benefits, i.e. positively affect cardiovascular deceases and act as suppressor of reactive oxygen species. On the other hand they can also conduce to cancer reveal because their estrogen-like molecular structure. Many of the anticancer mechanisms of isoflavones were detected by in vitro procedures, including cell cultures. Several new extraction procedures, such as accelerated solvent extraction, have been developed as an improvement of the traditional procedures (soxhlet and/or ultrasonication methods) and also applied on isoflavones extraction from biological materials. High performance liquid chromatography (HPLC) with various detection methods is common for isoflavone determination. Fast column in HPLC enables shorter times of determination. The goal of our work was to apply ultra fast column on HPLC/DAD determination of isoflavones with modified accelerated solvent extraction (ASE).