This instruction of simultaneous Aflatoxin and Ochratoxin (Afla-Ochra) determination in food and feed focuses on the enrichment step of extract using immunoaffinity column (IAC) and quantification with HPLC. Accepted laboratory extraction methods could be maintained. Full performance of the IAC column is given if pronounced criteria regarding organic solvent tolerance, elution process of analyte and working range of column is followed. Many pretreatment methods of Aflatoxin and Ochratoxin determination in food and feed show low sensitivity because of interfering substances if problematic matrices are applied. This method of content determination of Aflatoxins and Ochratoxins combines the high selectivity of an immunoaffinity column (IAC) with its potential to concentrate eluate and additional step of purification by HPLC column. Please notice that this instruction focuses on the handling with the IAC column. For the commodity extraction step a literature method is given. The given apparatus (e.g. HPLC system) might serve as example among other possibilities. Range: 0.02 - 3ng AFL 0.02 - 2ng OTA. Sensitivity: 0.02 - 3ng AFL 0.02 - 2ng OTA. ?