Quantification of mycotoxins in foodstuffs is extremely difficult as a limited amount of toxins are known to be presented in the food samples. Mycotoxins are secondary toxic metabolites, made primarily by fungal species, contaminating feeds and foods. Due to the presence in globally used grains, it is an unpreventable problem that causes various acute and chronic impacts on human and animal health. Over the previous few years, however, progress has been made in mycotoxin analysis studies. Easier techniques of sample cleanup and advanced chromatographic approaches have been developed, primarily high-performance liquid chromatography. Few extremely sophisticated and adaptable tools such as high-resolution mass spectrometry and gas chromatography–tandem MS/MS have become more important. In addition, Immunoassay, Advanced quantitative techniques are now globally accepted for mycotoxin analysis. Thus, this review summarizes these traditional and highly advance methods and their characteristics for evaluating mycotoxins. © 2020 The Authors. Food Science & Nutrition published by Wiley Periodicals, Inc.