Successful applications of toxins derived from Bacillus thuringiensis (Bt) strains in Bt-based bioinsecticides and more recently in Bt-plants in crop protection have enhanced the importance of analytical quantification of Cry toxin dosages for studies on various topics including environmental risk assessment (ERA), resistance management, quality control and regulatory compliance. It is essential to follow-up distribution and environmental degradation of these lectin type, crystalline (Cry) toxin proteins showing insect specificity at order level. Thus, Cry1Ab toxin produced by Bt-maize of genetic event MON 810 is specific to lepidopteran species. Widely used analytical methods for detection of Cry toxins are enzyme-linked immunosorbent assay (ELISA) systems. Reported Cry1Ab toxin concentrations in MON 810 maize show high variability: order of magnitude differences have been observed among various plant parts from different varieties, cultivated at different locations, and sometimes even within the same plant variety at a single location. Besides biological sources of variability, numerous analytical problems have been identified and are reported in this report, influencing the results of quantitative determination of Cry1Ab toxin and explaining the high variability among documented data on toxin content. Conclusions in every case refer to genetic event MON 810, but can be extended to other genetic events producing Cry1Ab toxin.