4.3.1. Immunoaffinity Column Method (Method A)

A corn sample (10 g) was extracted with 50 mL Phosphate Buffered Saline (PBS). After shaking for 60 min and centrifugation for 10 min, 35 mL of supernatant (extract A) was filtered through a glass microfiber filter. Subsequently, 35 mL methanol was added to the 15 mL remaining supernatant and the sample was shaken and centrifuged the same as extract A. After these two steps, 10 mL extract was diluted with 90 mL PBS and filtered through a glass microfiber filter (extract B). Extract B (50 mL) was passed over Myco6in1<sup>+</sup> (multiantibody IMA column, VICAM, Watertown, MA, USA) and washed with 20 mL PBS. At this step, 5 mL extract A was passed through the same Myco6in1+ column and washed with 10 mL water. The extract was eluted two times with 1.5 mL methanol:water (80:20, v:v) containing 0.5% acetic acid. The last eluted step was repeated and collected in another vial. Then, the extract was evaporated to dryness at 40–50 ◦C under a gentle stream of nitrogen. Both tubes were reconstituted with methanol:water (80:20, v:v) containing 0.5% acetic acid and combined into one vial.
