Extraction and Clean-up of Lupin Samples

Different categories of the treated lupin samples were, initially, homogenized by the use of a blender. Then, 200 mg of homogenized lupin samples was weighed and extracted with 1 mL of MeOH:H₂O 60:40 (v:v) by a Precellys Evolution homogenizer (Bertin Technologies SAS, Montigny-le-Bretonneux, France) with 3 cycles of 10 sat 7000 rpm with a 45 s stop between each cycle followed by centrifugation for 10 min at 4 °C at 11424 RCF. Then, 50 µL of the supernatant was collected and diluted in 1 mL with a final ratio of H₂O:MeOH 90:10 (v:v). The resulting solution was loaded onto a polymeric SPE cartridge for the clean-up step.