Benjawan Kongcheep1, Siwalai Wongviriyakit1, Watinee Kumpum1, Hunor Farkaš2#, Jog Raj2*, Anawach Phuengkasem2 and Marko Vasiljević2 1Betagro Science Center Co., Ltd. 136 Moo 9, Klong Nueng, Klong Luaung, Pathumthani, 12120, Thailand.
2PATENT CO. DOO. Mišićevo, operating under A&P Nutrition trademark on the international markets, Vlade Ćetković 1a, 24211 Mišićevo, Serbia.
*Corresponding author: jog.raj@apnutrition.com
#Presenting author: hunor.farkas@apnitrition.com
Mycotoxins workshop, June 01-03, 2026, Berlin, Germany
OBJETIVE
In this study, a multi-mycotoxin analysis method based on solid-liquid extraction (SLE) for the analysis of aflatoxins (AFB1, AFB2, AFG1, AFG2), deoxynivalenol (DON), fumonisin B1 (FB1), fumonisin B2 (FB2), ochratoxin A (OTA), T-2 toxin, and zearalenone (ZEN) in various agricultural (corn, wheat, barley, wheat bran, soya bean, sunflower seed) and food/feed matrices (compound feed for pigs and poultry and total mixed ration for dairy cows) was developed and validated using LC-MS/MS Triple Quadrupole (Waters Xevo TQ-XS) at Betagro Science Center, Thailand.
MATERIALS & METHODS
The samples were finely ground and homogenized using a laboratory mill, and 2.5 g of sample was weighed and placed in a conical tube.
To this tube, 10 ml of extraction mixture I (80 % acetonitrile:19.9 % water:0.1 % formic acid) was added, and the lid was closed.
2.5 g sample
10 ml extraction mix I
The tube was then placed on an orbital shaker at 230 rotations per minute (rpm) for thorough mixing (90 minutes at room temperature).
The tube was subsequently centrifuged at 4000 × g for 5 minutes at 4 °C, and the supernatant was transferred to another 50 ml tube.
Then, 500 µl of supernatant was pipetted and combined with 500 µl of diluent solvent (50 % acetonitrile:0.1 % formic acid) in a 15 ml tube, and the solution was mixed by vortexing for 30 s.
4000 × g, 5 min, 4 °C
To compensate for matrix effects during electrospray ionization, mixed 13C-labelled internal standards for aflatoxins (AFB1, AFB2, AFG1, AFG2), deoxynivalenol (DON), fumonisin B1 (FB1), fumonisin B2 (FB2), ochratoxin A (OTA), T-2 toxin, and zearalenone (ZEN) were pipetted (20 µl) into 1000 µl of the extract contained in the 15 ml tube, and the solution was mixed by vortexing for 30 s.
A total of 1020 µl of liquid was prepared by filtering the extract through a membrane syringe filter into a glass vial for LC-MS/MS analysis.
230 rpm, 90 min
13C-labelled internal standards 500 µl supernatant
1020 µl
RESULTS
The recoveries of 10 mycotoxins spiked in food/ feed and cereal matrices were recorded in the range of 93.6 %–114.2 % after internal standard correction (relative standard deviations [RSDs] below 14.2 %).
CONCLUSIONS
These data demonstrate that this is a robust method based on the dilute-and-shoot technique for the simultaneous analysis of 10 mycotoxins using LC-MS/MS.