In this experiment, a 5 g (accurate to 0.0001 g) ground sample was placed into a flask, and 25 mL of acetonitrile-water (86:14, v/v) was added to the flask. Next, the solution was shaken for 1 h using a mechanical shaker, filtered with quantitative filter paper and then filtered with a 0.22-μm-pore-size membrane filter (Jinlong Co., Ltd., Tianjin, China). The filtrate was placed into a sterile centrifuge tube and analyzed using the HPLC-MS/MS system. For the HPLC-MS/MS procedure, separation was carried out in a C18 column (2.1 mm×50 mm, 1.8 μm particle size; Agilent Technologies Inc., USA), and the column temperature was 33°C. For AFB1 and DON, chromatographic elution was performed with a mobile phase consisting of methyl alcohol (part A) and 0.2% formic acid-water in ultrapure water (v/v, part B). The gradient elution at a 0.2 mL/min flow rate started at 80% B and was decreased to 50% B after 2 min and 0% B after 6 min via a linear gradient mode. Next, 100% A at a 0.3 mL/min flow rate was held for 2 min, and then 80% B at a 0.2 mL/min flow rate was started at 8.01 min and held for 2 min. For ZEN, chromatographic elution was performed with a mobile phase consisting of methyl alcohol (part A) and 1 mmol/L ammonium acetate in ultrapure water (part B). An isocratic elution at a 0.2 mL/min flow rate started at 20% B and was held for 1.4 min. Next, 100% A at a 0.3 mL/min flow rate was started at 1.41 min and held for 2 min. Then, 20% B at a 0.2 mL/min flow rate was started at 3.42 min and held for 2 min. The injection volume was 2 μL. The mass spectrometer was operated using a turbo ion-spray ionization source configured for electrospray ionization (ESI) in positive and negative switching ion modes, and acquisition was conducted using multiple reaction monitoring with a dwell time of 100 ms. The ion voltages for the positive and negative ion modes were both 4,500 V. Resolutions at Q1 and Q3 were set to units. Argon gas (99% purity) was used in the ESI source and the collision cell. The mass parameters of the ESI detection mode, precursor ion, product ion, cone voltage, fragmentor voltage, cell accelerator voltage and collision energy for each mycotoxin are summarized in Table 1.

aAFB1, aflatoxin B1; ZEN, zearalenone; DON, deoxynivalenol.

Note: The content above has been extracted from a research article, so it may not display correctly.



Q&A
Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.



We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.