biocommaIntroduction
Mycotoxins are toxic secondary metabolites produced byfungi, primarily absorbed by humans through diet. Aspergillus,Penicillium, and Fusarium are the main genera generating themost common mycotoxins found in food, including aflatoxin,ochratoxin A, sterigmatocystin, patulin, fumonisin, zearalenone,deoxynivalenol, NIV, T-2 toxin, etc. Biocomma establishedan LC-MS/MS method using Copure® 302 MultifunctionalPurification Column for detection of 10 mycotoxins in wheatflour with good recovery and stability for your reference.
Experiment
Extraction
Weigh 5 g of sample into a 50 mL centrifuge tube. Add 20 mLof acetonitrile-water-acetic acid solution (80:19:1) and vortexto mix well. Then, extract by ultrasonic treatment for 20 min.Centrifuge at 6000 r/min for 10 min. The supernatant is readyfor purification.
Purification (Copure® 302 Multifunctional PurificationColumn)
Add 10 mL of prepared sample to a glass tube. Insert therubber head of the purification column into the tube, and pushto the bottom. Pipette 5 mL of the purified sample to a samplevial or EP tube. Evaporate sample to dryness at 40 ℃ . Diluteto 1 mL by adding acetonitrile-water solution (50:50). Vortex for30 seconds to dissolve the residue. After filter by microporousmembrane, the sample is ready for analysis.
Instrument Conditions
1.Chromatography Conditions
Equipment: UPLC-MS/MS (Thermo Scientific TSQ Endura)
Chromatographic column: Commasil® BEH T-C18 (2.1 mm×100mm, 3 μm)
Mobile phase: A:5mM Ammonium acetate solution (contains0.1% formic acid)
Mobile phase: B: Acetonitrile (contains 0.1% formic acid)
Mobile phase gradient: initial 90%A,40%A (0 min~2 min), 10%A(2 min~6 min), 10%A (6min~7 min), 90%A (7 min~8 min),90%A (8 min~10 min)
Flow rate: 0.3 ml/min
Column temperature: room temperature
Injection volume: 5.0 μL
2.Mass Spectrometry Conditions
Sheath gas pressure: 30 arb
Auxiliary gas pressure: 8 arb
Ion exchange tube: 300 ℃
Auxiliary air temperature: 350 ℃
Detection mode: MRM
Scan method: positive ion mode (ESI+) and negative ion mode(ESI-)
Table 1. Ion Selection Parameters (*Quantitative Ions)
Note: The detection of zearalenone is in negative ion mode(ESI-), and the others are in positive ion mode (ESI+).
Results
Table 2. Spiked Recovery of 10 Mycotoxins in Wheat Flour
Based on Table 2, the recovery rates of 10 Mycotoxins areall between 90-110% with RSD less than 10%, which meetthe standard of the experimental requirements. The overallevaluation of recovery rate is better than Brand A.
Figure 1. TIC of Wheat Flour Sample
① Before Purification
② Purified by Brand A
③ Purified by Copure® 302 Multifunctional Purification Column
Based on Figure 1, the interference of impurities in sample ③ is obviously reduced, therefore miscellaneous peaks are fewer inthe TIC chromatogram. The purification effect of Copure® 302 Multifunctional Purification Column is better than Brand A to meetexperiment requirements.
Ordering Information
