biocommaIntroduction
Aflatoxins are toxic and highly carcinogenic metabolites ofAspergillus flavus and Aspergillus parasiticus fungi. When cowsconsume contaminated feed, aflatoxin B1 will be transformedinto aflatoxin M1. A portion of aflatoxin M1 is excreted throughurine and milk, while other portions are stored in meat.Biocomma established a rapid LC method using Copure® 223Multifunctional Purification Plate for detection of aflatoxinsM1 and M2 in milk with good recovery and stability for yourreference. The recoveries were 90-110% with RSD less than5% at both 5 ng/g and 10 ng/g concentrations.
Experiment
Extraction
Weigh 4 g of sample into a 50 mL centrifuge tube. Add 10 mL ofAcetonitrile and vortex to mix well. Then, extract by ultrasonictreatment for 20 min. Centrifuge at 6000 r/min for 10 min. Thesupernatant is ready for purification.
Purification (Copure® 223 Multifunctional PurificationColumn)
Add 10 mL of supernatant to a glass tube. Insert the rubberhead of the purification column into the tube, and push to thebottom. Pipette 5 mL of the purified sample to a sample vial orEP tube. Evaporate sample to dryness and redissolve by 1 mLof initial mobile phase. Vortex for 30 seconds to dissolve theresidue. After filter by microporous membrane, the sample isready for analysis.
Instrument Conditions
Equipment: UltiMate 3000 (Thermo Fisher Scientific) with FLDdetector.
Chromatographic column: Agilent ZORBAX C18 (4.6 mm×250mm, 5 μm)
Mobile phase: A: Deionized water, B: Acetonitrile-methanol(50/50)
Mobile phase elution conditions: A: 70%, B: 30%
Flow rate: 1.0 mL/min
Column temperature: room temperature
Injection volume: 20.0 μL
Detection wavelength: excitation wavelength 360 nm, emissionwavelength 430 nm
Detector: FLD
Results
Table 1 Aflatoxin M Group Spike Recovery in Milk
Figure 1. Chromatograms of Spiked Samples Purified byDifferent Brands at 5 ng/g
① Purified by Brand A
② Purified by Copure® 223 Multifunctional Purification Column
③ Before Purification
Based on Figure 1, the miscellaneous peaks of sample ② arefewer than sample ① . Less interference of impurities indicatesbetter purification effects. The recovery rates of AflatoxinM1 and M2 are both between 90-110%, and the RSD valueis less than 5%, which meet the standard of experimentalrequirements.
Ordering Information
