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LC/MS Analysis of Aflatoxin M1 in Milk on Ascentis® Express HILIC after SPE using Supel™ Tox AflaZea, Quantifying Picogram Concentrations

LC/MS Analysis of Aflatoxin M1 in Milk on Ascentis® Express HILIC after SPE using Supel™ Tox AflaZea, Quantifying Picogram Concentrations

Conditions

sample preparation SPE (Solid Phase Extraction)
sample preparation reconstitut in 500 µl 20:80 acetonitrile:water, filter into 750 µl poly vial using PVDF filter, 0.2 µm
sample/matrix cow milk
extraction process centrifuge 45 g liquid milk for 60 min at 5000 rpm (RCF 4863 xg), cool at 0 C for 30 minutes, remove fat layer, transfer middle layer to collection flask,
SPE tube/cartridge Supel(TM) Tox AlflaZea, 6 mL (55314-U)
sample addition 10 mL, 2 drop/second using vacuum
washing 2 x 2 mL acetonitrile (collect with sample)
eluate post-treatment evaporate to dryness at 70 C using nitrogen
column Ascentis Express C18, 50 cm x 2.1 mm I.D., 2.7 µm particles (53822-U)
column temp. ambient
mobile phase [A] 0.1% (v/v) formic acid; [B] acetonitrile
gradient 90% A for 0.5 min; to 10% A in 0.5 min; held at 10% for 3 min; to 90% A in 0.10 min; held at 90% A for 3 min
flow rate 0.4 ml/min
injection 10 µl
detector MS/MS (MRM transitions)

Description

Analysis Note Aflatoxin M1 is the metabolite of Aflatoxin B1 and can be found in milk products when animals have been fed alatoxin-contminated feeds. Aflatoxins are positively associated with hepataocellular carcinomas. Aflattoxin M1 was detected in milk and quantified at concentrations fo 25 pg/mL. Sample preparation included a protein precipitation with solvent partitionng and sample clean-up using Supel Tox AflaZea SPE tubes. Analysis was perfomed via LC-MS using an Ascentis Express C18, 5 cm x 2.1 mm I.D., 2.7 μm HPLC column for the separation. Detection and quanitification of aflatoxin M1 at 25 pg/mL satisfies the current regulatory requirements for aflatoxin M1 for EU treaty participants as stated in EC 1881/2006., Sample pretreatment: centrifuge 45 g liquid milk for 60 min at 5000 rpm (RCF 4863 xg), cool at 0 °C for 30 minutes, remove fat layer, transfer middle layer to collection flask, add 20 mL acetonitrile to 50 mL centrifuge tube, add 20 mL of sample to acetonitrile, add contents of 1 Supel Q non-buffered tube, immedately disperse salt, quicky place on shaker for 5 minutes, centrifuge at 5000 rpm for 20 min, remove top layer (loading solution)
Categories Analytical Chromatography, Food & Beverage Analysis, SPE Applications, Mycotoxins, Beverage Testing, Milk
Featured Industry Food and Beverages
Legal Information Ascentis is a registered trademark of Sigma-Aldrich Co. LLC, Supel is a trademark of Sigma-Aldrich Co. LLC
suitability application for SPE, application for HPLC

Materials

     
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