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Key Documents

G006300

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

application for SPE, application for HPLC

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About This Item

technique(s)

HPLC: suitable
solid phase extraction (SPE): suitable

test parameters

sample preparation: SPE (Solid Phase Extraction)
sample/matrix: cow milk
SPE tube/cartridge: Supel Tox AlflaZea, 6 mL (55314-U)
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,
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
sample preparation: reconstitut in 500 μL 20:80 acetonitrile:water, filter into 750 μL poly vial using PVDF filter, 0.2 μm
column: Ascentis Express C18, 50 cm x 2.1 mm I.D., 2.7 μm particles (53822-U)
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
column temp.: ambient
detector: MS/MS (MRM transitions)
injection: 10 μL

suitability

application for HPLC
application for SPE

application(s)

food and beverages

Analysis Note

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)
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.

Legal Information

Ascentis is a registered trademark of Merck KGaA, Darmstadt, Germany
Supel is a trademark of Sigma-Aldrich Co. LLC

Analito

Descripción

    Aflatoxin M1 solution, 0.5 μg/mL in acetonitrile, analytical standard

Analytical column

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Related product

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SPE tube or plate

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standard

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