Analytical methods - Acrylamide

prEN 16618 Food analysis – Determination of acrylamide in food by liquid chromatography tandem mass spectrometry (LC-ESI-MS/MS) (under approval – publication date March 2015)

• Scope

This document specifies a method for the determination of acrylamide in bakery ware such as bread, toasted bread, crisp bread, butter cookies and biscuits, as well as potato products such as potato chips, potato crisps and potate pan cake and roasted coffee, by liquid chromatography in combination electrospray ionisation and tandem mass spectrometry (LC-ESI-MS/MS). This method has been validated in an interlaboratory study via the analysis of both naturally contaminated and spiked samples, ranging from 14,3 µg/kg to 9 083 µg/kg.

The limit of quantification (LOQ) depends on the type of instrument used and on the actual performance of the instrument. The majority of the laboratories participating in the validation study was able to determine acrylamide in a butter cookie sample at a level of 14,3 µg/kg. The validation by interlaboratory study showed that it can be expected to be in the range below 15 µg/kg and 30 µg/kg.

• Principle

- Extraction of acrylamide with water

- Addition of isotopic labelled acrylamide

- Centrifugation of the extract

- Cleanup of the supernatans on two SPE columns:

• SPE with silica-based C₁₈ groups as well as anion and cation exchangers: acrylamide is not retained
• SPE with a polymer-based phase: high capacity to bind acrylamide
  • Elution with 60 % methanol in water

- Evaporation of methanol

- LC-MS/MS analysis

• HPLC column with graphitised carbon as stationary phase

• Key steps
  • Never exceed 40 °C during extraction or the work-up process. Acrylamide has been found to be formed as an artefact in some analytical procedures for acrylamide.
  • Extraction
    • Make sure that particles are small enough before extraction (< 1 mm)
    • Extraction for bakery ware and potato product samples
      • Test portion: 2.0 g ± 0.01 g
      • Addition of 40 mL of water and 400 µL of internal standard solution
      • Shaking of the mixture by hand, by vortex mixer and finally on a mecha