VIDEO | Dissolving allergens

Research assistant Khyla Koegelenberg talks about the work done to optimize our sample preparation for the detection and quantification of allergenic proteins by LC-MS/MS.

The objective of this study was to optimize the sample preparation procedure, which aims to solubilize proteins from a food sample. The main target of the extraction procedure is to obtain the best protein quality and yield. Selecting an appropriate extraction method has to be the first step before further analysis. The success of the LC-MS/MS proteomics analytical method lies in a sample preparation procedure that allows efficient detection and quantification of allergens in the subsequent mass spectrometric analysis. The ideal sample preparation protocol would allow the total extraction of all protein components from the proteome of the samples to be analysed. Whilst keeping contamination by other molecules to a minimum and lastly enabling a high level of reproducibility across all types of food matrices.

When food has been heat-processed to some degree, protein aggregation or other chemical reactions such as the Maillard reaction can take place. This renders proteins less soluble. If your sample preparation is not optimized to overcome this, this may result in quantification errors or even false negatives.

The factors we considered in our quest to optimize our extraction procedure was the extraction buffer, pH, temperature and homogenization method.

This AOAC young scientist presentation by Khyla Koegelenberg highlights the challenges of selecting a sample preparation method that will work for all the different sample matrices, and addresses the factors to consider when optimizing the extraction method.