A liquid chromatography-tandem mass spectrometry method for simultaneous analysis of acrylamide and the precursors, asparagine and reducing sugars in bread

A LC-MS-MS method for simultaneous determination of acrylamide, asparagine, fructose, glucose and sucrose in bread was developed. The method is based on aqueous extraction by blending. After centrifugation the samples were cleaned up by solid phase extraction on C18 cartridges conditioned with 2 mL of methanol and 2 x 2 mL of water and subsequently flushed with sample solution before the actual analytical sample fractions were collected. Analytes were separated on a Hypercarb column (100 mm x 2.1 mm, 5 mu m) and detected by tandem MS with electrospray ionisation. Acrylamide and saccharides were ionised in positive mode. Asparagine in wheat bread was detectable at lower levels using negative ion mode. To compensate for matrix induced signal suppression D-3-acrylamide and N-15(2)-asparagine were used as internal standards for acrylamide and asparagine, respectively. Recoveries were in the range 93-112% for acrylamide spiked at 30, 250 mu g kg(-1) and 97-101% for asparagine spiked at 70, 140 mg kg(-1). Sorbitol was used as internal standard for fructose and glucose. Samples were diluted to obtain acceptable recoveries in saccharide analyses. LODs were 13 mu g kg(-1) for acrylamide and 2 mg kg(-1) for asparagine in wheat bread.