Asparagine is known to accumulate in wheat grain under conditions of sulphur deficiency, leading to increased levels of acrylamide formation during processing. Analyses of milling fractions and of the outer layers of the grain prepared by hand dissection showed that the highest levels of asparagine were present in the bran fractions and in particular the aleurone layer, when grain were grown with sufficient sulphur supply. However, even mild S deficiency resulted in disproportional increases in the asparagine contents of white flour fractions, implying that optimisation of yield in a conventional milling system is not an appropriate strategy for processing grain from sulphur-deficient crops.