Excessive consumption of free sugars (which typically includes a composite of glucose and fructose) is linked to an increased risk of developing chronic metabolic diseases including obesity, non-alcoholic fatty liver disease (NAFLD), type 2 diabetes, and cardiovascular disease. Although current knowledge of fructose metabolism has primarily come from studies involving liver metabolism, where it is implicated in impaired insulin sensitivity, increased fat accumulation and dyslipidaemia, we still have a limited understanding of how consumption of fructose, as part of a mixed meal, may alter hepatic fatty acid synthesis and partitioning. Moreover, the effect of dietary fructose in subcutaneous adipose depots, the largest human fat depot, has not been studied. A consistent finding in the literature is the effect of fructose on hepatic de novo lipogenesis (DNL). By undertaking a randomised cross-over study, we have investigated the effect of a mixed meal, high and low in fructose, in healthy men and women. Hepatic DNL was assessed using stable-isotope tracers. We found after consumption of the low fructose meal, there was no difference in fasting or postprandial hepatic DNL between men and women. In contrast, after consumption of the high fructose meal, despite fasting hepatic DNL being similar, postprandial DNL was significantly higher in women compared to men (effect of sex p < 0.05, time x sex interaction p< 0.05). The potential implications of this observation will be discussed. Moreover, we have started in vitro cellular studies to investigate the effects of fructose on subcutaneous adipocyte metabolism. Our preliminary findings suggest that fructose is metabolised by human adipocytes and is used as a substrate for DNL. This talk will discuss our recent findings and provide evidence for tissue-specific and sex-specific responses to fructose challenges.