Unlike other macronutrients such as protein, the amounts and types of fat in the human diet vary tremendously across cultures and over time have changed significantly within Westernized countries. Studies of the effect that fat sources, fat amounts and changes in fat intake have on human disease are extremely difficult to conduct with traditional dietary assessment methods for a number of reasons. These include the hidden nature of many fats, the variation in fatty acids contained in foods and feed and the sensitivity of individuals to questions about fat intake in their diets. For these reasons biomarkers of fat intake are particularly desirable. Fat and fat-soluble substances have the advantages over other nutrients of a long half-life and readily accessible storage depots (in the absence of starvation, undernutrition or eating disorders). Technological advances in quantitative measurements of individual fatty acids, with the help of gas chromatography and mass spectrometry (GCMS)((3)) and high performance liquid chromatography (HPLC), made possible the study of specific isomers of minor fatty acids from small tissue samples. Technological advances also opened the gateways to the study of fats that represent less than 1% of the total fat profiles, such as decosahexanoic acid (DHA), eicosapentanoic acid (EPA) and conjugated linoleic acid (CLA). Biological advances enhanced our appreciation of the differences between fats of differing chain lengths within a family, including the saturated fats. Challenges remain, such as assessing total fat intake, discriminating the contribution of endogenously produced fats, determining how to evaluate the importance of relative versus absolute contributions of fat and accounting for the factors that influence deposition and mobilization of individual fats within and between individuals. Factors that can influence deposition and mobilization include genetic variation, disease status, lifestyle differences (i.e., alcohol consumption and smoking), circulating apolipoprotein levels and the hormonal milieu of the individual and the source tissue.