Lipids are major sources of metabolic energy in sharks and are closely linked to environmental conditions and biological cycles, such as those related to diet, reproduction and migration. In this study, we report for the first time, the total lipid content, lipid class composition and fatty acid profiles of muscle and liver tissue of white sharks, Carcharodon carcharias, of various lengths (1.5–3.9 m), sampled at two geographically separate areas off southern and eastern Australia. Muscle tissue was low in total lipid content (<0.9% wet mass, wm) and was dominated by phospholipids (>90% of total lipid) and polyunsaturated fatty acids (34±12% of total fatty acids). In contrast, liver was high in total lipid which varied between 51–81% wm and was dominated by triacylglycerols (>93%) and monounsaturated fatty acids (36±12%). With knowledge of total lipid and dry tissue mass, we estimated the energy density of muscle (18.4±0.1 kJ g−1 dm) and liver (34.1±3.2 kJ g−1 dm), demonstrating that white sharks have very high energetic requirements. High among-individual variation in these biochemical parameters and related trophic markers were observed, but were not related to any one biological or environmental factor. Signature fatty acid profiles suggest that white sharks over the size range examined are generalist predators with fish, elasmobranchs and mammalian blubber all contributing to the diet. The ecological applications and physiological influences of lipids in white sharks are discussed along with recommendations for future research, including the use of non-lethal sampling to examine the nutritional condition, energetics and dietary relationships among and between individuals. Such knowledge is fundamental to better understand the implications of environmental perturbations on this iconic and threatened species.