Adipose tissue as well as other depots of fat (triglycerides) are increasingly being recognized as active contributors to the human function and metabolism. In addition to the fat concentration, also the fatty acid chemical composition (FAC) of the triglyceride molecules may play an important part in diseases such as obesity, insulin resistance, hepatic steatosis, osteoporosis, and cancer. MR spectroscopy and chemical-shift-encoded imaging (CSE-MRI) are established methods for non-invasive quantification of fat concentration in tissue. More recently, similar techniques have been developed for assessment also of the FAC in terms of the number of double bonds, the fraction of saturated, monounsaturated, and polyunsaturated fatty acids, or semi-quantitative unsaturation indices. The number of papers focusing on especially CSE-MRI-based techniques has steadily increased during the past few years, introducing a range of acquisition protocols and reconstruction algorithms. However, a number of potential sources of bias have also been identified. Furthermore, the measures used to characterize the FAC using both MRI and MRS differ, making comparisons between different techniques difficult. The aim of this paper is to review MRS-and MRI-based methods for in vivo quantification of the FAC. We describe the chemical composition of triglycerides and discuss various potential FAC measures. Furthermore, we review acquisition and reconstruction methodology and finally, some existing and potential applications are summarized. We conclude that both MRI and MRS provide feasible non-invasive alternatives to the gold standard gas chromatography for in vivo measurements of the FAC. Although both are associated with gas chromatography, future studies are warranted.