Non-alcoholic fatty liver (NAFL) has become a major threat to metabolic health. Animal studies have suggested that disturbances in choline metabolism may be linked to the development of NAFL. However, to date, human data on the link of choline and NAFL is scarce. The trimethylammonium (TMA) group of choline can be detected with 1H-MRS at 3.20 ppm. Here, we investigated the relationship between intrahepatic choline levels and hepatic lipid content on healthy overweight/obese subjects using 1H-MRS at 3 Tesla. Our results showed that high hepatic lipid content is associated with low choline content in the liver.