Purpose To investigate reproducibility of proton magnetic resonance spectroscopy (1H‐MRS) to measure hepatic triglyceride content (HTGC). Materials and Methods In 24 subjects, HTGC was evaluated using 1H‐MRS at 3.0 Tesla. We studied “between‐weeks” reproducibility and reproducibility of 1H‐MRS in subjects with fatty liver. We also studied within liver variability and within day reproducibility. Reproducibility was assessed by coefficient of variation (CV), repeatability coefficient (RC), and intraclass correlation coefficient (ICC). Results The CV of between weeks reproducibility was 9.5%, with a RC of 1.3% HTGC (ICC 0.998). The CV in fatty livers was 4.1%, with a RC of 1.3% HTGC (ICC 0.997). Within day CV was 4.5%, with a RC of 0.4% HTGC (ICC 0.999). CV for within liver variability was 14.5%. Conclusion Reproducibility of 1H‐MRS to measure HTGC for “between‐weeks” measurements and in fatty livers is high, which is important for follow‐up studies. Within liver variability displays a larger variation, meaning that liver fat is not equally distributed and during consecutive measurements the same voxel position should be used. J. Magn. Reson. Imaging 2009;30:444–448. © 2009 Wiley‐Liss, Inc.