ABSTRACT: BACKGROUND:Whole-organ, noninvasive techniques for the detection and quantification of nonalcoholic fatty liver disease features have clinical and research applications. However, the effect of time of day, hydration status, and meals are unknown factors with potential to impact bias, precision, reproducibility, and repeatability of chemical shift-encoded MRI (CSE-MRI) to quantify liver proton density fat fraction (PDFF). PURPOSE:To assess the effect of diurnal variation on PDFF using CSE-MRI, including the effect of time of day, the effect of meals and hydration status, as well as the day to day variability. STUDY TYPE:Prospective. SUBJECTS:Eleven healthy subjects and nine patients with observed hepatic steatosis. FIELD STRENGTH/SEQUENCES:A commercial quantitative confounder-corrected CSE-MRI sequence (IDEAL IQ) and an MR spectroscopy (MRS) sequence (multiecho STEAM) were acquired at 1.5T. ASSESSMENT:MRI-PDFF and MRS-PDFF estimates were compared across six visits (before and after a controlled breakfast, before and after an uncontrolled lunch, at approximately 4 pm, and then before breakfast on the following day) with three repeated measures for a total of 360 MRI-PDFF and MRS-PDFF measurements. STATISTICAL TESTS:Linear regression, Bland-Altman analysis, and mixed effect models were used to determine the bias, precision, and repeatability of PDFF measurements. RESULTS:No statistically significant linear trend was observed across visits for either MRI-PDFF or MRS-PDFF (P = 0.31 and 0.37, respectively). The repeatability was measured to be 0.86% for MRI-PDFF and 1.1% for MRS-PDFF over all six visits. For MRI-PDFF, the variability between all six visits (0.94%) was only slightly higher than within each visit (0.66%), with P < 0.001. For MRS-PDFF, the variability between all six visits was 1.29%, compared with 0.87% within each visit (P < 0.001). DATA CONCLUSION:Our results may indicate that it is not necessary to control for the time of day or the fasting/fed state of the patient when measuring PDFF using CSE-MRI. LEVEL OF EVIDENCE:2 Technical Efficacy: Stage 2 J. Magn. Reson. Imaging 2020;51:407-414.