Background and Aims Shift workers face an increased risk of cardiovascular disease (CVD), type-2 diabetes and obesity. Eating during the night is a likely contributing factor, as it coincides with the time at which postprandial metabolism is least efficient. In this pilot randomised crossover trial, we examine the effects of a short overnight fast on CVD risk markers (primarily postprandial triglyceride and glucose response) of night shift workers. Methods and Results Night shift workers with abdominal obesity underwent 4-week intervention and control periods, separated by ≥ 2 weeks washout. In the intervention period, an overnight fast (0100h-0600h) was implemented, by redistributing 24-hour energy intake. Usual dietary habits were followed in the control period. Outcomes between intervention and control were compared using mixed effects linear regression models. Nineteen adults completed the trial [13 females, mean (±SD) age 41 ± 10 year, BMI 30.7 ± 5.7 kg/m2]. Postprandial triglyceride and glucose response post intervention were not different to post control. The overnight fast was well-tolerated by participants with an adherence rate of 95%, assessed by weekly 24-hour dietary recalls. Exploratory analysis indicates lower mean body weight post intervention compared to post control (mean difference: -0.9 kg, 95% CI: -1.3 – -0.4). Conclusions Night shift workers who habitually ate during their night shifts were able to rearrange their meal times to maintain a small overnight fast, which may have promoted small weight changes. This warrants further investigation into the role of meal timing in mitigating the metabolic consequences of night shift work.