Background: There are several health and social problems which are associated with shift work. This is mainly due to negative working environment and impairment of biological rhythms during the shift work. Aims and Objectives: The present study was designed to study the impact of night shift working on sleep quality, calorie intake, and serum thyroid stimulating hormone (TSH) level among health-care workers. Materials and Methods: Sleep quality, calorie intake, and serum TSH level were assessed by the Pittsburgh sleep quality index (PSQI), 24-h dietary recall, and chemiluminescence immunoassay, respectively, among night shift and day shift workers. Results: The PSQI scores (P = 0.026) and serum TSH level (P = 0.023) showed statistically significant difference among day shift and night shift workers. There was no statistically significant difference in calorie intake among the day shift and night shift workers. Conclusion: Shift working has a negative impact on sleep quality and serum TSH level with increased risk of subclinical hypothyroidism.