Background This research was aimed to know and investigate the difference of female labour’s work fatigue level between the morning, evening, and night shifts in winding PT. Iskandar Indah Printing Textile Surakarta.

Methods This research is an observational cross sectional analytical approach, sample were 56 woman in Winding division. Sampling technique uses in this research was purposive sampling by determining the predefined characteristics. The data collection was done by measuring the labour fatigue level using Reaction Timer. The data analysis used statistic non parametric kruskal Wallis by using computer program SPSS 17.00 Version.

Results The result of statistic showed the difference of female labour’s work fatigue level between the morning (326,41 ± 79,52), evening (393,32 ± 83,20), and nights shift (483,00 ± 118,66) showed very significance value p = 0.001.

Conclusions The highest level of fatigue was night shift. To solved this problems, it could be recommended by giving some nutritions, providing shuttle facilities for female night shift workers and applying rotation shift patern with rota metropolitan (2–2) and rota continental (2–3). Design recommendations: as few night shifts as possible, no more than two to four consecutive night shifts, forward rotation: early-late-night, avoidance of the accumulation of working hours, as far as possible two consecutive days off at the weekend, early shift not to start too early, night shift not to end too late, and predictability of shift schedule.