The aim of the present study was to investigate the effect of time-of-day of training during Ramadan on soccer players’ chronotype and mood states. Thirty participants were randomly assigned to either a morning training group (MTG, trained between 0700 and 0830 hours, n = 10), an evening training group (ETG, trained between 1700 and 1830 hours, n = 10), or a control group (CG, did not train, n = 10). They completed two questionnaires before (BR) and at the end of (ER) Ramadan: the Horne and Östberg’s self-assessment questionnaire and the Profile of Mood States (POMS) questionnaire. The results showed that the MTG maintained the Horne and Östberg’s scores during ER compared with BR, with all players classified as “moderately morning type”. However, Horne and Östberg’s scores were reduced in the ETG (p < 0.05), with all players classified as “moderately evening type”, and in the CG, with six players classified as “neither type” and four as “moderately evening type”. There was not a significant effect of Ramadan on mood states. Only fatigue was increased and vigor was reduced in the ETG. Therefore, morning training did not affect the chronotype of the players during Ramadan. However, evening training reduced the Horne and Östberg’s scores. During ER, the participants of the MTG were classified as “moderately morning type” and the participants of the ETG were classified as “neither type” (n = 6) and “moderately evening type” (n = 4).