Background: Infantile colic is period of devastated and inexplicable crying in an apparently healthy infant, leads to drained, bothered and worried parents. It affects 30% of infants worldwide with equivalent frequency in male and female. Aim: To evaluate the impact of an educational program on mothers' knowledge regarding infantile colic syndrome. Methods: Quasi- experimental (pre/posttest) research design was used in all of the five maternal and child health care centers. Sample: Included 402 mothers. Two tools; Tool (1): A structure interview questionnaire sheet composed of three parts; part (I): Mothers' personal data; part (II): Infants' data and part (III): Mothers' knowledge about infantile colic syndrome. Tool (II): Infant colic scale, which included total of 22 items in five subscales. Results: It was found that 29.9% and 68.9 of mothers had university education and from rural area. There was improvement of mothers' knowledge in posttest mean=19.46 while it was 8.46 in pretest. There was statistical significance relation between mothers' total mean score of knowledge and education, occupation and residence in pretest. Conclusion and recommendations: There was improvement in mothers' knowledge after the implementation of the educational program. Availability of educational material in MCH to increase awareness regarding infantile colic is recommended.