Antenatal care is a detailed, systematic examination and follow-up of pregnant female that includes education, counseling, monitoring and therapy to address and treat minor issues and provide appropriate screening during pregnancy. Aim of the study: Evaluate Antenatal education guidelines’ effectiveness on maternity nurses’ performance (Knowledge and Practice). Design: A quasi-experimental design (pretest- post-test). Setting: study was conducted in obstetric skill lab of faculty of nursing, Benha university . Sample: A purposive sample was used to recruit 140 maternity nurses from Maternal and child health centers at kalioubia governorate.. Tools: two data collection tools: A self-administered questionnaire sheet and an observational practice checklist. Results: Highly statistically significant improvement in maternity nurse’s knowledge score, it changed from 51.4% to 85% at post-intervention. Maternity nurses’ practice mean score was satisfactory improved, as the mean score increased from 56.4% to 82.1% at the post-intervention phase. Highly positive correlation between total nurse’s knowledge and practice score regarding antenatal care was found. Conclusion: Provision of antenatal educational guidelines helped strengthen the awareness and practice of maternity nurses on the various aspects of the antenatal period. Recommendations: The care of pregnant women should be an integral part of the educational curriculum for nursing students.