The main objective of this study was to explore the major factors that hinder the participation of female students in learning handball in some selected preparatory schools, in South Wollo Zone, Amhara Regional state. The selected schools for this study are Motta, Sinan and Liyew Asress Zewdie preparatory schools. And the total numbers of female students in each school are 118, 106 and 98 respectively. Samples were taken from grade 12th females students, from the total of 322 female students, 178 female students were selected by using simple random sampling technique. To get supplementary information for the study area 4 subject teachers and 3 school principals were selected purposefully. The study was carried out with descriptive survey method and both qualitative and quantitative approaches were used. The quantitative data obtained through questionnaire was analyzed and interpreted by using SPSS version 16.0. The data found from questionnaire was analyzed quantitatively using descriptive statistics particularly percentiles, mean and standard deviation and inferential statistics (Chi-Square test). On the other hand, the data found from interview and observations were analyzed qualitatively using narrative descriptive method. The major finding indicated that the performance of the teachers’ ability to demonstrate during the lesson was poor, teacher’s interactions with their students and lack of interest and motivation had also seen as problems during the lesson. It was concluded that the participation of female students with handball lesson as subject matter were highly affected by the school with shortage of facilities and equipment, uncomfortable environment of the school, attitude of female student, teachers, parents, school societies. Therefore, based on the findings of the study, it is recommended that to allow female students to participate in handball learning physical education teachers, parents, school societies and school administrations must work cooperatively. Additionally, female students should to be aware of the importance of learning handball and physical activities.