The purpose of this study is to explore factors that influence practical work in chemistry for secondary schools in Wolaita Zone, Ethiopia. The study has identified teachers', learners' and school principals' perceptions to indicate the key factors that seem to inhibit the effective use of practical work in chemistry. The sample for the study comprised 56 chemistry teachers, 75 secondary school students, and 5 school principals. Data were collected using structured questionnaires, focus group discussion and interview. The collected data were analyzed using simple quantitative and qualitative analysis. The finding of the study revealed that factors that influencing practical work in chemistry of secondary schools are teachers' poor knowledge of practical work, full-time occupancy of chemistry teachers, absenteeism of the teacher at practical classes, late commencement of teaching a practical class; lack of awareness and motivation of school managements and unsafe working environments. Furthermore, lack of separate chemistry laboratory, lack of equipment in the laboratory, too short period allocated for practical work, low attitude students' toward practical work in chemistry, teachers' low level of expectation for the development of Information and communication technology (ICT) were the major factors that affect the practical work in chemistry. Finally, recommendations were forwarded based on the major finding in order to improve practical work in chemistry.