This study aims to determine the analytical thinking skills of biology education students from the application of the developed biology learning book. This research used two classes of student research subjects which are determined by purposive random sampling technique. Data was collected using questionnaire and essay test. The data were analyzed by percentage technique and t test. The results showed that the ability to set learning objectives is included in the very good category (91.84%), the ability to develop learning indicators is also good category (86.38%), the ability to design learning models is included in good category (84.62%), the ability to design instructional media is included in the good category (82.36%), and the ability to design evaluation is included in the very good category (92.46%).The analytical thinking skills of biology education students who are taught using biology learning book is higher than those who do not have developed book.