This research aims to develop biochemical practicum guidelines II and test the guidelines for small-scale practicum guides. This research was developed based on the Thiagarajan (4D) method. The practicum guide produced is a biochemistry practical guide II specifically for students, biochemical practicum guide II specifically for lecturers /assistants and multiple choice tests. The validation guide shows guidelines for biochemical practices specifically students have an average value of 88.89 with very feasible criteria and biochemical practicum guidance guide II specifically for lecturers / assistants has an average value of 91.11 with very feasible suggestions. The results of a small group biochemical practicum guide for students specifically had an average score of 85.94 with very decent criteria. This shows that the practicum guide for the results of the development is feasible to be used in biochemistry practicum II lectures.