Introduction An instructional workbook for students taking biochemistry in Libya was developed. The research aimed to understand the level of acceptability to selected local biochemistry faculty and their perception of the basis for recommending the proposed instructional workbook. The study also aimed to determine the effect of sex, age, length of service and educational attainment on acceptability and recommendation. Methods The study utilized the descriptive method of research to develop and validate an instructional material for biochemistry. Faculty from a private university teaching college-level biochemistry were given a copy of the instructional material whose topics included the cell, carbohydrates and proteins. The respondents were asked to answer a questionnaire to validate the instructional material based on objectives, content, usefulness, clarity, presentation, language and style, and evaluation. Results The respondents level of acceptance in terms of the learning objectives was high with a grand weighted mean of 3.55. Content, usefulness, clarity, presentation, evaluation, and language and style were rated as acceptable. Age, length of service and educational attainment had significant impact on the acceptability of the proposed instructional workbook in terms of all seven criteria, except for age which was not significant for language and style. Conclusion Faculty teaching biochemistry have rated as acceptable the proposed instructional workbook on cell, carbohydrates and proteins and recommend its use as a teaching tool. The proposed workbook may be considered for use as a standard tool for teaching biochemistry in Libya.