Objective Compare the differences of common knowledge，attitudes and motivations in voluntary blood donation between students, taking one comprehensive university and one medical university of Nanjing as examples. Methods Establishing questionnaires via document research, expert consultation and focus group discussion Chi-square test was used to compare the differences of common knowledge. T test was used to compare the differences of attitudes and motivations. Results 1.Students in comprehensive universities know less about blood donation than students in medical universities. 2. Students in medical universities score higher in attitudes to blood donation than students in comprehensive universities. No significant differences are found between them in feelings when they are donating blood. 3. Students in medical universities score higher in motivation of altruism than students in comprehensive universities. And among the students who didn’t donate blood, students in medical universities also score higher in motivation of self-efficacy and reward. Conclusions It’s necessary to strengthen the education of blood donation in comprehensive universities. Suitable broadcasting and recruiting strategies can be established on the base of differences between them in motivation and attitudes to donate blood.