Introduction: Many countries struggle to supply enough blood while maintaining their quality and safety. Increasing the number of regular donors is expected to increase the donor pool and blood safety. Thus, this study describes lapsed and regular blood donors’ characteristics, knowledge, motivation, and barriers concerning blood donation.Methods: This observational cross-sectional study has adopted an assisted self-administered questionnaire, which was distributed to blood donors at the Department of Transfusion Medicine, Hospital Sultanah Bahiyah, Kedah. A total of 328 participants consisting of 164 lapsed and 164 regular donors were selected. Logistic regression tests were used to determine the factors that predict lapsed donors. Results: Out of the selected 328 respondents, 54.3% were in the 25–39 age group, 66.2% were males, and 85.1% were Malays. Most of the respondents (88.4%) showed adequate blood donation knowledge, and 99.7% cited altruism as a motivator for blood donation. About 47.0% of the respondents claimed they lack enough time as their donation barrier. Donors who were younger in age, had a moderate blood donation knowledge (adjusted OR, 3.60; 95% CI, 1.34-9.64), didn’t know where to donate (adjusted OR, 2.79; 95% CI, 1.47-5.29), lack enough time (adjusted OR, 1.83; 95% CI, 1.04-3.24), and insufficient information about blood donation campaigns (adjusted OR, 2.19; 95% CI, 1.23-3.91) were more likely to lapse. Conclusion: Donor education, convenient time and location, and sufficient information about blood donation campaigns targeted at young donors are critical for preventing lapsed donors, which could subsequently increase the regular donor pool.