This study explores the KAP of clinical trial participants on the use of surplus biospecimens for future research in Indian oncology trials. A hospital-based prospective study was conducted among 112 oncology clinical trial participants. Non-probability purposive sampling technique was used to recruit the participants. This study captured their socio-demographic information and pre and post-KAP assessment by using self-administered questionnaires and its highlights ethical concerns and the lack of global guidelines. The main goal is to understand participants’ understanding and opinions on sample collection, utilization, and storage, as well as the ethical implications. SPSS software used for statistical analysis to evaluate pre and post-intervention KAP scores using frequency, percentage, mean difference, standard deviation, and applied paired t-test and Wilcoxon signed ranks test to assess Different KAP Levels. The study analyzed knowledge, attitude, and practice levels among oncology clinical trial participants using statistical tests. Pre-test to post-test comparisons showed significant increases in knowledge (p < 0.001, Wilcoxon Signed Ranks Test), attitude (p < 0.001, paired-samples t-test), and practice (p < 0.001), highlighting the intervention’s effectiveness. This study highlights the effectiveness of interventions in improving the knowledge, attitude, and practice (KAP) of oncology clinical trial participants in India regarding the use of surplus biospecimens, emphasizing the importance of standardized guidelines in global oncology trials.