Preparing maximum platelets from all daily blood donations is essential to maintain adequate stock of platelets that may be feasible by storing buffy coat overnight. The overnight stored buffy coat method is prevalent in Europe and was not studied much in India. Therefore, a comparative study was planned to compare RDPs prepared from fresh and overnight stored buffy coat methods. In this study total of eighty RDPs were prepared. Forty RDPs were prepared by each method by matching similar platelet counts in whole blood in pairs and avoiding other bias. Quality analysis was done in various hematological, biochemical, metabolic, and activation parameters and with National guideline. RDPs prepared by the overnight stored buffy coat method had statistically higher mean platelet counts, lesser WBC contamination, lesser glucose concentration, higher lactate concentration, lesser pH, and higher MPV & PDW than RDPs prepared by the fresh buffy coat method. P selectin levels were lesser in the overnight stored category. The platelet yield in RDPs (extraction from the whole blood) was more in the overnight stored category. Compared with the national guideline, 87.5% of units passed the quality criteria in the fresh category compared to 100% of units in the overnight category. RDPs prepared from overnight stored buffy coats not only had better quality parameters but also were in a less activated state. Because of better quality and feasibility in blood centers, the overnight stored buffy coat method may also be used along with the traditional preparation method.