Objective:To compare the platelet count, platelet concentration/yield, residual Red blood cells (RBCs) and White blood cells (WBCs) counts in platelet-rich plasma (PRP) samples prepared from the single- and the double-centrifugation protocols.Methods:It was a Cross-Sectional study, conducted at the Department of Hematology & Transfusion Medicine, The Children’s Hospital and UCHS, Lahore from October 2021 to January 2022 including 50 voluntary, healthy individuals of age 20-45 years of both genders, after taking informed consent. Complete blood count analysis of all participants was done initially by drawing 3ml blood in EDTA vial. From all the participants, 20 ml venous blood sample was taken in syringes containing tri-sodium citrate and then shifted to harvest tubes. Group-I comprised of PRP samples prepared by single- centrifugation method. While Group-II samples were prepared by Double-centrifugation method consisting of soft and hard spin. The platelet, RBC and WBC counts in prepared PRP samples were determined by using automated SYSMEX XP-100 hematology analyzer. Platelet yield or Platelet concentration (%) was calculated for samples using formula. The data analysis was done using SPSS version 23.Results:The mean PRP platelet count in Group-I was 594.6±157.4×103/μl whereas in Group-II was 923.06 ± 127.58×103/μl. In Group-I, the mean platelet concentration/yield in PRP was 175.75 ± 55.08% while in Group-II, it was 276.78 ± 112.7%. Significant difference was observed between the platelet counts and platelet concentration/yields from the PRP samples of two Group-s (p < 0.01). Significant difference between the WBCs count was also observed (p < 0.01) with higher WBCs in Group- I PRP. Residual RBCs were almost same among two Group-s.Conclusions:The double centrifugation protocol resulted in higher platelet quantity and yield with less contamination by red and white blood cells than did the single centrifugation protocol for PRP preparation. So, double centrifugation method is beneficial in preparation of autologous as well as allogenic PRP.