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Objective: To determine if there was any difference in SpO2 readings during exchange blood transfusion (EBT). Methods: A prospective cross-sectional study of neonates with severe neonatal jaundice requiring EBT was conducted. Oxygen saturation was recorded before, immediately and 15 minutes after EBT by using a pulse oximeter. Results: This study included 30 neonates with 20 males and 10 females. The age ranged from 1 to 12 days with a mean of (5.4 ±2.9) days. Pre-EBT SpO2 ranged from 90% to 98% with a mean value of (94.3 ± 2.2)%; SpO2 in the end of EBT ranged from 85% to 99% with a mean value of (94.1 ± 3.2)%; SpO2 at 15 minutes after EBT ranged from 77% to 99% with a mean value of (94.8 ± 4.1)%. There was no significant difference between SpO2 values at onset of EBT and either immediately or 15 minutes after EBT (P=0.770 and 0.422, respectively). SpO2 showed no significant difference between neonates who were infused with blood of different storage times (<24 h or ≥24 h) at the onset of EBT (P=0.584), immediately (P>0.999) and 15 minutes after EBT (P=0.887). Besides, SpO2 values were compariable in neonates with hematocrit <45% or ≥45% at the onset of EBT (P=0.284), immediately (P=0.118) and 15 minutes after EBT (P=0.868). Conclusions: EBT does not affect SpO2 in neonates.