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Overnight fasting (OF) is a dietary practice gaining popularity for its potential health benefits, including improved metabolic health and weight management. However, the impact of OF on hemostasis and blood clotting parameters, such as Activated Partial Prothrombin Time (aPTT) and kaolin sensitivity, remains largely unexplored. This study was carried out in the University of Benin and environs in Ovia North East local government area, Benin City, Edo state. It was a longitudinal study involving sixty (60) apparently healthy adults both sexes aged 18-30 years and divided into two phases; the fasting phase for 12 hours (test phase) and non-fasting phase (control phase) who maintained regular eating pattern. Blood samples from the fingertip and applied to the glucose strip indicator region of the glucose strip and accurately inserted into the glucometer for both random and fasting blood glucose analysis. About 5mls of blood samples were collected with minimal trauma via the cubital fossa using a sterile disposable syringe and needle. 4.5ml of blood was dispensed into 0.5ml of sodium citrate anticoagulated container in a ratio of 1:9. Result revealed a reduction in the activated partial thromboplastin time during the fasting phase when compared with the non-fasting phase although not statistically significant (p<0.05), and this reduction could be due to an increase in clot formation time by the activation of the intrinsic coagulation pathway associated with dehydration from Overnight Fasting it. In conclusion Overnight Fasting could serve as a scientific explanation of the risk of thrombosis in extended periods of food and water deprivation. In conclusion, this study provides valuable insights into the relationship between Overnight Fasting and coagulation dynamics, highlighting the need for further research to elucidate the mechanisms behind these changes and their clinical implications. Understanding the impact of OF on coagulation may help guide healthcare professionals and individuals in optimizing fasting strategies for improved health outcomes.