eng Oriental Scientific Publishing Company Biosciences Biotechnology Research Asia 0973-1245 2026-08-26 23 3 59618 article Asma Tabassum 1 Rajesh Medar 1 Bhagyalakshmi Manjappa 1 Department of Studies and Research in Biochemistry, Tumkur University, Tumkur, Karnataka, India The protein and fat structures of the DMWE (Donkey milk whey extract) are very close to that of the human breast milk when compared to that of the bovine milk, and therefore has a significant value in the functional food science. This study uses in vitro and in vivo models to examine DMWE's antithrombotic, antidiabetic, and antioxidant activities. DMWE was acquired after acid induced whey separation and subjected to biological tests. DMWE demonstrated a strong dose-dependent procoagulant effect that reduced plasma recalcification time in 100-500 µg concentrations, reducing plasma recalcification time to 36 s (P < 0.0001) at 100-500 µg.The mouse tail bleeding model significantly reduced the bleeding duration from 217 s to 40 s (P < 0.01).DMWE also experienced a substantial reduction in the APTT (activated partial thromboplastin time) and PT (prothrombin time) that witnessed the activation of intrinsic and extrinsic pathways. DMWE had a procoagulant activity but it was also found to have antiplatelet and thrombolytic potential in decreasing order of lysis of the clot under observation after 90 min at 37ºC. DMWE inhibited alpha-amylase (28-67%) and alpha-glucosidase (35-54%) with an IC₅₀ of 39 µg/mL and this is similar to acarbose. DPP-IV (Dipeptidyl Peptidase-IV) inhibition reached 63% with an IC₅₀ of 395 ± 1.856 µg/mL, compared to sitagliptin (57%, IC₅₀ 3.5 µg/mL). The antioxidant activity demonstrated 79% DPPH (2,2-Diphenyl-1-picrylhydrazyl) scavenging, with IC50 values of 185 µg/mL (DMWE) and 160 µg/mL (ascorbic acid). The biological benefits are linked to bioactive proteins and peptides such as lactoferrin and lysozyme, which have antioxidant, antibacterial, and metabolic activities. DMWE has tremendous opportunities as a functional nutraceutical to be implemented in the metabolic regulation and specialized nutrition. https://www.biotech-asia.org/vol23no3/evaluation-of-donkey-milk-whey-extract-equus-asinus-for-hemostatic-and-metabolic-applications/ Alpha-amylase inhibition; Alpha-glucosidase inhibition; Antiplatelet aggregation; Bioactive peptides; Lysozyme Milk whey extract; Thrombolytic activity