**Background:** Zivzik pomegranate (Punica granatum) is a fruit from Siirt, Turkey, known for its nutritional and antioxidant properties. This study aimed to evaluate the antioxidant capacities, enzyme inhibition effects (against acetylcholinesterase, α-glycosidase, α-amylase, and human carbonic anhydrase II), and antimicrobial activity of pomegranate juice and ethanol (EEZP) and water (WEZP) extracts from peel and seed. The chemical profile was determined using LC-MS/MS analysis of 53 phenolic compounds.
**Methods:** Antioxidant activities were assessed using ABTS•+ and DPPH• radical scavenging, Fe3+ reducing, Cu2+ reducing (CUPRAC), and Fe3+-TPTZ reducing (FRAP) assays. Total phenolic and flavonoid contents were measured spectrophotometrically. Enzyme inhibition studies were performed using Ellman's method for AChE, Tao et al. for α-glycosidase, Xiao et al. for α-amylase, and Verpoorte's method for hCA II. Antimicrobial activity against E. coli and S. aureus was tested by disk diffusion. LC-MS/MS quantified 53 phenolic compounds.
**Key Results:** The ethanol extract of peel (EEZP) had total phenolic content of 59.7 mg GAE/g and total flavonoid content of 88.0 mg QE/g. LC-MS/MS identified 30 compounds; major ones included ellagic acid (199.967 mg/g), catechin (27.664 mg/g), epigallocatechin gallate (25.600 mg/g), epicatechin (24.210 mg/g), nicotiflorin (23.535 mg/g), astragalin (20.551 mg/g), gallic acid (20.021 mg/g), epigallocatechin (19.148 mg/g), quinic acid (17.460 mg/g), and tannic acid (12.300 mg/g). Antioxidant assays: EEZP peel showed IC50 values of 5.9 μg/mL for ABTS•+ scavenging and 16.1 μg/mL for DPPH• scavenging. In reducing assays at 30 μg/mL, pomegranate juice had absorbance of 1.810 (Fe3+ reducing), 2.790 (Cu2+ reducing), and 2.230 (Fe3+-TPTZ reducing). Enzyme inhibition: EEZP peel had IC50 values of 7.3 μg/mL for α-glycosidase, 317.7 μg/mL for α-amylase, 19.7 μg/mL for AChE, and 106.3 μg/mL for hCA II. For comparison, acarbose (α-glycosidase standard) had IC50 22,800 μM; tacrine (AChE standard) had IC50 5.97 μg/mL; acetazolamide (hCA II standard) had IC50 8.4 μg/mL. Antimicrobial testing: E. coli and S. aureus were resistant to all four standard antibiotics (amoxicillin-clavulanic acid, gentamicin, ampicillin-sulbactam, ciprofloxacin). Some extracts showed small zones (7–10 mm) but were considered resistant per CLSI criteria.
**Clinical Implications:** The study demonstrates that Zivzik pomegranate extracts, particularly the ethanol extract of peel, possess strong antioxidant, anti-Alzheimer (via AChE inhibition), antidiabetic (via α-glycosidase and α-amylase inhibition), and antiglaucoma (via hCA II inhibition) potential. The high content of bioactive phenolics like ellagic acid, catechins, and flavonoids supports these activities. These findings suggest that Zivzik pomegranate could be developed as a natural therapeutic agent for managing oxidative stress, type 2 diabetes, Alzheimer's disease, and glaucoma. However, further in vivo studies and cytotoxicity assessments are needed to confirm safety and efficacy.