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Abstract In the recent years , there has been remarkable attention was given to the utilization of pomegranate by-products as an attempt to reduce the environmental contamination which is happened as a result of food industry process. Pomegranate and its by-products have gained a special position in the scientific research as a large quantities of bio-wastes are generated during the process of pomegranate juice extraction .These non-edible wastes are about 50% of pomegranate fruit .They don’t only offer a nutraceutial and biological effects , but also they can used as an additives in the technological applications . The biological effects and technological uses of these wastes are attributed to their chemical compounds which exert an antihepatotoxicity, antiatherogenic , antinephrotoxicity ,antidiabetic, antioxidant, antimicrobial ,antifungal , antioxidant effects , and also used as a food preservation agent in different products . Therefore , this study was conducted to determine the proximate composition and the bioactive compounds of pomegranate peel , seed and their pomace , study the effects of pomegranate peel , seed and their pomace in hepatotoxic rats by CCl4 , and evaluate the sensory and physical properties of cupcakes which replacing fat by pomegranate peel , seed and their pomace as a “fat replacer “. To achieve these aims , the design of the study was as follows: Pomegranate pomace was obtained then washed and dried as well as further biological and technological experiments. Sixty six male albino rats (200±5) g were divided into 2 main groups .All rats was fed on the basal diet one week for adaptation. The first main group (n=6): served as (normal control group ) was fed on the basal diet all experiment period. The second main group (hepatotoxic rats) (n=60) were divided into 10 subgroups (6 per each group) according to the following : Subgroup (1) : (n=6) rats (positive control group) fed the basal diet. Subgroup (2) : (n=6) rats fed (basal diet + 1.5% )the dried pomegranate peel . Subgroup (3) : (n=6) rats fed( basal diet + 3%) the dried pomegranate peel . Subgroup (4) : (n=6) rats fed (basal diet + 4.5%) the dried pomegranate peel . Subgroup (5) : (n=6) rats fed (basal diet + 1.5%) the dried pomegranate seed . Subgroup (6) : ( n=6) rats fed (basal diet + 3% ) the dried pomegrante seed . Subgroup (7) : (n=6) rats fed (basal diet + 4.5%) the dried pomegranate seed. Subgroup (8): (n=6) rats fed (basal diet + 1.5% )the dried pomegranate pomace. Subgroup (9): (n=6) rats fed (basal diet + 3% ) the dried pomegranate pomace. Subgroup (10): (n=6) rats fed (basal diet + 4.5% )the dried pomegranate pomace. At the end of experimental period (30 days), blood samples were collected and the serum was centrifuged for the following biochemical analysis : Liver functions : alanine transaminse (ALT) , aspartate aminotransferase (AST) , alkaline phosphatase (ALP) , total bilirubin (T.B) ,direct bilirubin (D.B), indirect bilirubin (ID.B), total protein (T.P) , albumin , globulin , and albumin to globulin ratio (A/G ratio). Antioxidant status in the liver tissues : malondialdehyde (MDA), superoxide dismutase (SOD) , , and total antioxidant capacity (TAC) . Lipid profile : total cholesterol (TC), triglycerides (TG) , high density lipoprotein (HDLc) , low density lipoprotein (LDLc) , and very low density lipoprotein (VLDLc) . Serum glucose . Atherogenic indices :atherogenic index (AI) , cardiac risk ratio (CRR) , castielli′s risk index (CRI) , atherogenic cofficient (AC) , and atherogenic fraction (AF) . Kidney function: creatinine , urea , and uric acid (U.A). The obtained results could be summarized as follows: a.Chemical analysis : 1.Proximate composition of pomegranate peel, seed , and pomace dried powder • Pomegranate seed had the highest significantly (P ≤ 0.05) content of crude protein , fat , and fiber, then pomegranate pomace and peel with values (13.84 , 27.16 , 33.21 g /100g ) , ( 9.41 , 15.16 , 21.43 g /100 g) , and (3.18 , 1.54 , 12.13 g /100g ) respectively. • Pomegranate peel had the highest significantly (P ≤ 0.05) content of moisture , ash , and total carbohydrates , then pomegranate pomace and seed .The values of moisture , ash , and total carbohydrates were (13.46 , 3.11 , 66.58 g / 100 g) , (10.18 , 2.14 , 41.86 g /100 g) , and (6.11 , 1.63 , 18.05 g /100g ) respectively 2.Total phenols , total flavonoids , anthocyanin ,and antioxidant activity (DPPH) of pomegranate peel, seed , and pomace dried powder • Pomegranate peel had a significantly (P ≤ 0.05) highest content of total phenols , total flavonoids , anthocyanin , and antioxidant activity (DPPH) which were ( 171.18 mg ⁄g , 43.84 mg ⁄g , 71.18mg⁄100 g , and 86.11% ) respectively. • The values of pomegranate pomace were higher than pomegranate seed which were (95.63 mg ⁄g, 26.11mg ⁄g , 33.37mg ⁄ 100g ) and (2.11mg ⁄g , 1.94 mg ⁄ g , 7.16 mg ⁄ 100 g ) respectively .However , pomegranate seed antioxidant activity (DPPH)% was higher than the antioxidant activity of pomegranate pomace which was (79.26 ) and (74.14 ) % respectively. b.Blood analysis : 3.Effect of pomegranate peel , seed , and pomace dried powder on the liver enzymes of normal and hepatotoxic rats • A significant reduction (P ≤ 0.05) in the serum levels of ALT, AST ,and ALP in the negative control group compared to positive control group . • An improvement was observed in the hepatotoxic rats which fed on 4.5 and 3% of pomegranate seed, followed by the hepatotoxic groups which fed on 4.5% and 3% of pomegranate pomaace , and finally the hepatotoxic rats which fed on 4.5 and 3% of pomegranate peel respectively . 4.Effect of pomegranate peel , seed , and pomace dried powder on total bilirubin , direct and indirect of normal and hepaotoxic rats • A significant reduction (P ≤ 0.05) in the serum levels of T.B, D.B and ID.B in the negative control group than positive control group. • An improvement was observed in the hepatotoxic rats which fed on 4.5 and 3% of pomegranate seed , followed by the hepatotoxic groups which fed on 4.5% and 3% of pomegranate pomaace, and finally the hepatotoxic rats which fed on 4.5 and 3% of pomegranate peel respectively . 5.Effect of pomegranate peel , seed , and pomace dried powder on total protein , albumin , globulin , and A/G ratio of normal and hepaotoxic rats • The highest elevation in the serum T.P, albumin , and globulin was observed in the hepatotoxic groups which treated with 4.5% of pomegranate seed. 6.Effect of pomegranate peel , seed , and pomace dried powder on antioxidant status of liver tissues of normal and hepatotoxic rats • The hepatotoxic rats which treated with 4.5 and 3% of pomegranate seed respectively showed an elevation in the TAC and SOD in the liver tissues , while MDA had an opposite trend , which its levels in the serum had decreased. 7.Effect of pomegranate peel , seed , and pomace dried powder on lipid profile and serum glucose of normal and hepatotoxic rats • Feeding the rats on the diet supplemented with 4.5% of pomegranate seed resulted in a marked reduction in the serum T.C , TG, LDLc , and VLDLc , while HDLc had an opposite trend which had an elevated levels in the serum. • Serum glucose had a marked reduction in the hepatotoxic groups after being treated with 4.5% of pomegranate seed. 8.Effect of pomegranate peel , seed , and pomace dried powder on atherogenic indices enzymes of normal and hepatotoxic rats • Atherogenic indices in the serum were improved gradually by adding pomegranate seed to the diet of hepatotoxic rats as a result of the improvement in the lipid profile . 9.Effect of pomegranate peel , seed , and pomace |