Browsing by Author "Dissanayake, K.S.M."

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    Comparison of Antioxidant Activity of Hydrolysate Products of Crude Collagen Extracted from Chicken Egg Shell Membrane Using Different Methods
    (Uva Wellassa University of Sri Lanka, 2018) Dissanayake, K.S.M.; Aruppala, A.L.Y.H.; Abeyrathne, E.D.N.S.
    Collagen is a highly valuable protein used in food industry. Egg shell membrane is a safe source for collagen. Extraction of collagen from chicken membrane and producing its hydrolysates were carried out using different methods. Objective of this study was to extract collagen from chicken egg membrane with simple and non-toxic method followed by hydrolysis to find out the functional properties of the hydrolysates. Shell membrane was separated by manual peeling by adding 0.5 M Acetic acid, 0.5 M Citric acid followed with extraction of collagen with pepsin digestion. pH of the extracted collagen was adjusted to 6.5 and hydrolyzed using protease with different time combinations (0, 3, 6, 9, 12, 24 hours) at 37 °C followed by heat inactivation at 100 °C for 15 min. Best hydrolysates were selected by 10% SOS-PAGE by visual observations. Selected hydrolysates were subjected to antioxidant activity by Thiobarbituric Acid Reactive Substances (TBARS) method and Diphenyl-l-picryhydrazyl (DPPH) radical scavenging activity. The highest collagen yield was observed from citric acid (0.15 g 1 0g-1) extraction than acetic acid (0.08 glOg-1) treatment (P > 0.05). SOS-PAGE did not show bands even 0 hours, so, 0 hour hydrolysates were selected for antioxidant testing. In DPPH analysis, citric acid extraction shows higher scavenging activity (96.79%) than acetic acid (72.37%) (P < 0.05).flowever in TBARS method also did not show significant difference among the treatments (P > 0.05) and it showed 0.00 mg 1.1level of Melonaldehyde. This concludes that collagen hydrolysates showed good antioxidant activity with citric acid extraction than acetic acid extraction comparing with the ascorbic acid as positive control which can be used as a natural antioxidant in food industry.
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    Comparison of the Functional Properties of Hydrolysates Produced from Chicken Egg Shell Membrane Collagen Crude Extraction Extracted with Different Methods
    (Uva Wellassa University of Sri Lanka, 2013) Dissanayake, K.S.M.
    Collagen is a highly valuable protein used in food industry. Egg shell membrane is a safe source for collagen. Extraction of collagen from chicken membrane and producing its hydrolysates were carried out using different methods. Objective of this study was to extract collagen from chicken egg membrane with simple and non-toxic method followed by hydrolysis to find out the functional properties of the hydrolysates. Shell membrane was separated by manual peeling by adding 0.5 M Acetic acid, 0.5 M Citric acid followed with extraction of collagen with pepsin digestion. pH of the extracted collagen was adjusted to 6.5 and hydrolyzed using protease with different time combinations (0, 3, 6, 9, 12, 24 hours) at 37°C followed by heat inactivation at 100°C for 15min.Best hydrolysates were selected by10% SDS-PAGE Gel electrophoresis by visual observations. Selected hydrolysates were subjected to Anti-bacterial activity, Metal (Fe") chelating activity and antioxidant activity by Thiobarbituric Acid Reactive Substances (TBARS) method and Diphenyl-l-picryhydrazyl (DPPH) radical scavenging activity. The highest collagen yield was observed from citric acid (0.15g/10g) extraction than acetic acid (0.08g/10g) treatment (P>0.05). SDS-PAGE did not show bands even 0 hours, therefore it was selected for antioxidant testing. In anti-bacterial test it showed inhibition for locally isolated E.coli and Salmonella with 0.625 ppm 'minimum concentration. In DPPH analysis, citric acid extraction shows lower scavenging activity (68.12%) than acetic acid (75.53%) (P>0.05). However in TBARS method also did not showed significant difference among the treatments (P>0.05) and it showed 0.00 mg/1 level of Melonaldehyde. Both acetic and citric extractions showed less Fe" chelating activity around 2% (P>0.05). This concludes that collagen hydrolysates showed good antioxidant activity with citric acid extraction than acetic acid extraction comparing with the ascorbic acid as positive control which can be used as a naturalanti-bacterial agent for foods and antioxidant agent in food industry which does not contains iron as Fe'. Key words: Collagen, Hydrolysates, Antioxidant activity, Pepsin, Protease
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    Determine the Effect of Functional Properties on Chicken Patty Incorporated with Salt Extracted Bioactive Compounds from Pterygoplichthys pardalis (Scavenger Fish)
    (Uva Wellassa University of Sri Lanka, 2019) Dissanayake, K.S.M.; Ediriweera, T.K.; Abeyrathne, E.D.N.S.
    Pterygoplichthys pardalis is a tropical and invasive fresh water fish. Based on literature, the extracted compounds from this fish consist of anti-oxidative, antimicrobial and Fe2+ chelating activities. Hence, the objective of this study was to determine the effect of these functional properties on chicken patty, incorporated with salt extracted bioactive compounds from Pterygoplichthys pardalis. Female fish were collected from a local reservoir and slaughtered in the field. Gonads (excluding GI tract and mucus) were separated within 3 hours and stored at 4C. Separated parts were followed for extraction of proteins with distilled water (1:4) and then 10% (w/v) NaCl solution (1:4) and lyophilized. Extracted protein samples were incorporated to the preparation of chicken patty (Chicken meat, Salt, Spices) with 0, 0.5, 1, 1.5 and 2% (w/w) levels. Then TBARS assay, DPPH assay, Fe2+ chelating activity and Total plate count were done for the product for Day 0, 2, 4, 7 and 14. According to TBARS and DPPH assay 2% (w/v) incorporation level gave the highest antioxidant activity and Fe2+ chelating activity (p<0.05). Microbial counts of meat patty with 2% (w/v) incorporation level was suitable for 7 days compared with the control (p<0.05). As conclusion, 2% (w/v) incorporation level of salt extraction from P. pardalis can be used as a natural antioxidant, antimicrobial and metal chelating agent in chicken patty. However further studies needed to check for the maximum level of incorporation.
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    Evaluation of Antioxidative and Antimicrobial Activities of a Prawn Based Dipping Incorporated with Bioactive Fish Protein Hydrolysates from Pterygoplichthys pardalis
    (Uva Wellassa University of Sri Lanka, 2019) Ediriweera, T.K.; Dissanayake, K.S.M.; Abeyrathne, E.D.N.S.
    Currently, the world is moving towards functional foods instead of traditional nutrient based foods. “PrawRy” is a value-added prawn and dairy based pre-cooked instant powdered food, which can be processed as a dipping, sauce, salad dressing or as a pudding. Due to its raw materials it is indisputably high in nutrition and its functionalities can be promoted by incorporating bioactive natural compounds. In the present study the “PrawRy dipping”, which was made by cooking for 10 minutes with 1:2 ratio of powder mix (Prawn, milk, butter, garlic) and water, was enriched with pre-proved bioactive salt extracted internal organ (except GI tract) protein (protein: NaCl 10% (w/v) =1:4) hydrolysates of Pterygoplichthys pardalis. Fish protein hydrolysate (FPH) was prepared followed by incorporation of it in to “PrawRy dipping” at 0, 0.5, 1, 1.5, and 2% (w/w) and kept under refrigerated conditions. The bioactivities were then analyzed at 0, 2, 4 and 7 days of storage. Antioxidative properties were evaluated by TBARS, DPPH scavenging assays and Fe (II) chelating activity. Total plate count was detected to check the microbial growth in the final product. According to the results obtained, significant differences were obtained between treatments against non-treated product (P<0.05) in both DPPH scavenging and TBARS assays, Total plate count and in Fe (II) chelating activity assay. Further, significant differences were obtained between days against non-treated product (P<0.05) in TBARS assay, Total Plate Count, Fe (II) chelating activity assay while no such difference was demonstrated in DPPH scavenging assay (except in day 0). The highest antioxidative activity and resistance against microbial growth was demonstrated by 2% (w/w) FPH incorporated “PrawRy dipping”. The study concluded that the “PrawRy dipping” can be enriched with antioxidative and antimicrobial properties by incorporating salt extracted 2% (w/w) protein hydrolysates of P. pardalis as a value-added functional food.
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    Extraction of Crude Collagen from Thunnus albacares (Yellowfin Tuna) Skin and Determination of Antioxidant and Metal Chelation Activities of Its Hydrolysates
    (Uva Wellassa University of Sri Lanka, 2019-02) Thilanja, G.P.D.D.S.; Dissanayake, K.S.M.; Abeyrathne, E.D.N.S.
    Collagen is a dominant protein in connective tissues and highly valuable in food industry. Fish processing byproducts are good alternative source for collagen. The objective of this study was to develop a simple non-toxic method to extract crude collagen from Yellowfin tuna skin and to check functional properties of its hydrolysates. Extraction procedures were conducted using acetic acid and citric acid with 0.5 M concentrations. Based on 8% SDS-PAGE gel, type I collagens were identified. Enzymatic hydrolysis was done with Protease, Trypsin and Pepsin enzymes with different time combinations (0 h, 3 h, 6 h, 9 h, 12 h and 24 h) at 37 °C after adjusting to its optimum pH level. Best hydrolysate was selected and subjected to antioxidant activity by Diphenyl-1-picryhydrazyl (DPPH) radical scavenging activity and Metal (Fe2+) chelating activity. Proximate analysis was conducted for raw skin to determine moisture, ash, crude protein, crude fat content and 59.44±0.013%, 1.91±0.37%, 28.55±1.19%, 6.83±0.30% values were obtained respectively. Hydrolysates produced after incubating for 0 h at 37 °C followed with heat inactivation was selected as the best. Hydrolyzed produced using citric acid showed lower scavenging activity (63.62%) compared to acetic acid (85.07%) (p<0.05). In both acetic and citric extractions Fe2+ chelating activity did not show significant difference among the treatments (p>0.05). According to the collagen hydrolysates incubated at 0 h at 37 °C showed good antioxidant activity with acetic acid extraction with Pepsin enzyme. This conclude that collagen hydrolysates produced using acetic acid and Pepsin showed good antioxidant activity comparing with the ascorbic acid as positive control and it could be deserved to use as good alternative source as a natural anti-oxidant in food industry.
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    Extraction of Crude Skin Collagen from Pterygoplichthys pardalis and Determination of Antioxidant and Metal Chelation Activities of Its Hydrolysates
    (Uva Wellassa University of Sri Lanka, 2019-02) Dilhani, E.H.P.U.; Dissanayake, K.S.M.; Abeyrathne, E.D.N.S.
    Collagens have a great demand in the food industry and fish skin is a safe alternative source of collagen. Pterygoplichthys pardalis is a freshwater fish which threatens endemic fish and inland aquaculture, and has no economic benefit. Objective of this study was to extract crude collagen from P. pardalis skin with simple and non-toxic method followed by identifying the antioxidant properties of its hydrolysates. Proximate composition was determined in raw fish skins with and without bony plates separately. Acid and Pepsin soluble collagens were extracted from P. pardalis skin. As with the pretreatment process of citric acid (CA) and EDTA were tested to decalcify the fish skin. Three different concentrations were used with CA as 1.2, 2.2 and 3.2 kgm-3 and for EDTA as 0.1, 0.2 and 0.3 M. Selected crude collagens were subjected to the hydrolysis using Pepsin, Protease and Trypsin enzyme after adjusting to its optimum pH with different time combinations (0, 3, 6, 9, 12 and 24 h) at 37 °C followed by heat inactivation at 100 °C for 15 min. Extracted crude collagen and best hydrolysates were selected by 8% and 15% SDS-PAGE respectively. Antioxidant activity of the best hydrolysates was evaluated using DPPH scavenging assay and metal chelation activity by Fe (II) chelating activity. All treatments were replicated (n=3). Raw fish skins with and without bony plates contained 44.29±3.69%, 58.79±1.05% moisture, 16.40±0.93%, 5.38±1.61% ash, 26.75±8.93%, 26.89±3.25% crude protein respectively. Extracted collagens with CA treatment showed higher yield compared to EDTA treatment (p<0.05). The Antioxidant properties were not significantly different (p>0.05) but metal chelation activities of selected best hydrolysates were higher in CA than EDTA treatment (p<0.05). These results conclude that collagen hydrolysates produced from P. pardalis with all three enzymes with 0 h at 37 °C followed by heat inactivation have good antioxidant and metal chelating properties.