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Institute of Home and Food Sciences
Synopsis for MS. Food Science and Technology

Title: “Extraction and Application of Fruit By-product Phytochemicals for the
Development of Functional Fish Meat Products”
Name: Hassan Mazhar

Registration No.: 2012-GCUF-02299

The food industry generates large amount of wastes or by-products annually around the world
from a variety of sources. In that food wastes or by-products are an excellent source of
nutraceuticals, bioactive, inherently functional and possess many components that are good for
human health. Food wastes or by-products conversion to the functional food ingredients is the
healthy trends in the food industry. For this purpose, ultrasound technology assisted process will
be applied for the extraction of phytochemicals from fruit by-products (Papaya and Orange peel)
with distilled water, aqueous methanol and aqueous ethanol as the solvent. The ranges of
selected processing variables will be extraction temperature, sonication amplitude level,
water/waste ratio and sonication time conditions for maximum yield of phytochemicals from
fruit waste using Box-Behnken Design (BBD). Afterwards, the resultant extracts will be filtered
and subjected to drying process. The extracted phytochemical samples will be analyzed for
chemical composition (moisture, protein, fat, total phenols, cyanogenic glycosides and tannin
contents). Natural vegetable phytochemical extracts will be supplemented at different
concentration to fish meat for preparation of different meat products i.e. patties, balls/pops and
finger/sticks. The un-cooked and fried fish meat products will be tested for oxidative stability
(peroxide and free fatty acid values) and consumer acceptability at different intervals during 60-
days storage at refrigerator and −18°C. The data of phytochemical yield and product
development obtained for each treatment will be subjected to statistical analysis to determine the
level of significance by using the software package.


Dr. Dr. FAISALABAD Institute of Home and Food Sciences Synopsis for Ms. Muhammad Imran (Chairman) ________________ 2. Food Science and Technology Title: “Extraction and Application of Fruit By-product Phytochemicals for the Development of Functional Fish Meat Products” 1. Dr. Zarina Mushtaq (Member) ________________ 2 . GOVERNMENT COLLEGE UNIVERSITY. Probable Duration (Research): 06 Months Supervisory Committee: 1. Muhammad Kamran Khan (Member) ________________ 3. Date of Initiation (Research): 1-10-2017 3. Date of Admission: 1-09-2016 2.

2005). Undoubtedly. and other helpful nutrients that have shown to be protective against oxidation. avoidance of organic solvents as its action also works in GRAS solvents. reduced risk of coronary heart disease and stroke. vitamins and minerals. discoloration. 2011). food preservation. thus limiting further exploitation. Fish meat is a valuable source of high biological value protein. ultimately decreasing consumer confidence in the product and costing the industry over $700 million annually. reduction in extraction time. as well as certain types of cancer (Williamson.. which can potentially enhance the extraction of heat 3 . Fish meat is devoid of antioxidants and phytochemicals. e. Introduction Diet is an important factor affecting human health and well-being. Therefore one good possibility would be to generate bioactive ingredients that could be applied to design stable fish foods.. Ultrasound is applied to impart positive effects in food processing such as improvement in mass transfer. O'Shea et al. assistance of thermal treatments and manipulation of texture and food analysis (Knorr et al. The action of ultrasound is due to cavitation. Meat is an important component of a healthy and well balanced diet due to its nutritional richness. The major advantages of ultrasound are minimum effect on extractable materials. Due to increasing production of plant food processing. 2013). omega fatty acids. These reactions lead to off-flavor. membrane structure and tumor initiation in biological system (Muik et al.1.g. fragmentation and mass transfer resulting in high yield of extracted materials and fast rate of extraction. 2001. 2009). maximize quality and ensure the safety of food products. Ultrasound is one of the emerging technologies that was developed to minimize processing. Lipid and color oxidation are major causes of quality deterioration in meat products during storage. and loss of nutritive value. which generates high shear forces and micro bubbles that enhances surface erosion.. Jansen et al. innovative and rapidly growing part of the overall food market. Intake of such food products with oxidized lipid constituents can modify DNA. by-products disposal represents a growing problem since the plant material is usually prone to microbial spoilage. proteins. 2012. functional foods represent an important.. (Schieber et al. Epidemiological studies have pointed out that consumption of functional foods imparts health benefits.

.sensitive bioactive and food components at lower processing temperatures and potentially in large industrial scales (Awad et al. One of Most significant category of natural antioxidant is phenolic compound which is our concern. Objectives:  Extraction of bioactive compounds from fruit and vegetable waste/by-products by using ultrasound technology  Develop a range of functional/healthier un-cooked and fried fish meat products using extracted bioactive compounds  Characterization of functional fish meat products for oxidative stability and consumer acceptability at different storage intervals 2. This can be costly for the manufacturer and also may have a negative impact on the environment. and much validation is extracted on the potency of antioxidant and prevention of diseases (Scalbert et al. Research has shown that these by-products can have a high nutritional value. The focus of this review is on the nutritional and functional 4 . such as dietary fiber or in more recent times. Fresco et al. that they could be used as a food ingredient due to their functional abilities such as gelling and water binding. the preparation and processing procedures can lead to one third of the product being discarded. 2012). it can be the most viable approach to extract the phytochemicals from fruit and vegetable waste using the innovative technology. It has also been suggested. Therefore. 2006). (2012) producers are striving to create products which contain a value added factor. Due to possible health hazards of some artificial antioxidants growing attention has been paid to recognize natural antioxidants which are possibly more economically feasible and effective antioxidants.. 2005. Review of Literature Norah O'Shea et al. The extracted phytochemicals can be used in meat to further enhancing the meat acceptability and improving the functional stability during storage. In the fruit and vegetable industry. phytochemicals. The production and addition of such nutrients can be quite costly for the producer..

beetroot. carotenoids and of the by-products of food processing and their potential applications as nutritional new ingredients in foods. aonla. broccoli. mint. consumers are increasingly aware of diet related health problems. lotus stem. P < 0. Different extraction methods (i. Charanjit Kaur et al. Liu (2003) In general. supercritical fluid extraction (SFE) and microwave assisted extraction) have been developed for the recovery of bioactive compounds and essential oils. Furthermore. yam. black carrots. black carrots. fenugreek.05). kachnar and ginger had high phenolic contents.6578. chenopodium. aonla. fruits and plant by products are known to contain a wide variety of phytochemicals. (2001) There is a rapidly growing body of literature covering the role of plant secondary metabolites in food and their potential effects on human health. By-products of plant food processing represent a major disposal problem for the industry concerned. such as polyphenols. The total phenolics of the extracts was determined spectrophotometrically according to the Folin–Ciocalteau procedure and ranged from 34 to 400 mg (100 g)−1 on a fresh weight basis. Wang and Weller (2006) Extraction is the first key step to isolate natural bioactive compounds from plants and materials.e.8% in long melon. The purpose of this review is to highlight the potential of selected by- products as a source of functional compounds. but they are also promising sources of compounds which may be used because of their favourable technological or nutritional properties. The anti- oxidant activity expressed as per percent inhibition of oxidation ranged from a high of 92% in turmeric extracts to a low of 12. Anti-oxidant activity correlated significantly and positively with total phenolics (r2=0. The results indicate that vegetables containing high phenolics may provide a source of dietary anti-oxidants. solvent extraction. therefore demanding natural ingredients which are expected to be safe and health-promoting. 5 . ginger. More than 5000 individual phytochemicals have been identified in fruits. fenugreek. (2002) The anti-oxidant activity of extracts from 36 vegetables was evaluated by using a model system consisting of β-carotene and linoleic acid. Brussels sprouts. vegetables and grains. Mint. expelling extraction process. Schieber et al. coriander and tomato. Other vegetables found to have high anti- oxidant activity (>70%) were kachnar.

assisted extraction is adjustable to be utilized with polar and non-polar solvents in various temperatures. (2015). which bear the C6–C3–C6 structure. several disadvantages like extra solvent amount in solvent extraction. The intensity of ultrasound power creates extra vibration in sample molecules and facilitates the recovery of target compounds from solid material to the liquid solvent phase. and in fish. ubiquitous in plants are an essential part of the human diet. (2006) Phenolic compounds. Fruits. Some of these by-products have been the subject of investigations and have proven to be effective sources of phenolic antioxidants. account for more than half of the over eight thousand different phenolic compounds. These compounds posses an aromatic ring bearing one or more hydroxyl groups and their structures may range from that of a simple phenolic molecule to that of a complex high- molecular weight polymer. Samaram et al. vegetables and beverages are the major sources of phenolic compounds in the human diet. Therefore. The antioxidant activity of phenolic compounds depends on the structure. phenolic-rich extracts have shown antioxidant activities comparable to that of synthetic antioxidants. low yield in expelling process. high yield in short extraction time beside the utilization of low solvent amount are remarkable advantages of ultrasound-assisted extraction technique. Flavonoids. Nagendran Balasundram et al. Moreover. massive investment in supercritical fluid extraction (SFE) and the requirement for the aqueous phase in microwave assisted extraction indicates the demand of comprehensive extraction method to recover different target compounds in economic condition. which could be valuable natural sources of antioxidants. meat and poultry products. ultrasound. The food and agricultural products processing industries generate substantial quantities of phenolics-rich by- products. Practical aspects of extraction and production of sufficient amounts of natural antioxidants from most of these sources remain to be elucidated. Samaram et al.Ultrasound-assisted extraction is a new simple technique for the recovery of oil and bioactive compounds from different sources. and are of considerable interest due to their antioxidant properties. (2013) However. When tested in edible oils. in particular the number and positions of the hydroxyl groups and the nature of substitutions on the aromatic rings. 6 .

3.2. Analysis of fruit phytochemical extracts The phytochemical extracts will be analyzed for chemical composition. Extraction of phytochemicals from fruit by-products Ultrasound (model VCX 750. the resultant extracts will be filtered and subjected to drying process. Total polyphenols Total polyphenols will be measured by using Folin-Ciocalteu method following the protocol of Singleton et al. 3. protein and fat contents Moisture content will be analyzed by using air forced draft oven (Model: DO-1-30/02. Sonics & Materials..3. Procurement of raw materials Fruit by-products (Papaya and Orange peel) will be collected from local fruits and processing industries in Faisalabad. Crude fat will be estimated by AACC (2000) Method No. sonication amplitude level. dust and other foreign debris. For the intention. Moisture.1. aqueous methanol and aqueous ethanol as the solvent.3.03 (AOAC 2006). Protein will be calculated by the Method No. Inc. Materials and Methods 3. (1999). Pakistan. 30-10.3. Response surface methodology will be used to establish the optimum conditions for the sonication assisted extraction of phytochemicals.1. USA) technology assisted process will be applied for the extraction of phytochemicals from fruit by-products with distilled water. water/waste ratio and sonication time will be established based on a Box-Behnken experimental design. The selected raw materials will be cleaned to remove the adhered dirt.3. A quadratic polynomial regression model describing extraction efficiency as a function of four independent variables such as extraction temperature. 3. Pakistan). 50 µL extract will be added to 250 µL of Folin-Ciocalteu reagent with 750 µL of 20% sodium carbonate solution and the volume will be 7 . 990. 3. Afterwards.2. PCSIR.

absorbance will be recorded at 765 nm with UV/Visible Spectrophotometer against control. gently boiling for 1 hour and will be filtered using Whatman No.5 g of the sample extract and 100 mL of distilled water will be added in a conical flask.4. The used volume of AgNO 3 will be noted for each individual extract sample. balls/pops and finger/sticks. Standard tannic acid solution will be prepared from which a standard curve wil l be drawn (absorbance versus concentration in mg/cm 3). 3. Distilled water will be used as blank regarding the calibration curve. Functional meat product development and analysis Natural fruit phytochemical extracts will be supplemented at different concentration to fish meat for preparation of different meat products i. the flasks will be allowed to stand in a water bath at 250 °C for 20 min and the optical density will be measured at 700 nm through spectrophotometer.4. 50 mL aliquot will be put into each flask.02 M AgNO 3 by microburette. From this curve. 8 mL of 6 M NH 4OH and 2 mL of 5% KI solution will be transferred into the distillate solution before titration against 0. Determination of cyanogenic glycosides The cyanogenic contents in extract samples will be estimated by alkaline titration according to the method outlined in AOAC (1990) Method No.. 10 mL of saturated sodium carbonate solution and will be diluted to 100 mL with distilled water. The filtrate will be diluted to 100 mL and then be cooled. distillation process will be carried out by connecting the flask to a vapor distilling apparatus and distillate will be collected in a flask containing 20 mL of 2.115.3. 3. After thorough mixing. 0.3. The sealed flask will be given rest (3 h) to get proper hydrolysis process of the mixture.5% NaOH solution until a marked volume. 2005). Un-cooked 8 .e. After 2 hours. 3. Then. For the greenish blue color development. This will be followed by 5 mL Folin–Dennis reagent. patties.3. The partial/parfrying of the products will be carried out to determine the lipid stability. 26. 20 g of extract will be taken in kjeldhal flask following the addition of 200 mL distilled water and will slowly be mixed the sample. the concentrations for each sample will be used for the tannin content calculation (Nwinuka et al.made upto 5 mL with distilled water. 44 filter paper. Estimation of tannin For tannins.

5. odor. The sensory analysis will be performed at different storage intervals for experimental treatments. The order of presentation will be balanced to avoid carry-over effects. The average of the three runs will be reported as the measured value with standard deviation. 3. Samples will be presented to participants in sensory booths under white lighting. 2008).and fried meat products will be vacuum sealed in plastic bags and then will be stored at refrigerator and −18 °C in a freezer for a storage period of 60-days. Sensory evaluation of meat products Experienced and untrained assessors will carried out the sensory analysis of meat product samples according to the instructions given by Meilgaard et al. respectively. Statistical analysis The behavior of the Box-Behnken Model will be explained by the following the quadratic equation. Consumers will also rate their expected liking and intention to purchase. The sample analysis for storage stability and consumer acceptability will be carried out in triplicate and the significant differences will be calculated among means at a probability level of 5%. Ca 5a‒40). The oxidative stability of oils extracted from fish meat products will be assessed by measuring peroxide value (AOCS 1998 Method No. Each panelist will receive the samples assigned with random three‒digit code numbers. Each panelist will be asked to list their preference on a 9‒cm comparison line (1 = dislike extremely to 9 = like extremely). flavor. 3. The data of phytochemical yield obtained for each treatment will be subjected to statistical analysis to determine the level of significance by using the software package (MATLAB) according to the method described (Montgomery. Cd 8‒53) and free fatty acid value (AOCS 1998 Method No. 9 . Each judge will give the written informed consent after explanation of risks and benefits of participation prior to the study.6. The panelists will be provided informative instructions and brief definitions of attributes such as color. (2007). off-flavor and overall acceptability. The Duncan’s multiple range (DMR) test will be used to estimate the level of significance that existed between the mean values for product data analysis.

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