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QUALITY OF GRAPE AND APPLE FRUITS AND THEIR PROCESSED PRODUCTS By Faten Anwar Abd-Eldaim Ahmed B.Sc. Agric. Sci. (Home Economic), Fac. Agric., Cairo Univ., 2000 M.Sc. Agric. Sci. (Food Science) Fac .Agric. Minia Univ., 2006 THESIS Submitted in Partial Fulfillment of the Requirements for the Degree of DOCTOR OF PHILOSOPHY In Agricultural Sciences (Food Science) Department of Food Science Faculty of Agriculture Cairo University EGYPT 2016 Name of Candidate: Faten Anwar Ahmed Abd Eldaim Ahmed - Degree: Ph.D. Title of Thesis: Effect of Pesticide Residues on the Quality of Grape and Apple Fruits and their Processed Products Supervisors: Prof. Dr. Abd EL-Rahman Mohamed Khalaf-Allah Prof. Dr. Nagwa Mohamed ElShimi Prof. Dr. Emam Abd Elmobdy Abdel Rahim Dr. Alaa Aldin Mohsen Mohamed Department: Food Science Approval: 30 / 7 /2016 ABSTRACT The present study aimed to estimate penconazole and chlorfenapyr residue in grape, apple fruits and their products, to determinate the rates of fungicides dissipation, half-life values (RL ) and pre-harvest intervals (PHI) and analyzed 50 physical, chemical and biologically. Applications of penconazole and chlorfenapyr pesticides were carried out on grape and apple fruits grown in the open field at Kalyoubia Governorate, samples were collected after (one hour, one day, 3, 5, 7, 9, 11, 15 and 21 days) of application. The pesticide residues were analyzed by GLC and HPLC. The PHI time was 3 and 10 days after penconazole application for grape and apple for fresh fruits or juices product. The PHI time was 21 day after the chlorfenapyr application for grape and apple, that for fresh fruits or juices product. The penconazole residue in grape and apple RL was 2. 62 and 1.11 days, 50 respectively, while the RL for chlorfenapyr was 2. 86 and 2. 64 days for grape 50 and apple, respectively. Viscosity, total acidity and T.S.S. observed slightly alterations but not significant for fresh, juice or jam samples. The tested pesticides were influenced (slightly increased) on minerals contents for grape and apple samples. The slightly increased in ash by pesticides application was confirmed by the increases in all minerals. Vitamins (A, B , B and C) content were slightly 1 2 decreased by pesticide application. Total phenols and flavonoids were decreased in their juice or jam. Also, grape contained more amounts than apple. The decrease values were lower in juice than those of jam for both fruits. Also, reducing and non-reducing sugars were decreased. The sensory parameters had some variations relative to each other under the conditions of penconazole and chlorfenapyr applications. Feeding rats on the polluted products of fruits showed some toxicity of penconazole or chlorfenapyr. Blood glucose, urea, creatinine and uric acid content were increased by the pesticide ingestion. Albumin, globulin and Hb were decreased relative to control. AST and ALT were increased in plasma. Also, the highly elevation of plasma bilirubin is confirmed by the decreasing in Hb. The observations were the same for all organs under study (liver, kidneys, heart and spleen). The ingestion of penconazole and chlorfenapyr increased plasma lipids profile fraction, but decreased protein profile fractions. Liver function and kidneys function were inhibited under the ingestion of penconazole or chlorfenapyr. Keywords: Pesticide, residues, Penconazole, Chlorfenapyr, MRLs, RL , 50 PHI, Minerals, Vitamins, Phenols, Flavonoids, Blood glucose, Urea, Creatinine, Uric acid , Albumin, Globulin, AST, ALT. INTRODUCTION There has been interesting for the inclusion of fruits and vegetables in the human diet, mainly for the health benefits associated with their consumption (Liu et al., 2000 and Martin et al., 2002). A major benefit from a higher intake of fruits may be due to the increased consumption of vitamins , minerals and dietary fiber other constituents that may lower the risk of cancer and heart disease as well as prevent degenerative disease including antioxidant compounds such as carotenoids, flavonoids and other phenolics (Grassmann et al., 2002). These compounds are found ubiquitously in edible plants and are important constituents of the human diet. Epidemiologic studies that analyze the health implications of dietary components must estimate the intake in sample population. Therefore, the availability of appropriate and complete food composition datais crucial. In this study edible portion of two popular fruit crops: Grape (Vitis vinifera L.) native to the Mediterranean region, central Europe and southwestern Asia, from morocco and Portugal north to southern Germany and east to northern Iran (Robinson, 2001). In Egypt grapes are grown from Aswan in the south to Alexandria in the north there are also numerous hieroglyphic references from ancient Egypt. The grape is eaten fresh, processed to make juice and jam or wine also dried to produce raisins (Berkowitz, 1996). Apple (Malus sylvestris) is one of the most popular and favorite among the health conscious, fitness lovers who family believes in the concept of health is wealth. This wonderful is packed with rich phyto-nutrients that in the true sense, indispensable for optimal health (Kalinowska, 2014). Grape and apple have low calorized content and relatively high nutritive value; they contain carbohydrate, organic acids, fibers (pectins), tannins, aromatic substances and enzymes. These substances determine nutritive value and taste of fruits. These fruits are also rich in many minerals and vitamins essential for the appropriate functioning of the human organism, and rich in phenolic compounds, characterized by relatively high antioxidant activity. These fruits influence peristaltic movements slow down the absorption of carbohydrates, increase lipid breakdown in the human organism, enhance the sensation of satiety, reduce the levels of total cholesterol, blood triglycerides and homocystein concentrations and have a protective action on blood vessels (Leong and Shui, 2002). Due to seasonal gluts and highly perishable nature, considerable postharvest (0- 4 °C), they can only be stored for 2-4 weeks (Pantastico, 1975). Processing of fruits into value added products is the best option to control the huge losses. Fresh fruits are utilized for the preparation of jam and squash on a limited scale though they could be canned, frozen, dried and made into juice, sauce and confectionary (Vinson et al., 2001 and 2005). Penconazole (Triazole) formulated as (conazol 10 % EC) Penconazole is of great importance for the control of some fungal disease such as Erysiphaceaeventurai SPP and of the pathogenic Ascomycets, Basidiomyces and Deuteromycetes especially on cucurbit, apples, grapes, pomefruit, ornamentals and vegetables. Chlorfenapyr formulated as challenger super 24 % SC. This pesticide is using for control of many species of insects and mites such as leprosis mite, colorde potato beetle, Diamond-back moth, Heliothis SPP, Helicovepa SPP and Tetranychus SPP especially on fruits and vegetables, its using in resistance management programs for control the various cotton pests (Pesticide Manual, 2000). Diseases are common occurrence on plants, often having asignificant economic impact on yield and quality, thus managing diseases considered an essential component of production for most crops. However, equally important is the fact that grapes and apples have little or no inherent resistance to several fungal diseases (Mike and Mizuho, 2009). The diseases affect leaves, shoots and fruit. Leaf diseases can drastically reduce photosynthesis and in extreme cases, can cause defoliation. Also, fungal disease can vendor fruit unusual and can very easily cause severe losses in yield (Miles et al., 2010). Thus, pesticides are needed to control a disease during the establishment and development of grape or apple, also to increase productivity and to reduce blemishes (Green and Spilker, 1986). Pesticide residues could affect the ultimate consumes especially when these commodities are freshly consumed. The total dietary intakes of pesticide residues that remain on agricultural commodities are known these residues. The levels of pesticides residues are controlled by the maximum residue limits (MRLs) which are established by each country (Zawiyah et al., 2007). The Aim of Study: 1-The pre-harvest intervals (PHI) for grape and apple with tested pesticides (penconazole – chlorfenapyr). 2-The persistence of penconazole and chlorfenapyr on grape and apple fruits under the normal field conditions. 3-The effect of processing methods (juice and jam) on the persistence of pesticide residues. 4-The effect of pesticide treatments on the chemical composition of grape and apple. 5-The biological evaluation on the experimental rats that to treatment the intoxicated animal by pesticides (penconazole- chlorfenapyr) using evaluating of liver and kidney functions, blood hemoglobin, plasma sugar, lipid profile, protein profile. REVIEW OF LITERATURE For centuries, pesticides have been used in agriculture to enhance food production by eradicating unwanted insects and controlling disease vectors. The use of pesticides caused severe environmental and health hazards to organisms by used recomanded dosesticides inpest control. Considerable importance is given to functional food which a part from their basic nutritional functions, provide physiological benefits and play an important role in disease prevention or slow the progress of chronic disease there has been much more interest in the grape and apple as a medicinal and nutritional product because of its multi-functional and their great benefit in the human diet due to their several groups of substances that are useful in disease risk treatments. Grape in the non – climacteric fruit, botanically a true berry that grows on the perennial and deciduous woody vines of genus Vitis. Grapes can be eaten raw or used for making jam, juice, jelly, vinegar, wine grape seed extracts, raisins and grape seed oil. Grapes are also used in some kinds of confectionery. In Egypt the quantity exported in the year 2006 reached to 68, 296 tonnes (Heia, 2005). It is important to produce high quality table grapes by producing crop with low residue levels by applying a crop protection program that optimizes the use of plant protection products according to integrated crop management (ICM) integrates pest management (IPM) principles. In the end, the partners have been working together first to meet the demand for high – quality table grapes don’t exceed the maximum residue levels established by codex or European Union and second to bring down the level of residues of plant protection products in the final produce without compromising crop quality. The fruits and their processing products 1- Grape Grape is a fruiting berry of the deciduous woody vines of the botanical genusVitis. Grapes can be eaten raw or they can be used for making wine, jam, juice, jelly, grape seed extract, raisins, vinegar and grape seed oil. Grapes are a non-climacteric type of fruit, generally occurring in clusters.The cultivation of the domesticated grape began 6.000 – 8.000 years ago in the Near East. The earliest archeological evidence for a dominant position of wine-making in human culture dates from 8,000 years ago inGeorgia (Mukherjee, 2003). Grapesarerichin polyphenolic phytochemical compound anti-oxidant such as resveratrol whichreduces stroke risk by altering the molecular mechanisms inside the blood vessels (Bombardelli et al., 1995). Black rot, downy mildew and powdery mildew are the most destructive fungus disease of grapes, as well as insect pests such as: grape berry moth, grape flea beetle and grapevine beetle (Pesticide Manual, 2000). 2- Apple Apple fruit is one of the most popular and favorite fruits among the health conscious, fitness lovers who firmly believe in the concept of “health is wealth.” This wonderful fruit is packed with rich phyto-nutrients that, in the true sense, indispensable for optimal health. The antioxidants in apple have several health promoting and disease prevention properties, and thereby, truly justifying the adage, “an apple a day keeps the doctor away (Kalinowska, 2014). Often called as the "nutritional powerhouse", the potential health benefits they bring, with links to better and best health, together with prevention of many diseases. The vitamins present in apple are the key in maintaining red blood cells and the nervous system in good health (Coelho et al., 2012). Apple scab is the most serious disease. As a result, many of fungicide sprays can be applied each season to control the disease. Anystis baccarum is a commonly occurring predatory mite in the orchards. The effects of three commonly used

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DOCTOR OF PHILOSOPHY. In. Agricultural Sciences. (Food Science). Department of Food Science. Faculty of Agriculture. Cairo University. EGYPT. 2016 Title of Thesis: Effect of Pesticide Residues on the Quality of Grape and Apple Fruits and their hieroglyphic references from ancient Egypt.
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