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E- Immunoassays Immuoassays are finding increasing use in the food industry for the qualitative and quantitative analysis of food products. Immunoassays specific for low molecular weight carbohydrates are developed by attaching the carbohydrate of interest to a protein, and then injecting it into an animal. With time the animal develops antibodies specific for the carbohydrate molecule. These antibodies can then be extracted from the animal and used as part of a test kit for determining the concentration of the specific carbohydrate in foods. Immuoassays are extremely sensitive, specific, easy to use and rapid. Analysis of Polysaccharides A wide variety of polysaccharides occur in foods. Polysaccharides can be classified according to: 1- Their molecular characteristics (e.g., type, number, bonding and sequence of monosaccharides) 2- Physicochemical characteristics (e.g., water solubility, viscosity, surface activity) 3- Nutritional function (e.g., digestible or non-digestible). Indigestible polysaccharides form part of a group of substances known as dietary fiber. which also includes lignin (which is a polymer of aromatic molecules). Consumption of many types of dietary fiber has been shown to have beneficial physiologically functional properties for humans, e.g., prevention of cancer, heart disease and diabetes. Analysis of Starch Starch is the most common digestible polysaccharide found in foods. In natural foods, such as legumes, cereals or tubers, the starch granules are usually separated from the other major components by drying, grinding, steeping in water, filtration and centrifugation. The starch granules are water-insoluble and have a relatively high density (1500 kg/m3) so that they will tend to move to the bottom of a container during centrifugation. Analysis methods Once the starch has been extracted there are a number of ways to determine its concentration: 1- Specific enzymes are added to the starch solution to breakdown the starch to glucose. The glucose concentration is then analyzed using methods described previously (e.g., chromatography or enzymatic methods). The starch concentration is calculated from the glucose concentration. 2- Iodine can be added to the starch solution to form an insoluble starch-iodine complex that can be determined gravimetrically by collecting, drying and weighing the precipitate formed. or titrimetrically by determining the amount of iodine required to precipitate the starch. 3- If there are no other components present in the solution that would interfere with the analysis, then the starch concentration could be determined using physical methods, e.g., density, refractive index or polarimetry. Analysis of Fibers Over the past twenty years or so nutritionists have become aware of the importance of fiber in the diet. Consumption of fiber helps protect against colon cancer, cardiovascular disease and constipation. Adequate intake of dietary fiber is therefore beneficial to good health. Dietary fiber is defined as plant polysaccharides that are indigestible by humans. Plus lignin, the major components of dietary fiber are cellulose, hemicellulose, pectin, and hydrocolloids. Crude Fiber Method The crude fiber method gives an estimate of indigestible fiber in foods. It is determined by sequential extraction of a defatted sample with 1.25% H2SO4 and 1.25% NaOH. The insoluble residue is collected by filtration, dried, weighed and ashed to correct for mineral contamination of the fiber residue. Filth detection Examination of whole foods for insect and rodent contamination: Insect tunneling and webbing. Whole larvae and adults. Rodent contamination: rodent excreta. Animal hairs Filth recovery methods 1- Sedimentation in heavier than water liquids: This procedure can be used to separate rodent excreta pellet fragments from cereals. In a liquid with a specific gravity near 1.49 the pellet fragments tend to settle out while much of the cereals float. 2- Sieving: in certain instances, a size separation offers a rapid means of separating filth from food. 3- When gasoline is mixed with an aqueous mixture containing insects or insect fragments, the insects float up with the gasoline layer. Fruits The water content of fruits varies from 60- 90%, fruits undergo a series of progressive changes from the unripe stage, to ripe stage, to the rotten and fermentative stages. The ripening point of a fruit is considered to be that point at which the sugar content is a maximum. In oranges and grapefruit, maturity is based on a minimum invert sugar-citric acid ratio (eight to one). Apples are considered ripe when the last trace of starch disappears. The browning of fruit flesh on exposure to air is due to the oxidation of the tannin in the fruit by an oxidase enzyme in the presence of the oxygen of the air. Maturity test for oranges and grapefruit Since the colour of grapefruit and oranges is inadequate as a measure of maturity, a more objective standard is used. This is based on the ratio of total solids to acid present in the juice, this is often termed the sugar: acid ratio as the major part of the total solids is sugar. Procedure: Determine the acidity by titration with 0.1 N sodium hydroxide solution and express the acidity in terms of percentage of citric acid. Determine the total solid content of the fruits. The maturity ratio is then the percentage of total solids divided by the percentage of citric acid. The minimum ratio for orange is 8: 1, and for grapefruit is 7: 1. Maturity test for apples 1- The firmness of the fruit. 2- The starch content. 3- The soluble solids. 4- The development of red colour. 5- The colour of the seeds. Starch test: As apples mature, the amount of starch decreases and in some varieties it is negligible. Procedure: 1- Dip slices of apple into a solution of iodine in potassium iodide for 1 minute. 2- Use the depth of the blue staining as an index of the amount of starch present. Soluble solids: As apples mature, the soluble solids content increases. Colour of seeds: As apples mature, the seeds turn brown to black in colour. Frozen fruits and fruit products Frozen fruits are made by freezing a properly prepared fruit ingredient or mixture of fruit ingredients specified in fill of container. May be added an optional sweetening ingredient: Sugar, any mixture of sugar with dextrose or corn syrup. An optional liquid packing medium: Heavy syrup, medium syrup, light syrup, corn syrup. Vegetable Products The analysis of these products includes the estimation of moisture, ash, metallic constituents, sugars, acids,…. Maturity in frozen vegetables This method depends upon the determination of the specific gravity of the sample by means of the difference in weight in air and in a liquid of known specific gravity. Canned Peas Prepared from: A variety of Pisum sativum . Water Optional ingredient: salt, sugar, dextrose, spice, flavoring, adjusting pH to 8. Quality: the standard quality for canned peas is: 1- Not more than 4% by count of the peas in the container are spotted or discoloured. 2- Normally coloured, not artificially coloured. 3- Weight of pea pods and other harmless extraneous vegetable material is not more than 0.5- 1% of the drained weight of peas in the container. 4- Weight of pieces of peas is not more than 10%. 5- The alcohol insoluble solids is not more than 23.5%. Spices, Flavors and Condiments Spices are aromatic vegetable substances used for the seasoning of food. Some of the determinations of spices includes; moisture, ash, nitrogen, heavy metals, reducing sugars and starch. Volatile and non-volatile extract: Extract 2 g of the sample in continuous extractor for 20 hours with anhydrous ether. Evaporate the ether to a small volume. Transfer to a small weighed beaker. Evaporate at room temperature, allow to stand over sulphuric acid for 18 hours in a desiccator. Weigh and calculate the gain in weight as percentage of total ether extract. Heat the extract gradually and then at 110o C until successive weighings show no loss. The difference in weight equals the volatile ether extract while the residue is the non-volatile ether extract. Alcohol extract: Shake 2 g of the sample in a 100 ml volumetric flask with 95% alcohol for 8 hrs. Filter and evaporate 50 ml of the filtrate to dryness in a dish. Dry at 100o C, the residue is the alcohol extract.