Practical 2 : Food Test

Introduction : For objective 1 The Iodine test is used to test for solution dissolved in an aqueous solution with starch producing a purple black color. the presence of starch. of potassium iodide that Iodine reacts

This reaction is the result of the formation of polyiodide chains from the reaction of starch and iodine. The amylose, or straight chain portion of starch, forms helices where iodine molecules assemble, forming a dark purple/black color. The amylopectin, or branched portion of starch, forms much shorter helices and iodine molecules are unable to assemble, leading the color to be of an orange/yellow hue. As starch is broken down or hydrolyzed into smaller carbohydrate units, the purple-black color is not produced. Therefore, this test can determine completion of hydrolysis when a color change does not occur. Benedict's reagent is used as a test for the presence of reducing sugars. This includes all monosaccharides and the disaccharides, lactose and maltose. Even more generally, Benedict's test will detect the presence of aldehydes (except aromatic ones), and alpha-hydroxy-ketones, including those that occur in certain ketoses. Thus, although the ketose fructose is not strictly a reducing sugar, it is an alpha-hydroxyketone, and gives a positive test because it is converted to the aldoses glucose and mannose by the base in the reagent. To test for the presence of monosaccharides and reducing disaccharide sugars in food, the food sample is dissolved in water, and a small amount of Benedict's reagent is added. During a water bath, which is usually 410 minutes, the solution should progress in the colors of blue (with no glucose present), green, yellow, orange, red, and then brick red or brown (with high glucose present). A colour change would signify the presence of glucose. The common disaccharides lactose and maltose are directly detected by Benedict's reagent, because each contains a glucose with a free reducing aldehyde moiety, after isomerization. Sucrose (table sugar) contains two sugars (fructose and glucose) joined by their glycosidic bond in such a way as to prevent the glucose isomerizing to aldehyde,

or the fructose to alpha-hydroxy-ketone form. Sucrose is thus a non-reducing sugar which does not react with Benedict's reagent. Sucrose indirectly produces a positive result with Benedict's reagent if heated with dilute hydrochloric acid prior to the test, although after this treatment it is no longer sucrose. The acidic conditions and heat break the glycosidic bond in sucrose throughhydrolysis. The products of sucrose decomposition are glucose and fructose, both of which can be detected by Benedict's reagent. The biuret test is a chemical test used for detecting the presence of peptide bonds. In the presence of peptides, a copper(II) ion forms a violet-colored complex in an alkaline solution.[1] Several variants on the test have been developed. The biuret reagent is made of potassium hydroxide (KOH) and hydrated copper (II) sulfate, together with potassium sodium tartrate. The reagent turns from blue to violet in the presence of proteins, blue to pink when combined with shortchain polypeptides. The lipid is a group of macromolecules that are insoluble in water. The major kinds of lipids in biological world include fats, phospholipids and steroids. The fats or triacylglycerols are ,ade up of glycerol ( an alcohol ) and fatty acids ( a monocarboxylic acid ) which are bonded together by esther linkages. The lipids are greasy and would turn the area of dry lipids on a brown paper into semi-transparent.

For objective 2 Vitamin C is found in green vegetables, frits, and potatoes. It is essential for a healthy diet. The chemical name for name vitamin C is acsor bic acid. Ascorbic acid is a good reducing agent and therefore it is easily oxidized. Methods for the detection of vitamin C involve titrating it against a solution of an oxidizing agent.

Objective 1 : To test the presence of starch, reducing sugars, non-reducing sugars, proteins and lipids in food samples

Materials and Apparatus : Iodine solution, Benedict solution, sodium hydrogen carbonate solution, 20% sodium hydroxide solution, hydrochloric acid, 1% copper (II) sulphate solution and food samples (unknown to students). ( A milk, B honey, C peanut butter, D ribena/raisin juice, E mayonnaise ). The food sample can be varies. Test tubes, test tube holders, beakers, Bunsen burner, dropper, wire gauze, tripod stand, white tile and filter paper.

Procedure : 1) Five samples of food labeled A, B, C, D and E are prepared. 2) Food tests are carried out to determine presence of starch, reducing sugars, nonreducing sugars, proteins and lipids in these food samples.

Result : Test For Nonreducing sugar Procedure 1. 2ml of food sample A was poured into a boiling tube. Three drops of hydrochloric acid were added. 2. The mixture was heated in boiling water bath for 5 minutes. Observation 1. Brick red precipitate were found in food samples B, C, D and E 2. Food samples A remained blue in colour. Inference 1. Non reducing sugar is present in food sample B(peanut jam), C(Ribena/Raisin juice), D(honey) and E(mayonnaise) 2. Non reducing sugar is absent in food sample A(milk)

3. The boiling tube was removed from water bath and the mixture was cooled under running tap. 4. The acid in the mixture was naturalized by sodium hydrogen carbonate powder until effervescence stopped. 5. Then, Benedictss test was conducted on the mixture. 6. Any colour change was observed in the mixture. 7. Steps 1 to 6 were repeated using food samples B,C,D and E

Starch

1. 1cm3 of each food samples are being placed in respective test tube, labelled A,B,C,D, and E. 2. Using a syringe, 1cm3 of distilled water were added into each test tube. 3. The diluted solution of the food samples were then being stirred by using a glass rod.

1. Peanut butter changes the colour of iodine solution from brownish to dark blue. 2. Milk, honey, Ribena and Mayonnaise did not cause colour changes to the iodine solution.

1. The colour changes of iodine solution from brownish to dark blue indicates the presence of starch 2. Only peanut butter contained starch.

4. Next, 2 drops of each mixture were placed on a white tile, using a dropper. 5. On each samples, another two drops of iodine were added, to test the presence of starch. 6. The changes from the food test were observed.

Reducing sugar

1. 2ml of each food samples are poured into a test tube respectively. All the test tube is labeled A, B, C, D and E. 2. 1ml of Benedicts solution is added to each food sample in the test tube. 3. The mixture is shaken and then heated by placing all the test tube in a boiling water bath. 4. Any change in colour in each mixture is observed. 1. 2 ml of each food sample is poured into a test tube. 2. 1ml of 20% sodium hydroxide solution is then added to each of the food sample and shaken well. 3. A few drops of 1 % copper (II) sulphate solution are then added slowly to each of the mixtures 4. Each mixture is shaken well and allowed to stand

1. A brick-red precipitate is formed in food samples labelled B, C and E. 2. The other food samples remain blue in colour.

1. The colour changes indicates the presence of reducing sugars. 2. Milk, Mayones, and Honey has reducing sugars in it.

Protein

1. Food sample A & C turns purple. the other food sample turns blue in colour.

1. Food sample A & C contains protein

5. Any colour change in the food sample is observed Lipid 1. A small amount of each of the food samples is rubbed on a piece of filter paper. 2. Then the filter paper is dried. 1. The filter paper that was rubbed against food sample B,C and E shows a translucent mark. 2. The filter paper of the food sample A and D remain opaque. 1. Food sample B,C and E contain lipid. 2. Food sample A and D do not contain lipid.

3. The filter paper is held against the light. 4. All observations are recorded.

Objective 2 : To determine the vitamin C content in various fruit juices.

Problem : Do imported fruits contain more vitamin C than local fruits?

Materials and Apparatus : 1.0% dichlorophenolindophenol solution (DCPIP), 0.1 % ascorbic acid solution, freshly prepared lime juice, pineapple juice and orange juice specimen tubes, syringes with needles ( 1 ml and 5 ml ), beakers, gauze cloth and knife.

Procedure : 1. 2. 3. 4. 1 ml of 1.0% DCPIP solution is placed in a specimen tube using a 1 ml syringe. The 5 ml syringe is filled with 0.1% ascorbic acid solution. The needle of the syringe is placed into the DCPIP solution. The ascorbic solution is added drop by drop to the DCPIP solution. The mixture is gently stirred with the needle of the syringe. The ascorbic acid solution is continuously added until the DCPIP solution is decolourised. The volume of ascorbic acid solution used is recorded. 5. Steps 1 to 4 are repeated using freshly squeezed lime juice, pineapple juice and orange juice. The volume of fruit juice required to decolourise the DCPIP solution in each case is recorded. 6. The results are tabulated. The percentage and the concentration of vitamin C in each of the fruit juices are calculated using the formulae given.

Results : Solution/fruit juice Volume of solution/fruit needed to decolourise 1 ml of DCPIP solution 1 2 3 Average 1.0 1.0 1.0 1.0 3.2 3.0 3.1 3.1 Percentage of vitamin C Vitamin C concentration in fruit juice in fruit juice

0.1 % ascorbic acid Lime juice

Pineapple juice

4.9

4.9 4.8 4.9

Orange juice

5.0

5.3 5.2 5.2

Calculations : Volume of 0.1% ascorbic acid used to decolourise DCPIP = x cm3 Volume of fruit juice used to decolourise DCPIP = y cm3 x cm3 0.1% ascorbic acid ( concentration of 1 mg/cm3 ) can decolourise 1 cm3 of DCPIP and y cm3 of fruit juice concentration of k mg/cm3 can decolourise 1 cm3 of DCPIP So, The quantity of ascorbic acid in x cm3 of ascorbic acid 0.1% = quantity of ascorbic acid in y cm3of fruit juice Thus, x cm3 X 1 mg/cm3 = y cm3 X k mg/cm3 k mg/cm3 = X mg/cm3

= mg/cm3

Questions : 1. Explain the need for our diet to contain : carbohydrates, sodium chloride, fresh fruit and milk. Carbohydrates are the starches and sugars that provide the energy our bodies need to function and grow. Salt helps maintain the fluid in our blood cells and is used to transmit information in our nerves and muscles. It is also used in the uptake of certain nutrients from our small intestines. The body cannot make salt and so we are reliant on food to ensure that we get the required intake. Fruits will carve your appetite and cease your sugar cravings. Fruits will also give you that boosted energy that most sugary snacks give you. Milk is used for formation of bone and tooth and is needed in muscle and nerve coordination. By consuming milk, we can prevent from getting osteoporosis when we become older.

2. How do vegetarians ensure that they receive a balanced diet? Vegetarian diets are very much indeed balanced. In fact, recent studies have shown that we should be eating more vegetables and fruits, rather than the other food groups. Vegetarians can live a very healthy life, just as much as the next person and even more. Cutting out meat from your diet doesn't mean that a person eats everything they used to, but with no meat. It doesn't; vegetarians find supplements for what meat gives us via fruits, vegetables, soy, legumes and nuts. A vegetarian eats a more abundant amount of things grown directly from our earth, which is actually better when compared to the diet of a carnivore. Vegetarianism involves the practice of following a diet that includes fruits, vegetables, cereal grains, nuts, and seeds, with or without dairy products or eggs. A vegetarian does not eat meat, including game, poultry, fish, crustacea and shellfish, and may also abstain from by-products of animal slaughter such as animal-derived rennet and gelatin. Various foods or treats, such as cake, chocolate, chips, gum, marshmallows and gummy candies, often contain unfamiliar animal ingredients, and may especially be a concern for vegetarians due to the likelihood of such additions. While some vegetarians are unaware of animal-derived rennet's role in the usual production of cheese and may therefore unknowingly consume the ingredient, others of the diet are not bothered by its consumption. Often, however, animal-derived products, such as certain cheeses, gelatin or other animal-derived ingredients, are scrutinized by vegetarians prior to purchase or consumption.

3. The body cannot digest fibre. Why is it still important in our diet? Fiber is important because it has an influence on the digestion process from start to finish: Because it demands that food be more thoroughly chewed, fiber slows down the eating process and helps contribute to a feeling of being full, which in turn can help prevent obesity from overeating. Fiber makes food more satisfying, probably because the contents of the stomach are bulkier and stay there longer. Fiber slows digestion and absorption so that glucose (sugar) in food enters the bloodstream more slowly, which keeps blood sugar on a more even level. Fiber is broken down in the colon (the main part of the large intestine) by bacteria (a process called fermentation), and the simple organic acids produced by this breakdown helps to nourish the lining of the colon.These acids also provide fuel for the rest of the body, especially the liver, and may have an important role in metabolism.

4. A slighty overweight friend decide to go on a crash diet. She/he tells you that she/he is only going to eat grapefruit and drink black coffee for two weeks. What advice would you give to your friend? Black coffee contains a lot of caffeine while grapefruit contains vitamin C. Black coffee can negatively affect young adults, children and the elderly who are more sensitive to caffeine, and those with compromised immune systems and hypertension. Although caffeine's effect on heart disease remains under speculation, those with heart rate fluctuations should be moderate in their consumption. The oils in the ebony beans can also contribute to elevated blood cholesterol levels, especially the "bad" LDL count. Aside from that it effects blood vessel tone and can cause irregular heartbeat. The grapefruit diet is a short-term fad diet . Specifics of the diet vary widely, but it usually involves eating half a grapefruit or drinking grapefruit juice with each meal and significantly reducing caloric intake, often to below 800 calories per day. The diet is based on the claim that grapefruit has a fat-burning enzyme or similar property. The grapefruit diet is a low-carb diet. It suggests that grapefruit helps burn body fat when eaten with foods high in dietary fat, which is why the grapefruit diet encourages consumption of meat, eggs and other foods that are rich in fat and protein. Hence, I will tell my friends to stop the crash diet and if she wants to go on a diet, she should meet dieticians to help her in consuming balanced intake of nutrition.

5. Eskimos who eat mainly eat, seal oil and whale meat rather than beef and butter have a low incidence of heart disease. How do you explain this? Fish, seal oil and whale meat has lower cholesterol than the beef and butter. So, the Eskimos have low in fats especially in triglycerides and lipoproteins and in cholesterol. The large amount of fatty marine food that the Eskimos were eating somehow protected them from heart disease.

Discussion : For objective 1 1. When a reducing sugar is heated with Benedicts solution, the reducing sugar reduces the blue copper(II) sulphate in Benedicts solution to form a brick-red precipitate of copper(I) oxide. 2. (a) (b) A brick-red precipitate indicates the presence of reducing sugar. The final colour of the solution depends on how much of this precipitate is formed. Therefore, the colour gives an indication of how much reducing sugar is present. A brick-red precipitate indicates that a large amount of reducing sugar is present while an orange or a green precipitate indicates the presence of a lesser amount of reducing sugar. If the original pale blue colour of the solution remains, this indicates that reducing sugar is not present.

(c)

3. The different food samples are first heated with dilute hydrochloric acid to hydrolyse or break down the disaccharide compound (sucrose) to its constituent monosaccharides (glucose and fructose) 4. The mixture of food sample and hydrochloric acid is first neutralized by adding sodium hydrogen carbonate powder before adding Benedicts solution

For objective 2 1. 0.1% ascorbic acid solution contains 0.1 g ascorbic acid in 100 ml of solution. 2. The vitamin C (ascorbic acid) content in fruit juices can be determined by using the DCPIP solution (dichlorophenolindophenol). Vitamin C reduces the blue DCPIP solution to a colourless solution.

3. (a)

(b)

During the investigation, the DCPIP solution should not be shaken vigorously when the ascorbic acid solution and the fruit juices are being added. This is because oxygen from the atmosphere oxidizes the reduced DCPIP solution and turns it blue again. If this happens, more fruit juice is needed to reduce the DCPIP solution. Therefore, the actual vitamin C content in the fruit juice cannot be known.

4. Lime juice contains the highest concentrations of ascorbic acid. 5. If the vitamin C content in a fruit juice is high, less volume of the fruit juice is required to decolourise the fixed amount of DCPIP solution. 6. During the extraction of juices from the fruits, the fruits should not be pounded to hard, as the heat produced will destroy part of the vitamin C in the juices. 7. The fruit juices cannot be prepared a day earlier before the investigation or exposed to air for a day because the vitamin C in it easily destroyed by oxidation.

Conclusion : For objective 1 1. 2. 3. 4. 5. Non reducing sugar present in peanut jam, ribena/raisin juice and mayonnaise Starch present in peanut butter. Reducing sugar present in milk, mayonnaise and honey. Protein present in milk and peanut butter. Lipid present in honey, peanut butter and mayonnaise.

For objective 2 1. Lime juice contains a higher concentration of vitamin C compared to pineapple and orang juice.