Practical 2: Food Test Objective: To test the presence of starch, reducing sugars, non-reducing sugars, proteins and lipids

in food samples. Theory: Nutrition is the supply of food required by organisms and cells to stay alive. In science and human medicine, nutrition is the science or practice of consuming and utilizing foods. Nutritional science studies how the body breaks food down (catabolism) and repairs and creates cells and tissue (anabolism) - catabolism together with anabolism is known as metabolism. Nutritional science also examines how the body responds to food. The human body requires seven major types of nutrients. A nutrient is a source of nourishment, an ingredient in a food, for example protein, carbohydrate, fat, vitamin, mineral, fiber and water. Macronutrients are nutrients we need in relatively large quantities. Micronutrients are nutrients we need in relatively small quantities. Carbohydrates are molecules that consist of carbon, hydrogen and oxygen atoms. Carbohydrates include monosaccharides (glucose, fructose, galactose), disaccharides (maltose, sucrose, lactose), and polysaccharides (starch). Nutritionally, polysaccharides are more favored for humans because they are more complex molecular sugar chains and take longer to break down - the more complex a sugar molecule is the longer it takes to break down and absorb into the bloodstream, and the less it spikes blood sugar levels. Spikes in blood sugar levels are linked to heart and vascular diseases. Proteins are molecules that contain nitrogen, carbon, hydrogen and oxygen. Simple proteins, called monomers, are used to create complicated proteins, called polymers, which build and repair tissue. When used as a fuel the protein needs to break down, as it breaks down it gets rid of nitrogen, which has to be eliminated by the kidneys. Lipids (for example fats) are molecules that consist of carbon, hydrogen, and oxygen atoms. Fats are triglycerides - three molecules of fatty acid combined with a molecule of the alcohol glycerol. Fatty acids are simple compounds (monomers) while

triglycerides are complex molecules (polymers). Lipids supply as much as four times the amount of energy as carbohydrates and proteins. Hypothesis: Starch is present in peanut, potato while reducing and nonreducing sugar might be found in all food samples. Protein is present in peanut, potato, milk and lychee. Lipid is found in peanut, potato and honey syrup. Variables: a) Manipulated: Types of food samples b) Responding: Colour changes of food samples c) Fixed: Volume of the reagent used Materials: Iodine solution, Benedict solution, sodium hydrogen carbonate solution, 20% sodium hydroxide solution, hydrochloric acid, 1% copper(II) sulphate solution, pounded peanut, potato, milk, honey syrup, jelly, lychee. Apparatus: Boiling tubes, test tube holders, beakers, Bunsen burner, dropper, wire gauze, tripod stand, filter paper. Procedure: 1. Four boiling tube are prepared and labeled with A,B,C and D. 2. 2ml of pounded peanut is poured into each boiling tube. 3. In boiling tube A, starch test is carried out. Three drops of iodine solution are added into the boiling tube. Any colour changes in the mixture is observed and recorded. 4. In boiling tube B, reducing sugar test is carried out. 1ml of Benedicts solution is added to the pounded peanut (diagram 1a). The mixture is shaken and then heated by placing the boiling tube in a boiling water bath (diagram 1b). Any colour changes in the mixture is observed and recorded. 5. In boiling tube C, non-reducing sugar test is carried out. A few drops of dilute hydrochloric acid are added to the pounded peanut. The mixture is then heated in a boiling water bath for five minutes (diagram 2a).

6. The boiling tube is removed and cooled under a running tap. The acid in the mixture is neutralized by adding sodium hydrogen carbonate until effervescence stops (diagram 2b). Benedicts test is carried out to test the presence of non-reducing sugar in the mixture (diagram 2c & 2d). Any colour changes in the mixture is observed and recorded. 7. In boiling tube D, proteins test is carried out. 1ml of 20% sodium hydroxide solution is added to the pounded peanut and shaken well. A few drops of 1% copper(II) sulphate solution are added slowly to the mixture. The mixture is shaken well and allowed to stand. Any colour changes in the mixture is observed and recorded. 8. A small amount of pounded peanut is rubbed on a piece of filter paper. The filter paper is then dried and held against the light. Any observations are recorded. 9. Steps 1 to 8 are repeated by replacing the pounded peanut with potato, milk, honey syrup, jelly and lychee.

Results: Food Samples Test for Starch Observation Mixture from yellow blue Reducing sugar The pale blue Pounded peanut Inference

turns Pounded peanut brownish- contains starch to dark

mixture does not does not contains change colour Non-reducing Pounded Peanut sugar An precipitate formed mixture. Protein Mixture turns Pounded peanut in reducing sugar

orange Pounded peanut is contains non-

the reducing sugar

from pale blue to contains protein purple Lipid Filter shows translucent mark paper Pounded peanut a contains lipid

Food Samples

Test for Starch

Observation Mixture from yellow blue

Inference contains

turns Potato brownish- starch to dark

Reducing sugar Potato

An orange-yellow Potato precipitate formed mixture in

contains

is reducing sugar the

Non-reducing sugar

The

pale

blue Potato does not non-

mixture does not contains change colour

reducing sugar

Protein

Mixture

turns Potato

contains

from pale blue to protein purple Lipid Filter shows translucent mark paper Potato a lipid contains

Food Samples

Test for Starch

Observation Mixture

Inference does not

remains Milk

brownish-yellow Reducing sugar An precipitate formed mixture Milk Non-reducing sugar The pale in

contains starch contains

orange Milk

is reducing sugar the

blue Milk

does

not non-

mixture does not contains change colour

reducing sugar contains

Protein

Mixture

turns Milk

from pale blue to protein purple Lipid Filter paper Milk does not

remains opaque.

contains lipid.

Food Samples

Test for Starch

Observation Mixture

Inference syrup

remains Honey

brownish-yellow

does not contains starch

Honey syrup

Reducing sugar

An orange-yellow Honey precipitate formed mixture in is contains

syrup

the reducing-sugar

Non-reducing

green Honey

syrup

sugar

precipitate formed mixture in

is contains

small

the amount of nonreducing sugar

Protein

Colour

of

the Honey

syrup

mixture turns to does not contains orange Lipid Filter shows translucent mark protein paper Honey syrup

a contains lipid

Food Samples

Test for Starch

Observation Mixture

Inference does not

remains Jelly

brownish-yellow Reducing sugar

contains starch contains

An orange-yellow Jelly precipitate formed mixture in

is reducing sugar the

Jelly

Non-reducing sugar

The

pale

blue Jelly

does

not non-

mixture does not contains change colour

reducing sugar does not

Protein

Mixture turns to Jelly browninshorange

contains protein

Lipid

Filter

paper Jelly

does

not

remains opaque.

contains lipid.

Food Samples

Test for Starch

Observation Mixture

Inference

remains Lychee does not contains starch contains

brownish-yellow Lychee Reducing sugar

An orange-yellow Lychee precipitate formed in

is reducing sugar the

mixture Non-reducing sugar The pale blue Lychee does not non-

mixture does not contains change colour

reducing sugar contains

Protein

Mixture

turns Lychee

from pale blue to protein purple Lipid Filter paper Lipid does not

remains opaque.

contains lipid.

Discussion: 1. The food sample in solid needed to be cut into very small pieces or smashed (similar with chewing mechanism) before adding a little distilled water in order to provide a more concentrated solution for testing. 2. In Benedicts test, the reducing sugar is heated and it reduces the copper(II) sulphate in Benedicts solution to form a red precipitate of copper(I) oxide. 3. The presence of reducing sugar is not only determined by the presence of brick-red precipitation, but also with orange or green precipitate. A brick-red precipitation indicates the presence of a large amount of reducing sugar; while an orange or green precipitate indicates the presence of a lesser amount of reducing sugar. If the original pale blue colour remains unchanged, this indicates that reducing sugar is absent. 4. Different food samples are first heated with dilute hydrochloric acid to hydrolyse the non-reducing sugar (sucrose) to its constituent monosaccharides (glucose and fructose) which are reducing sugars. The mixture is neutralised by adding sodium hydrogen carbonate before carry out Benedicts test.

Questions: 1. Explain the need for our diet to contain: carbohydrates; sodium chloride; fresh fruits and milk. Carbohydrates are the main energy source for our body, they are the energy that gets used first (before protein, fat and alcohol). It is recommended that about 45-60% of our energy intake should come from carbohydrates. Sodium chloride or salt is important because they maintain the acid-base balance and water balance of our body. It is also an important component in blood plasma. Fresh fruits provide our body vitamins which helps to prevent heart diseases, stroke, and obesity. Other than that, fruit also contains fibre that cant be broken down, which aids peristalsis. Milk contains many beneficial nutrients such as calcium, phosphorus, protein and other, which helps to build strong bones and prevent osteoporosis (especially among women).

2. How do vegetarians ensure that they receive a balanced diet? Vegetarians don't eat dairy products, meat of animals and eggs. Vegetarians need to eat a lot of other calcium-rich foods. Calcium-rich breakfast cereals, soy milk, and orange juice are good choices. Other foods that have calcium include certain legumes, certain leafy green vegetables, nuts, seeds, and tofu. Vegetarians can also take a daily calcium supplement. Vegetarians can also eat a wide variety of food to ensure they receive a balanced diet. Eating a wide variety of foods-especially legumes and grains-will give you the protein you need. Vegetarians who don't eat fish or eggs need to find other good sources of omega-3 fatty acids, such as hemp seeds, flaxseeds, pumpkin seeds, walnuts, certain leafy green vegetables, soybean oil, and canola oil.

3. The body cannot digest fibre. Why is it still important in our diet? Although our body cant digest fibre, it is vey important for its ability to prevent or relieve constipation. This prevents the build-up of toxic substances in rectum which can lead to bowel cancer. Foods

containing fibre can also provide other health benefits as well, such as helping to maintain a healthy weight and lowering your risk of diabetes and heart disease.

4. A slightly overweight friend decides 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 your friend? The black coffee contains lots of caffeine while the grapefruit is rich in vitamin C. Metabolic rate increased significantly during the 3 hour after caffeine ingestion. Caffeine in coffee stimulates the metabolic rate and this is accompanied by greater oxidation of fat in normal weight subjects, which resulting in weight loss over the time. Despite of this fabulous effect, caffeine can cause health problems and negative side effects that for some are simply not worth the slight increase in metabolism. Consuming too much caffeine can cause restlessness, poor sleep, anxiety and irritability in some people, as well as headaches, abnormal heart rhythms or other problems. Grapefruit is chalk full of vitamin C, which helps keep the immune system strong and fights illnesses. Folic acid is also present in grapefruit. Folic acid, like vitamin C, helps protect your heart form heart disease and other ailments. When combined with potassium, folic acid and vitamin C become powerhouses for the human body. There have been many studies on the grapefruit that show it having excellent effects on the human body. It increases vitamin levels, supports healthy immune systems by utilizing vitamin C. Grapefruit is extremely good for the heart and arteries as well as for the body's muscles. It balances cholesterol and prevents fatty cells from collecting in the body, allowing the metabolism to work more effectively. So it was hypothesized that the grapefruit encourages fat loss. Ill advice my friend to stop taking crash diet and advise her to see a dietician if he/she wants to go on a healthy diet.

5. Eskimos who eat mainly fish, seal oil and whale meat rather than beef and butter have a low incidence of heart diseases. How do you explain this? This is because fish, seal oil and whale meat contains lower cholesterol than the beef and butter. There is evidence that omega-3 fatty acids, primarily those found in oily fish, when eaten as part of a healthy diet, can help maintain a healthy heart and reduce risk of heart disease. Studies suggest omega-3 fatty acids may lower triglycerides, which are a type of fat in the bloodstream. Omega-3 fatty acids may also slow down the growth of plaques in the arteries and reduce inflammation throughout the body.

Conclusions: Pounded peanut and potato provides lots of energy because peanut contains starch, non-reducing sugar, protein and lipid while potato contains starch, reducing sugar, protein and lipid. Milk contains small amount of reducing sugar and protein which is important for human development and growth. Honey syrup contains reducing-sugar, small amount of non-reducing sugar and lipid. Jelly contains reducing sugar only, thats why it is sweet. Lychee is a nutrition fruit, it contains reducing sugar and protein only. Hypothesis is accepted.

References: WebMV. (2014). Low cholesterol in fish. Retrieved from

http://www.webmd.boots.com/cholesterol-management/features/lowcholesterol-diet-fatty-fish WebMV. (2014). Vegetarians diet- How can vegetarians eat a balanced diet. Retrieved from http://www.webmd.com/food-recipes/tc/vegetarian-

diets-how-can-vegetarians-eat-a-balanced-diet WebMV. (2014). Caffeine. Retrieved from http://www.webmd.com/vitaminssupplements/ingredientmono-979CAFFEINE.aspx?activeIngredientId=979&activeIngredientName=CAF FEINE

Photos of the results:

Boiling tube A to D (from left to right) containing honey syrup and chemical reagent.