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

in food samples. Theory: All living things contain organic macromolecules: Lipids, proteins, carbohydrates. These organic molecules are made up of only a small number of elements: carbon, hydrogen, oxygen, and to smaller amounts nitrogen, phosphorus and sulfur. They are called as macronutrient and be needed in large amounts for our body. Carbohydrates include sugars and starch. For sugars, it can be present as all monosaccharides such as glucose and fructose and some disaccharides such as maltose. It can be classified into two types, that is reducing sugars and non-reducing sugars. Reducing sugar is the sugar that forms an aldehyde or ketone in the presence of an alkaline solution while as non -reducing sugars do not possess to this group. For starch, Starch is a mixture of two polysaccharides, amylose and amylopectin. The relative amounts of the two polysaccharides vary widely in different types of starch. Amylose molecules consist of long straight chains of glucose units. Amylopectin has a complicated branched structure. Besides, proteins are complex, specialized molecules. The building blocks of proteins are amino acids. There are 20 different amino acids that combine to form polypeptides. The functional group of amino acids are the carboxyl group ( -COOH ) and the amino group (NH2). The carboxyl group and amino group of the adjacent amino acids can be linked together to form a peptide linkage through dehydration reaction. Lipids consist of glycerol and fatty acids "tails". The fatty acid "tails" are long chains of carbon and hydrogen that contribute to the non-polar behavior of fats - they don't mix with water.

Therefore, to differentiate all the class food, food test is needed. Food test is several types of simple test, easily performed in the laboratory, used to identify the main classes of food. Benedicts test is used to test for reducing sugars. The yellow, green and orange color of precipitate also indicates the presence of reducing sugar. For nonreducing sugars, it will give negative result on the Benedict's test. For starch, it can be tested by iodine test. The two polysaccharides combine with iodine to give the characteristic blueblack colour of the standard iodine test. Biuret test can used to test for the presence of protein. A solution that contains proteins will give violet coloration to the Biuret reagent The lipids are greasy and able to turn the area of dry lipids on a filter paper into a semi-transparent.

Hypothesis: Food sample A contains lipids, Food sample B contains protein, Food sample C contain reducing sugar, Food sample D contains starch, Food sample E contains reducing sugar and Food sample F contains non-reducing sugars.

Variables:

a) Manipulated a) Responding a) Manipulated

: Types of food samples : Changes of colour of solution : Volume of reagent be used

Materials:

Food sample A-Pounded groundnut, Food sample BMilk powder, Food sample CHoney, Food sample Dpotato, Food sample E-Jelly, Food sample F-Mandarin oranges, Iodine solution, Benedicts solution, sodium hydrogen carbonate solution, 20% sodium hydroxide solution, hydrochloric acid, 1% copper(II) sulphate solution and filter paper

Apparatus : Test-tubes, test-tube holders, beakers, Bunser burner, dropper, wire gauze, tripod stand, white tile, filter paper.

Procedure: 1. Six food samples labelled as A, B, C, D, E and F are prepared. 2. Food tests are carried out to test for the presence of starch, reducing sugars, non-reducing sugars, proteins and lipids in these food samples. A. Testing for the presence of starch ( Iodine Test ) 1. 2ml of each of the samples is poured into separate test tubes. 2. Each test tube is labelled with A, B, C, D, E and F. 3. Three drops of iodine solution are added to test tube A. 4. The colour change in test tube A is observed and recorded in a table. 5. Steps 3 to 4 is repeated with other test tube.

B. Testing for the presence of reducing sugar ( Benedicts Test ) 1. 2 ml of each of the samples is poured into separate test tubes. 2. Each test tube is labelled with A, B, C, D, E and F. 3. 1 ml of Benedicts solution is added to test tube A. 4. The mixture is shaken. 5. The test tubes are heated in a water bath until the mixture is brought to a boil. 6. The colour change in the mixture is observed and recorded in a table. 7. Steps 3 to 6 is repeated with other test tube.

C. Testing for the presence of non-reducing sugar 1. 2 ml of each of the samples is poured into separate test tubes. 2. Each test tube is labelled with A, B, C, D, E and F. 3. Four drops of dilute hydrochloric acid are added to test tube A. 4. The mixture is heated in a water bath for about 5 minutes.

5. The test tube is removed from the water bath and cooled under tap water. 6. The acid is neutralized by adding sodium hydrogen carbonate solution until the effervescence stops. 7. Benedicts test is conducted. 8. 1 ml of Benedicts solution is added to each of the test tubes. 9. The mixture is shaken. 10. The test tubes are heated in a water bath until the mixture is brought to a boil. 11. The colour change in mixture is observed and recorded in a table. 12. Steps 3 to 11 is repeated with other test tube.

D. Testing for the presence of protein ( Biuret Test ) 1. 2 ml of each of the samples is poured into separate test tubes. 2. Each test tube is labelled with A, B, C, D, E and F. 3. 20% sodium hydroxide solution is added in excess to the food samples and shaken. 4. A few drops of 1% copper (II) sulphate solution are added slowly to the mixture. 5. The mixture is shaken and allowed to stand. 6. The colour change in the mixture is observed and recorded in a table. 7. Steps 3 to 6 is repeated with other test tube.

E. Testing for the presence of lipid 1. A small amount of food sample A is rubbed on a piece of filter paper. 2. The filter paper is dried using a hair dryer. 3. The filter paper is held against light and the observation is recorded. 4. Steps 1 to 3 are repeated with other food sample.

Observation:
Test for nonreducing sugar

Food Sample

Test for starch

Test for reducing sugar The solution remains blue colour

Test for protein

Test for lipids

Inference

Pounded Groundnut

The solution change to a little dark blue colour

The solution A purple stays solution is unchanged. formed.

Presence of translucent spots.

Groundnut contains starch, protein and lipids.

Milk powder

A brownish white solution is formed.

A pale green solution is formed.

A cloudy blue solution is formed.

A violet solution is formed.

Absence of translucent spots.

Milk powder contains reducing sugar and protein only.

Honey

The solution remained brownish yellow colour.

An orange precipitate is formed.

A light green solution is formed.

An orange solution is formed.

Presence of translucent spots.

Honey contains reducing sugars, nonreducing sugars and lipids.

Food Sample

Test for starch

Test for reducing sugar An orange precipitate is formed.

Test for nonreducing sugar The solution stays in blue colour.

Test for protein

Test for lipids

Inference

Potato

A dark blue solution is formed.

A blue solution is formed.

Presence of translucent spots.

Potato contains starch, reducing sugar and lipids.

Jelly

Solution remained brownish yellow.

An orange precipitate is formed.

The solution stays in blue colour.

A little purple solution is formed.

Presence of translucent spots.

Reducing sugar, protein and lipid are present in jelly.

Mandarin Oranges

Solution remained yellowish orange colour.

Solution remaineds its blue colour.

A reddish orange precipitate is formed.

A cloudy blue solution is formed.

Absence of translucent spots.

Mandarin oranges contain nonreducing sugar.

Discussion : From the experiment, we can identify all the food class and nutrient by undergoing the food test experiment. Different type of reagent can identify different type of nutrients. Benedicts solution is used to test for presence of reducing sugars. Benedicts solution contains copper sulphate. When testing with the food that containing reducing sugars, reducing sugars will reduce copper (II) ions present in the blue copper sulphate solution to insoluble brick-red copper (I) oxide which is a precipitate. The solution will change from a blue solution to brick red precipitate. The final results are not only in brick-red precipitate only. It can form in orange or light green precipitate. The orange or green precipitate indicates the presence of a lesser small amount of reducing sugar. Unlike some other indicators, Benedicts solution does not work at room temperature. It must be heated first. Iodine test can test for the presence of starch. The two polysaccharides in starch will combine with iodine to give the characteristic blue-black colour of the standard iodine test. The negative result is the solution remains unchanged of yellow colour. For protein test, Biuret test is usually be used. When mixed with dilute alkaline copper sulphate solutions, nitrogen atoms in the peptide bonds form a violet complex with copper(II) ions. Therefore the solution will change from a blue colour to a violet and show a positive result. The lipids are greasy. When test with a filter paper, with presence of lipids, it able to turn the area on a filter paper into a semi-transparent. From all the food test above, we can know that one type of food sample contains not only one type of food nutrients only. It consists of many nutrients that are needed in our body. Besides, the food that we have chosen for the experiment must be naturally. This is because industry manufacture food will cause the result inaccurate.

Question: 1. Explain the need for our diet to contain: carbohydrates; sodium chloride; fresh fruits and milk. We need the diet contain carbohydrates is because carbohydrates are the main food that provide about 45% of the bodys energy needs to ensure the day's work. Sodium chloride is needed for our body to maintain the pH of our stomach and water balance. Fresh fruits are rich in vitamin and contain lots of fibre which helps to keep your body healthy. By consuming it, constipation and diverticular disease are less likely to develop. Milk are rich in protein that are classified as 'glow' food and used for formation of bone and tooth. Besides, protein is also needed in muscle and nerve coordination.

2. How do vegetarians ensure that they receive a balanced diet? Meat and other animal-based foods possess a full profile of essential amino acids, most plant-based foods are deficient in one or more aminos and thus not considered complete protein sources. Vegetarians need a combination of plant-based protein sources to get all the essential amino acids and ensure that they receive a balanced diet.

3. The body cannot digest fibre. Why is it still important in our diet? Fibre consists mainly of cellulose. Even though we can't digest cellulose, it's still an important part of your diet. The cellulose fibers from vegetables can help to scrub out your intestines and keep them clean

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 to your friend? Eating grapefruit is good for health and it will give the required calories for survival. Grapefruit is a good source of vitamin C, pectin fiber and contain beneficial antioxidant lycopene. With a normal grapefruit diet, it will be able to help the bodys metabolism burn fat. However drinking black coffee only is not good and is only a stimulant. Black coffee contains a lot of caffeine. Too much caffeine will cause unstable blood

sugar, resulting in fatigue and drowsiness. Caffeine will also speed up metabolism and cause the heart to beat abnormally. Therefore, I will suggest my friend to seek for a dieticians for further information and stop the crash diet that will harm the body system and affect the healthy.

5. Eskimos who eat mainly fish, seal oil and whale meat rather than beef and butter have a low incidence of heart disease. How do you explain this? Fish, seal and whale meat are very rich in omega-3, which is the healthiest polyunsaturate. We can get some in beef and butter as well, but none at all in margarine, corn oil and sunflower oil. Omega 3 will reduces blood clots and inflammation thus lowering the risk of heart disease and stroke

Conclusion: Hypothesis is accepted. Food sample A contains lipids, Food sample B contains protein, Food sample C contain reducing sugar, Food sample D contains starch, Food sample E contains reducing sugar and Food sample F contains nonreducing sugars.

Rujukan

Anonymous. (2005). Food tests. Didapatkan Februari 9, 2014, daripada BioToics:

http://www.biotopics.co.uk/nutrition/footes.html Anonymous. (2013). Food Chemistry Testing. Retrieved Februari 11, 2014, from SCIENCE Company: http://www.sciencecompany.com/-w152.aspx Foo Yuen Kooi. (2003). STPM BIology Volume 1. Bangi: Penerbit Pelangi. Liew Shee Leong, Lee Soon Ching dan Chan Cheok Lean. (1984). Biologi STPM Jilid 1. Petaling Jaya: Penerbit Fajar Bakti Sdn. Bhd. Wai, M. C. (2003). Biologi SPM Tingkatan 4-5. Petaling Jaya: Sasbadi Sdn. Bhd.