PHYSIOLOGY OF DIGESTION Aegina Festin, Princess Allyza Mondala, Veronica Raquepo and Ansley Yee University of the Philippines

Manila
Abstract. To understand the process of physical digestion of macromolecules, and the concept of chemical digestion of carbohydrates, two experiments were conducted. In the first experiment, a volunteer was asked to bring a favourite food, and with a dry mouth, imagine its taste at first, before smelling it then finally eating it. The other experiment tested the presence of starch (Lugols test) and maltose (Benedicts test) in different solutions (C1) distilled water, (C2) salivary amylase, (C3) salivary amylase with HCl, and (C4) salivary amylase heated in boiling water for five minutes. In Lugol's test, C1 became dark blue indicating no reaction. C2 had the lightest color; C3 was slightly darker than C2 since HCl destroyed some amylase so less starch was digested; C4 was darker than C3 but lighter than C1 due to enzyme denaturation when heated. In Benedict's test, C2 has amylase and starch. The salivary amylase broke down the latter into maltose and it became red orange after reacting to Benedicts solution after heat application. CuSO4 containing Cu2+ was reduced into Cu+ by the reducing sugar (maltose) turning the solution orange. C3, like C2 has salivary amylase that turned starch into maltose. HCl in C2 destroyed some of the salivary amylase resulting to less ptyalin concentration and a lighter orange coloration. Heat applied to C4 caused denaturation of the enzyme so little starch was converted into so there was less color change. Lastly, C1 showed no reaction since starch is a non-reducing sugar, therefore no maltose was produced.

INTRODUCTION The digestive system is responsible in the breaking down of food into simpler particles so that nutrients in it could be absorbed by the body. This process of breaking down food is called digestion. It involves two major processes, namely physical digestion, where large pieces of food are ground into smaller particles, and chemical digestion, where large biopolymers are broken down into simpler forms with the help of enzymes that are released in the digestive tract. Enzymes are protein catalysts which speed up the rate of chemical reactions. Temperature and pH are the factors affecting enzyme activity. Low temperature slows down a chemical reaction because the overall kinetic energy decreases. However, a high temperature also slows down a chemical reaction because high temperature changes the structure of enzymes. At a specific pH, the enzymes function is at its optimum. Thus, a change in pH will decrease the catalytic activity of enzymes. This experiment focuses on the physical digestion of macromolecules and the chemical digestion of carbohydrates. It has the following objectives:

To describe the process of physical digestion of macromolecules; To be familiar with biochemical tests for starch; To determine the effects of enzymes, pH, and temperature on the digestion of carbohydrates.

Experiments were conducted at RH 323 of the College of Arts and Sciences, University of the Philippines, Manila last May 9, 2013 to observe and understand the process of physical digestion of macromolecules and chemical digestion of carbohydrates. MATERIALS AND METHODS A. Physical Macromolecules Digestion of

A volunteer from the group was asked to bring her favorite food. After making sure that the volunteers mouth was as dry as possible, the first thing to do was for her to imagine her favorite food and how it tastes like. The food was then held near her nose for her to smell the aroma and imagine its flavor. The volunteer took a bite of the food while the other members observed which specific parts of her mouth were used to grind the food. Observations were also noted from the start of the activity up to swallowing. B. Chemical Carbohydrates Digestion of

water was placed in C1, 3.0 mL salivary amylase in C2 and C3 with C3 having an addition of 30 drops HCl. 3.0 mL salivary amylase was also placed in C4 and then placed in boiling water for 5 minutes. When all the setups were ready, 5.0 mL starch solution was then placed in each test tubes and incubated in a 37C water bath for an hour. After incubation, another set of four tubes were used and labeled C1 to C4. The incubated solutions were split evenly among the test tubes. One set was used for Lugols test or for the presence of starch. For the Lugols test, about three drops of Lugols iodine solution were added to each test tubes and color change of the solutions ranging from blue to black were noted. For the test of presence of maltose or Benedicts test, the other set of test tubes were used. 5.0 mL of Benedicts reagent was added to each test tube and then placed in boiling water for 10 minutes. Color changes from greenish to yellow to orange and red were noted. RESULTS Upon seeing the food, the volunteers mouth started to get moist with saliva from the salivary glands after then he even experienced stomach churnings. Smelling the food even intensified the sensations as he swallowed the accumulated saliva in his mouth for so many times signifying intensified release of

Four test tubes labeled C1, C2, C3 and C4 were used. 3.0 mL distilled

salivary amylase in the mouth, churnings of stomach were also contemporary. During the process of eating the food, the saliva moistens and the teeth then tears and chops the food at the same time. The food then turned moistened and grinded and rounded like a ball before swallowing and getting into further digestive tract. Lugols iodine solution is named after a French physician, Jean Lugol; it consists of iodine and potassium iodide in water. It can be used for testing the presence of starch in a solution. Benedicts reagent is named after an American chemist, Stanley Rossiter Benedict; it is a solution of copper (II) sulphate (CuSO4), tartaric acid and sodium hydroxide. It is used as a test for the presence of reducing sugar (sugars with a free aldehyde or ketone group) including glucose, lactose, and fructose, but not sucrose. All monosaccharides are reducing sugars; they all have a free reactive carbonyl group. Some disaccharides have exposed carbonyl groups and are also reducing sugars. Other disaccharides such as sucrose are non-reducing sugars and will not react with Benedict's solution. Starches are also non-reducing sugars.

Set-up Number C1 (water + starch) C2 (amylase + starch) C3 (amylase + HCl + starch) C4 (amylase + heat + starch)
Lugols Test: + = starch is present - = starch is absent

Lugols Test + -

Benedicts Test +++ ++

Benedicts Test - = Maltose is absent + = Maltose is present in small amt, ++ = Maltose is present in moderate amt. +++ = Maltose is present in large amount

Table1. Test reactions to Carbohydrate Indicators

DISCUSSION Physical digestion is, basically, "mashing" or breakdown of food by physical means. The food is ground up (by the teeth), mashed into a paste (by the stomach), and so forth. It is mixed with other foods in the process, but no new molecules are produced. Digestion begins when you smell something irresistible or when you see a favorite food you know will taste good. Just by smelling or thinking about how delicious that Tapsilog is going to taste, you begin to salivate and the digestive process kicks in, preparing for that first scrumptious bite. The presence of food stimulates salivary gland. This flow of saliva is set in

motion by a brain reflex that's triggered when we sense food or even think about eating. In response to this sensory stimulation, the brain sends impulses through the nerves that control the salivary glands, telling them to prepare for a meal. These nerve impulses from the brain also start the stomach churning. Churning is due to the muscular contraction in the stomach which helps mix food substances. Also on involved in the physical digestion is peristalsis which involves muscle contraction in the GI tract to push the food down leadin to a slight change in orientation and shape. Lugols Test After the Lugols Iodine solution was input into C1 which contained water and starch, the solution turned into dark blue because starch was present due to no reaction occurred. Whereas in C2, all the starch was digested so the solution had the lightest color among the set-ups. In C3, HCl was present so it destroyed some amylase resulting to less starch digested. The color of the solution in this test tube was a little darker compared to C2. It has a mixed hue of maroon and black. The solution contained in C4, on the other hand, was darker compared to the C3s but lighter than the C1s. This was due to the denaturation of the enzyme when amylase was heated. The iodine ions in Lugol's solution, once it is added to starch, most likely lodge in the coil spacing of starch (specifically, the helix structure of amylase). The iodine in the helix structure changes the electron configuration such that the compound

structure absorbs light in the appropriate regions to make it appear black/purple. Benedicts Test Test tube C2 has amylase and starch in it and so the salivary amylase broke down the latter into maltose. This gave rise to the red orange coloration of the solution because it reacted to the Benedicts solution after heat was applied. The CuSO4 containing Cu2+ was reduced into Cu+ by the reducing sugar (in this case, the maltose) turning the solution into orange. In test tube C3, like C2, the salivary amylase turned the starch into maltose hence the change in color of the solution. This set up had HCl in it, destroying some of the salivary amylase resulting to a less ptyalin concentration. Less ptyalin concentration means a lighter orange coloration of the solution. The half-reaction for Benedicts test for reducing sugars can be shown as: 2 Cu2+ + 2 e2 Cu+

Each copper (II) ion, Cu+2, is reduced to a copper (I) ion, Cu+1, by an electron from the reducing sugar. The reducing sugar is oxidized as a result of giving up its electron. A brickred precipitate, copper (I) oxide, Cu2O, may appear in the bottom of the tube. The more reducing sugar present in the mixture, the more precipitate will form. C4, on the other hand, the heat applied in the amylase caused denaturation of the enzyme and so, little

starch was converted into maltose and resulted to a less change of color of the solution. In C1, starch is a non-reducing sugar so there was no reaction between this and the Benedicts reagent, hence no color change and no maltose was produced.

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