Locker Number: 50 Date Submitted: November 22, 2011 Members: 1. Calumpang, Gino I. 2. Flores, Czarriel 3. Potencio, Valaine C.

Activity Number 2 BIOCHEMICAL PROCESSES

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Objectives: 1. To be able to define what Biochemical processes derived from the performed experiment. 2. To be able to elucidate some biochemical processes used in the experiment, and their definition. 3. To be able to list some of the essentialities of the biochemical processes. 4. To be able to know basic concepts of the biochemical processes.

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Introduction:

Biochemical processes that are used in the experiment are the following: Diffusion, Osmosis, Dialysis and Surface Tension. Diffusion is the movement of a fluid from an area of higher concentration to an area of lower concentration. Diffusion is a result of the kinetic properties of particles of matter. The particles will mix until they are evenly distributed. Diffusion is the tendency of the solute to spread throughout the solution. Small molecules and ions move with sufficient velocity to distribute themselves throughout the solvent rapidly. On the other hand, macromolecules move slowly because of their high molecular weight. The rate at which a substance diffuses across a uniform crosssectional area depends not only on the molecular size and shape but also on the concentration gradient of the substance. In the absence of any other influencing factor, particles of matter move spontaneously from a region of high concentration toward one of lower concentration.

Fig. 1: Representation of Diffusion Osmosis is the passage of a solvent through a semi-permeable membrane. Such a membrane is permeable only to the solvent, not to the solute. Therefore, osmosis is the movement of solvent molecules through a selectively permeable membrane into a region of higher solute concentration, aiming to equalize the solute concentrations on the two sides. It may also be used to describe a physical process in which any solvent moves, without input of energy, across a semi permeable membrane (permeable to the solvent, but not the solute) separating two solutions of different concentrations. Although osmosis does not require input of energy, it does use kinetic energy and can be made to do work

Fig. 2: Representation of Osmosis Dialysis is a common biochemical method of separation and purification by selective passage of ions and small molecules through a semi-permeable membrane that will not allow proteins to pass through. Therefore, dialysis is the process of

separating molecules in solution by the difference in their rates of diffusion through a semi permeable membrane Surface Tension is the tendency of a liquid surface to contract. the cohesive forces among the liquid molecules are responsible for this phenomenon of surface tension. In the bulk of the liquid, each molecule is pulled equally in every direction by neighboring liquid molecules, resulting in a net force of zero. The molecules at the surface do not have other molecules on all sides of them and therefore are pulled inwards. This creates some internal pressure and forces liquid surfaces to contract to the minimal area. Surface tension is responsible for the shape of liquid droplets. Although easily deformed, droplets of water tend to be pulled into a spherical shape by the cohesive forces of the surface layer. In the absence of other forces, including gravity, drops of virtually all liquids would be perfectly spherical.

Fig. 2: Representation of Surface Tension

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Data and Interpretation: A. Diffusion Sample Reagent Added Time to produce a uniform color 2 min. and 2 seconds Explanation

Distilled Water

10% CuSO4

Diffusion is the movement of a fluid from an area of higher concentration t o an area of lower concentration. Therefore, it took 2 minutes and 2 sec for Copper Sulphate to achieve equilibrium in diffusion with distilled water. Diffusion is the movement of a fluid from an area of higher concentration t o an area of lower concentration. Therefore, a shorter time of 5 seconds were spent to achieve equilibrium.

10% Prussian blue

5 seconds

B. Osmosis

Sample

Reagent Added 0.9% NaCl

Observation Blood cells are moving fast.

Explanation Osmosis is a special case of passive transport. These blood cells have been placed in 0.9% NaCl.

Defibrinated Blood

0.1 M NaCl

Cells are moving slowly.

Osmosis is a special case of passive transport. These blood cells have been placed in 0.1 M NaCl. 0.585 grams NaCl is dissolved in 100 mL solution in preparing 1.0 M of NaCl. Therefore, it is less concentrated than with the 1.0 M NaCl. Thus, salt there can be a hypertonic solution. But lesser than the latter one. Osmosis is a special case of passive transport. These blood cells have been placed in 1.0 M NaCl. 5.85 grams NaCl is dissolved in 100 mL solution in preparing 1.0 M of NaCl. Therefore, it is more concentrated and it is not moving. Thus, salt there can be a hypertonic solution. The control group in an experiment is the group that nothing is done to. The control is left alone so that the other variables may be compared to it. The control is the variable that is usually the "normal"

1.0 M NaCl

Cells are not moving and are scattered and separated.

Control

Cells are of great number and are tightly compressed .

C. Dialysis Name of Test Test for Proteins Sample / Reagents Dialysate 25% Trichloroacetic acid Dialysate Fehlings Reagent Observation White precipitate was formed. Explanation Proteins are present due to the formation of white precipitate. Reducing sugar is present due to the formation of red and yellow precipitate.

Test for sugars

There is the formation of red and yellow precipitate.

D. Surface Tension Sample Distilled Water Reagents Added Chloroform Coalition Time 10.84 sec. or 11 sec. Explanation The cohesive forces among the liquid molecules are responsible for this phenomenon of surface tension. In the bulk of the liquid, each molecule is pulled equally in every direction by neighboring liquid molecules, resulting in a net force of zero.

Distilled Water

Chloroform Soap Solution

56 sec.

This explains that water and chloroform per se had lesser time of coalition compared to the one with soap. Remember that soap reduces, or even destroys the surface tension of water. Thus, it did take great coalition time when soap was introduced to the set-up.

Sample Bile Solution

Reagents Added Sulfur Powder

Observation The sulfur powder dissolved.

Explanation Sulfur is hydrophobic enough to float on the water. Therefore when it dissolved, it remained floating. The sulfur power was dissolved due to the principle like dissolves like. Sulfur exists as S8. S8 is 8 sulfur atoms covalently bonded in a ring. Each S atom has two lone pairs of electrons. The entire molecule is nonpolar.

Sulfur Powder

The powder did not dissolve and they join together at the center.

Sulfur is hydrophobic enough to float on the water. Sulfur exists as S8. S8 is 8 sulfur atoms covalently bonded in a ring. Each S atom has two lone pairs of electrons. The entire molecule is nonpolar. "Like dissolves like" so water which is polar will not dissolve S8 which is nonpolar. S8 dissolve in liquids like toluene, which is also nonpolar.

Questions: 1. What are the types of transport processes by which metabolites pass through biomembrane? Answer: Diffusion and Osmosis are the types of transport processes by which metabolites pass through biomembrane. Diffusion describes the spread of particles through random motion from regions of higher concentration to regions of lower concentration. The time dependence of the statistical distribution in space is given by

the diffusion equation. The concept of diffusion is tied to that of mass transfer driven by a concentration gradient. Osmosis is the movement of solvent molecules through a selectively permeable membrane into a region of higher solute concentration, aiming to equalize the solute concentrations on the two sides. It may also be used to describe a physical process in which any solvent moves, without input of energy across a semipermeable membrane (permeable to the solvent, but not the solute) separating two solutions of different concentrations. Although osmosis does not require input of energy, it does use kinetic energy and can be made to do work. 2. What is the effect of: a. hypotonic b. hypertonic c. isotonic salt solution on the red blood cells? Answer:

1. If the medium is hypotonic a dilute solution, with a higher water concentration than the cell the cell will gain water through osmosis. 2. If the medium is isotonic a solution with exactly the same water concentration as the cell there will be no net movement of water across the cell membrane. 3. If the medium is hypertonic a concentrated solution, with a lower water concentration than the cell the cell will lose water by osmosis.

1. How does the kidney maintain the bodys internal environment? Answer:

All organ systems (Brain, Heart, Lungs, Intestines, Liver, Muscles, Nerves and all other tissues (with the millions of natural chemical reactions that occur in them) could function only in an appropriate, very restrictive and sensitive 'internal environment'. Kidneys are the primary organs which sense any alterations in such internal environment and take appropriate corrective measure to restore it to normal. Components of Internal Environment: BLOOD PRESSURE OXYGEN DELIVERY TO TISSUES / ANEMIA WATER CONTENT OF THE BODY INTERNAL ENVIRONMENT INTERNAL CHEMICAL ENVIRONMENT HORMONES 1. Blood Pressure: All organs need to be supplied with blood at certain pressure. Maintaining such a perfusion pressure is dependent on Blood Pressure. Kidneys play a major role in maintaining and regulating Blood Pressure. When kidneys fail, such a regulation is not available and patient's Blood Pressure will increase, causing Hypertension. The resulting hypertension could further affect various organs. The heart is further stressed, since it has to pump against such a high pressure within the blood vessels. 2. Oxygen delivery to the organs and tissues: All organs and tissues need Oxygen. It is delivered to the those organs by the blood. Red blood cells (RBC) in the blood are the 'box cars' to transport the oxygen. If there is not enough RBCs then the oxygen delivery system will be inadequate. Kidneys monitor such 'trafficking of oxygen.' If kidneys sense that less oxygen is being transported, then they try to add more 'box cars' (RBCs) to the blood to carry more oxygen. The kidneys do it by producing more of a 'bone marrow stimulant' called Erythropoietin. Erythropoietin stimulates bone marrow to produce more RBCs. When the kidneys are diseased, such a regulatory function of the kidneys to ensure that there are enough 'box cars' (RBCs) to carry the oxygen is impaired. Consequently, there are not enough RBCs in the blood causing anemia.

normal. When the water content of the body increases the organs and tissues get 'water logged'; and when the water content is low the tissues and organs are 'dehydrated'. Both these abnormalities adversely affect the body and causes malfunction of various organs and tissues. Kidneys constantly monitor the water content of the body and maintain at normal levels to the extent feasible by limiting or increasing the elimination of water from the body in the form of urine. When the kidneys are diseased, such a regulatory function of the kidneys to ensure that the 'humidity' (Hydration) of the internal environment is maintained, is impaired. Consequently, more fluid could be lost via urine even when the body is dehydrated, further worsening the functions of all organs and tissues; or more fluid is retained due to decreasing urine volume, and cause dysfunction of organs and tissues as they get water logged. Such excess retention of water causes swelling of the body (legs, arms, face and abdomen). Such swelling is called edema. Since heart is the organ to pump the fluid (lymph and blood) around, such excess fluid retention could also cause heart failure (congestive heart failure). 4. Internal chemical environment: For proper various organs and tissues, the internal chemical environment monitored, regulated and maintained within a narrow variability. Some of the chemicals (Acid-Base, electrolytes) and ranges they need to be maintained in are: function of need to be range of the narrow

SODIUM POTASSIUM CHLORIDES BICARBONATE CALCIUM MAGNESIUM PHOSPHORUS

135 TO 145 mEq/L 3.5 TO 5.5 mEq/L 100 to 110 mEq/L 24 to 26 mEq/L 8.6 to 10 mgs/dl 1.6 to 2.4 mgs/dl 3.0 to 5.0 mgs/dl

URIC ACID pH CREATININE BUN Blood Urea Nitrogen)

2.5 to 6.0 mgs/dl 7.4 0.8 TO 1.4 MGS/DL 15 to 20 mgs/dl

Kidneys are the primary organs that monitor, regulate and maintain such a delicate and fragile internal chemical environment. They do this by adding or removing chemicals from the blood. At other times they add or remove hormones to and from the blood. For example, if the Calcium level is low, kidneys convert pro-vitamin D to active vitamin D. (For practical purposes, the only organ that generates the active vitamin D is the kidney). The active vitamin D then transfers calcium from the bone to the blood. It also makes the intestines absorb more calcium from the food and the blood calcium level is thus maintained on an ongoing basis. When the kidneys are affected this mechanism suffers and the blood levels of calcium could decrease (Hypocalcaemia) causing other complications. 5. Hormones: Many hormones that are secreted by various glands in the body (like Insulin, Thyroid etc.,) are removed by the kidneys after such hormones have served their purpose or when they are in excess.

2. Explain how soap lowers the surface tension of fats. Answer: Soaps and detergents are formed by special molecules, which have a hydrophilic head, which therefore loves to remain in water and a hydrophobic tail, which avoids water and loves fat substances . Because of their hydrophobic tail, a part of the molecules of detergent collects to the water surface forming a monomolecular layer , it lowers the surface tension of the water and makes easier its penetration into the fabrics to be cleaned. Within the water, the molecules of detergent collect themselves in micelles and membranes, little aggregates of molecules united by their hydrophobic tail. When they meet dirt, these molecules surround the particles and insert their tail in them. The hydrophilic heads attract the dirt toward water and with the

agitation of the liquid they contribute to remove the dirt from the fabric. The crown of hydrophilic heads carries the particles of dirt in the water , where they end up in suspension and then they are rinsed away. Hence, the dirt water contains also greasy particles which have been emulsified. For the same reason, the detergents aid the formation of emulsions. The substances which lower the surface tension of a liquid are called surfactants. The lowering of the surface tension of the water allows the formation of soapy membranes Soaps molecules' two ends make it able to break through the surface tension of water. The end of the detergent molecule which attaches to fat (grease) tries to avoid being around water molecules. It is known as hydrophobic, meaning water fearing. By attempting to move away from the water molecules, the hydrophobic ends of the detergent molecules push up to the surface. This weakens the hydrogen bonds holding the water molecules together at the surface. The result is a break in the surface tension of the water.

3. What is the role of bile salts during fat digestion? Answer: Bile is a complex fluid containing water, electrolytes and a battery of organic molecules including bile acids, cholesterol, phospholipids and bilirubin that flows through the biliary tract into the small intestine. There are two fundamentally important functions of bile in all species: Bile contains bile acids, which are critical for digestion and absorption of fats and fat-soluble vitamins in the small intestine. Many waste products, including bilirubin, are eliminated from the body by secretion into bile and elimination in feces.

Bile acts to some extent as a surfactant, helping to emulsify the fats in the food. Bile salt anions have ahydrophilic side and a hydrophobic side, and therefore tend to aggregate around droplets of fat (triglycerides and phospholipids) to form micelles, with the hydrophobic sides towards the fat and hydrophilic towards the outside. The hydrophilic sides are positively charged due to the lecithin and otherphospholipids that compose bile, and this charge prevents fat droplets coated with bile from reaggregating into larger fat particles. Ordinarily, the micelles in the duodenum have a diameter of around 14-33 m.

The dispersion of food fat into micelles thus provide a largely increased surface area for the action of the enzyme pancreatic lipase, which actually digests the triglycerides, and is able to reach the fatty core through gaps between the bile salts. A triglyceride is broken down into two fatty acids and a monoglyceride, which are absorbed by the villion the intestine walls. After being transferred across the intestinal membrane, fatty acids are reformed into triglycerides, then absorbed into the lymphatic system through lacteals. Without bile salts, most of the lipids in the food would be passed out in feces, undigested. Since bile increases the absorption of fats, it is an important part of the absorption of the fat-soluble substances, such as the vitamins D, E,K and A. Besides its digestive function, bile serves also as the route of excretion for bilirubin, a byproduct of red blood cells recycled by the liver. Bilirubin derives from haemoglobin by glucuronidation. The alkaline bile also has the function of neutralizing any excess stomach acid before it enters the ileum, the final section of the small intestine. Bile salts also act as bactericides, destroying many of the microbes that may be present in the food.

References: http://en.wikipedia.org/wiki/Diffusion http://chemistry.about.com/od/chemistryglossary/a/diffusiondef.htm http://en.wikipedia.org/wiki/Dialysis_(biochemistry) http://wiki.answers.com/Q/What_is_the_control_group_in_an_experiment http://en.wikipedia.org/wiki/Bile#Physiological_functions http://www.vivo.colostate.edu/hbooks/pathphys/digestion/liver/bile.html

How Does Detergent Break a Surface Tension? | eHow.com http://www.ehow.com/howdoes_5452223_detergent-break-surface-tension.html#ixzz1eF4JIqpn http://answers.yahoo.com/question/index?qid=20081123033646AAvta2i http://mbabu.com/Nephrology.htm#WHAT DO KIDNEYS DO?