URINE INGREDIENT ANALYSIS

Name: Yunita Punarisma Class: XI IA 3/ 15

BIOLOGY LABORATORY STELLA DUCE SENIOR HIGH SCHOOL YOGYAKARTA

I.

IntroductionUrine is a liquid product of the body that is secreted by the kidneys by a process called urination and excreted through the urethra. Cellular metabolism generates numerous waste compounds, many rich in nitrogen, which require elimination from the bloodstream. This waste is eventually expelled from the body in a process known as micturition, the primary method for excreting water-soluble chemicals from the body. These chemicals can be detected and analyzed by urinalysis. Amniotic fluid is closely related to urine, and can be analyzed by amniocentesis. Urine formation begins with the movement of plasma ultra filtrate into the kidneys. This plasma ultra filtrate is an essentially protein-free fluid which passively passes from the glomerular capillaries into the Bowman's space. This process is driven by Starling forces. Glomerular filtration is followed by reabsorbtion of water and solutes from the different parts of the renal tubules, then by the secretion of selected solutes into the renal tubules.

II.

Problems:a. What are the physical and chemist feature in urine? b. What are the substances kinds that implied in urine?

III. Objectives:a. To detect physical feature and chemist feature in urine. b. To detect kinds of substances that implied in urine.

IV. Literature

Blood Cleaning by the Kidneys

This section is about processes performed by the kidneys in order to filter (clean) blood. These are: 1. Glomerular Filtration also called "Ultra-filtration", 2. Tubular Reabsorption also called "Selective Re-Absorption" and 3. Tubular Secretion. (1) Glomerular Filtration Blood enters the kidney via the renal artery. This separates many times (Renal Artery -> Segmental Arteries -> Interlobar Arteries -> Arcuate Arteries -> Interlobular Arteries -> Afferent Arterioles), eventually forms manyafferent arterioles, each of which delivers blood to an individual kidney nephron. The diameter of the afferent (incoming) arteriole is greater than the diameter of the efferent arteriole (by which blood leaves the glomerulus). The pressure of the blood inside the glomerulus is increased due to the difference in diameter of the incoming and out-going arterioles. This increased blood pressure helps to force the following components of the blood out of the glomerular capillaries:y y y y

Most of the water; Most/all of the salts; Most/all of the glucose; Most/all of the urea.

The above are filtered in preference to other components of blood based on particle size. (Water and solutes of relative molecular mass less than 68,000 form the filtrate.) Blood cells and plasma proteins are not filtered through the glomerular capillaries because they are relatively larger in physical size. The water and salts that have been forced out of the glomerular capillaries pass into the Bowman's Capsule and are called the glomerular filtrate. This glomerular filtrate is formed at a rate of above 125 cm3 per minute in humans. This volume is approx. 20% of the plasma delivered during that time. (Again: It contains all the materials present in the blood except blood cells and most proteins - which are too large to cross the basement membrane of the glomerulus.)

(2) Tubular Reabsorption Only about 1% of the glomerular filtrate actually leaves the body because the rest (the other 99%) is reabsorbed into the blood while it passes through the renal tubules and ducts. This is called tubular reabsorption and occurs via three mechanisms. They are:y y y

Osmosis Diffusion, and Active Transport.

Reabsorption varies according to the body's needs, enabling the body to retain most of its nutrients. The processes of tubular reabsorption occur in the following order : In the PCT Most of the volume of the filtrate solution is reabsorbed in the proximal convoluted tubule (PCT). This includes some water and most/all of the glucose (except in the case of diabetics). Most of the energy consumed by the kidneys is used in the reabsorption of sodium ions (Na+), which are solutes - that is, they are dissolved in the water component of the filtrate solution. As the concentration of Na+ in the filtrate solution are high (about the same as the concentration of Na+ in blood plasma), Na+ moves from the tubular fluid into the cells of the PCT. In the cases of many Na+ ions this occurs with the help of symporters. Symporters simultaneously facilitate passage through the PCT membrane of both Na+ and other substances/solutes. Other such substances that are reabsorbed with Na+ in this way include glucose (an important type of sugar), amino acids, lactic acid, and bicarbonate ions (HCO3-). These then move on through cells via diffusion and/or other transport processes. A short way to summarize the above is to say that solutes are selectively moved from the glomular filtrate to the plasma by active transport. (However, almost all glucose and amino acids, and high but variable amounts of ions, are reabsorbed again later - see the next section, below). Following the movement of solutes (including Na+), water is then also reabsorbed by osmosis. About 80% of the filtrate volume is reabsorbed in this way. As this part of the reabsorption process is not controlled by the proximal tubule itself, it is sometimes called obligatory water reabsorption. In the Loop of Henle: the remaining water (together with the dissolved salts and urea) passes from the PCT into the descending limb of Henle. It then passes along the Loop of Henle, and up the ascending limb of Henle. The different permeability properties of the two limbs of the Loop of Henle, together with their counter flow arrangement, allows a countercurrent multiplication to generate a high solute concentration in the tissue fluid of the medulla (that is, outside of the tubules). The highest solute concentrations are generated deep in the medulla. This is explained as follows:

1. Descending Limb of Loop of Henle The epithelium lining of the descending limb of Henle is relatively permeable to water - but much less permeable to the salts Na+ and Cl-, and to urea. Therefore water gradually moves from the descending limb and into the interstitium (surrounding the tubules) as fluid flows through this part of the system of renal tubules. 2. Thin Ascending Limb of Loop of Henle The thin ascending limb of Henle differs from the descending limb in that it is impermeable to water (so the water that is inside the tubule at this stage generally remains inside it), but is highly permeable to Na+ and Cl-, and somewhat permeable to urea. Therefore while the tubular fluid flows back towards the renal cortex, Na+ and Cl- (which are more concentrated in the tubular fluid than in the interstital fluid) diffuse from the tubules into the interstitium. Some urea also enters the tubules at this stage - but the loss of NaCl from the tubular fluid greatly exceeds the gain in urea. 3. Thick Ascending Limb of Loop of Henle the thick ascending limb of Henle (and its continuation into the first part of the DCT), reabsorbs NaCl from the tubular fluid via a different transport process from that of the thin ascending limb of Henle. The overall effect of the processes outlined above is that the concentration of the fluid inside the renal tubules that form the Loop of Henle is highest at the deepest part of the renal medulla, and is less concentrated in the renal cortex. This is what is meant by the "concentration gradient" of the Loop of Henle. The term "counter-current" is also used in descriptions of the Loop of Henle - and refers to the tubular fluid flowing in opposite directions along the descending and ascending limbs (as indicated by the thin red arrows in the diagram above. In the DCT The water, urea, and salts contained within the ascending limb of Henle eventually pass into the distal convoluted tubule (DCT). The DCT reacts to the amount of anti-diuretic hormone (ADH) in the blood:y

The more ADH is present in the blood, the more water is re-absorbed into it. This happens because the presence of ADH in the blood causes the cells in the last section of the DCT (and associated tubules and collecting ducts) to become more permeable to water, therefore they allow more water to pass from the tubular fluid back into the blood. This results in more concentrated urine. The opposite is also true, i.e. if the level of ADH in the blood is reduced then the cells in the latter sections of the DCT (and associated tubules and collecting ducts) becomes less permeable to water therefore less water is able to pass from the tubular fluid back into the blood - which results in less concentrated urine.

y

The amount of ADH in the blood may be affected by conditions such as diabetes insipidus, or by consumption of diuretics* in the diet (*substances that occur in some foods and drinks).

(3) Tubular SecretionThe third process by which the kidneys clean blood (regulating its composition and volume) is called tubular secretion and involves substances being added to the tubular fluid. This removes excessive quantities of certain dissolved substances from the body, and also maintains the blood at a normal healthy pH (which is typically in the range pH 7.35 to pH 7.45). The substances that are secreted into the tubular fluid (for removal from the body) include:y y y y y y y

Potassium ions (K+), Hydrogen ions (H+), Ammonium ions (NH4+), Creatinine, urea, some hormones, and Some drugs (e.g. penicillin).

Tubular secretion occurs from the epithelial cells that line the renal tubules and collecting ducts. It is the tubular secretion of H+ and NH4+ from the blood into the tubular fluid (i.e. urine - which is then excreted from the body via the ureter, bladder, and urethra) that helps to keep blood pH at its normal level. The movement of these ions also helps to conserve sodium bicarbonate (NaHCO3). The typical pH of urine is about 6. Urine formed via the three processes outlined above trickles into the kidney pelvis. At this final stage it is only approx. 1% of the originally filtered volume but includes high concentrations of urea and creatinine, and variable concentrations of ions. The typical volume of urine produced by an average adult is around 1.5 - 2.0 dm3 per day.

Urine physical characteristics: Volume is one of the physical characteristics of urine. Other physical characteristics that can apply to urine include color, turbidity (transparency), smell (odor), pH (acidity - alkalinity), and density.y

Color: Typically yellow-amber but varies according to recent diet and the concentration of the urine. Drinking more water generally tends to reduce the concentration of urine, and therefore cause it to have a lighter color. (The converse is also true.)

y

Smell: The smell (or "odour", which is the more clinical term, American spelling "odor") of urine may provide health information. For example, urine of diabetics may have a sweet or fruity odour due to the presence of ketones (organic molecules of a particular structure). Generally fresh urine has a mild smell but aged urine has a stronger odour, similar to that of ammonia. Acidity: pH is a measure of the acidity (or alkalinity) of a solution. The pH of a substance (solution) is usually represented as a number in the range 0 (strong acid) to 14 (strong alkali, also known as a "base"). Pure water is "neutal" in the sense that it is neither neither acid nor alkali, it therefore has a pH of 7. The real significance of pH in terms of physical chemistry is that pH is a measure of the activity of hydrogen ions (H+) in a solution. The pH of normal urine is generally in the range 4.6 - 8, a typical average being around 6.0. Much of the variation is due to diet. For example, high protein diets result in more acidic urine, but vegetarian diets generally result in more alkaline urine (both within the typical range 4.6 8). Density: Density is also known as "specific gravity". This is the ratio of the weight of a volume of a substance compared with the weight of the same volume of distilled water. Given that urine is mostly water, but also contains some other substances dissolved in the "water", its density is expected to be close to, but slightly greater than, 1.0. This is true - the density of normal urine is in the range 0.001 to 0.035.

y

y

V. Hypothesis:a. Physically, urine`s color should be yellow (pH=4,8 7,5) b. Substances, that may implied in urine: y y y y y y y y NaCl KCl C6H12O6 Protein Ion ClIon PO43+ Ion SO42Ion H2O

y y VI.

2% Urea 2% other metabolism product

Tools and Materials: Tools: y y y y y Reaction tube Pipette Chemistry glass Bunsen Lamp Test Tube Clamp

Materials: Urine 250 ml every student Liquid AgNO3 Liquid HCl Benedict Fehling Paint color indication Acetate acid Ammonium oxalate Sugar Water Label

VII.

Procedures: (1) Color and Smell Test: Take 5 ml urine and pack into reaction tube.

Place in an available place. Check the color urine of available standard urine`s color. Note the result. Smell the urine and note the result. (2) pH measurement: Take 5 ml of urine and pack it into reaction tube. Insert paper pH or pH stick into urine during 30 second. Lift pH stick and check the color with color indicator in box pH of pH stick. Note the result. (3) Dilution measurement: Take 5 ml urine and pack into reaction tube. Keel over reaction tube up to flat Count the time urine that gets down to genuinely finished by animate stopwatch moment urine begin to drip and kill stopwatch when urine genuinely finished. Note the result. (4) Urine ingredient analysis: Glucose Analysis Sugar solution test with solution Benedict as color change standard that happened. Its way, make sugar solution from one sugar tablespoon that melted into half water glass. Put 20 gram of sugar solution drops into reaction tube. Drop the sugar solution with 5ml solution of Benedict, shake it until dissolved. Clamp the reaction tube with pincers. Heat the end of tube reaction above Bunsen lamp`s flame. Every second u take the reaction tube up, so it won`t burnt. Not until boiling, in 3-5 minutes. Attention the color changes. If it`s contains glucose, then it should appeared a red brick color.

The, test urine with the same procedure started from second point, we only change the glucose solution with urine. Better be students are checking their own urine narrowly. Protein analysis: Put 5 ml of urine into the reaction tube. Add 5 ml of Biuret. Attention the color`s changing (if reddish or purple colors mean it is positively contains protein) Note the result. Ion NH4 analysis: Put 2 ml of urine. Add 5 drops of urine into Na2CO3 and placed the litmus paper above the tube. Heat carefully If the red litmus color changed into blue, means there was some reaction 2 NH4Cl + Na2CO3 (NH4)2CO3 + H2O 2 NaCl + (NH4)2CO3 2 NH4OH + H2CO3

Ion Ca test: Add 1 ml of acetic acid and 1 ml of ammonium oxalate into 5 ml of urine. Attention the appearing of colorless sediment (calcium oxalate) Note the result. Ion Mg test: Solution that contained a calcium oxalate from the result of the test above was continued by heating it until boil. Filter it in hot state with filter paper.

Take the filtrate and add 2 ml of ammonium until smell something typical. Attention the ammonium magnesium ammonium phosphate (MgNH4PO4) Ion Cl test: Add 3 ml of AgNO3 into 5 ml of urine. Attention the sediment. Note the result. Ion PO4 test: Add ammonium molybdate then add 1 ml of concentrated HNO3. Heat until boiled. Attention the yellow sediment. Note the result. Ion SO4 test: Add 3 ml of BaCl2 and 2 ml of sloppy HCl into 5 ml of urine. Attention the sediment Note the result. (5) Pregnancy test: Patch all urine in a clean and dry place. Ripped the sachet and take the test`s tool. Put the end of absorbent test tool in urine container in 5 seconds (don t run over the narrow sign that located in the test`s tool) The test result can be read in 3 minutes. Procedure to read the test result: Positive result is when there appeared two color lines or if the color line of the test area (T) was darker of equal than in control area (C). Negative result is when there appeared one color line or if the color line of the test area (T) was lighter than the red color line in control area or (C)

VIII.

Observation data: A. Physical and chemist characteristic of urine.

NameFani Feli Tami

Color+++ + ++

pH6 6 6

Smell++ + +++

Dilution20 s 10 s 10 s

B. Substances that contained in urine. No 1 2 3 4 5 6 7 8 Substance tested Glucose Protein NH4+ Ca+ Mg+ ClPo4SO4Fani + + Feli + + + Tami + + Explanation Unchanged Unchanged Litmus turned into blue No sediment No sediment Color changing and sediment No yellow sediment White sediment

C. Pregnancy test. Fani Feli Tami -

IX.

Discussion

Physical and chemist characteristic of urine. Color of urine: Fani got +++, Feli +, and Tami ++. The more + they have means darker urine`s colors they have. Colors of humans are all different to each others. Each of them has their own information of what substances that contained in their body. The three main causes of abnormal urine color are: Foods you have eaten, Drugs you have taken, Health issues. The most common colors are: y Orange: There are many foods that may cause an orange urine color. Blackberries can give your urine orange hue. Beets may give your urine a red or orange urine color.

Rhubarb can cause an orange urine color as well. Senna herbs have also been known to cause an orange color in urine. Other foods or medications with orange dye may cause an orange urine color. There are some serious health issues to consider that may cause orange urine. Dehydration causes a darker urine color that may sometimes appear to be orange urine. Jaundice can cause bile to show up in your urine. The bile can give urine an orange color. y Light yellow: A light yellow urine color typically signifies very healthy urine. The light straw yellow urine will have a low odor, few bubbles in the urine, little to no foam in urine, and have a clear urine consistency. So, we can conclude that the healthiest urine above three people is Feli`s urine. pH of Urine: between three people, they have the same pH of urine in 6. Means they have acidity of solution. y Urine pH is used to classify urine as either a dilute acid or base solution. Seven is the point of neutrality on the pH scale. The lower the pH, the greater the acidity of a solution; the higher the pH, the greater the alkalinity. The glomerular filtrate of blood is usually acidified by the kidneys from a pH of approximately 7.4 to a pH of about 6 in the urine. Depending on the person's acid-base status, the pH of urine may range from 4.5 to 8.

Smell of urine: Fani got ++, Feli got +, and Tami got +++. Actually, smell doesn t give you any information accurately from your body. It is mostly the result of hat you have eaten. But if you have smelled some sting odor frequently, you better check to doctor to see hat happened. Urine`s dilution: Fani had 20 s, Feli 10 s, and Tami 10 s. The faster urine to flow, the dilute urine that they have. Te more dilutes urine means you are not thirsty and have no dehydration. It is good, because you also have a good kidney. Urine`s composition Glucose: we test them by using Benedict that had been heated with urine fro 3-5 minutes. We have the result all negative. We didn t find any color changing in reaction tube. Means they have no glucose in

their urine. This is good, cause glucose containing in urine means they have some disease that caused by glucose contents inside. It prevents them from being diabetes disease. Protein: in protein test, same like glucose, we didn t find any color`s changing in reaction tube. Proteins are large molecules which help make up our muscles, important parts of our immune system, and many other portions of our bodies. Most proteins are also too large to pass through the filtering system of the kidney. And since they are not supposed to pass into the kidney, there is no mechanism for proteins to be reabsorbed if they make it in there. Therefore, if protein is detected in the urine, it means there is something going on with the filter (called the glomerulus) that is allowing the proteins to pass. So, we can conclude that students have no problems with their internal excretion organs. NH4+: we only have 1 student that positive contained ammonia in her urine test, who was Feli. As the literature shown above, ammonia should contain in urine. Because, the third step of kidney`s procedure (tubular secretion) the ammonia is removed from the blood flow. It is also caused the odor of the urine. The students that have no ammonia in their urine may have some trouble with their tubular secretion. Ca+: we didn t find any sediment inside the reaction tube. Means the students have no calcium oxalate in their body. Calcium oxalate crystals in the urine are the most common constituent of human kidney stones, and calcium oxalate crystal formation is also one of the toxic effects of ethylene glycol poisoning. The urine picture can be shown below as

my opinion supports. Mg+: in our experiment, we again had no found any sediment that indicates any magnesium inside the urine. Though actually, magnesium in urine means that the person is in health conditions. So magnesium in our body is so important because Magnesium is needed for more than 300 biochemical reactions in the body. It helps maintain normal muscle and nerve function, keeps heart rhythm steady, supports a healthy immune system, and keeps bones strong. Magnesium also helps regulate blood sugar levels, promotes normal blood pressure, and is

known to be involved in energy metabolism and protein synthesis. There is an increased interest in the role of magnesium in preventing and managing disorders such as hypertension, cardiovascular disease, and diabetes. Dietary magnesium is absorbed in the small intestines. Magnesium is excreted through the kidneys.

Cl-: we have all students positively have Chlorine in their test urine. Because the color changing and sediment appearing. And it should be appearing because, the reabsorption do remove chlorine from blood because it has no use for body anymore. We can conclude that there was no problem with their glomerulus. PO4-: we have all negative result for phosphate contents in the reaction. Because we didn t see any sediment inside the reaction tube. It is good, because phosphate contents indicate a crystal in kidney that can cause kidney stone. SO4-: we have all positive result, because there was white sediment in reaction tube. Sulfate should be removed from the blood from reabsorption.

Pregnancy test: in this test, we have all negative result because appeared one color line or if the color line of the test area (T) was lighter than the red color line in control area or (C) X. Conclusion: 1. Urine`s physical and chemist feature: y Physical: urine`s color is yellow - dark yellow, it`s dilution, and has a typical smell from ammonia. Chemist: urine has pH between 4.8 and 7.5. Otherwise, is the health pH one.

y

2. Urine`s composition: glucose and calcium oxalate only exist when the person got some infection or problems with their excretion organs. y y y y NH4+ ClSO4Mg+

y XI. References: y y y

PO4 3-

http://en.wikipedia.org/wiki/Sulfate http://en.wikipedia.org/wiki/Urine http://id.wikipedia.org/wiki/Fosfat

Guide,

Yogyakarta, May 3rd, 2010 Student,

Mr. Gun

Yunita Punarisma