Locker no: 26 Date Performed: February 24, 2011Member: Maila H. Aurestila

Josephine Ivy T. Jumala Ralph Renon T. Casamayor

Activity 11URINE

I. Introduction

Urine is a liquid waste excreted by the body. It is usually clear and transparent and has an amber color. Normally the average amount of urine excreted in 24 hours is from 40 to 60 ounces (about 1,200 cubic centimeters). It usually contains 960 parts of water and 40 parts of solid matter. Abnormal urine may contain sugar, albumin, bile pigments and abnormal quantities of its normal components (Urine, 2003).

Here are some physical characteristics of urine: Colour: Typically yellow-amber but varies according to recent diet and the concentration of the urine; Smell: Generally fresh urine has a mild smell but aged urine has a stronger odour, similar to that of ammonia; Acidity: 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; Density: also known as "specific gravity". Specific gravity of normal urine has a usual range of 1.015-1.025

An array of test is performed in urine which is known as urinalysis. This is commonly used to diagnose different kind of diseases, such as diabetes and ketunuria (Urine and Urinalysis (Composition of Urine), 2011).

In this experiment we will perform different test to analyze the urine sample composition or condition, thus the following test were performed: Test for urea, Uric acid test, Creatinine test, Test for the presence of glucose, Test for the presence of Acetone bodies, and Test for the presence of albumin. The physical characteristics of the urine sample were also examined.

The following objectives were formulated to serve as guide to the experiment:

1.) To analyzed and determine the different composition of the urine samples used2.) To determine the implications of the results to the health of the person who donated

the urine

II. Results and Discussion

Physical Characteristics

Description

Color Orange-yellowOdor Aromatic odor

Transparency TransparentpH 6.0

Actual Sp. Gr. 1.020

Corrected Sp. Gr.

The results above shows the physical characteristics that were obtained through different test and observations performed.

Color

Based on our observations, the color of the urine sample is yellow-orange. Compared to the normal urine, which has “amber yellow” in color, is due to the pigment called urochrome (urobilin + urobilinogen).

Studies show that the changes in the normal color of urine may be caused by disorders, diseases, recent food intakes and many other causes. Since the urine sample used has yellow-orange in color, it shows that it is not nomal. Based on resources, dark yellow or orange urine can be caused by recent use of laxatives or consumption of B complex vitamins or carotene. Orange urine is often caused by phenazopyridine (used in the treatment of urinary tract infections), rifampin, and warfarin.

Odor

The result shows that the odor of the urine sample is aromatic, which means that it is normal. Based on resources, urine may develop an ammoniacal odor, if stored longer, which is due to hydrolysis or urea to ammonia.

Transparency

The normal urine is clear and transparent when it is freshly voided. Based on our results, it shows that the transparency of the urine sample is normal because it is clear and transparent. If urine has a cloudy phase, it may contain different substances like mucus, leukocytes and ephithelial cells, which may have been caused by current diseases. It may also contain calcium phosphate or ammonium-magnesium phosphate if the urine is alkaline.

pH

Based on our results, the urine sample has a pH of 6.0. The pH level is obtained through pH paper. This shows that the urine sample has a normal pH level. The pH of normal urine is generally in the range 4.6 - 8, a typical average being around 6.0.

The pH level of urine may vary because of the food and water intake, drugs used or diseases that are currently present in the human body.

A high urine pH (alkaline urine) may indicate: Gastric suction, Renal or Kidney failure, Renal or Kidney tubular acidosis, Urinary tract infection, Vomiting

A low urine pH (acidic urine) may indicate: Chronic obstructive pulmonary disease (such as emphysema), Diabetic ketoacidosis, Diarrhea, Starvation

Specific Gravity

The specific gravity of urine may vary, depending on the solid and fluid intake of the person. Specific Gravity may be influenced by the macromolecules or pathological conditions that are present in the urine sample. The specific gravity of normal urine has a usual range of 1.015-1.025. Based on our results, it shows that the urine sample used has a 1.020 sp. Gr. This somehow shows that the urine sample contains substances that resulted to a high specific gravity, within the normal range.

Normal Constituents

Reagents Added Results

Urea(a)

(b)

Phenol RedDil. Acetic

Pink to Yellow

1% Urease YellowBaryta Mixture

AlcoholFormation of round

crystals

Uric Acid(a)

(b)

10% Na2CO3Benedict’s uric

Conc. HCl

Bluish green transparent + bluish

green cloudy

Crystals formed

CreatinineSat. Picric

10% NaOHYellow to red

The normal urine is composed mostly of water. It is also composed of organic molecules that are derived from the normal waste of the body. The constituents normally present in urine are the following: Urea, Uric Acid and Creatinine.

Test for Urea

Test A

Normal urine contains Urea. Urea is an organic (i.e. carbon-based) compound whose chemical formula is: CON2H4 or (NH2)2CO. It is also known as carbamide. Urea is derived from ammonia and produced by the deamination of amino acids. The amount of urea in urine is related to quantity of dietary protein.

Urea may break down into ammonia and carbon dioxide due to factors like bacteria and others. The ammonia present will result to the increase of pH of the urine. By using the phenol red, as a pH indicator, we have obtained the pH of the urine through observations. It shows that the urine sample turned to pink when it reacted to phenol red. The result shows that the solution is basic. After adding diluted Acetic acid, the solution turned to yellow. This shows that the urine sample is not very basic, because it immediately turned into acidic color, which is yellow.

When 1% urease is added, the urine sample turned to yellow. Urease is an enzyme which catalyzes the hydrolysis of urea to ammonia and carbon dioxide. The result shows that when urease is added, there is no increase in pH. This only shows that the urine contains less urea which resulted to a lesser amount of ammonia produced. If there is low ammonia produced, the pH of the urine will be lower than the basic pH.

Test B

The quantitative determination of urea, founded on the property that its solutions undergo decomposition in closed vessels at a temperature from 1200 to 2400. The carbonic acid which is thus formed is combined with baryta and the amount of urea is calculated from that of carbonate of baryta. The formation of crystals indicates the amount of urea present in the urine samples.

Test for Uric Acid

Test A

Uric acid is an organic (i.e. carbon-based) compound whose chemical formula is: C5H4N4O3. Due to its insolubility, uric acid has a tendency to crystallize, and is a common part of kidney stones.

The first test involves the test for the presence of uric acid in the urine sample. Using the benedict’s uric acid reagent, the presence of uric acid can be determined. Benedict’s uric acid reagent is also known as phosphotungstic acid. Uric acid has the ability to reduce phosphotungstic acid to a blue-phosphotungstate complex. Since our urine sample turned to blue, then uric acid is present in the urine.

Test B

The addition of a concentrated acid may trigger or accelerate the formation of uric acid crystals in the urine. The result shows that crystals were formed on the urine sample. Uric acid crystals may indicate that the person’s kidney is still functioning properly, due to the high amount of crystals produced. Because if only few crystals were produce the kidney was not able to remove this waste from the body.

Test for Creatinine

Creatinine is a waste product that the muscles produce at a steady rate as part of normal daily activity. The bloodstream carries creatinine to the kidneys, which filter it out of the blood, and it is passed out of the body in urine. This gives doctors information about how well the kidneys are functioning. In the Jaffe reaction, creatinine is treated with an alkaline picrate solution to yield a bright orange-red complex. This means that if the sample yields an orange –red complex then the result is positive for the presence of creatinine.

Creatinine is removed from the body entirely by the kidneys. If kidney function is abnormal, creatinine levels will increase in the blood (because less creatinine is released through your urine).

Based from our results creatinine is present in the urine sample. Thus the kidney of the person with the urine sample is functioning well.

Pathological Constituents

Results with Benedict’s Reagent

GlucoseNormal Sample Trace

w/ 0.2% Glucose Tracew/ 0.5% Glucose Tracew/ 1.0% Glucose Green Yellow (+)

Urine also contains different organic compounds that are used by the body. The glucose, which is urine constituent, may also be present in the normal urine, but in very small amount, that it is difficult to detect.

Based on our observations, the urine sample used contains glucose. Benedict’s test has been used to detect the presence of glucose. Benedict’s test is known to be a general test for carbohydrates. The results were compared to different samples containing different concentrations of glucose. This shows that our normal urine sample contains traces of glucose.

There are many indications if sugars, particularly glucose, are present in our urine. The presence of glucose in urine may be a sign of diabetes mellitus, pregnancy etc.

Results w/ Rothera’s testAcetone Bodies

Normal Sample Yelloww/ Acetone Purple color produced

The test above shows the clinical test for the presence of ketone compounds in the urine. By using sodium nitroprusside and ammonium hydroxide, ketone may be detected.

Based on our observations, it shows that our urine sample do not contain acetone or any ketone compounds. The presence of purple color in the test indicates the presence of ketone compounds. Since, a yellow color is produced; this means that our urine sample is negative of ketone substances.

Results with Heat Coagulation Test

AlbuminNormal Sample No coagulation

w/ 0.2% albumin ++w/ 0.5% albumin +++w/ 1.0% albumin ++++

We have learned that protein can be denatured by applying high amount of heat energy. Protein may be denatured, and may appear as a cloudy or a coagulate substance after heating. Albumin, which is a protein substance, is also a constituent of urine. Albumin is present in normal urine in very small amount.

In previous experiments, it has been shown that heat coagulation may be used to determine the presence of albumin. It is also been proven that an increase in albumin content may also increase the degree of coagulation. High amounts of albumin present in the urine, indicates that the kidney already has problems.

The result above shows that our urine sample contains no albumin because it did not produce coagulates or cloudy substances.

III. Conclusion

In this experiment, we tested for the different characteristics and composition of the urine sample. The different physical characteristics of urine are color, odor, transparency, pH, and specific gravity. Among these characteristics the urine sample was all normal except for the characteristic of color. Instead of amber yellow, the urine sample is color orange yellow, this indicates that the person might have ingested Vitamin B complex, or may have certain diseases.

Based from our results the presence of Urea, Uric Acid and Creatinine, in the urine sample is positive. The kidney if functioning properly will yield these substances out with the urine, because they are waste materials which needed to be excreted.

For the presence of glucose, the urine sample used is positive for the presence of glucose but only a trace; this can be compared to the two other samples, 0.2 % and 0.5% glucose, which also resulted in a trace. Thus the urine sample might contain 0.2 % to 0.5% glucose.

The presence of a purple color indicates the presence of ketone bodies. The urine sample yielded a yellow color thus, the result is negative.

The last test is the test for the presence of albumin in the urine, this will be indicated by the coagulation within the sample, after being heated. The greater the albumin present in the urine, the greater the degree of the coagulation. The sample used did not exhibit the presence of albumin, thus the kidney is still functioning.

IV. Suggestions and Recommendations

In this experiment, we have noticed that there were too many tests and our urine sample was not enough, thus we would like to recommend to students who will do the same experiment to drink lots of water, as to provide adequate amount of urine for the different tests.

V. Answers to Questions

1.) What is the end product of protein metabolism in men?

According to Gita Patel (2011) Digestion breaks protein down to amino acids. If amino acids are in excess of the body's biological requirements, they are metabolized to glycogen or fat and subsequently used for energy metabolism. If amino acids are to be used for energy their carbon skeletons are converted to acetyl CoA, which enters the Krebs cycle for oxidation, producing ATP. The final products of protein catabolism include carbon dioxide, water, ATP, urea, and ammonia.

2.) Describe the Urea Cycle.

Living organisms excrete excess nitrogen as ammonia, uric acid, and urea. The urea cycle takes place in the mitochondria and the cytosol. There are five enzymes involved and these are carbamoyl phosphate synthetase (CPS I), ornithine transcarbamoylase, argininosuccinate synthetase, arginase and argininosuccinase. Two ATP is also required. The urea cycle ultimately yield 6 ATPs (Lady-chiquita, 2011). 

3.) What is the end product of purine metabolism?

In 1997, Carol N. Angstadt, tell us that purines consist of a six-membered and a five-membered nitrogen-containing ring, fused together. The end product of purine catabolism in man is uric acid. Uric acid is formed primarily in the liver and excreted by the kidney into the urine.

4.) What are the ketone bodies? How are they formed?

Ketone Bodies are chemicals that the body makes when there is not enough insulin in the blood and it must break down fat for its energy. The three ketone bodies are acetoacetate, beta-hydroxybutyrate and acetone, although beta-hydroxybutyrate is not technically a ketone but a carboxylic acid. Ketone bodies can poison and even kill body cells (Ketone bodies, 1998).

In liver mitochondria, some acetyl-CoA is converted to acetoacetate, beta-hydroxybutyrate, and acetone, collectively called ketone bodies (Croston, PhD, n.d.)

5.) What are the conditions leading to ketosis?

Ketosis is a condition in which levels of ketones (ketone bodies) in the blood are elevated. Ketones are formed when glycogen stores in the liver have run out. The ketones are used for energy. Ketones are small carbon fragments that are the fuel created by the

breakdown of fat stores. Ketosis is potentially a serious condition if ketone levels go too high (Nordqvist, 2010). 

6.) What is a “threshold substance”? Give examples.

A threshold substance is a substance excreted by the urine only when its plasma concentration exceeds a certain value (biology-online.org, 2005). Examples are sodium chloride, amino acids, potassium, creatine, and ascorbic acid.

7.) What is Bence Jones Proteins?

Bence Jones proteins are small proteins (light chains of immunoblobulin) found in the urine. Bence Jones proteins are considered the first tumor marker (Pagana, K. D., 1998).  It is a low-molecular-weight, heat-sensitive urinary protein found in multiple myeloma, which coagulates when heated to 45°–55°C and redissolves partially or wholly on boiling.

VI. References

Angstadt, C. N., PhD. (1997). Purine Metabolism. Retrieved on March 6, 2011 athttp://library.med.utah.edu/NetBiochem/pupyr/pp.htm

Croston, G., PhD. (n.d.) Ketone bodies pathway. Retrieved on March 6, 2011 at http://www.biocarta.com/pathfiles/ketonebodiesPathway.asp

Ketone Bodies (1998). Retrieved on March 6, 2011 at http://www.paralumun.com/diabetesketone.htm

Lady-chiquita (2011). Urea Cycle. Retrieved on March 6, 2011 at http://hubpages.com/hub/Urea-Cycle

Nordqvist, C. (2010). Ketosis. Retrieved on March 7, 2011 at http://www.medicalnewstoday.com/articles/180858.php

Pagana, K. D. (1998). Mosby's Manual of Diagnostic and Laboratory Tests. St. Louis: Mosby, Inc., Retrieved on March 5, 2011 at http://medical-dictionary.thefreedictionary.com/Bence+Jones+protein+test

Patel G. (2011). Retrieved on March 5, 2011 at

 http://www.faqs.org/nutrition/Met-Obe/Metabolism.html#ixzz1G1BMPxm9

Threshold Substance (2005). Retrieved on March 6, 2011 at http://www.biology-online.org/dictionary/Threshold_substance

Urine, (2003). Retrieved on March 5, 2011 at http://www.wordiq.com/urine

Urine and Urinalysis (Composition of Urine), (2011). Retrieved on March 5, 2011 at http://www.ivy-rose.co.uk/HumanBody/Urinary/Urinary_System_Composition_Urine.php