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URINE

Sterile, liquid by-product of the body that is secreted by the kidneys. Cellular metabolism generates numerous by-products that require elimination from the bloodstream. These by-products are eventually expelled from the body in a process known as micturition.

Activity 26: Physical Properties of Urine

COLOR straw to amber *Colorless to straw indicates low specific gravity and large quantity *Amber indicates high specific gravity and small quantity

ODOR aromatic - depends on diet and drugs used Cloudiness: -freshly voided is clear (usually) but cloudy sometimes due to the following sediments: Amorphous phosphates form white cloud/precipitate Amorphous urates form a white/pink cloud of sediment Epithelial cells and mucus when large in amounts Pus makes urine turbid Fat render urine turbid Bacteria - produce uniform cloudiness

Abnormal (Pathological Coloration) Reddish amber increased in urobilinogen Brownish yellow/green indicate bile pigments Red to smoky brown due to blood and blood pigments Milky due to large amounts of pus, bacteria/fat Brownish black indicate melanin

pH -4.8-7.5 (Average: 6) 24 Hr. Specimen are less acidic than freshly passed and may become alkaline after standing Pathological Significance: Acidity - increased in acidosis, fevers, diet with excess proteins Alkaline in chronic cystitis and urine retention due to decomposition of urine in bladder

SPECIFIC GRAVITY (24 Hr. Specimen) -1.015 to 1.025 COMPOSITION -H2O: about 1000-1500g -Solids: about 55-70g in 24 hours Nitrogenous organic compounds Non-nitrogenous organic substances Inorganic salts Traces of Fe, Cu, Zn, Cl..etc.

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Average volume: 1-2 L a day

1.

Why does urine become turbid on standing? flocculate of molecules composed of nucleoprotein mucoid precipitation of calcium phosphate decomposition of urea to form ammonia fat globules or pus cells infection in the urinary tract

ABNORMAL

Polyuria = a condition of excessive production of urine Oliguria = decreased in quantity Anuria = total suppression of urine

2. Explain the alkaline fermentation of urine.

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3. What is the importance of determining the specific gravity of urine? rough estimate of the total solids in the urine provide information of the state of hydration of the patient - if highly concentrated, the patient is dehydrated determine the presence of renal disease helps distinguish between renal failure and dehydration determines the concentrating ability of the kidneys

Ammoniacal fermentation the conversion of the urea of the urine into ammonium carbonate, through the growth of bacteria. CON2H4 + 2H2O (NH4)2CO3 Note: Whenever urine is exposed to the air in open vessels for several days it undergoes this alkaline fermentation.

4. Why must the examination be done on a 24-hour specimen? if the complete examination of the constituents of the urine is needed notably better than that exceeding 24-hours because: - microorganisms may grow - convert urea to ammonium carbonate giving a more pungent ammoniacal odor than normal - nitrogenous constituents are altered by the loss of nitrogen as volatile ammonia

5.

What are the substances responsible for the normal color of urine? urochrome gives the normal yellow color is due to a pigment metabolite arising from the body's destruction of hemoglobin. uroerythrin urinary pigment that gives a redish yellow color to deposits of urates. urobilinogen when oxidized gives a darker or deeper yellow color

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6. Outline the origin of the color of urine and feces.

Hemoglobin Heme Biliverdin + 2H (reduction) Bilirubin +2H Mesobilirubinogen D-Urobilinogen +2H -2H
Stercobilinogen (L-urobilinogen)

+ Protein (Globin)

Activity 27: Constituents of Urine

Urobilin

-2H (auto-oxidation) D-Urobilin

-2H
Stercobilin (L-urobilin)

A.1. Sulfate white ppt. A.2. Chloride white ppt. A.3.b. Alkali Phosphate white ppt.

1 Urine Constituents

3.b.

What is triple phosphate?

- Ammonium magnesium phosphate

A.4.a. Addition of 1 ml. of ammonium thiocyanate - Formation of red color A.4.b. With 5 drops of potassium ferrocyanide - Blue solution

Sources of phosphate in urine: - breakdown of tissue, foods rich in phosphoproteins, phospholipids and nucleoproteins

3.a. Triple Phosphate

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How does urea become a component of urine?

It is the pricipal and product of protein metabolism and constitutes about 80% - 90% total nitrogen excretion.

B.1. Urea Crystals

B.2.a. Uric Acid Crystals

B.2.b. Result: Formation of Black spot Uric acid - Reducing property How does it become a component of urine?

-Uric acid is the final end product of purine oxidation in the body

3.a. Weyls Reaction Formation of Red color

3.b. Jaffes Reaction Dark orange - red solution

How does creatinine become a component of urine? - As a product from metabolism in the muscle tissues.

How does it become a component of urine?

- C8H6NOSOK -The potassium salt of indoxylsulfate (indican) found in urine is a result of bacterial action on tryptophan in the bowel.

4. Obermeyers Test blue color

5.a. Hippuric Acid Crystals

Odor:

6. Oxalic Acid Crystals

Irritating odor

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C.PATHOLOGICAL

Result brick red ppt. White ring b/n two liquids Red purple Blue green color
Acetone Albumin

7. Pigments Result White ppt. What pigments are usually present in urine?
Glucose brick red ppt.

1 . Glucose 2. Albumin

(Hellers Test) white ring

- Urochrome, urobilinogen, and uroerythrin

3. Acetone

8. Purine Bases Cloudy soln w/ white ppt.

Blood blue green color

4. Blood

(Legals Test) Red purple

What are the different microscopic constituents of urine?

Uric Acid

Calcium oxalate

Hippuric Acid

Ammoniummagnesium phosphate

Calcium phosphate

Cystine

Tyrosine

Cholesterol

Epithelial cells

Epithelial cells

Squamous cells

Leukocytes

D. Uric Crystals

Hyaline casts

Granular casts

Granular casts

Epithelial casts

Pus casts

Blood casts

Mucus threads

Crystals

Cells and Casts

Questions: 1. What pathological condition is indicated by? a. Glucose -normally cannot be detected in urine -its presence in appreciable amount is termed as glycosuria -alimentary glycosuria (glycuresis) occurs after intake of CHO -pathological conditions of the kidney such as glomerulonepthritis -increased amount of glucose in blood is called hyperglycemia -hyperglycemia and glycosuria is found in: diabetes mellitus b. Albumin its presence is called albuminuria; properly termed as proteinuria. 2 types: Functional - no diseased organ and the excretion is only slight and temporary due to strenuous exercises, cold baths, fever Pathological maybe either renal, pre-renal and post renal c. Ketones (end product of fatty acid metabolism) in urine made up of: -Aceto-acetic acid -Butyric acid -Hydroxybutyric acid -Acetone the first three are normal products of fat metabolism they are produced in the liver are utilized or destroyed in the extrahepatic cells **as long as production compensates destruction, no Ketonuria is produced; but when production is too great- ketonemia then ketonuria develops d. Blood its presence is called hematuria. Found whenever there is lesion in the kidney or any part of the urinary tract Hemoglobinuria presence of hemoglobin in urine due to: destruction of RBC liver cannot change all the hemoglobin into bile pigments

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2. What is meant by creatinine coefficient and what is its significance? Creatinine the anhydride of creatine creatinine bears a direct relation to the muscle mass of the individual expressed as CREATININE COEFFICIENT (amount of creatinine in mg excreted in 24 hours per kg of body weight) average adult male coefficient is 20 26; female is 14 22 ** Significance: decrease value indicates muscle wasting due to prolonged negative nitrogen balance; seen in starvation, diabetes, muscle dystrophy

3. Why is lactose found in urine of lactating women? Lactosuria appears after the 3rd month of pregnancy and increases in incidence up to the 35th week and following delivery -chiefly of mammary origin even prior to lactation -caused by high level of lactose of mammary origin in the maternalhoodstage -lactose is excreted by glomerular filtration and is not reabsorbed by the tubules so that all the filtered lactose appears in the urine

4. What is the significance of large amounts of indican in urine? Indican salts of indoxyl sulfate derived from indole which in turn arises from the action of putrefying bacteria on tryptophan and CHONS containing it. -occurs in large intestines if any indole is absorbed -it undergoes a series of detoxification transformations in the liver and indoxyl is formed -it is conjugated with sulfate and neutralized to yield a salt -indican is eliminated in the urine and can be detected by OBERMEYERS TEST -an increase in the amount is indicative of increased putrefaction in the large intestines

5. What is phosphaturia and is its significance? -normally kidneys excrete between 1 and 5 grams of phosphoric acid in the form of phosphates but at times it becomes excessive . Such abnormal excretion and precipitation is called PHOSPHATURIA 2 types of phosphate excreted: a. Alkaline phosphate those combined with alkali metals like Na, K, and NH4+ b. Earthy phosphate combined with earth metals like Ca, Mg SIGINFICANCE: -the amount is increased in hyperthyroidism -in the early stages of pulmonary tuberculosis -diseases on the bones like rickets/osteomalacia -index for the extent of phosphate metabolism

BLOOD

Activity 28: Substances in Blood

Specialized bodily fluid that delivers necessary substances to the body's cells such as nutrients and oxygen and transports waste products away from those same cells

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Erythrocytes (Red Blood Cells) -7-8um diameter -Biconcave disc -without nuclei -lifespan: 120 days

Blood performs many important functions within the body: Supply of oxygen to tissues Supply of nutrients such as glucose, amino acids, and fatty acids Removal of waste such as carbon dioxide, urea, and lactic acid Immunological functions, including circulation of white blood cells, and detection of foreign material by antibodies Coagulation Regulation of body pH Regulation of core body temperature

Leukocytes (White Blood Cells) -Have nucleus -Most live for a few hours to a few days

Thrombocytes (Platelets) -2-4 um diameter -no nucleus -Lifespan: 5-9 days

A. Inorganic substances 1. Reagent: Ash + dil. HCl + potassium ferrocyanide Result: prussian blue solution Constituent of blood present: iron Equation: 4Fe+3 + 3Fe(CN) -4 6 Fe4[Fe(CN)6]3 2. Reagent: blood + water + heat to boiling Result: redish brown solution Reagent: dil. HNO3 + AgNO3 Result: white ppt. formed Constituent of blood present: chloride Equation: AgNO3 + Cl AgCl + NO3

3. Reagent: dil. HNO3 + ammonium molybdate Result: formation of yellow brown ppt. Constituent of blood present: phosphate Equation: H2PO4 + 12MoO4 + 3NH4 + 22H3O (NH4)3PO4.12MoO4 + 34H2O 4. Reagent: ammonium oxalate Result: formation of white ppt Constituent of blood present: calcium Equation: Ca+2 + C2O4 CaC2O4

B. Hemoglobin Blood exposed to air: dark red color Conclusion: formation of oxyhemoglobin took place upon exposure of blood to air C. Glucose in Blood occurs after the separation from coagulated proteins Reagent: Benedicts reagent Result: brick red ppt.

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Questions:
1.

2. Structure of the Hemoglobin molecule conjugated CHON with a Mw of 68000 made up of CHON globin to which heme groups are attached globin contains high amount of histidine, lysine and little of isoleucine (for the specificity of hemoglobin) its buffering property and the CO2 carrying capacity iron containing pigment heme is specially for the pigmentary property

Inorganic constituents of blood and its function Calcium plays a role in blood clotting Phosphorus for the maintenance of c id-base balance and calcium equilibrium Chlorine in the form of NaCl- plays a role in osmotic pressure Magnesium 18 parts magnesium gives rise in rate vasodilation and hyperirritability of the nervous system Iron essential component of hemoglobin, muscle myoglobin Copper important in bone formation and cellular respiration

3. Methemoglobin and its color a derivative in which iron is in the ferric state produced by the oxidation of hemoglobin as when potassium ferricyanide is added to blood color: red-brown

5. Importance of blood sugar determination a. important in the detection of diabetes mellitus and hyperglycemia b. to diagnose toxic or inflammatory condition of the liver due to: galactosemia presence of lacic acid acidosis thymine deficiency

4. Carboxyhemoglobin and its color CO combines with heme portion of hemoglobin to form carbon monoxide hemoglobin called also carboxyhemoglobin and carbonyl hemoglobin Color: bright cherry red

c. adjust insulin dosage d. understand effects of various foods

A. Guaiac Test -bluish green color B. Benzidine Test -Bluish green color C. Hemin Test -Red brown rods
Guaiac Test and Benzidine Test bluish green color

Activity 29: Tests for Blood

Hemin Test red brown rods

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1. What is the chemical name and structural formula of HEMIN?

Synonyms: Ferriheme chloride, Ferriprotoporphyrin chloride 2. Describe a method on detecting blood stains on cloth.
Hemin Test -this test is quite satisfactory and the technique is simple and gives equally reliable result with fresh blood from clothes on stains as long as that it is not been exposed to the sun or high temperatures for a long period of time

IUPAC Name: chloro[3,7,12,17-tetramethyl-8,13-divinylporphyrin2,18-dipropanoato(2)]iron(III)

Activity 30: Plasma

Plasma - straw colored liquid - A watery, protein rich fluid that forms the matrix of the blood - 91.5% water and 8.5% solutes, most of which are proteins

A. Preparation of Oxalated Plasma -light yellow viscous fluid

B. Separation of Plasma Proteins and Fibrinogen 1. Plasma + distilled water + saturated ammonium sulfate sol. = light yellow solution with bubbles suspended

3. Filtrate from 2 + water + saturated ammonium sulfate sol. = cloudy light yellow solution with bubbles and white ppt.

4. Precipitate + water and perform the Biuret test = purple solution

2. Precipitate + water and perform the Biuret test = purple solution

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5.Filtrate from 4 contains albumin + distilled water and perform the Biuret test =purple solution 6.Heat the remaining filtrate from 4 =cloudy solution with white ppt. *Substance precipitated: albumin C. Serum 1.Serum clear yellow fluid 2. Fibrin color: red - appearance: thread like appearance 3. Millons Test red color

Questions: 1. What is the difference between plasma and serum?

Basis 1. How to be obtain? Plasma When whole blood is mixed with an anticoagulant Fluid part of blood that contains water soluble solutes 2. Components Composed of proteins, fibrinogen and clotting factors Preferred in hematology Serum When blood is allowed to clot

Fluid portion left after blood has clotted

Doesnt contain fibrinogen

3. Preference

Preferred in clinical chemistry

2. What are plasma proteins? Give the function of each. 3. What is meant by icterus index? What is its significance? Icterus index calculation of bilirubin in plasma or serum. For the determination of latent jaundice, symptom of liver disease characterized by yellow skin due to deposition of bile pigments.

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