BIOCHEMISTRY OF

URINE
BY: JEROME S. MONTANO, RMT
TOPICS
• URINE FORMATION
• URINALYSIS
• PHYSICO-CHEMICAL COMPOSITION
• OSMOLALITY
• BIOCHEMICAL ANALYTES
URINE FORMATION
FUNCTIONS OF THE KIDNEY-
Controlling the blood volume and composition by:
1. Eliminating the wastes by filtering the blood plasma
2. Regulating the blood pressure and blood volume
3. Regulating fluid osmolarity
4. Acid-Base balance by regulating pCO2
5. Secretion of hormones such as Rennin, Erythropoietin,
Calcitrol and Gluconeogenesis
6. Detoxification of free radicals and drugs
URINE FORMATION
URINE- A liquid containing multiple waste products of metabolism, especially urea and other
nitrogenous compounds, that are filtered from the blood by the kidneys. Urine is stored in the urinary
bladder and excreted from the body through the urethra.
URINE FORMATION
1. GLUMERULAR FILTRATION- creates a plasma like filtrate of the blood
2. TUBULAR REABSORPTION- removes useful solutes from the filtrate and returns them in the
blood (water, glucose, amino acids and ions)
3. TUBULAR SECRETION- removes additional waste from the blood and adds them to the filtrate
(H, K, Creatinine and Drugs)
4. WATER CONSERVATION- removes water from the urine and returns it to the blood thus
concentrating the waste
URINE FORMATION
URINE FORMATION
URINE COMPOSITION
URINALYSIS
• Routine urine examination
• Most useful tool for clinician as an indicator for health and diseases
• Used in renal metabolic disorders
• Divided into four main group of examination
1. Physico-Chemical exam- Color, appearance, volume, SG, odor
2. Chemical examination- pH, sugar, protein, ketone bodies,
bilirubin, urobilinogen, occult blood, Nitrite and Ascorbic acid
3. Microscopic Exam
4. Bacterial screening
PHYSICO-CHEMICAL EXAMINATION
COLOR
• Normal color of urine ranges from pale yellow to deep amber
• Pigment responsible for the normal coloration is UROCHROME
*Neon Yellow- cause by Vitamin B, but may indicate also liver
diseases
*Port wine color- may indicate porphyria
*Blue or green color- presence of bilirubin, UTI
*Red or pinkish- medications from Rifampin, phenazopyridine,
laxatives
- blood in urine due to injury, obstruction, infection,
kidney disease, cancer
- poisoning from lead or mercury
*Brown urine- severe muscle injury, liver disorder, kidney
disorder
*Milky urine- UTI and other bacterial infection
* Dark yellow- severely dehydrated
PHYSICO-CHEMICAL EXAMINATION
COLOR
PHYSICO-CHEMICAL EXAMINATION
APPEARANCE
• Normal appearance of urine should be clear, turbidity of urine may
indicate presence of bacteria, crystals, cells, excess proteins and even
bilirubin.
Examples of substances that can cause turbidity
1. Amorphous phosphates- white and cloudy on standing of alkaline
urine
2. Amorphous urates- Pink and cloudy in acid urine
3. Pus cells
4. Bacteria- uniformly cloudy, does not settle at the bottom even after
centrifugation
PHYSICO-CHEMICAL EXAMINATION
VOLUME
• Normal volume is 750 –2000ml/24Hrs
• OLIGOURIA- less than 750ml in 24Hrs
- dehydration
-  infection
- renal stones or obstruction
- kidney failure
• POLYURIA- over 2000ml in 24Hrs
- Diabetes mellitus
- hypertension
- nephrotic syndrome
- alcohol or drug intake
- endocrinal disorder
• ANURIA- less than 100ml or absence of urine as seen in kidney failure and stenosis
PHYSICO-CHEMICAL EXAMINATION
ODOR
• Healthy urine may have a mild smell but generally does  not have a foul odor
• In some cases, an unusual or strong urine odor may be  due to benign conditions that are
not harmful, such as  eating certain foods or taking certain medications and  volatile acids

1. PUNGENT ODOR- due to the production of ammonia, is typical of most bacterial urinary
tract infection
2. SWEET OR FRUITY ODOR-  production of ketones in the urine
3. MAPLE SYRUP ODOR- Maple syrup disease- inherited disorder in which body is unable
to process certain protiens
4. MOUSY ODOR- Phenylketonuria- inherited disorder that causes amino acids to build up
in the body
5. SWEATY FEET ODOR- Isovaleric acidemia
6. RANCID BUTTER OR FISHY ODOR- Hypermethioninemia- excess methionine in the
blood
PHYSICO-CHEMICAL EXAMINATION
SPECIFIC GRAVITY
• Specific gravity reflects kidney's ability to concentrate
• measures your kidneys’ ability to balance water content and excrete waste
• urine specific gravity is generally considered normal in the ranges of 1.005 to 1.030.
Twenty four hours specimen is 1.015 to 1.025

HIGH SPECIFIC GRAVITY- can indicate that you have extra substances in your urine, such as:
Glucose, protein, bilirubin, red blood cells, white blood cells, crystals, bacteria

LOW SPECIFIC GRAVITY- may indicate the presence of diabetes insipidus, a disease caused
by impaired functioning of antidiuretic hormone, glomerulonephritis, pyelonephritis, and
other renal abnormalities
OSMOLALITY
OSMOLALITY
• Determination  of  urinary  osmolality  is  more  important  than  specific  gravity  because it gives
more accurate  reflection of the concentration of dissolved substances
• It is the measure of the moles of dissolved particles (un 
dissociated molecules and ions) contained in a kilogram  (Kg) of a solvent
• The  normal  osmolality  of  random  urine  specimen  varies  from  40­-1350  mOsm/kg
(milliosmole)
• The  normal  osmolality  of  24  hours  urine  specimen  varies  from  500­-800  mOsm/kg

HIGH URINE OSMOLALITY- congestive heart failure, dehydration, high glucose, acute kidney injury
LOW URINE OSMOLALITY- excessive fluid intake or over-hydration, kidney failure, renal tubular
necrosis
BIOCHEMICAL ANALYTES
GLUCOSE
GLYCOSURIA- condition in which a person's urine contains more sugar, or glucose, than it should. It
typically occurs due to high blood sugar levels or kidney damage
- common symptom of both type 1 diabetes and type 2 diabetes
- Indicative also of Pregnancy and kidney disorder
- The normal amount of glucose in urine is 0 to 0.8 mmol/L (millimoles per liter)

KETONE
Urine ketone testing is most often necessary in people with type 1 diabetes who:
• have blood sugar levels over 300 milligrams per deciliter (mg/dL)
• are sick
• have symptoms of diabetic ketoacidosis (DKA), an acute complication of diabetes
BIOCHEMICAL ANALYTES
KETONE
• Monitoring ketone levels in urine is important if you have type 1
diabetes. Ketones are more commonly seen in the urine of
people with type 1 diabetes than in people with type 2 diabetes
• Normal or trace levels of ketones in urine are less than 0.6
millimoles per liter (mmol/L)

SMALL TO MODERATE KETONE- ketone level of 0.6 to 1.5

mmol/L (10 to 30 mg/dL)
MODERATE TO LARGE KETONE- ketone level of 1.6 to 3.0
mmol/L (30 to 50 mg/dL) 
VERY LARGE KETONE- ketone level greater than 3.0 mmol/L (50
mg/dL)- indicative of Diabetic Ketoacidosis
BIOCHEMICAL ANALYTES
BILIRUBIN
• Bilirubin is a byproduct of the breakdown of hemoglobin
• Normal urine has no bilirubin
• Presence may be an indication of liver disease, bile duct
obstruction or hepatitis
• Since the bilirubin in samples is sensitive to light, exposure of
the urine samples to light for a long period of time may result in
a false negative test result

BILIRUBIN- yellowish substance made during the body's normal

process of breaking down red blood cells. Bilirubin is found in bile,
a fluid in your liver that helps you digest food. If your liver is
healthy, it will remove most of the bilirubin from your body. If your
liver is damaged, bilirubin can leak into the blood and urine.
Bilirubin in urine may be a sign of liver disease
BIOCHEMICAL ANALYTES
BILIRUBIN
• Normal range is between 0.3 and 1.2 milligrams per
deciliter (mg/dL)
• The test is usually done in conjunction with other tests for liver
disease as part of a hepatic function panel. It can also be done to:
1. Diagnose hepatitis, cirrhosis, or other liver diseases
2. Find blockages in structures carrying bile from your liver
3. Monitor an existing liver disorder
4. Diagnose disorders related to red blood cell production problems
5. Test a patient who has history of drinking large amounts of
alcohol
6. Test a patient who has a suspected drug toxicity
BIOCHEMICAL ANALYTES
BILIRUBIN
QAULITATIVE TESTS FOR BILIRUBIN IN URINE

1. FOAM TEST- urine is shaken and the formation of yellow

foam will indicate presence of Bilirubin
2. GMELIN’S TEST-  a chemical test used for detecting the
presence of Bilirubin in urine. Nitric acid is used as
the oxidizing agent and the presence of Blue, green and
violet rings are seen if bilirubin is present
BIOCHEMICAL ANALYTES
BILE AND BILE SALTS
• Bile salts appear in the urine when there is an obstruction to the
biliary tract, leading to an increase in the blood's bile acids
• Serum bile salts may rise even without the biliary obstruction if
liver cell damage usually removes reabsorbed bile salts from the
portal blood
• Bile salts are positive in urine in other liver diseases
• Cholestasis, retention of bile, and bile salts lead to Itching
(Pruritus). In many cases of jaundice, itching is caused by the
bile salts in blood circulation
• The level of bile salt is correlated with itching
• Normal result of bile salt test is <10 micromolar/L
BIOCHEMICAL ANALYTES
BILE AND BILE SALTS
QUALITATIVE TESTS FOR BILE SALTS IN URINE

1. HAY’S TEST- specific test used for the qualitative detection of

bile salts in urine
Principle: Bile salts have a property of lowering the surface
tension of the fluid. If Bile salts present in urine and sulfur powder
is added to the urine in the test tube, the sulfur particles will sink.
In normal cases it does not sink rather, it floats on the surface of
the fluids
- use of Sulfur powder or “flower of Sulfur”

Positive result: Sulfur will sink down to the bottom of the

test tube
BIOCHEMICAL ANALYTES
BILE AND BILE SALTS
QUALITATIVE TESTS FOR BILE SALTS IN URINE

2. SMITH’S TEST- The presence of bile salt in the given urine

sample is indicated by the formation of a green colored ring
at the intersection of both the layers
Reagent used: Smith’s reagent composed of methylene
iodide and a zinc reagent

3. PETTENKOFER’S TEST- The presence of bile salt in the given

urine sample is indicated by the formation of a red ring is
between both layers.
Reagent used: concentrated H2SO4
BIOCHEMICAL ANALYTES
BLOOD
• Presence of blood may indicate infection, trauma to the urinary tract or
bleeding in the kidneys
• Hematuria- presence of blood in the urine
• False positive readings most often due to contamination with
menstrual blood
• ormal result is 4 red blood cells per high power field (RBC/HPF) or
less when the sample is examined under a microscope
INCREASE RED BLOOD CELLS IN THE URINE
• viral infection
• Inflammation of the kidney or bladder
• blood disorder
• Bladder or kidney cancer
INCREASE WHITE BLOOD CELLS IN THE URINE
• bacterial urinary tract infection. This is the most common cause of a
high white blood cell count in urine.
• Inflammation of the urinary tract or kidneys
BIOCHEMICAL ANALYTES
PROTEIN
• rough estimate of the amount of albumin in the urine
• Normally, there will be no protein or a small amount of protein in
the urine. When urine protein is elevated, a person has a condition
called proteinuria
• random urine sample, normal values are 0 to 14 mg/dL. For a 24-
hour urine collection, the normal value is less than 80 mg per 24
hours

NORMAL PROTEIN IN URINE is seen during stress, exercise, fever,

aspirin therapy, or exposure to cold
HIGH PROTEIN IN URINE is seen chronic kidney disease
BIOCHEMICAL ANALYTES
CREATININE
CREATININE- chemical waste product of creatine, an amino acid
made by the liver and stored in the liver. Creatinine is the result of
normal muscle metabolism
- As a waste product, creatinine is filtered out of the blood by
the kidneys and removed from the body in urine
•  Normal range is 955 to 2,936 milligrams (mg) per 24 hours for
males, and 601 to 1,689 mg per 24 hours for females
• This test can find out whether your kidneys are working normally
or to see if treatment for kidney disease is working

• HIGH CREATININE LEVEL- kidney failure, kidney infection, late

stage muscular dystrophy, kidney disease, myasthenia gravis,
kidney stones or other urinary tract obstruction, diabetes
BIOCHEMICAL ANALYTES
UROBILINOGEN
UROBILINOGEN- formed from the reduction of bilirubin. It is
formed in the intestines by bacterial action on bilirubin. About half
of the urobilinogen formed is reabsorbed and taken up via the portal
vein to the liver, enters circulation and is excreted by the kidney

• Normal urine contains some urobilinogen. If there is little or no

urobilinogen in urine, it can mean your liver isn't working
correctly. Too much urobilinogen in urine can indicate a liver
disease such as hepatitis or cirrhosis
• Normal range from 0.1-1.8 mg/dl (1.7-30 µmol/l)
BIOCHEMICAL ANALYTES
NITRITE
• Nitrite formed by gram negative bacteria converting urinary
nitrate to nitrite
• Presence of nitrite in urine is normal but elevated results may
mean presence of bacterial infection in the urinary tract
• A positive nitrite result signifies that bacteria capable of this
conversion (eg, Escherichia coli, Klebsiella, Proteus, Enterobacter,
Citrobacter, Pseudomonas) are present in the urinary tract
THANK YOU!

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