HISTORY AND IMPORTANCE

 Analyzing Urine – beginning of laboratory medicine

o Edwin Smith Surgical Papyrus – historical study of urine
 Examining a bladder-shaped flask of urine
 Hippocrates (5th century BCE) – wrote Uroscopy
 1140 AD – color charts were developed that described the significance of 20 different colors
 Frederick Dekkers – discovered albuminuria by boiling urine (1694)
 Pisse Prophets (charlatans) – compromised the credibility of urinalysis
o Thomas Bryant (1627) – wrote a book about the pisse prophets
 Thomas Addis (17th century) – examination of urinary sediment with the invention of
microscope
 Richard Bright (1827) – introduced the concept of urinalysis as part of a doctor’s routine patient
examination
 2 Unique Characteristics of a Urine Specimen:
1. Urine is a readily available and easily collected specimen
2. Urine contains information
 Clinical and Laboratory Standards Institute (CLSI) – defines urinalysis as “the testing of urine with
procedures commonly performed in an expeditious, reliable, accurate, safe, and cost-effective
manner.”

URINE FORMATION

 Urine – ultrafiltrate of plasma formed by the kidneys

 Filters 170,000 mL of essential substances

URINE COMPOSITION

 Urine – 95% Water and 5% Solutes

Primary Composition in Normal Urine

Component Comment
Urea Primary organic component. Product of protein
and amino acid metabolism. (Liver)
Creatinine Product of creatinine metabolism by muscles
Uric Acid Product of nucleic acid breakdown in food and
cells
Chloride Primary inorganic component. Found in
combination with sodium (table salt) and many
other inorganic substances
Sodium Primarily from salt, varies by intake
Potassium Combined with chloride and other salts
Phosphate Combines with sodium to buffer the blood
Ammonium Regulates blood and tissue fluid acidity
Calcium Combines with chloride, sulfate, and phosphate
Creatinine – also a product of purine metabolism

URINE VOLUME

 Urine excretion = Body’s hydration

 Factors that influence urine volume:
1. Fluid Intake
2. Fluid Loss from nonrenal sources
3. Variations in the secretion of antidiuretic hormone
4. Need to excrete increased amounts of dissolved clots
 Daily Urine Output – 1,200 to 1,500 mL; 600-2,000mL is considered normal
 Oliguria – a decrease in urine output
o May lead to Anuria (cessation of urine flow)
 Clinical Significance: Severe acute nephritis, Hg poisoning, obstructive uropathy,
kidney failure
o May be seen in Blood Transfusion Reaction; Renal Schemia, Stone, Disease
o Clinical Significance: Dehydration, Renal insufficiency, poorly compensated heart
disease, calculi formation, kidney tumors
 Nocturia – increase in the nocturnal excretion of urine (>500 mL at night)
 Polyuria – increase in daily urine volume (>2.5 L/day in adults; >2.5-3.0 mL/kg/day in children)
o Often associated with diabetes mellitus and diabetes insipidus
 Diabetes Mellitus - ↑specific gravity, ↓insulin = ↑glucose
 Diabetes Insipidus - ↓specific gravity, ↓ADH
o May be artificially induced by diuretics, caffeine, or alcohol (all suppresses ADH)
 Polydipsia – compensates fluid loss; increased ingestion of water (can be seen in DM)
 Diuresis – transitory increase in urine volume
 Dysuria – ex. UTI, cystitis

SPECIMEN COLLECTION

 Gloves – should be worn at all time when handling urine

 Containers – must be clean, dry, leak-proof and disposable
o Should have a wide mouth (for female)
o Should have a wide flat bottom to prevent overturning
o Should be made of a clear material
o Capacity – 50 mL (allows 12 mL)
o Sterile containers – if 2 hours have passed the specimen collection
 Labels
o Should be labeled with the patient’s name, identification number, date and time of
collection
o Should be attached to the container and not on the lid
  Requisitions – this form can be manual or computerized)
o May include method of collection, type of specimen, possible interfering medications,
and the patient’s clinical information.
o The time the specimen is received in the laboratory should be found in this form

SPECIMEN REJECTION

 Unacceptable situations include:

1. Specimens in unlabeled containers
2. Nonmatching labels and requisition forms
3. Specimens contaminated with feces or toilet paper
4. Containers with contaminated exteriors
5. Specimens of insufficient quantity
6. Specimens that have been improperly transported

SPECIMEN HANDLING

 Specimen Integrity
o Should be delivered to the laboratory within 2 hours
 Cannot be delivered within 2 hours = should be refrigerated/preserved

Changes in Unpreserved Urine

Analyte Change Cause
Color Modified/Darkened Oxidation/Reduction of
metabolites
Clarity Decreased Bacterial growth and
precipitation of amorphous
material
Odor Increased Bacterial multiplication causing
breakdown of urea to ammonia
pH Increased Breakdown of urea to ammonia
by urease-producing
bacteria/loss of CO₂
Glucose Decreased Glycolysis and bacterial use
Ketones Decreased Volatilization and bacterial
metabolism
Bilirubin Decreased Exposure to light/photo
oxidation to biliverdin
Urobilinogen Decreased Oxidation in urobilin
Nitrite Increased Multiplication of nitrate-
reducing bacteria
Red and white blood cells and Decreased Disintegration in dilute alkaline
casts urine
Bacteria Increased Multiplication
Trichomonas Decreased Loss of motility, death
  Specimen Preservation
o Refrigeration (2⁰C to 8⁰C) – most routinely used method of preservation
o The specimen must return to room temperature before chemical testing by reagent
strips

Urine Preservatives
Preservatives Advantages Disadvantages Additional Information
Refrigeration Does not interfere with Precipitates Prevents bacterial
chemical tests amorphous phosphates growth for 24 hours
and urates Maintains an acid pH
up to 8 hours
Boric Acid Prevents bacterial Interferes with drug Keeps pH at about 6.0
(cloudy specimen) growth and and hormone analyses Can be used for urine
metabolism culture transport
Preserves protein and Does not interfere with
formed elements well routine analysis other
than pH
Phenol Does not interfere with Causes an Odor change
routine tests
Toluene Does not interfere with Floats on surface of
routine tests specimens and clings to
pipettes and testing
materials
Thymol Preserves glucose and Interferes with acid
sediments well precipitation tests for
protein
Formalin Excellent sediment Acts as a reducing Rinse specimen
(formaldehyde) preservative agent, interfering with container with formalin
chemical tests for to preserve cells and
glucose, blood, casts
leukocyte esterase, and
copper reduction
Sodium fluoride Good preservative for Inhibits reagent strip Prevents glycolysis
drug analyses tests for glucose,
blood, and leukocytes
Commercial Convenient when Check tablet
preservative tablets refrigeration not composition to
possible. determine possible
Have controlled effects on desired tests
concentration to
minimize interference
Urine Collection Kits Contains collection
(Becton, Dickinson, cup, transfer straw,
Rutherford, NJ) culture and sensitivity
 (C&S) preservative
tube, or UA tube
Light gray and gray C&S Sample stable at room Do not use if urine is Preservative is boric
tube temperature (RT) for below minimum fill line acid, sodium borate
48 hours; prevents and sodium formate.
bacterial growth and Keeps pH at about 6.0
metabolism
Yellow UA Plus tube Use on automated Must refrigerate within Round or conical
instruments 2 hours bottom, no
preservative
Cherry red/yellow Stable for 72 hours at Must be filled to Preservative is sodium
Preservative Plus tube RT; instrument- minimum fill line. propionate, ethyl
compatible Bilirubin and paraben, and
urobilinogen may be chlorhexidine; round or
decreased if specimen conical bottoms
is exposed to light and
left at RT
Saccomanno Fixative Preserve cellular For cytology studies
elements

TYPES OF SPECIMENS

Types of Urine Specimens

Type of Specimen Purpose Additional Information
Random Routine Screening Most commonly received specimen.
May be collected at any time
First Morning Routine Screening Ideal screening specimen.
Pregnancy Tests Concentrated specimen.
Orthostatic Protein Evaluates Orthostatic Proteinuria
24-hour (or timed) Quantitative Chemical Must begin and end with an empty bladder
Tests, Hormone Studies
Catheterized Bacterial Culture Passes hollow tube (catheter) through the
urethra
Midstream Clean-Catch Routine Screening Alternative to Catheterized Specimen
Bacterial Culture
Suprapubic Aspiration Bladder urine for
bacterial culture
Cytology

 Prostatitis Specimen
 o Three-Glass Collection
 Male mid-stream clean-catch -> first urine is not discarded, but collected in a
sterile container -> Midstream portion is collected in another sterile container ->
Prostate is then massaged so that prostate fluid will be passed with the
remaining urine into a third sterile container
 Prostatic Infection = third specimen will have WBC/high-power field count and
bacterial count 10 times that of the first specimen
o Stamey-Mears Test – 4 glass method: Initial Voided Urine (VB1), Midstream Urine (VB2),
Expressed Prostatic Secretions (EPS), and a post-prostatic massage urine specimen (VB3)
 Midstream Clean-Catch – cleanse glans penis (for male) and urinary meatus (for female) with
antiseptic towelette (or with soap and water)
 Pediatric Specimens – a soft, clear plastic bag with hypoallergenic skin adhesive to attach to the
genital area is needed
 Drug Specimen Collection
o Chain of Custody (COC) – process that provides this documentation of proper sample
identification from the time of collection to the receipt of laboratory results
o May be “witnessed” or “unwitnessed”
 “Witnessed” – collection of 30 to 45 mL of urine
o Must be taken within 4 minutes to confirm the specimen hasn’t been adulterated
o Blueing agent added to the toilet water reservoir in unwitnessed collection
o Temperature – 32.5⁰C to 37.7⁰C
 2 hours post prandial & Fasting Specimen – to diagnose DM
 Glucose Tolerance Test (GTT) – may include fasting, half-hour, 1-hour, 2-hour, and 3-hour
specimens
 12-hour – for Addis count
 Afternoon specimen (2-4 pm) – urobilinogen determination