CHAPTER TWO

Collection And Preservation Of

Urine Specimen

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 LEARNING OBJECTIVES
 Upon completing this chapter, the learner will be able
to:
 Describe procedures and materials used for proper collection of
urine specimens.
 Compare methods of urine collection.
 Suggest appropriate urine collection method depending on
testing ordered.
 Explain what changes occur in unpreserved urine over time
 Explain urine preservation methods.
 Select appropriate types of urine preservation depending on
testing ordered.

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 Collection of Urine Specimen
Accurate urinalysis begins with the proper collection technique.
Improper collection may invalidate the results.
Sample should be delivered as soon as possible to the lab.
If delay is necessary the appropriate preservatives should be
added.
Avoid samples that is contaminated with feces.
Collect Urine Volume Needed for Testing(1o-15ml)
But collection of a larger volume is encouraged to ensure
sufficient urine for additional or repeat testing.
Written criteria for the collection, preservation, handling,
storage, and labeling of urine must be available.

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 Urine Containers
Specimens must be collected in clean, dry, leakproof containers.

Disposable containers should be used because they eliminate

the chance of contamination due to improper washing.

Containers for routine urinalysis should have a wide mouth to

facilitate collections.

They should be made of a clear material to allow for

determination of color and clarity.

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 The recommended capacity of the container is 50 mL, which
allows 12 mL of specimen needed for microscopic analysis.
Additional specimen for repeat analysis, and enough room for the
specimen to be mixed by swirling the container.
Individually packaged sterile containers with secure closures should be
used for microbiological studies of urine.

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 Labels
 All specimens must be labeled immediately after collection with

 Patient’s last and first name

 Identification number

 The date and time of collection; and

 additional information

 such as the patient’s age and location,

 the health-care provider’s name, and

 the preservative used, if any, as required by facility protocol.

 Labels must be attached to the container, not to the lid, and should
not become detached if the container is refrigerated or frozen.

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 Requisition Form
The information on the form must match the information on

the specimen label.

Additional information on the form can include:

Method of collection or type of specimen
Possible interfering medications
Patient’s clinical information.
The time the specimen is received in the laboratory should
be recorded on the form.

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 Specimen Rejection
 Situations leading to specimens that should be rejected can

include:

1. Specimens in containers that are unlabeled or improperly

labeled

2. Labels and requisition forms that do not match

3. Specimens contaminated with feces or toilet paper

4. Specimens of insufficient quantity

5. Wrong type/amount of preservative

6. Specimens that have been transported improperly

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 Types of Specimens
 The type of specimen selected, the time of collection, and

the collection technique are usually determined by the

tests to be performed

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 1. First Morning Specimen
Immediately on rising from sleep collects a urine specimen.
Retained in the bladder for approximately 8 hours.
Most concentrated and bladder incubated.
Preferable for:
Routine screening
Nitrite tests
Protein tests
Pregnancy tests
Orthostatic protein

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 2. Random Urine Specimen
 For ease and convenience, routine screening most often is
performed on random urine specimens.
 It is collected at any time.
 it may show erroneous results resulting from dietary intake or
physical activity just before collection.
 Preferable for:
 Routine screening

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 3. Postprandial Specimen
 Postprandial specimens are those collected 2 to 3 hours after

eating.

 Preferable for:

 screen for carbohydrate metabolism disorders.

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 4. 24-Hour (or Timed) Specimen
 A carefully timed specimen must be used to produce accurate

quantitative results.

 Preferable for:

 Quantitative chemical analysis

 Clearance tests

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 24-HourUrine Specimen Collection Procedure
[Link] the patient with written instructions, and explain the collection
procedure.

2. Provide the patient with the collection container and preservative, if required.

3. Day 1: 7 a.m.: Patient voids and discards specimen; collects all urine for the next
24 hours.

4. Day 2: 7 a.m.: Patient voids and adds this urine to the previously collected urine.

5. Specimen is transported to the laboratory, where the entire 24-hour specimen is

thoroughly mixed and the volume is measured and recorded.

6. The required amount of urine (approximately 50 mL) is aliquoted for testing.

7. The remaining specimen is discarded.

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 5. Midstream Clean-Catch Specimen
A choice for obtaining non contaminated specimens.

It is easy to perform and it provides a sample that can be used

for bacteriologic examination as well as for routine urinalysis

Preferable for:
Bacterial and fungal cultures
Routine screening
Cytology

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 6. Catheterized Specimen
Insertion of a sterile tubing of the bladder to obtain a suitable
specimen.
Used if the patient is having difficulty voiding.
Used to avoid vaginal contamination, especially during
menstruation.
The possibility of introducing organisms into the bladder which
may, in turn, cause infection,
Preferable for:
Bacterial culture

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 7. Suprapubic Aspiration
Collecting urine directly from the bladder by puncturing.
completely free of extraneous contamination.
Avoids vaginal and urethral contamination.
It can also be useful in getting urine from infants and small
children.
Preferable for:
For bacterial culture
Cytology

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 8. Three-glass collection
All portions, beginning, middle, and final portion of the void,
are collected in three separate containers.
The prostate is massaged prior to collection in the third
container.
Urinary tract infections will show increased white blood cell
counts and bacteria in the second and third containers,
while prostate infections will demonstrate white blood cell
counts and bacteria higher in the third container than in the
second.
Preferable for:
Prostatic infection

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 9. Pediatric Specimens
 Collection of pediatric specimens can present a challenge.

 Soft, clear plastic bags are available.

 Hypoallergenic skin adhesive to attach to the cleaned

genital area of both boys and girls

 Care must be taken to a void fecal contamination.

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 Handling and Preservation
 Urine specimens should be delivered to the laboratory
immediately after collection.

 However, this is not always possible; if a delay in specimen

transportation is to be 2 hours or longer;

 precautions must be taken to preserve the integrity of the

specimen.

 Protecting it from the effects of light and room temperature

changes.

 A variety of changes can occur in unpreserved urine.

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 Changes Occurring to Unpreserved Urine
Potential
Change Observation Mechanism

Color Darkens Oxidation or Reduction of solutes

Clarity Decreases Increased turbidity due to proliferation of bacteria or precipitation of chemical substances

Odor Ammonical, foul Bacterial conversion of urea to ammonia (and bacterial proliferation)
smelling
PH Increase Bacterial conversion of urea to ammonia; loss of CO2
Glucose Decrease Consumed by cells and/or bacteria

Ketones Decrease Volatilization and bacterial conversion

Bilirubin Decrease Photo-oxidation to biliverdin by light exposure

Urobilinogen Decrease Oxidation to urobilin

Nitrite Increase Bacterial conversion of dietary nitrates

Blood cells Decrease Lysis and/or disintegration,

Casts Decrease Lysis and/or disintegration,

Bacteria Increase Exponential proliferation of bacteria

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Trichomonas Decrease Loss of characteristic motility and death
 Preservatives
 Refrigeration:

 Used most routinely is refrigeration at 2°C to 8°C.

 It decreases bacterial growth and metabolism.

 If the urine is to be cultured kept refrigerated until cultured, up to 24

hours.

 The specimen must return to room temperature before chemical

testing by reagent strips.

 Refrigeration also can cause precipitation of amorphous urate and

phosphate crystals

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  Boric acid
 preserves formed elements but interferes with the pH reading.

 Boric acid is the preservative used in tubes used to preserve urine for
culture and sensitivity.

Boric acid tube used for urine culture

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  Chlorhexidine:

 Prevents bacterial growth and is useful as a glucose

preservative.

 Although specimens transferred to this tube for transport are

stable for 72 hours,

 If not protected from light will yield

erroneous bilirubin and Urobilinogen results.

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  Formalin

 Formalin (1 drop/30-mL urine) is a good preservative for

urinary sediment

 But if used in too large a concentration it will precipitate

protein

 give a false-positive test for reducing substances.

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  Preservative Tablets

 Commercially available preservative tablets (1 tablet/30-mL

urine), usually act by releasing formaldehyde.

 At this concentration the formaldehyde will not interfere with

the test for reducing substances.

 But higher concentrations will result in false positives.

 Formaldehyde increases the specific gravity by 0.005/1

tablet/30 ml.

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  Toluene

Toluene (2-mL/100-mL urine) preserves, ketones, proteins, and

reducing substances,

But it is not effective against bacteria already present in the

urine.

In addition, toluene is flammable

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  Thymol

 Thymol (one small crystal) is an adequate but rarely used

preservative for most urinary constituents.

 Thymol interferes with the acid precipitation test for protein.

 But it does not interfere with reagent strip tests for protein.

 Chloroform

 It has been used for inhibiting bacterial growth.

 It is not recommended for the routine specimen because it causes

changes in the characteristics of the cellular sediment

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 References
1. Graff's Textbook Of Urinalysis And Body Fluids Third Edition.

2. Urinalysis And Body Fluids By Susan King Strasinger Seventh

Edition.

3. Fundamentals Of URINE & BODY FLUID ANALYSIS By

Nancy A. Brunzel Fourth Edition.

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