CHAPTER TWO
Collection And Preservation Of
Urine Specimen
Objectives
At the end of this chapter the students will enable to :
Identify factors affecting the quality of a specimen.
List the basic procedures and prequations of urine
collection.
Describe types of urine specimens.
Identify the commonly used preservatives and
differentiate the advantages and disadvantages of their
use.
Chapter two
2.1- Collection of urine specimen
2.2 -Preservation of urine specimen
2.3- Types of Examination in Routine Urinalysis
Collection of Urine Specimen
In order to make Urinalysis reliable the urine must be
properly collected.
Improper collection may invalidate the results of the
laboratory procedures, no matter how carefully and skillfully
the tests are performed.
Sample should be delivered as soon as possible to the lab
after collection
If delay is necessary the appropriate preservatives based on
test types should be added.
instruct the patient to bring the mid-stream urine
Avoid samples that is contaminated with blood in case of
manusteral women
The container should be dried and clean
Urine Containers
types of containers used for collection of urine
Urine bottles
Disposable plastic containers
Poystrine containers
Waxy coated paper
Plastic caps
URIN-TEK tube
Note : The container should be wide mouth, clean ,dry, leak
proof
Urine Containers cont’d…
The container should be labeled with :
The test order
The patient’s name
Time and date of collection
The preservative added
NOTE :Specially samples that comes from ward
should contain all the above information in addition to
the above the labeling on the sample container must
matched with the request paper
Types of Specimen
First Morning Specimen : obtained during the first
urination of the day
Most concentrated
Bladder incubated
preferable for:
Nitrite
Protein
Microscopic examination
Types of Specimen cont’d…
Random Specimen - a spciemen obtained at any time
during examination.
Most convenient
Most common
Preferable for
Chemical Screen
Microscopic examination
Types of Specimen cont’d…
Postprandial : a specimen obtained 2 hours after
meal.
Good for glucose determination
24- Hour specimen - obtained within 24 hours.
Necessary for quantitative tests, especially for
quantitative determination of protein.
Types of Specimen cont’d…
Collection of 24 hour Urine Specimen
Procedure
1. Instruct patient to completely empty his bladder and
discard his urine exactly at the beginning of the 24 hour
time collection (let say at 6:00 a.m.).
2. Collect all urine voided during the following 24 hours,
including that voided exactly at the end of the 24 hour
period in a container (at 6:00 a.m.) of the following
(second) day.
3. Provide the patient with appropriate type and amount of
preservative.
[Link] container should be labeled
Types of Specimen cont’d…
Clean Catch Urine can be collected by two ways:
A) Mid- stream Specimen - a specimen obtained from the
middle part of the first urine.
commonly used for routine urinalysis.
important for bacteriological urine culture.
B) Catheterization is the process of passing a tube through the
urethra to the bladder for the withdrawal of urine (it may
introduce urinary tract infection).
Types of Specimen cont’d…
The best method of ‘clean catch’ urine collected as follows:
[Link] genital area should be cleaned with soap and water and
rinsed well. This is to keep off bacteria on the skin from
contaminating the urine specimen.
[Link] patient should urinate a small amount and this is
discarded.
[Link] urine that comes next, the mid-stream specimen,
should be collected into a sterile container of 30 to 50 ml.
4. After obtaining the specimen the patient continues to
urinate and this is discarded.
Types of Specimen cont’d…
Sources of Errors
Bacteriologically or chemically contaminated specimen
such as detergent remnants in the container.
Wrong type/amount of preservative.
Partial loss of specimen or inclusion of two-morning
specimen in the 24 hr collection.
Inadequate mixing of specimen before examination.
Careless measuring of the 24 hr volume.
Wrong type of urine specimen, Such as using random
sample for urine culture
Inappropriate storage condition .***
Preservation of Urine Specimen
Urine specimen after voided in a container, should be
examined immediately, or within maximum delay of 30
minutes.
If not the specimen should be properly preserved to
prevent those changes that occur in old urine sample.
Urine preservation cont’d…
Changes that occur in old urine specimen
Bacteria will grow, and change the pH into alkaline.
This environment will convert urea into ammonia;
In Acidic urine glucose will be converted into acids and
alcohol
Concentration of glucose will decrease, because of
utilization of the glucose by the bacteria
Acetones will evaporated as a result Ketones
concentration will decrease
Urine preservation cont’d…
Bilirubin will converted into biliverdin by light
Urobilinogen change into urobilin, and concentration will
also decrease by light
Nitrate concentration either increased if bacteria reduce
nitrites, or decrease if it is converted to ammonia, which
is volatile; and urine pH will become alkaline
Turbidity will increase, because of bacterial growth,
crystal formation, and precipitation of amorphous
substances
Red blood cells, leucocytes, and casts will layzed
Urine preservation cont’d…
Methods used
A. Physical methods
B. Chemical methods
Urine preservation cont’d…
A. Physical method of urine preservation
Refrigeration
used for short period of time
serves for 6 to 8 hours
Limitation:
turbidity due to amorphous may affect microscopical
exam.
freezing
Good for bilirubin and urobilinogen
Limitation:
increase [Link]
destroy formed elements
Urine preservation cont’d…
B. Chemical Methods
Different chemical are available to preserve urine for
long period of time
The choice of the chemical, depend on the required
preserved substance in the urine sample.
Urine preservation cont’d…
Chemical Preservatives
Toluene
It preserves acetone, protein and other reducing
substances like glucose; it is added into the urine
sample by making thin layer over the urine
The disadvantage is risk of fire accidents
Thymole
It preserves most urine constituents; it is added into the
urine by making small crystal of 5mm diameter for a
100ml of urine.
But it causes false positive results for protein tests
Urine preservation cont’d…
Chloroform
It preserves urine aldosterole level
Has higher specific gravity than urine it settles at the
bottom of the urine sample thus needs frequent mixing
Formaldehyde
It preserves most of the formed elements; [one drop per 30
ml ]
The disadvantage of this chemical preservative is that it
interferes with urine glucose determination
Boric acid
It preserves most chemicals and formed elements
But the problem is that it precipitate uric acid
Exercises:
Answer the following questions:
[Link] type of specimen would be appropriate for both routine
urinalysis and bacteriological culture?
2. List important points that should be labeled on the 24 hr
urine collecting bottle.
3. Discuss about source of errors in the collection of urine.
4. What kind of changes take place in old urine specimen?
5. Discus about physical and chemical preservatives of urine
with their respective advantage and disadvantages.
6. Mention limitation of chemical tests
7. Asume, you are instructed to bring your urine sample for
student demonstration? Are you voluntary?, if you say yes,
show the important procedures during collection ,if No reason
out it
Types of Urinalysis Examination
1. Physical Examination of Urine
2. Chemical Analysis of Urine
3. Microscopic Examination of Urine