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Lab 14: Urinary System (Part II)

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Title: Lab 14: Urinary System (Part II)


1
Lab 14 Urinary System (Part II)
  • Dr. Kim Wilson

2
INTRODUCTION
  • The analysis of urine has 2 purposes. One is to
    detect body disturbances, such as endocrine or
    metabolic abnormalities, in which the kidneys
    function normally but excrete abnormal amounts of
    metabolic end products specific for a particular
    disease. The second purpose is to detect disease
    conditions within the kidneys or urinary tract
    themselves that may adversely affect the urinary
    system's ability to function efficiently in the
    maintenance of fluid, electrolyte acid-base
    balance. Urine may be analyzed both physically
    chemically using relatively simple tests in order
    to check for abnormalities. In this lab, a few
    of these physical chemical tests will be
    conducted on a sample of your own urine. For
    reference, you should review information
    pertaining to urinalysis in your textbook and in
    your lab manual (Unit 21).

3
PART 1 APPEARANCE AND PHYSICAL PROPERTIES OF
URINE
  • INSTRUCTIONS
  • Obtain a clean specimen cup and collect a sample
    of your urine. We won't be testing the specimen
    for the presence of micro-organisms, however,
    obtain a "midstream" specimen since it yields the
    best results.
  • Pour part of your specimen (approx. 10 ml) into a
    test tube and cap it. This portion will be
    allowed to set for 1 hour and then centrifuged in
    order to microscopically examine its sediments.
    Use the remaining portion of your specimen to
    perform the following physical and chemical tests.

4
PART 1 APPEARANCE AND PHYSICAL PROPERTIES OF
URINE
  • A. COLOR
  • Freshly voided urine is often described as having
    a straw or amber colored appearance. However,
    the normal color (which is due to the presence of
    a pigment known as urochrome) may range
    considerably from person to person. Concentrated
    urine, resulting from loss of water in severe
    sweating, will be dark yellow a light amber
    yellow is more characteristic of dilute urine.
    Certain deviations from the normal amber color
    may be the result of disease, whereas other color
    changes may have no pathological significance.
    For example, a brownish yellow or green color may
    be the result of bile pigments, suggestive of
    liver disease. Likewise, a smoky brown or
    reddish-brown colored urine usually indicates
    disease, but may (on occasion) be the result of
    ingestion of plant dyes found in beets or
    rhubarb.
  • Record color on Results Sheet.

5
PART 1 APPEARANCE AND PHYSICAL PROPERTIES OF
URINE
  • B. CLARITY (CLOUDINESS)
  • A freshly voided urine sample should be clear or
    transparent. If urine is cloudy when voided or
    becomes cloudy after standing, the sample should
    be checked (by microscopic examination chemical
    tests) for the presence of mucous, pus, bacteria,
    sperm, phosphate, or urate crystals.
  • Record clarity on Results Sheet.
  • C. ODOR
  • Normal, freshly voided urine has a
    "characteristic" odor which is believed to be due
    to the presence of volatile acids. Urine that
    has been standing for a long time at room
    temperature develops an ammonia-like odor which
    is due to the decomposition of urea by bacteria
    in the specimen. The urine of patients with
    diabetes mellitus may have an acetone odor due to
    the presence of ketones. The urine of patients
    with urinary tract infections may be
    foul-smelling, especially when the infecting
    organism is a coliform bacillus. Abnormalities
    in urine odor may simply be due to the ingestion
    of certain foods for instance, garlic and
    asparagus can give urine characteristic odors.
  • Record odor on Results Sheet.

6
PART 1 APPEARANCE AND PHYSICAL PROPERTIES OF
URINE
  • D. REACTION (pH)
  • The pH of urine is a measure of its hydrogen ion
    concentration. A pH below 7 indicates acid
    urine a pH above 7 indicates alkaline urine.
    Freshly voided urine from persons not on special
    diets is usually acid and has a pH of
    approximately 6.0 however, normal kidneys are
    capable of producing urine that can vary from a
    pH of 4.5 to a pH of 8. These variations seen in
    pH may be related to diet, medication, or
    disease. For example, persons on high protein
    diets produce a more acid urine whereas persons
    on diets high in vegetables, milk or other dairy
    products produce an alkaline urine. Persons with
    diabetes mellitus (uncontrolled) excrete urine
    containing large amounts of acid whereas persons
    with urinary tract infections or persons taking
    certain antibotics will excrete an alkaline
    urine.
  • INSTRUCTIONS
  • 1. Place a few ml's of fresh urine into a small
    cup. Dip a strip of pH test paper into the
    sample 2 or 3 times.
  • 2. Tap the pH paper on the edge of the cup to
    remove excess urine. Wait one minute
    record the pH on the Results Sheet.

7
PART 2 CHEMICAL COMPOSITION OF URINE
  • A. GLUCOSE
  • Glucose is the sugar most commonly found in
    urine, although other sugars such as lactose,
    fructose, or galactose may also be found under
    certain conditions. The presence of detectable
    amounts of glucose in urine is known as
    glycosuria, and it may either occur in normal or
    abnormal conditions. For example, glucose may
    normally spill into the urine following
    consumption of a heavy meal, especially one that
    contains large amounts of carbohy-drates, or in
    conjunction with emotional stress. The most
    common abnormal condition resulting in glycosuria
    is diabetes mellitus. There are several
    different tests available which check for the
    presence of glucose in urine.

8
INSTRUCTIONS
  • Test 1 Glucose Determination Using Clinitest
    Reagent Tablets.
  • 1. Place 10 drops of water 5 drops of urine in
    a test tube.
  • 2. Add 1 Clinitest tablet. Note Handle
    Clinitest tablets with extreme
    caution, using gloves. These tablets contain
    caustic soda which is highly
    sensitive to moisture from air or water.
    Excessive moisture may cause chemical burns
    or bottle explosion.
  • 3. Watch while a complete boiling reaction takes
    place. (Boiling occurs because the
    concentrated sodium hydroxide in the
    tablet generates heat.) Do not shake the test
    tube during the boiling or for the
    following 15 seconds after the
    boiling has stopped.
  • 4. After the 15 seconds waiting period, gently
    shake the test tube to mix the
    contents. Compare the color in the tube to
    the standard color chart (for 5 drop
    method, not 2 drop method).
  • 5. Record results as Negative, Trace, 1, 2,
    3, or 4 on the Results Sheet.

9
Test 2 Glucose Determination Using Diastix
  • 1. Pour a small amount of urine into a small
    cup.
  • 2. Dip the reagent end of the strip in urine (do
    not touch the reagent end with your hands).
    Remove immediately.
  • 3. While removing, draw the edge of the strip
    against the rim of the cup to remove excess
    urine.
  • 4. Compare the reagent side with the
    corresponding color chart (on bottle) at
    exactly 30 seconds.
  • 5. Record results on Results sheet.

10
Test 3 Glucose Determination Using Benedict's
Test
  • 1. Place 5 ml of Benedict's solution in a test
    tube.
  • 2. Add exactly 8 drops of urine and boil the
    solution for 2 to 5 minutes.
  • 3. If the solution remains blue, the test is
    negative green is recorded as , yellow as
    , orange as , and red as .
  • 4. Record results on Results sheet.

11
B. KETONES
  • Normally the body completely metabolizes fats to
    carbon dioxide and water. Whenever there is
    inadequate carbohydrate intake or a defect in
    carbohydrate metabolism, the body metabolizes
    increased amounts of fatty acids. When this
    increase is large, fatty acid utilization is
    incomplete and intermediate products (Ketones)
    appear in the blood and are excreted in the
    urine. Diabetes mellitus is the most important
    disorder in which ketonuria (presence of ketones
    in urine) occurs. As with glucose, there are
    several different tests available which check for
    the presence of ketones.

12
INSTRUCTIONS
  • Test 1 Ketone Determination Using Acetest
    Reagent
  • Tablets
  • 1. Place an Acetest tablet on a clean dry piece
    of paper and put 1 drop of urine on the tablet.
  • 2. At 30 seconds, compare the color formed with
    the color chart.
  • The color will remain unchanged or
    cream-colored in a negative test, but will
    vary from pink to purple, depending on
    the amount of ketone bodies present, in a
    positive test.
  • 3. Record results on Results Sheet.

13
Test 2 Ketone Determination Using Ketostix
Strips
  • Place a small amount of urine in a small cup.
  • Dip the reagent end of the strip in urine (do not
    touch the reagent end with your hands). Remove
    immediately.
  • 3. While removing, draw the edge of the strip
    against the rim of the cup to remove excess
    urine.
  • 4. Exactly 15 seconds after removing the strip
    from the specimen, compare the
    reagent side with the corresponding color chart
    (on bottle). The color will remain unchanged or
    appear cream colored (from wetting) if negative
    if positive, the color will vary from pink to
    purple depending upon the amount of ketones
    present.
  • 5. Record results on Results Sheet.

14
C. PROTEIN
  • Normally, between 40 and 80 mg of protein is
    excreted daily, but as much as 100 - 150 mg per
    day may be considered within normal limits.
    Typically however, this normal protein isn't
    detected in the urine. Proteinuria refers to an
    increased amount of protein in the urine and is
    one of the most important indicators of renal
    disease however, this condition may also
    intermittently occur in the absence of disease -
    usually as a result of strenous physical exercise
    or severe emotional stress. Several tests are
    available for measuring urinary protein.

15
INSTRUCTIONS
  • Test 1 Protein Determination Using Albustix
    Reagent
  • Strips.
  • 1. Place a small amount of urine in a small cup.
  • 2. Dip the reagent end of the strip in the urine
    (do not touch the reagent end with your hands).
    Remove immediately.
  • 3. While removing, draw the edge of the strip
    against the rim of the cup to remove excess
    urine.
  • 4. Immediately after removing the strip from the
    urine, compare the reagent side with the
    corresponding color chart (on bottle).
  • 5. Record results on Results Sheet.

16
D. BILE SALTS
  • Bile salts are an essential component of bile.
    Normally the majority these
  • substances are recycled in the body with only
    small amounts lost in urine (the
  • loss usually isn't detectable). Hay's Test for
    bile salts involves the addition of
  • sulfur to urine. Since bile salts lower the
    surface tension of the water in urine, if
  • bile salts are present, the sulfur will sink.
  • INSTRUCTIONS
  • Hay's Test for Bile Salts in Urine
  • 1. Place a small amount of urine in a small cup.
  • 2. Sprinkle a little finely powdered sulfur on
    the surface of the urine. If the sulfur sinks
    at once, bile salts are present in the amount
    of 0.01 or more. If the sulfur sinks
    only after gentle agitation, bile salts are
    present in the amount of 0.0025 or more.
    If the sulfur remains floating even after
    gentle shaking, bile salts are absent.
  • 3. Record results on Results Sheet.

17
E. COMBINATION TESTS
  • INSTRUCTIONS
  • Combistix Method (Determination of glucose,
    protein, pH)
  • 1. Place a small amount of urine in a small cup.
  • 2. Dip the reagent end of the Combistix strip in
    urine ( do not touch the reagent end with your
    hands). Remove immediately.
  • 3. While removing, draw the edge of the strip
    against the rim of the cup
    to remove excess urine.
  • 4. The yellow part of the strip is the indicator
    for protein. It should be read immediately and
    compared with the color chart (on bottle).
  • 5. The aqua part of the strip is the test for
    glucose. This should be read after 30 seconds by
    comparing the strip to the color chart (on
    bottle).
  • 6. The orange part of the strip is the pH
    indicator. It should be read immediately and
    compared with the color chart (on bottle).
  • 7. Record results on Results sheet.

18
PART 3 THE MICROSCOPIC EXAMINATION OF SEDIMENT
OBTAINED FROM
URINE
  • INSTRUCTIONS
  • 1. Obtain the urine which has set in the test
    tube for 1 hour.
  • 2. Read the Discussion (below) and complete the
    Procedure (below).

19
PROCEDURE
  • 1. Place 10 ml of fresh urine in a centrifuge
    tube and centrifuge for 5 minutes.
    (Precaution Make certain the centrifuge is
    balanced with another tube containing 10 ml of
    urine or water.)
  • 2. Pour out all the urine possible (do this
    quickly without shaking the tube or inverting it
    for more than a moment).
  • 3. Add 2 drops of Sedi-Stain to the sediment in
    the tube. Thoroughly mix the contents of the tube
    with the stain by tapping briskly on the
    bottom of the tube with the index finger.
  • 4. Transfer 1 drop of the stained sediment onto a
    clean glass slide.
  • 5. Cover the drop of sediment with a cover glass
    and examine it under the microscope using both
    low and high power objectives.
  • 6. Identify as many of the substances in the
    sediment as possible by comparing them with
    Figures 1 - 4 (see attached page - this
    handout), as well as with the urine chart (on the
    materials table in the lab). Make a drawing of
    your observations (white paper, high power). Be
    certain to properly label your drawing.
  • 7. Include your drawing in your lab report.
  • ANSWER THE QUESTIONS FROM THE QUESTIONS SHEET.
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