Name: Section:

EXPERIMENT NO. 3b

THE PROPER TECHNIQUE OF PERFORMING CHEMICAL EXAMINATION OF URINE USING REAGENT STRIP
AND CONVENTIONAL METHODS

General Instructions:

1. All answers MUST be in capital letters except for units of expressions.

2. Answers should be brief and concise.

3. Avoid erasures and alterations.

Criteria:

Content 50%

Accuracy 20%

Organization of thoughts 20%

Neatness 10%

1. Why is the timing of reagent strip readings considered critical when reporting its results? (5 points)

The timing of reagent strip readings is considered critical when reporting its results because the
reactions on the strip can change over time, affecting the accuracy of the results. Reagent strips, also
known as dipsticks, are used to test various substances in bodily fluids, such as urine or blood. These
strips contain specific chemical reagents that react with the substances of interest, producing color
changes or other measurable responses. The timing is critical for several reasons Reaction Time, Reagent
Stability, Color development.

2. What is the most convenient as well as the most reliable method in the chemical examination of the
urine and why? (5points)

The most convenient and reliable method for chemical examination of urine is the use of reagent strips
(dipsticks). Reagent strips are widely used in clinical settings and laboratories simple to use and do not
require specialized equipment or extensive sample preparation. They are designed for quick and easy
testing, making them highly convenient for healthcare professionals, laboratories, and even at-home use.
3. List all the parameters of the reagents strip and state the reaction principle of each. (10 points)

The parameters commonly tested by reagent strips, along with their reaction principles, include:

pH (Acid-Base Balance):

Reaction Principle: pH indicator dyes on the strip change color in response to the acidity or alkalinity of
the urine. The strip is usually labeled with a color chart to compare and determine the pH level.

Protein:

Reaction Principle: Protein tests on reagent strips typically use tetrabromophenol blue or other indicator
dyes that change color in the presence of proteins. The intensity of the color change corresponds to
protein concentration.

Glucose (Sugar):

Reaction Principle: Glucose oxidase or glucose dehydrogenase enzymes on the strip catalyze the
oxidation of glucose. This reaction produces hydrogen peroxide, which then reacts with a chromogen,
leading to color change. The intensity of the color change correlates with glucose concentration.

Ketones:

Reaction Principle: Reagent strips use nitroprusside or similar compounds to detect acetoacetic acid, a
type of ketone. The reaction between acetoacetic acid and the reagent results in a color change,
indicating the presence and concentration of ketones.

Bilirubin:

Reaction Principle: Bilirubin reagent strips contain diazo reagents that react with bilirubin to form
azobilirubin. This reaction produces a color change, indicating the presence of bilirubin.

Urobilinogen:

Reaction Principle: Urobilinogen reagent strips use p-dimethylaminobenzaldehyde as a reagent to react

with urobilinogen. The reaction forms a colored compound, and the intensity of the color change
corresponds to urobilinogen concentration.

Blood (Hematuria):

Reaction Principle: Hemoglobin or myoglobin in the urine reacts with a peroxidase-like reagent on the
strip in the presence of hydrogen peroxide. This reaction produces a color change, indicating the
presence of blood in the urine.

Leukocytes (White Blood Cells):

Reaction Principle: Leukocyte esterase on the strip reacts with esterase enzymes released by white blood
cells (leukocytes) in the urine. The reaction leads to a color change, indicating the presence of white
blood cells, which may suggest infection or inflammation.
Specific Gravity:

Reaction Principle: Reagent strips often measure specific gravity using a pad that contains chemicals
sensitive to solute concentration. The pad's color change is used to estimate the specific gravity of the
urine, reflecting its concentration.

Nitrite:

Reaction Principle: Nitrite reagent strips detect the presence of nitrate-reducing bacteria in urine. These
bacteria convert nitrate to nitrite, which reacts with a reagent on the strip to produce a color change,
indicating the potential presence of a urinary tract infection.

4. Give one commonly occurring condition associated with each urine reagent strip parameter and
discuss shortly. (30 points)

pH (Acid-Base Balance):

Condition: Urinary Tract Infection (UTI)

Discussion: UTIs can lead to an abnormally acidic pH in the urine. The presence of bacteria in the urinary
tract can produce acidic byproducts, resulting in lower pH levels. Symptoms of UTIs include frequent
urination, burning sensation during urination, and cloudy or foul-smelling urine.

Protein:

Condition: Kidney Disease (Proteinuria)

Discussion: Proteinuria is the presence of excess protein in the urine, which can be an early sign of
kidney disease. The kidneys should filter out waste products while retaining essential proteins in the
blood. When the kidneys are damaged or not functioning properly, they may allow protein to pass into
the urine.

Glucose (Sugar):

Condition: Diabetes

Discussion: Diabetes is a condition characterized by elevated blood sugar levels. If the blood glucose
levels are consistently high, the excess sugar can spill over into the urine, causing glycosuria. Glucose in
the urine is a common early sign of diabetes mellitus.
5. List the conventional methods of urine chemical examination and its positive reaction (20 points).

Clinitest:

Test: Detects reducing substances (e.g., glucose) in the urine.

Positive Reaction: A color change from blue to various shades of green, yellow, orange, or brown
indicates the presence of reducing substances.

Sulfosalicylic Acid Test:

Test: Detects the presence of protein (albumin) in the urine.

Positive Reaction: The formation of a cloudy or turbid appearance in the urine indicates the presence of
protein.

HCl Acidification Test:

Test: Detects the presence of xanthine or uric acid crystals in urine sediments.

Positive Reaction: The formation of crystals (yellow or red-brown) upon acidification indicates the
presence of xanthine or uric acid.

Heat Test:

Test: Detects the presence of bilirubin in the urine.

Positive Reaction: A green color develops when bilirubin is present in the urine after heating, usually
indicating liver or gallbladder issues.

Acetest Tablet Test:

Test: Detects the presence of ketones in the urine.

Positive Reaction: A color change of the reagent tablet to purple indicates the presence of ketones, such
as acetoacetic acid.

Nitric Acid Test:

Test: Detects the presence of tyrosine or homogentisic acid in urine sediments.

Positive Reaction: The formation of a dark color upon addition of nitric acid indicates the presence of
tyrosine or homogentisic acid crystals.
6. Choose one conventional method of urine chemical examination and discuss its principle and
procedure. (10 points)

Sulfosalicylic Acid Test, which is used to detect the presence of protein (specifically, albumin) in the
urine.

Principle:

The Sulfosalicylic Acid Test is based on the principle that when urine contains proteins, particularly
albumin, the addition of sulfosalicylic acid causes protein precipitation, resulting in a turbid or cloudy
appearance. This turbidity is due to the formation of protein-sulfosalicylic acid complexes. The degree of
turbidity is directly proportional to the amount of protein present in the urine.

Procedure:

Begin by collecting a fresh urine sample in a clean, dry container. It's important to work with a well-
mixed sample.

Label a test tube or small plastic cup with the patient's information and the date and time of collection.

Using a pipette or dropper, transfer a small volume (usually 1-2 mL) of the urine sample into the labeled
container.

Add a few drops (2-3 drops) of the sulfosalicylic acid reagent to the urine sample in the test tube.

Gently mix the contents of the test tube by swirling or using a glass rod. Ensure thorough mixing.

Observe the reaction immediately and compare the appearance of the urine to a standard reference. A
positive reaction is indicated by the formation of turbidity or cloudiness in the urine.

The degree of turbidity can be graded as trace, 1+, 2+, 3+, or 4+ based on the amount of protein present.
A higher numerical value indicates a higher concentration of protein.

Record the results in the patient's medical records or on a laboratory report.