ACTIVITY13

CARBON DIOXIDE DETERMINATION

Unit Intended LearningOutcomes

At the end of the activity, the students are expected to:

Demonstrate appropriately the specimen preparation.

Perform properly the method in determining carbon dioxide.
Analyze correctly the test results with clinical signs and symptoms.

Pre-laboratory Discussion

The total CO2 content includes the serum bicarbonate as well as the available form of CO2 (i.e.
dissolve CO2 and carbonic acid). Generally, the serumbicarbonate(HCO3-), composesabout 95% of
thetotal CO2 content:thuswe can use thismeasurementasanexcellentestimatorof
serumbicarbonate.The total CO2 contentnormally equals 23 to 30 mEq/L in serum.

The kidneys and lungs maintain daily acid-base balance. Understanding this normal physiology allows us
to appreciate abnormalities. This discussion refers to bicarbonate rather than total CO 2 content, as we
measure total CO2 content as a surrogate for bicarbonate.

Bicarbonate and carbonic acid constitute the major buffer pair in body fluids. Carbonic acid dissociates
into hydrogen ion and bicarbonate with a dissociation constant of 7.9 x 10.7. Carbonic acid maintains an
equilibrium with water and CO 2. We usually describe dissociation constants and hydrogen ion
concentration as negative logarithm. Thus, the negative logarithm of the dissociationconstant equals 6.1.
This value is called the pK2. Normal hydrogen ion concentration equals 40 nano-equivalents/Liter,
corresponding to pH 7.4.
 Teaching and Learning Activities

Pre-Analytical Phase

1. Disinfect the working area using 10% sodium hypochlorite.

2. Prepare all the materials/equipment needed.
3. Wear all personal protective equipment (PPE) before starting the activity.

Analytical Phase

Materials

1. Serological pipettes 1 mL and 5 mL

2. Test tubes 10 x 100 mm

Reagents

1. Carbon dioxide Base Reagent

2. Carbon dioxide Indicator
3. Carbon dioxide Standard, 26 mmol/L

Procedure

Reagents Standard Test Sera/Control

Carbon dioxide Standard 0.1 mL ----------
Test Sera/Controls ---------- 0.1 mL
Carbon dioxide Base Reagents 3 mL 3 mL

1. Do not mix to prevent the loss of CO2.

Reagents Standard Test Sera/Control

Carbon dioxide Indicator 1 drop 1 drop

2. Mix well and read % Transmittance/Absorbance of each tube at 550 nm against water blank.

Calculation of Results

OD of Unknown Value of the

Value of the test/unknown =--------------------------------× Standard 26
mmol/L OD of Standard mmol/L
Note:

1. All glasswares should be acid cleaned and rinsed with deionized water.
2. Use vacuumcollectiontubes and avoidprolongedcentrifugation to preventthe loss of CO 2
3. Run the test within one (1) hour after withdrawal of the specimen.

Post-Analytical Phase

1. Segregate and dispose all wastes into proper trash bins.

2. Disinfect the working area using 10% sodium hypochlorite.
3. Remove all the PPE properly after the activity.
Name: _ Date: _
Section: Group: Score: _

Review Questions

1. What is the ratio of bicarbonate to carbonic acid when kidneys and lungs are functioning properly?

2. Write the Handerson-Hasselbach equation. What is the importance of this equation in blood gas
determinations?

3. Completethe table below of the followingblood gas abnormal conditions:(State if

“higher”or “lower”)

Conditions pH HCO3- PCO2 Typical Causes

1.
Metabolic acidosis 2.
3.
1.
Metabolic alkalosis 2.
3.
1.
Respiratory acidosis 2.
3.
1.
Respiratory alkalosis 2.
3.
 Assessment Criteria for Passing

Unit Intended Learning Outcomes
1. Demonstrate appropriately the specimen
preparation.
2. Perform properly the method in determining
carbon dioxide.
3. Analyze correctly the test results with clinical
signs and symptoms.
To achieve this unit, the learner must:
a. Wear PPE such as gloves, mask, hair net,
and lab gown properly.
b. Collect accurately 4-5 mL of bloodin
venipuncture procedure.
c. Ensure proper labelling of the specimen by
writing the name, age, sex, date & time of
collection on the tube.
a. Demonstrate the correct pipetting technique
by aspirating 0.1 mL, 1 mL, and 3 mL volume
of samples andreagents.
b. Set and read the absorbance of
spectrophotometer against water blank at
550 nm.
a. Interpret laboratory test result.
b. Correlate laboratory results with clinical signs
and symptoms.
c. Ensure proper waste disposal in the following
containers: sharps, biodegradable, non-
biodegradable, and infectious waste.

Rubrics

Weight 1- 2 - Developing 3 - Satisfactory 4 - Exemplary Score

Unsatisfactory
Pre-Analytical (35%)
Score ×
Wore 1 out of 4 Wore 2 out of Wore 3 out of Wore 4 out of 1.25
PPE (5%) =
PPE 4 PPE 4 PPE 4 PPE
Score ×
1-2.5 mL of 2.6 to 3.9 mL 4 to 5 mL of 3.75
Venipuncture < 0.9 mL of =
blood of blood blood
(15%) blood collected
collected collected collected

Labelled the Labelled the Labelled the Labelled the Score ×

3.75
tube with only tube with only tube with only tube with =
Specimen labelling
1 information 2 information 3 information complete
(15%)
about the about the about the patient’s
patient patient patient information
Analytical (45%)
 All aspirated Score × 5 =
All aspirated Only 1 volume Only 2
volume of
volume of of either volumes of
samples and
samples and sample or either sample
reagents were
Pipetting reagents were reagent or reagent
exactly the
techniques (20%) less than the needed for the needed for the
required
required analysis was analysis were
amount
amount needed aspirated aspirated
needed for
for analysis correctly correctly
analysis
Performed Score ×
Performed Performed Performed 6.25
incorrect =
incorrect correct setting correct setting
setting of
setting of of wavelength of wavelength
Setting the wavelength of
wavelength of of the device of the device
spectrophotometer the device and
the device and and reads the and reads the
and reads the reads the
read the absorbance absorbance
correct absorbance absorbance
absorbance without water against water
(25%) against water
without water blank at 550 blank at 550
blank at 550
blank nm nm
nm
Post-Analytical (20%)
Interpretation Interpretation Score ×
Interpretation 3.75
Interpretation of lab results of lab results =
and
Laboratory Results and correlation was correct was incorrect
correlation of
(15%) of lab results while the while the
lab results
were incorrect correlation correlation
were correct
was incorrect was correct
2 types of 3 types of Score ×
4 types of All wastes 1.25
wastes were wastes were =
Waste Disposal wastes were were disposed
disposed to disposed to
(5%) not disposed to to appropriate
appropriate appropriate
appropriate bag bag
bag bag
Total Score: