Clinical Chemistry 2 - Blood Gases, PH, Buffer

Systems

Principles of Medical Laboratory Science (Holy Name University)

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Blood Gases, pH, & Buffer 4/16/2 CC
System LEC 1 2

BLOOD GASES, PH, BUFFER SYSTEMS ⊟ Body produces much greater quantities of H+, but
The information on the patient’s acid-base balance and blood gas the body controls and excretes H+ in order to
homeostasis are used to assess patients in life-threatening maintain pH homeostasis.
situations. ○ Any values of H+ outside of normal range
cause alterations in rates of chemical
Definitions: reactions and metabolic processes leading to
Arrhenuis’s definition: alterations in consciousness, neuromuscular
+ Acid: a substance that increases the concentration irritability, tetany, coma, death
of hydrogen ion/H+ when dissolved in water ○ Sources of H+ ions:
+ Base: a substance that increases the ✓ Breakdown of phosphorus-containing proteins
concentration hydroxyl ions/OH- when dissolved releases phosphoric acid
in water. ✓ Anaerobic respiration of glucose produces
lactic acid
Bronsted & Lowry’s definition: most widely accepted and most ✓ Fat metabolism produces fatty acids
clinically relevant and ketones
× Acid: a substance that donates a proton in a reaction ✓ Loading and transport of carbon dioxide in the
× Base: a substance that accepts a proton in reaction blood as bicarbonate release H+

Lewis definition: ⊟ The logarithmic pH scale expresses H+ concentration

↣ Acid: molecule or ion that accepts a pair of electrons pH = log 1/concentration of H+ = -log concentration of H+
to form a covalent bond
↣ Base: a molecule that donates a pair of electrons to form ⊸ Arterial blood pH is 7.4; equivalent to an
a covalent bond H+ concentration of 40 nmol/L
⊸ Venous blood pH is 7.35
The relative strengths of acids and bases, their ability to ⊸ Because pH is a negative log of H+ concentration,
dissociate in water, are described by their dissociation an increase in H+ decrease pH and a decrease in H+
constant/ionization constant/ K value. increases pH
------------------------------------- ⊸ Acidosis: pH below 7.34
pKa ⊸ Alkalosis: pH above 7.44
∶ Defined as the negative log of the ionization constant ⊸ Chemically speaking, pH of 7.35 is not acidic if
∶ Is also the pH in which the protonated and one considers a neutral pH is 7.0
unprotonated forms are present in equal o However, at pH 7.35 concentration is higher
concentrations than optimal for most cells; this is called
∶ Strong acids have pKa values less than 3.0; strong bases physiologic acidosis.
have pKa values greater than 9.0 Suffixes
∶ For acids, raising the pH above the pKa will cause ⊸ -osis refers to a process in the body
the acid to dissociate and yield a H+ (hydrogen) ⊸ -emia refers to corresponding state in the
∶ For bases, lowering the pH below pKa will cause the base blood (alkalemia, acidemia)
to release OH- (hydroxyl ion) ⊸ pH controlled by systems that regulate the production &
retention of acids and bases; this include:
Buffer o buffers
~ Is a combination of a weak acid or a weak base and o respiratory centers and lungs
its salt; a system that resists changes in pH o kidneys
~ The effectiveness of a buffer depends on the pKa of the o buffer systems (chemical buffer system) act
buffering system and the pH of the environment faster than lungs & kidneys (physiological
~ The bicarbonate-carbonic acid system with a pKa of 6.1 buffering system); physiological buffers have
is one of the principal buffers in the body(H2CO3 ⟷ more buffering power.
HCO3- + H+)
~ The reference value for blood plasma pH is 7.40 Buffer System: Regulation of H+
~ If pH of 100 ml of water is 7.35 and one drop of 0.05 ⊙ Body’s first line of defense against extreme changes
mol/L of HCl is added, pH will change from 7.35 to in H+ concentration; resist changes in pH when a
7.0 strong base or acid is added (bind H+ when pH drops,
~ To change 100ml of blood to form pH of 7.35 to 7.0, release H+ when pH rises)
25ml of 0.05 mol/L of HCl is needed; for 5.5L of blood in
the average body, more than 1,300 ml of HCl is needed. Bicarbonate-Carbonic Acid Buffer System
⋮ Consist of a weak acid (carbonic acid/ H2CO3) and its salt
Acid-Base Balance or conjugate base (bicarbonate/ HCO3-)
Maintenance of H+
⊟ Normal concentration of H+ in the ECF
(extracellular fluid) is 36-44 nmol/L (pH 7.34-7.44)
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Blood Gases, pH, & Buffer 4/16/2 CC
System LEC 1 2
⋮ H2CO3 (carbonic acid) is a weak acid since it does not and diffuse into plasma; to maintain electroneutrality
completely dissociate into H+ and HCO3- (same
⋮ When a strong acid is added, the HCO3- (bicarbonate)
will combine H+ to form H2CO3
HCl + NaHCO3 ⇢ H2CO3 + NaCl
⋮ When a strong base is added, H2CO3 will combine
with OH- to form H2O and HCO3-
NaOH + H2CO3 ⇢ NaHCO3 + H2O
⋮ A smaller change of pH result from adding acid or base
⋮ Buffering power of this system is directly related to the
concentration of buffering substances
○ When all available HCO3- (carbonic acid)
(referred to as alkaline reserve) is used up,
the buffer system becomes ineffective and
blood pH changes
○ HCO3- concentration in ECF (extracellular
fluid) is 25 mEq/L

Buffer system is important because:

1. H2CO3 dissociate into H2O and CO2
allowing CO2 to be eliminated by lungs and
H+ as H2O
2. Changes in CO2 modify ventilation/
respiratory rate
3. HCO3- concentrations can be altered in
the kidneys
4. Counter effects of non-volatile acids (H+A-)
by binding the dissociated hydrogen ion
(H+A- + HCO3 + A-) ; resultant H2CO3 then
dissociates and H+ is neutralized by the
buffering system of hemoglobin.

⟡ Consists of sodium salts of HPO4 –

Phosphate Buffer System

(monohydrogen phosphate) and H2PO4-

⟡ NaH2PO4 acts as a weak acid, Na2HPO4 acts as a

(dihydrogen phosphate)

weak base
HCl + Na2HPO4 ⇢ NaH2PO4 + NaCl

⟡ Play a role in plasma and RBC; involved in the

NaOH + NaH2PO4 ⇢ Na2HPO4

exchange of sodium ion in the urine H+ filtrate

⟡ Buffer system in plasma

Plasma Proteins (esp. imidazole group of histidine):

⟡ Most circulating proteins have a net negative charge

and are capable of binding H+

Lungs and Kidneys: regulation of Acid-base balance

⤌ Carbon dioxide, the end product of aerobic
metabolism, diffusion out of tissues and enters plasma
and RBC
⤌ In the plasma, small amounts of CO2 is dissolved or
combined with proteins to form carbamino
compounds
⤌ Most of CO2 combines with H2O to form H2CO3,
which quickly dissociates to H+ and HCO3- ; reaction
accelerated by carbonic anhydrase in RBC membranes
CO2 + H2O ⇠ ca ⇢ H2CO3 ⟷ H+ + HCO3-
⤌ Chloride shift: HCO3- concentration increase in RBC
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Blood Gases, pH, & Buffer 4/16/2 CC
System LEC 1 2
number of positive and negative ions on each side of H+; HCO3- is reabsorbed into blood together
RBC membrane), chloride diffuses into RBC. with Na+

⟡ Kidney excretes little unbuffered H+

⤷ Maximum urine pH is 4.5

⟡ Most of urinary H+ is combined with

In the lungs, process is reversed:
⤏ Inspired O2 diffuses from alveoli into blood and is
bound to hemoglobin forming oxyhemoglobin or ammonia (NH3) and monohydrogen
O2Hb phosphate (HPO4-)
⤏ The H+ that was carried on the reduced
hemoglobin is released to recombined with HCO3-
to form H2CO3, which dissociates into H2O and
CO2; CO2 diffuses into alveoli and is eliminated
through ventilation
⤏ Net effect is a minimal change in H+
concentration between venous and arterial
blood
⤏ Ventilation affects pH of blood:
⤏ When lungs do not remove CO2 at the rate of its
production (ex: decreased ventilation or disease),
CO2 accumulates causing an increase in H+
concentration
⤏ When lungs remove CO2 faster than production
(ex: hyperventilation), H+ concentration is
decreased.

⤏ A change in H+ concentration of blood that results

from non-respiratory disturbances cause the
respiratory centers to respond by altering rate of
breathing to restore blood pH
○ When pCO2 or when blood H+
concentration rises, respiratory rate and
depth increases
○ When blood pH rises, respiratory rate
drops and respiration becomes shallower
○ Thus, lungs and the buffer system are the
first line of defense to change acid-base
status.

Kidneys:
⤷ Acts slowly but surely (takes hours to days)
to compensate for acid-base imbalances
⤷ Can rid body of acids generated by cellular
metabolism/nonvolatile acids (uric acid, phosphoric
acid, lactic acid, ketones)
⤷ Also regulates blood levels of alkaline substances
and renew chemical buffers
⤷ Main role is to reabsorb HCO3- from the
glomerular filtrate (if HCO3- is lost in urine, excess

⟡ Glomerular filtrate contains same HCO3-

acid gain in the blood results)

⟡ Reabsorption takes place in proximal tubules

levels as plasma

⟡ HCO3 is not directly transported across

tubular membrane; instead, Na+ in the
glomerular filtrate is exchanged for H+ in
tubular cells; H+ combines with HCO3- in
the filtrate to form H2CO3 which is
converted to H2O and CO2 by carbonic
anhydrase; CO2 diffuses into tubular
membranes and reacts with H2O to reform
H2CO3, which dissociates into HCO3- and
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Blood Gases, pH, & Buffer 4/16/2 CC
System LEC 1 2

and is excreted as ammonium (NH4) and

Acid-Base Disorders
dihydrogen phosphate (H2PO4--) ⧺ Acidemia – when blood pH is less than the

⟡ Blood plasma HCO3- levels > 26-30 mmol/L;

⤷ Factors that affect reabsorption of (bicarbonate) HCO3-: reference range; reflects excess acid or H+ concentration
⧺ Alkalemia – when blood pH is greater than reference
occurs when HCO3- is given Intravenously, range; excess base
excess amounts of lactate or acetate; HCO3 ⧺ Primary respiratory acidosis or alkalosis – disorder

⟡ Low plasma levels of HCO3- : occurs

will be excreted by kidneys caused by ventilatory dysfunction / changes in the
pCO2 or carbon dioxide pressure (respiratory
with diuretic use, chronic nephritis; component)
HCO3- reabsorbed by kidneys. ⧺ Non-respiratory acidosis or alkalosis – disorder resulting
from change in the bicarbonate levels (renal / metabolic

⟡ To assess acid-base homeostasis, components of the

Assessment of Acid-Base Homeostasis component)

bicarbonate buffering system are measured and ⧺ Whenever an imbalance occurs, the body tries to
calculated; inferences can be made pertaining to restore acid-base homeostasis by compensation

⟡ In the bicarbonate buffer system, the dissolved CO2

other buffer systems ⧺ Compensation is done by altering the function or
component that is not primarily affected by
(dissolved CO2) is in equilibrium with CO2 gas which the pathological process.
can be expelled by the lungs; thus the bicarbonate ○ If imbalance is of non-respiratory origin,
buffer system is referred to as an open system and the body compensates by altering ventilation

⟡ Lungs participate rapidly in regulation of the blood

dissolved CO2 is the respiratory component. ○ For disturbances of the respiratory
component, kidneys compensate by

⟡ Kidneys, the non-respiratory / metabolic

pH through hypo- and hyperventilation. selectively excreting or reabsorbing anions
and cations.
component, controls the bicarbonate concentration. ○ Lungs can compensate immediately but
response is short-term and often

⟡ Expresses acid-base relationship in a

Henderson-Hasselbalch Equation incomplete
○ Kidneys are slower to respond (2-4 days)
mathematical formula but response is long-term and complete
pH = pKa + log cA-/cHA ⧺ Fully compensated – implies that pH has returned to
○ pKa is the pH at which there is normal range (20:1 ratio has been restored)
equal concentrations of protonated ⧺ Partially compensated – implies pH is
and unprotonated species approaching normal
○ A- the proton acceptor or base (HCO3-)

⟡ In the plasma at 37oC body temperature, pKa of ⟡ Decrease in (bicarbonate) HCO3- (<24 mmol/L) resulting
○ HA is the proton donor or weak acid (H2CO3) 1. Primary Non-Respiratory Acidosis /Metabolic Acidosis

⟡ The equilibrium between H2CO3 (carbonic acid) and

the bicarbonate buffering system is 6.1 in a decrease in pH

⟡ Concentration of H2CO3 is proportional to the ⟡ Direct administration of acid-producing substance:

CO2 in plasma is 1:800 Causes:

⟡ In plasma at 37oC, the value for the combination of ⟡ Excessive formation of organic acids: ex:
partial pressure exerted by the dissolved CO2 ex: ammonium chloride, calcium chloride

⟡ Reduced excretion of acid: ex: renal tubular acidosis

the solubility constant for pCO2 and the factor to diabetic ketoacidosis, starvation

⟡ Excessive loss of HCO3- ex: diarrhea, drainage

convert mmHg to mmol/L is 0.0307 mmol/L/mm Hg
○ Temperature and solvent affect the constant;
if either changes, the solubility constant also from biliary, pancreatic or intestinal fistula

⟡ pH and pCO2 are measured in blood gas analysis

changes

⟡ HCO3- can be calculated as: ⟡ Hyperventilation (increase rate and depth of

Body compensates by:

⟡ Substituting normal values, the equation in healthy

pH = pKa + log cHO3- / (0.0307 x pCO2)
⟡ Secondary compensation: kidneys begin to
breathing) to blow off CO2

retain bicarbonate

2. Primary Respiratory Acidosis

individuals reads: ﹢ Results from a decrease in alveolar ventilation /
pH = 6.1 + log 24 mmol/L / (0.0307 mmol/L/mm Hg x 40 mm Hg) hyperventilation leading to decreased elimination of CO2
= 6.1 + log (24 / 1.2) by the lungs
= 6.1 + log (20) Causes:
= 6.1 + 1.3 ﹢ Chronic obstructive pulmonary disease (COPD)

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Blood Gases, pH, & Buffer 4/16/2 CC
System LEC 1 2
= 7.40 ﹢ Bronchopneumonia
﹢ Drugs: barbiturates, morphine, alcohol

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Blood Gases, pH, & Buffer 4/16/2 CC
System LEC 1 2
﹢ Mechanical obstruction or asphyxiation (strangulation, ↣ Partial pressure of each gas is equal to the BP at a
aspiration)
particular altitude times the appropriate percentage
﹢ Decreased cardiac output: ex: CHF or congestive heart
for each gas
failure (less blood going to lungs for gas exchange)
↣ Vapor pressure of water (47 mm Hg at 37oC) must be
accounted for calculating partial pressure for each gas;
Compensation:
﹢ Kidneys increase excretion of H+ and increase in the body, these gases are always fully saturated with
reabsorption of HCO3 water
↣ Partial pressure of oxygen in the body at sea level:
3. Primary Non-Respirator Alkalosis/ Metabolic alkalosis (760 mmHg – 47 mm Hg) x 20.93% = 149 mm Hg at 37oC
＝ Results from a gain in HCO3- leading to an increase in pH ↣ Partial pressure of CO2 in the body at sea level:
Causes: (760 mm Hg – 47 mm Hg) x 0.03% = 2 mm Hg at 37oC
＝ Excess administration of sodium bicarbonate
＝ Ingestion of bicarbonate-producing salts: ex: sodium 2. Adequate ventilation
lactate, citrate, acetate ∶ During inspiration, the thoracic cavity expands creating
＝ Excessive loss of acid: es: vomiting, nasogastric e negative pressure gradient causing air to enter alveoli
suctioning, prolonged use of diuretics that cause H+ ∶ During inspiration, airways are still filled with air retained
excretion from the previous expired breath; this is called dead
space air; dead space air dilutes the air being inspired;
Compensation: air being inspired is also fully saturated with water
＝ Depress respiratory centers causing hypoventilation vapor causing inspired air to have only a pO2 of 110mm
Hg (instead of 149 mm Hg)

⟡ Results from increased rate of alveolar

4. Primary Respiratory Alkalosis ∶ Three factors influence pO2 in alveoli:
○ Percentage of oxygen in inspired air
ventilation causing increases loss of CO2 ○ Amount of pCO2 in the expired air

⟡ Hypoxemia
Causes: ○ Ratio of the volume of inspired air to

⟡ Chemical stimulation of respiratory centers by drugs:

the volume of dead space air
∶ Percentage of oxygen in inspired air / fraction of inspired

⟡ Increase in environmental temperature

ex: salicylates oxygen/ FiO2 can be increased by breathing gas

⟡ Fever
mixtures of up to 100% oxygen

⟡ Hysteria
∶ Conditions that influences pCO2 in expired air; increased

⟡ Pulmonary emboli
metabolism, hyperthermia produce more CO2 than

⟡ Pulmonary fibrosis
can be eliminated
∶ Ratio of volume of inspired air to the volume of dead
space air is usually constant: but people with shallow

⟡ Kidneys excrete HCO3- in the urine and reclaim H+ to

Compensation: breaths have less fresh air entering the lungs than those
breathing deeply

⟡ Paper bag breathing – inhaled back the loss CO2

the blood
3. Gas exchange between the lungs and arterial blood
during hyperventilation Influenced by:
~ Destruction of alveoli (emphysema)
Oxygen and Gas Exchange ~ Pulmonary edema
Evaluation of patient’s oxygen status is done by measuring pCO2 ~ Airway blockage (asthma, bronchitis)
in blood gas analysis (together with pCO2 or carbon dioxide ~ Inadequate blood supply (pulmonary embolism,
pressure and pH) pulmonary hypertension, Congestive heart
failure)
For adequate tissue oxygenation, the following are necessary: ~ Diffusion of carbon dioxide and oxygen (oxygen
1. Available atmospheric oxygen diffuses 20 timed slower than carbon dioxide)
↣ Amount of O2 available in atmospheric air depends
on barometric pressure (BP) 4. Loading of oxygen onto hemoglobin
↣ At sea level, BP is 760 mm Hg ∶ Each hemoglobin molecule can combine reversibly
↣ Dalton’s law states that the total atmospheric pressure with four oxygen molecules
is the sum of individual gas pressures ∶ Actual amount of oxygen loaded onto hemoglobin
↣ One atmosphere exerts 760 mm Hg pressure and is dependent on:
made up to 78.1% nitrogen, 20.93% oxygen, 0.03% o Availability of oxygen
carbon dioxide, 1% inert gases; percentage of each gas is o Concentration of interfering substances
the same at all altitudes (carbon monoxide)
o pH
o temperature of blood

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System LEC 1 2
o Levels of carbon dioxide pressure and ⊸ Hemoglobin oxygen binding capacity; the maximum
2,3- diphosphoglycerate amount of O2 that can be carried by Hb in a given
∶ Normally more than 95% of functional hemoglobin (Hb quantity of blood; 1 gram of hemoglobin carries 1.39
capable of reversible binding oxygen) will bind oxygen ml of O2
∶ Increasing FiO2 further saturates hemoglobin; but once
⋯ Total hemoglobin is 15 g/dL; when hemoglobin
hemoglobin is 100% saturated, the increased oxygen in
is 100% saturated with O2, the O2 capacity is
alveoli only increases concentration of dO2 in the
20.8 ml O2/100 ml of blood
arterial blood ⊸ Oxygen content: is the total O2 in the blood;
∶ Prolonged administration of high concentrations of O2 determined from the sum of O2 bound to Hb (O2Hb)
causes oxygen toxicity or decreased ventilation and the amount of O2 dissolved in plasma (pO2)
that leads to hypercarbia ⋯ For every mm Hg of pO2, 0.00314 ml of O2 will
be dissolved in 100 ml of plasma at 37oC; if
Hemoglobin in blood exists either as: pO2 is 100 mm Hg; 0.3ml of O2 is dissolved
∶ Oxyhemoglobin (O2Hb) – with bound O2
in every 100 ml of plasma
∶ Deoxyhemoglobin (HHb; reduced Hb) – Hb not bound
⋯ Amount of dissolved O2 is usually not clinically
to O2 but capable of binding O2 when available
significant; but in instances when total
∶ Carboxyhemoglobin (COHb) – with bound CO
hemoglobin is low in hyperbaric conditions,
∶ Methemoglobin (methHb) – unable to bind O2
dissolved O2 becomes an important source of
because iron is oxidized
oxygen to tissues
⋯ When 97-99% of available hemoglobin is
5. Adequate hemoglobin or Hb
saturated with O2 and total hemoglobin is 15
6. Adequate transport (cardiac output)
g/dL, the oxygen content per 100 ml of
7. Release of oxygen to the tissue
plasma is 20.5ml.

*Any disturbance in these conditions result to poor tissue

Hemoglobin-Oxygen Dissociation
⟡
oxygenation
Hemoglobin must release O2 to the tissues
Assessing Oxygen Status ⤏ Increased H+ concentrations and pCO2 levels
in tissues change the molecular
Parameters commonly used to asses a patient’s oxygen status:
configuration of oxyhemoglobin facilitating
1. Oxygen saturation (SO2: O2 sat)

⟡
oxygen release
⊸ Represents the ration of O2 that is bound to
Oxygen dissociates from hemoglobin in a
hemoglobin compared to the total amount of
characteristic fashion; graphed with pO2 on the x-axis
hemoglobin capable of binding O2

⟡
and percent saturated O2 on the y-axis; sigmoid curve
⊸ Blood gas instruments uses algorithms to calculate SO2
Hemoglobin holds on to O2 until oxygen tension in
from pO2, pH and temperature of sample; but does
tissues is about 60mm Hg; below 60 mm Hg, O2 is
not account for presence of other hemoglobin types

⟡
released rapidly
that do not reversibly bind O2
The position of the curve reflects the affinity that
hemoglobin has for O2 and affects rate of
2. Measured fractional/ percent oxyhemoglobin (FO2Hb)

⟡
⊸ Ratio of the concentration of oxyhemoglobin to dissociation
the concentration of total Hb Factors affecting position and shape of
oxygen dissociation curve:
3. Trends in oxygen saturation measured by transcutaneous pulse ⤏ H+ ion activity
oximetry (SpO2) ⤏ Carbon dioxide pressure or pCO2 and CO levels
⊸ Pulse oximeters pass light od two or more wavelength ⤏ body temperature

⟡
through the tissues in the capillary bed of the toe, ⤏ 2,3-DiPhosphoGlyceric acid
finger or ear Shift to the right
⊸ Measures O2Hb and HHb; newer machines can now ⤏ In actively metabolizing tissues, there is
measure COHb and metHb increased temperature, increases pCO2,
⊸ Accuracy compromised by poor perfusion and increased 2,3-DPG and increased H+ (low
severe anemia pH)
⤏ Hemoglobin has decreased affinity for

⟡
4. Amount of O2 dissolved in plasma (pO2) O2 allowing release of O2 to tissues
⊸ Healthy adult breathing room air will have a pO2 of Shift to the left
90- 95 mm Hg ⤏ In the lungs, temperature, pCO2, H+ and
⊸ With a blood volume of 5L, only 13.5 ml of O2 will be 2,3- DPG decrease relative to tissue levels
available from pO2 in plasma: 1,000 ml of O2 is enhancing O2 binding to hemoglobin and
carried as O2Hb CO2 and H+ oxygen uptake
by hemoglobin

⟡ Hemoglobin in tissues (high CO2 and H+) release

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Blood Gases, pH, & Buffer 4/16/2 CC
System LEC 1 2
O2; release of O2 from hemoglobin accelerates
uptake of

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System LEC

⟡
1 2

⟡
⟡
Hemoglobin in the lungs CO2 and uptake O2
Electrochemical Cells – formed when two opposite
Elevation of CO (smoking or CO exposure) causes a shift electrodes are immersed in a liquid that conduct
to the left with loss of the sigmoid curve making the current.

⟡ HbF or fatal hemoglobin causes a shift to the left

release of O2 bound to hemoglobin more difficult
Measurement of pO2
Measurement ＋ pO2 electrodes, called Clark electrodes, measure the
Spectrophotometric Determination of Oxygen Saturation amount of current flow in a circuit that is related to the
Actual percent of oxyhemoglobin (O2Hb) can be amount of O2 being reduced at the cathode
determined spectrophotometrically using a CO-oximeter ＋ A gas permeable membrane covering the tip of the
designed to directly measure various hemoglobin species. electrode selectively allows O2 to diffuse into an
⤚ Each hemoglobin species measured will depend on electrolyte and contact the cathode; electrons are
number and specific wavelengths incorporated into drawn from the anode surface to the cathode surface to
the instrument; ex: a two wavelength system can reduce O2; a small constant polarizing potential is
measure only O2Hb and HHb expressed as a fraction applied between the anode and cathode; a small
or percentage of total Hb constant polarizing potential is applied between anode
⤚ Instruments should have a minimum of four and cathode; a microammeter placed between the
wavelengths; instruments with more than four anode and cathode measures the movement of
wavelengths can recognize dyes, pigments, turbidity, electrons (Current)
other hemoglobin species and abnormal proteins ＋ Four electrons are drawn for every mole of O2 reduced
(interferences) making it possible to determine pO2
⤚ Primary purpose of determining O2Hb is to
assess oxygen transport from the lungs Sources of error:
⤚ Measured O2Hb values reflect the patient’s true ＝ Build up of protein material on the surface of the
status because calculated oxygen saturation or SO2 membrane: retards diffusion and slows electrode
and O2Hb values will differ in the presence of response
＝ Bacterial contamination within measuring chamber:
dyshemoglobin consume O2
⤚ Samples are collected under anaerobic conditions ＝ Incorrect calibration
and mixed immediately with heparin; samples must
＝ Sample collection & handling: contamination of sample
be analyzed immediately to avoid consumption of with room air, delay in analysis
oxygen by metabolizing cells
Pulse Oximeters
Potential errors: ＝ Uses transcutaneous electrodes placed directly on the
⤚ Faulty instrument calibration skin for continuous measurement of pO2
⤚ Spectral interfering substances: substances that ＝ Measurement depends on oxygen diffusing from
absorb light at the wavelengths used capillary bed through the tissues to the electrodes
＝ pO2 measured by this method only reflects arterial pO2;
these two values are not equivalent; arterial pO2
Blood Gas Analyzers
affected by O2 consumption at electrode site, heating
⧺ Uses electrodes (microelectrochemical or
effects of electrodes, possible hypoperfusion from
microelectrochemical sensors) as sensing devices
cardiovascular instability
to measure oxygen pressure or pO2, pCO2 and pH
⧺ pO2 measurement is amperometric: meaning that the
Measurement of pCO2 and pH
amount of current flow is an indication of the oxygen
~ An increased concentration or activity of the ions
present
leads to an increase in the force exerted by these
⧺ pCO2 and pH are potentiometric: a chance in voltage
ions
indicates the activity of the analyte
⧺ calculate the following paramaters: bicarbonate, total ~ Potentiometric measurements measure how much
CO2, base excess, oxygen saturation or SO2 force/energy/potential a given ion possesses
⧺ analyzers may include hemoglobin, hematocrit, ~ Requires: reference electrode, an electrode responsive
electrolytes, metabolites glucose, lactate, creatinine, to an ion being measured, voltmeter (measures the
BUN) and CO-oximetry potential difference between the two electrodes)

⟡ Cathode – the negative electrode; a site to which Optical sensors

∶ Certain fluorescent dyes will react predictably
⟡
cations tend to travel, site at which reduction occurs
Reduction – is the gain of electrons by a particle with specific chemical such as O2, CO2 and H+
∶ Dye is separated from the sample by a membrane,
⟡
(atom, molecule or ion)
Anode – the positive electrode; the site to which analyte diffuses into the dye causing either and increase
or a quenching of fluorescence proportional to the
⟡
anions migrate; site at which oxidations occurs
Oxidation – is the loss of electrons by a particle amount of dye

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∶ Optical technology is applied in in-dwelling blood gas
heparin can dilute in sample; evacuated collection tubes
systems (fiber optic catheters with sensors placed
not appropriate
within patient’s arterial system)
o Mix thoroughly with anticoagulant; mix immediately
before analysis to resuspend cells
o Slow filling cause by mismatch of syringe and needle
Calculated parameters size; small needles cause bubbles to form (hemolyzed
Instruments include algorithms to perform calculations of other RBCs, affect pO2 and pCO2)
parameters from pCO2 and pH o Air trapped in syringe must be expelled immediately
~ Bicarbonate (HCO3-): based on Henderson-Hassel after blood draw
equation o Transport time should be minimal to reduce cell
~ Carbonic acid concentration: calculated using the metabolism (O2 and glucose used up, CO2 and
solubility coefficient of CO2 in plasma at 37 deg C; lactate produced)
solubility constant to convert pCO2 to H2CO3 is o Place syringe in an ice water slurry to minimize
0.0307; temperature and an increase in lipids change metabolism; but may increase pO2 due to oxygen from
the solubility constant water entering pores of the plastic syringe
~ Total carbon dioxide content (ctCO2): is bicarbonate plus o Low temperatures causes increased oxygen solubility in
dCO2 (Carbonic acid) plus CO2 associated with proteins blood and a shift to the left in the oxyhemoglobin curve
(carbamates) (more oxygen combining with Hb); causes falsely
~ Base excess: used to assess the non-respiratory elevated pO2
component; calculated from pH, pCO2 and Hb; a
positive value/ base excess indicates an excess of
⟡ pH- to determine whether a patient’s blood is
In the ABG report
bicarbonate or a relative deficit of non carbonic acid
suggestive of non- respiratory alkalosis; a negative
⟡ PaCO2 and PaO2- the partial pressures of CO2
within physiological range
value/base deficit indicates a deficit of bicarbonate or
relative deficit of non carbonic acids suggestive of non
⟡ HCO3- indicates how much HCO3 is in the blood (and
and oxygen in- arterial blood, respectively
respiratory acidosis

⟡ Base excess (or deficit) - a measure of the excess or

is therefore available as a buffer)
Correction for temperature
⊸ Values for pH, pCO2 are temperature dependent; deficiency of base in thee blood; by definition, it is the
all measurements done at 37 deg C amount of base (in mmol) that would correct one liter
⊸ Most analyzer software perform correction of blood to a normal pH (if an excess, this is the
⊸ Must use reference ranges for the patient’s temperature amount of base needed to be removed for a normal

⟡ Lactate- the end product of anaerobic glycolysis (a

for proper result interpretation; but reference range is pH, or if a deficit, the amount required to be added)
controversial and difficult to find
rise indicates poor oxygenation and perfusion of

⟡ Other parameters commonly found on ABG reports

QUALITY ASSURANCE tissues)
o Proper patient identification
o Correct labeling are: hemoglobin, glucose and electrolytes (Sodium,
o Other information needed for interpretation: potassium, chloride and ionized calcium)
FiO2, ventilation (on room air or supplemental
O2), body temperature
⟡ ABGs can be interpreted using a stepped approach
INTERPRETATION OF RESULTS
o Collecting personnel
o Choice of site: radial, brachial, femoral or
temporal arteries
⟡ The pH should be assessed first. A pH of less than 7.35
Step 1- check the pH
o Venous blood gas samples: can be used if pulmonary
function or O2 transport is not being assessed indicated acidosis and a pH greater than 7.45
o Capillary blood: to assess pH and pCO2; capillary
indicated alkalosis
pO2 does not correlate well with arterial pO2
o Central venous (pulmonary artery blood samples: used

⟡ Having determined if the patient is acidotic or alkalotic,

Step 2- check theHCO3 and PaCO2
to assess O2 consumption (Calculated from O2
content of arterial and pulmonary artery blood and
check the HCO3 and the PaCO2 to classify the results
cardiac output)
as follows:
✓ Metabolic acidosis: patients who are acidotic
Collection and handling: collection device, form and concentration
and have a HCO3 <22 (base excess <-2)
of heparin used, speed of syringe filling, maintenance of
✓ Respiratory acidosis: patients who are
anaerobic environment, mixing of samples, transport and storage acidotic with a PaCO2>6
time before analysis ✓ Metabolic alkalosis: Patients who are alkalotic
o 1-3 ml self-filling plastic disposable syringe containing
with a HCO3> 28 (base excess >+2)
heparin; dry/ lyophilized heparin better since liquid

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Blood Gases, pH, & Buffer 4/16/2 CC
System LEC 1 2
✓ Respiratory alkalosis: patient who are alkalotic
with a PaCO2 <4.7

It is possible for patients to have a mixed respiratory

and metabolic alkalosis disturbances exist simultaneously. If the
two processes oppose each other, pH derangement will be
minimised (See step 3). However two processes that cause pH to
move in the same direction may lead to profound acidosis or
alkalosis.

Step 3- check for compensation

Check to see if the patient is compensating for his or
her acid-base imbalance. Patients may partially or fully
compensate for an acid-base imbalance by the “opposite”
mechanism; for example metabolic acidosis will be compensated
for with respiratory alkalosis. This may create some apparently
normal results amongst some deranged ones. When interpreting
acid-base status, it is important always to take the clinical context
into account. For example, if presented with ABG results showing
a normal pH, low PaCO2 and low HCO3 in a diabetic patient with
high levels of ketones in urine the most likely primary disorder is
metabolic acidosis (Diabetic ketoacidosis), rather than respiratory
alkalosis

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