Dr Manas Krishna Borgohain,

Assistant Professor,
Deptt. Of Physiology,
JMCH
Oxygen is transported from lungs to the tissues in two methods:
 physically dissolved in the plasma, and,

 in combination with hemoglobin (Hb) in the red cell.

Approximately 98% of the oxygen is transported in combination with

hemoglobin and the remaining 2% in the physically dissolved form.

In Physically Dissolved Form
• In the physically dissolved form in plasma, only about 2% of oxygen is
transported. The amount of physically dissolved oxygen in the blood can be
predicted from Henry’s law.

• Henry’s law states that the amount of gas that dissolves in a liquid at a given
temperature is proportional to the partial pressure of the gas.
• Thus, the quantity of dissolved oxygen in arterial blood is calculated from the
following equation:

Dissolved O2 (ml/dl) = oxygen solubility  partial

pressure of O2 in arterial blood (PaO2).

= 0.003 (ml /dl of blood per mm Hg)  95 mm Hg (normal PaO2 is 95 mm Hg)
= 0.3 ml/dl of blood
Thus, at PaO2 of 95 mm Hg, the dissolved O2 is 0.3 ml/dl.

• That means, in a normal healthy adult, 0.3 ml of O2 is transported in dissolved

form in 100 ml of blood

• As cardiac output is 5 liters /min at rest, oxygen transported in dissolved form at rest

is about 15 ml/min.

• Therefore oxygen transported in dissolved form alone is grossly inadequate to meet

the oxygen requirement of the body, which is about 250 ml/min at rest.
In Combination with Hemoglobin

• More than 98% of O2 is transported in combination with Hb.

• This becomes possible due to the binding affinity of

hemoglobin for oxygen.

• Hemoglobin that binds with oxygen is called oxyhemoglobin

(HbO2) and the hemoglobin that does not bind with O2 is called

deoxyhemoglobin (deoxy-Hb) or reduced-hemoglobin.

Oxyhemoglobin Formation

• The hemoglobin molecule is a protein made up of four subunits that

are bound together. Each subunit consists of a molecular group known

as heme and a polypeptide attached to the heme. The four

polypeptides of a Hb molecule are combinely known as globin.

• Each heme group contains one atom of iron (Fe++) to which oxygen

binds
Structure of Hb molecule
• Since each iron atom can bind one molecule of oxygen, a single Hb molecule can

bind four molecules of oxygen.

• Hb binds with oxygen only when the iron is in ferrous (Fe++) state. The Fe++ iron in

Hb is oxidized to ferric (Fe+++) iron to form methemoglobin.

• Thus, methemoglobin can’t bind oxygen. Methemoglobin formation occurs under the

influence of various compounds like nitrites or sulfonamides. Methemoglobin is also

formed spontaneously.

• However, the enzyme methemoglobin reductase is present in red cell that reduces

methemoglobin to Hb. Therefore, normally only about 1.5% of total Hb is

methemoglobin.
 • Deficiency of methemoglobin reductase (a genetic

defect) increases the methemoglobin concentration

and decreases oxygen carrying capacity.

• Oxygen binds rapidly and reversibly to hemoglobin to

form oxyhemoglobin(HbO2): O2 + Hb  HbO2

R & T STATES OF Hb : During this process of HbO2 formation, the heme

remains in ferrous state. Thus, this process is oxygenation, not

oxidation. As there are four subunits in Hb molecule, Hb reacts rapidly (in

less than 0.01 s) with four molecules of oxygen to form HbO8.

1. When oxygen is not bound with Hb (deoxyhemoglobin) the four

subunits of Hb are tightly bound with each other. This configuration of

Hb is called Tense or T state. In this T configuration, affinity of Hb for

oxygen is less.
2. When, first oxygen molecule binds with Hb, the four subunits enter into

the relaxed or R state that exposes more oxygen binding sites.

Therefore, in R configuration, affinity of Hb for oxygen is greatly

increased by 200 to 500 times.

3.When PO2 is very less, most of Hb molecules are in T state and have

low oxygen affinity.

4.When PO2 is very high, the Hb molecules are in R state and have high

oxygen affinity
 Conversion of Hb molecule from T state to R state. As O2 is
added, salt bridges are successively broken and finally 2-3,BPG is
expelled

5. Thus, the quantity of oxyhemoglobin formation is a function of partial

pressure of oxygen in the blood.
• In pulmonary capillaries, where PO2 is high, the reaction favours to
form more oxyhemoglobin, and in tissue capillaries, where PO2 is low,
the reaction favours formation of deoxyhemoglobin that helps oxygen
to be unloaded from hemoglobin and becomes available to the cells.
O2 Carrying Capacity of Hb
• Each gram of hemoglobin binds with 1.34 ml of oxygen. The maximum amount of oxygen
that can be carried by hemoglobin is called the oxygen carrying capacity.

• In a healthy individual, the oxygen carrying capacity of arterial blood is about 20 ml of O2 per
100 ml of blood, considering the hemoglobin concentration of 15 g% (1.34 ml  15 g = 20.1
ml O2/dl blood).

• OXYGEN CONTENT of Hb (HbO2 content) is the amount of oxygen actually bound to

hemoglobin, whereas OXYGEN CAPACITY of Hb (HbO2 capacity) is the amount of oxygen
that can potentially bind with Hb.

• The percentage saturation of hemoglobin with oxygen (SO2) is the ratio of the quantity of
oxygen actually bound (oxygen content) to the quantity that can be potentially bound
(oxygen capacity) multiplied by 100.
Oxygen-Hemoglobin Dissociation Curve:

• Oxygen-hemoglobin (Oxy-Hb) equilibrium curve (dissociation or association curve)

explains the relationship between partial pressure of oxygen (PO2) in blood with oxygen

saturation of Hb. Oxy-Hb equilibrium curve is an S-shaped curve over the range of PO2

from 0 to 100 mm Hg

• The sigmoid shape of the curve results from hemoglobin affinity for oxygen at various PO2

levels.

• As PO2 rises, the Hb saturation progressively increases.

• However, the saturation is not linear with increase in PO2 for which

the curve becomes sigmoid shaped.

• The S-shaped oxyhemoglobin equilibrium curve enables oxygen to

saturate hemoglobin under high partial pressures in the lungs and to

give up large amounts of oxygen with small changes in PO2 at the

tissue level.
 Oxygen-hemoglobin dissociation curve
Note at PO2 of 27 mm Hg, Hb saturation of oxygen is
50% (P50)
• The curve is divided into two major phases: the steep phase and the plateau phase.

• The Steep Phase (unloading or dissociation zone)

• The curve has a steep slope between PO2 of 10 and 60 mm Hg. During this phase of the curve,

combination of oxygen with Hb increases very rapidly as the PO2 increases from 10 to 60 mm Hg.

Oxygen saturation of Hb is about 90% at PO2 60 mm Hg.

• Significance of Steep Phase

• In the steep phase, oxygen saturation of Hb is very high.

• Less increase in PO2 leads to greater percentage saturation of Hb and therefore, facilitates oxygen

loading.
 ➢ Also, change in steep portion of the curve in reverse direction (that is, from 60 to 10 mm Hg)

causes unloading of oxygen in the tissues.

➢ A small decrease in PO2 in the tissue results in unloading of large amount of oxygen to the

tissues.

➢ The steep phase allows large quantities of oxygen to be released from hemoglobin in the

tissue capillaries where a lower capillary PO2 prevails. This is especially achieved by shifting the

curve to right (shift occurs mainly on the steep phase) by increased H+ or by increased CO2.

• The Plateau Phase (loading or association zone)

➢ The curve begins to plateau at PO2 around 60 mm Hg and flattens at PO2 of 70 mmHg

➢ The increase in PO2 above 60 mm Hg produces only a small increase in oxygen binding.
➢ Increase in PO2 from 60 to 100 mm Hg in the plateau region of the curve illustrates that

oxygen saturation and content remain apparently constant over a wide alteration in

alveolar PO2.

➢ Significance of Plateau Phase

➢ There are two significances of plateau phase:

➢ 1. In different situations, like at high altitude or in pulmonary diseases in which a moderate

hypoxia (decrease in PO2 from 95 to 60 mm Hg) occurs, the total amount of oxygen carried

by Hb decreases only by 5−10%, since Hb saturation is about 90% at PO2 of 60 mm Hg.

➢ Thus, plateau in the curve provides a safety factor through which even a significant

decrease in lung function can allow normal saturation of Hb.

2. Oxygen saturation and content remain fairly constant in spite of wide fluctuations in alveolar
PO2(PAO2).

▪ For example, if PAO2 rises from 100 to 120 mm Hg, hemoglobin saturation increases only

slightly (97 to 98%).

▪ This is the reason why oxygen content cannot be raised appreciably by hyperventilation or

by breathing 100% oxygen because Hb is already completely saturated with oxygen at PO2 of

100 mm Hg. This is true only for normal people at sea level.

▪ If a person has low arterial PO2 due to lung disease or for his ascension to high altitude,

hyperventilation or breathing 100% oxygen increases Hb saturation with oxygen as they have

more deoxy-Hb initially.

The P50

The P50 is the level of PO2 at which 50% of the hemoglobin is saturated with oxygen.

1. P50 assesses the binding affinity of hemoglobin for oxygen.

2. In adults at sea level, the normal P50 occurs at a PO2 of 27 mm Hg.

3. Alteration in the P50 value has a greater impact on the steep phase of the curve.

4. If the P50 is high, it signifies the decrease in affinity of Hb for oxygen, which is

seen in right-shift of the oxygen-hemoglobin equilibrium curve.

5. Conversely, if P50 is low, it signifies shift of the curve to the left, in which affinity of

Hb for oxygen is more

Effect of P50 on Oxy-Hb dissociation
curve.
Factors Affecting Hb-Binding Affinity with Oxygen.

➢ The important factors are temperature, arterial PCO2, arterial pH and 2,3-
DPG. A rise in PCO2, , a fall in pH, and a rise in temperature all shift the
curve to the right.

➢ The effect of carbon dioxide and hydrogen ions on the affinity of hemoglobin
for oxygen is known as the Bohr effect.

➢ A shift of the oxyhemoglobin equilibrium curve to the right is physiologically

advantageous at the tissue level, as affinity of O2 for Hb is lowered
(increased P50).
➢ A rightward shift enhances the unloading of oxygen for a given PO2 in the tissue, and
a leftward shift increases the affinity of hemoglobin for oxygen, thereby lowering the
ability to release oxygen to the tissues.

➢ A simple way to remember the functional importance of these shifts is that an

exercising muscle is warm and acidic and produces large amounts of carbon dioxide
(high PCO2), all of which favor the unloading of more oxygen to metabolically active
muscles.
➢ Increase in temperature shifts the Oxy-Hb dissociation curve to
right and decrease in temperature shifts the curve to the left.

➢ In other words, high temperature decreases the affinity of Hb for

oxygen.

➢ This helps in release of oxygen in metabolically active tissues

in which temperature is more.
Effect of temperature on Oxy-Hb dissociation curve
pH

➢ Alteration in blood pH shifts the Oxy-Hb dissociation curve. Christian Bohr and Neils Bohr in 1904
demonstrated that respiratory acidosis shifts oxy-Hb dissociation curve to right. Since then, the
decrease in oxygen affinity in acidosis is known as Bohr effect.

➢ A decrease in pH shifts the curve to right and increase in pH shifts the curve to left.

➢ When blood passes through capillaries, CO2 enters red cell that decreases intracellular pH and
shifts the Oxy-Hb dissociation curve to right.

➢ It has been noted that under normal physiological conditions, binding of about 0.7 mole of H+
causes Hb to release 1 mole of oxygen.

➢ Thus, when blood passes through tissue capillaries, the acidic environment facilitates release
of oxygen from Hb into the tissues.
Effect of pH on Oxy-Hb dissociation curve
Carbon Dioxide

➢ During cellular metabolism CO2 is released into circulation that increases

generation of H+ and decreases pH. This shifts the curve to right. This helps in
release of oxygen from Hb.

➢ The shift to right in acidosis (Bohr effect) is partly due to effect of decrease
in pH and partly to the direct effect of CO2 on Hb.

➢ CO2 combines with unprotonated amino groups on Hb (Hb-NH2) to form

carbamino groups which changes the conformation of Hb. This results in
decrease in oxygen affinity of Hb.
2,3-DPG
➢ Red blood cells lack mitochondria. Therefore, anerobic glycolysis occurs in red cells. During
glycolysis, large quantities of 2,3-diphosphoglycerate (2,3-DPG), an organic phosphate
compound is produced as the metabolic intermediary. Thus, 2,3-DPG level is much higher in
red cells than in other cells.
➢ 2,3-DPG significantly affects affinity of hemoglobin for oxygen.
➢ This destabilizes the interaction of Hb with oxygen.
➢ Thus, binding of 2,3-DPG with Hb shifts the Oxy-Hb dissociation curve to right and
facilitates the release of oxygen.
➢ At the tissue level, an increase in 2,3-DPG facilitates unloading of oxygen from the red
cell.
➢ Hypoxia stimulates glycolysis resulting in increased production of 2,3-DPG.
➢ Red cell 2,3-DPG increases in anemia, exercise and hypoxic conditions like high altitude,
chronic lung disease etc. Therefore, in these conditions, Oxy-Hb dissociation curve shifts to
right
Factors that affect 2,3-DPG in Red Cells:

1. pH: Acidosis inhibits red cell glycolysis and therefore decreases 2,3-
DPG concentration.

2. Type of Hb: The γ chains of fetal Hb have less avidity for 2,3-DPG
than β chains of adult Hb. Therefore, fetal Hb has higher oxygen
affinity. This provides an advantage to fetus to extract oxygen from
maternal blood in the placenta.
3. Fetal Hb shifts Oxy-Hb dissociation curve to left.

4. Hormones: Growth hormone, thyroxine and testosterone stimulate the synthesis of 2,3-DPG.

5. Altitude: At high altitude, 2,3-DPG concentration increases substantially in red cells. This

increases the availability of oxygen to the tissues.

6. PO2: Hypoxia increases 2,3-DPG production.

7. Procedure of storage of blood: The 2,3-DPG concentration in red cells of blood stored in

blood bank decreases. Therefore, transfusion of stored blood decreases oxygen transport,

especially when transfused into hypoxic patients. However, if blood is stored in citrate-

dextrose-phosphate solution this effect is less than when stored in acid-citrate-dextrose

solution
Effect of 2,3-diphosphoglycerate (2,3-DPG)
on Oxy-Hb dissociation curve
➢ Myoglobin

➢ Myoglobin is the Hb pigment in muscle.

➢ Though it is similar to Hb in structure, myoglobin binds one molecule of O2 per

mole.

➢ It shifts Oxy-Hb dissociation curve to left and the curve becomes hyperbolic
(loses sigmoid shape) (Fig. 70.8).

➢ This helps in picking up of O2 from blood.

➢ Myoglobin is more in regularly exercising muscles especially that are trained in

isometric exercise. This helps the muscle to draw oxygen from myoglobin,
especially when blood flow ceases for a longer duration during sustained muscle
contraction.
Effects of myoglobin and fetal hemoglobin (Hb)
on Oxy-Hb dissociation curve
➢ Effect of Carbon Monoxide (CO)

➢ The binding affinity of CO with Hb is 210 times more than oxygen. Moreover,
CO interferes with oxygen transport by competing for the same binding sites on
hemoglobin.

➢ CO binds to hemoglobin to form carboxyhemoglobin (HbCO).

➢ When the blood is 60% saturated with CO (carboxyhemoglobin) the oxygen

content is reduced to less than 10 ml/dl.

➢ As the partial pressure of CO approaches 1 mm Hg, Hb is fully saturated with CO.

However, arterial PO2 may be normal as oxygen diffusion gradient remains
normal.

➢ But , oxygen content is greatly reduced as it can’t bind to Hb. This grossly
decreases the oxygen carrying capacity.
➢ Moreover, CO also shifts the Oxy-Hb dissociation curve to the left, which

decreases oxygen release to the tissues.

➢ Therefore, severe tissue hypoxia occurs in CO poisoning, which is fatal if

not treated immediately (for details refer ‘Hypoxia’).

➢ Normally, in healthy individuals, CO occupies 1–2% of Hb binding sites.

➢ However in chronic cigarette smokers, traffic personnel and in residents

of crowded traffic areas, CO concentration increases in plasma and CO

occupies about 10% of Hb

FACTORS THAT SHIFT OXYGEN-Hb DISSOCIATION CURVE

Factors that shift the curve to right:

➢ Increased temperature

➢ Decreased pH

➢ Increased PCO2

➢ Increased 2,3-DPG

➢ Hypoxia

➢ Shift of the curve to right increases P50 (decreased affinity

of Hb for oxygen), which facilitates oxygen release.

➢ Factors that shift the curve to left:

➢ Decreased temperature

➢ Increased pH

➢ Decreased PCO2

➢ Decreased 2,3-DPG

➢ Fetal Hb

➢ Carbon monoxide

➢ Shift of the curve to left decreases P50 (increases affinity of Hb for

oxygen) that facilitates oxygen uptake and prevents oxygen release.
O2 DELIVERY TO TISSUE

Diffusion of O2 from Capillaries to Tissue Fluid

o When the arterial blood reaches the peripheral tissues, diffusion of oxygen occurs from
the peripheral

o Capillary blood into the tissue fluid.

o This occurs along the concentration gradient of oxygen from area of higher level to
lower level. The PO2 in systemic capillary blood is95 mm Hg, whereas PO2 in the
interstitial fluid that surrounds the tissue cells is about40 mmHg.

o Thus, there is a great pressure gradient due to considerably large pressure

difference of O2 between the systemic capillary blood and the tissue fluid that causes
oxygen to diffuse rapidly from the capillary blood into the interstitial fluid.
o Therefore, PO2 falls rapidly in the capillary to about 40 mm Hg. Thus, the PO2 of the

blood leaving the tissue capillaries (PO2 at venous end of capillary) and entering the

systemic veins is 40 mm Hg

o From tissue fluid, oxygen rapidly enters the cell as in the intracellular fluid thePO2is

about 23 mmHg (ranges between 7 to 40 mmHg), as oxygen is constantly utilized by

cell. Especially in metabolically active tissues, this gradient is more as mitochondrial

oxygen utilization is more.

o Diffusion of oxygen to tissues depend on two important factors: i)rate of blood flow to the

tissue and ii) the rate of tissue metabolism. Oxygen delivery is more if the blood flow is

more and the metabolic activity is more

Delivery of oxygen from capillary blood to tissue.
Bohr Effect

Any factor that shifts Oxy-Hb dissociation curve to right , decreases affinity of oxygen for
Hb.

1. At the tissue level, CO2 enters the blood and shifts the curve to right. This helps in
unloading of oxygen from Hb and facilitates tissue oxygenation. This phenomenon
is called Bohr Effect as was initially described by Christian Bohr (1855-1911), who
has showed experimentally the influence of CO2 tension on blood binding of oxygen.

2. This occurs mainly due to decreased pH in the tissue.

3. Increased temperature of blood in the tissue also contributes.

4. In tissue, acidification of blood decreases affinity of O2 to Hb, as deoxygenated Hb

binds H+ more actively than that of oxyhemoglobin.

5. The PCO2 level in the tissue rises, the curve shifts to right and P50 rises. Thus, Bohr’s
effect helps in unloading of O2 and loading of CO2
Importance of Oxygen Saturation and Content

Oxygen Saturation

➢ The Oxygen saturation is the ratio of the amount of oxygen bound to Hb to the

maximum amount of oxygen that can bind Hb (100% oxygen capacity). At 100%

oxygen capacity, heme groups of Hb are fully saturated with oxygen.

Oxygen Content

➢ The oxygen content of blood is the volume of oxygen contained in unit volume of

blood, which includes the oxygen bound to Hb and also dissolved in plasma. As the

dissolved oxygen is negligible, the oxygen content depends on concentration of Hb and

oxygen binding capacity of Hb

Oxygen Extraction

➢ Oxygen extraction is the amount of oxygen taken up by the tissues from the

blood.

➢ This is an index of oxygen consumption of the tissue. It is better quantified in terms of

OXYGEN EXTRACTION RATIO (OER).

➢ OER is also called as oxygen coefficient ratio, is the amount of oxygen extracted

by the tissue divided by the amount of oxygen delivered.

➢ In metabolically more active tissues like cardiac muscle, OER is as high as 85% at

rest.
 APPLIED ASPECTS
Measurement of O2 Saturation of Hb

Pulse Oximetry

➢ Pulse oximetry is a noninvasive method of measurement of oxygen saturation of Hb.

Oxygen saturation is continuously measured in hospitalized patients, especially patients in

intensive care unit by this method. Instrument used is the pulse oximeter.

➢ The probe of the oximeter is attached usually to the finger-tip or ear lobule where the

pulsating blood vessels are accessible externally. Red and infrared light are transmitted

through the vascular bed, and pulsatile, non pulsatile and total absorbances are

calculated.
 ▪ The pulsatile component of absorbance represents the arterial
oxygenated blood and the non pulsatile component represents
the deoxygenated capillary and venous blood.

Abnormalities of Tissue Oxygenation

Less Oxygenation

▪ Less oxygenation of tissue could be due to less oxygen in the

atmosphere decreasing oxygen content in blood, decreased Hb level
in blood decreasing oxygen saturation, or decreased capacity of
tissue to utilize oxygen resulting in decreased oxygen extraction.
Excess Oxygenation & Metabolism

Pulmonary Damage by Free Radicals

o Though tissue oxygenation is essential for life, excess or inappropriate oxygenation and oxygen
metabolism is harmful for the tissues.

o During synthesis of ATP, molecular oxygen is reduced to form water in mitochondria.

o The reduction of oxygen is accomplished by addition of four electrons by the mitochondrial

electron transport system.

o However, leak in the electron transfer system allows oxygen to accept less than four electrons that
form free radicals.

o Free radicals cause damage to the tissues. Lung is frequently damaged by free radicals.
o Pulmonary capillaries are mainly damaged that results in pulmonary edema.

Reactive Oxygen Species & Antioxidants

o A free radical is an atom or a molecule with an unpaired electron in its outermost orbit.

o Superoxide radical (O2−) and hydroxyl radical (OH∙) are commonly produced free

radicals in the body. Hydrogen peroxide also can generate hydroxyl radical.

o Superoxide ion reacts with NO to form peroxy nitrite, which is also a free radical. These

free radicals are combinely known as reactive oxygen species (ROS). ROS cause tissue

damage and promote tissue degeneration. They are sometimes called pro-oxidants, as

they are produced during the process of tissue oxidation

o However, there are antioxidants in the body that prevent the body from

oxidative damage. Antioxidants are mainly enzymes such as superoxide

dismutase, catalase and peroxidases that neutralize ROS. Imbalance

between ROS and antioxidants by either more production of ROS or

decreased formation of protective enzymes results in oxidative stress

(oxidative tissue damage).

o ROS are also produced during inflammations, which occurs mainly due

to respiratory burst of neutrophils. Reperfusion injury deteriorates condition

by generating local oxidative stress