Other Buffers

The other buffer systems in the blood are the protein and phosphate buffer systems.

These are the only blood buffer systems capable of buffering respiratory acid-base disturbances
as the bicarbonate system is ineffective in buffering changes in H+ produced by itself.

The phosphate buffer system is NOT an important blood buffer as its concentration is too low

The concentration of phosphate in the blood is so low that it is quantitatively unimportant.

Phosphoric acid is triprotic weak acid and has a pKa value for each of the three dissociations:

pKa1 = 2 pKa2 = 6.8 pKa3 = 12

The three pKa values are sufficiently different so that at any one pH only the members of a
single conjugate pair are present in significant concentrations.

At the prevailing pH values in most biological systems, monohydrogen phosphate (HPO4-2) and
dihydrogen phosphate (H2PO4-) are the two species present. The pKa2 is 6.8 and this makes the
closed phosphate buffer system a good buffer intracellularly and in urine. The pH of glomerular
ultrafiltrate is 7.4 and this means that phosphate will initially be predominantly in the
monohydrogen form and so can combine with more H+ in the renal tubules. This makes the
phosphate buffer more effective in buffering against a drop in pH than a rise in pH.

Note: The ‘true’ pKa2 value is actually 7.2 if measured at zero ionic strength but at the typical
ionic strength found in the body its apparent value is 6.8. The other factor which makes
phosphate a more effective buffer intracellularly and in urine is that its concentration is much
higher here than in extracellular fluid.

Haemoglobin is an important blood buffer particularly for buffering CO2

Protein buffers in blood include haemoglobin (150g/l) and plasma proteins (70g/l). Buffering is
by the imidazole group of the histidine residues which has a pKa of about 6.8. This is suitable for
effective buffering at physiological pH. Haemoglobin is quantitatively about 6 times more
important then the plasma proteins as it is present in about twice the concentration and contains
about three times the number of histidine residues per molecule. For example if blood pH
changed from 7.5 to 6.5, haemoglobin would buffer 27.5 mmol/l of H+ and total plasma protein
buffering would account for only 4.2 mmol/l of H+.

Deoxyhaemoglobin is a more effective buffer than oxyhaemoglobin and this change in buffer
capacity contributes about 30% of the Haldane effect. The major factor accounting for the
Haldane effect in CO2 transport is the much greater ability of deoxyhaemoglobin to form
carbamino compounds.
Isohydric Principle

All buffer systems which participate in defence of acid-base changes are in equilibrium with
each other. There is after all only one value for [H+] at any moment. This is known as the
Isohydric Principle.

It means that an assessment of the concentrations of any one acid-base pair can be utilised to
provide a picture of overall acid-base balance in the body. This is fortunate as the measurement
of the concentrations of all the buffer pairs in the solution would be difficult. Conventionally, the
components of the bicarbonate system (ie [HCO3] and pCO2) alone are measured. They are
accessible and easy to determine. Blood gas machines measure pH and pCO2 directly and the
[HCO3] is then calculated using the Henderson-Hasselbalch equation.

Buffering in different sites

Respiratory disorders are predominantly buffered in the intracellular compartment. Metabolic

Various buffer systems exist in body fluids (see Table) to minimise the effects of the addition or
removal of acid from them.

In ECF, the bicarbonate system is quantitatively the most important for buffering metabolic
acids. Its effectiveness is greatly increased by ventilatory changes which attempt to maintain a
constant pCO2 and by renal mechanisms which result in changes in plasma bicarbonate.

In blood, haemoglobin is the most important buffer for CO2 because of its high concentration and
its large number of histidine residues.

Deoxyhaemoglobin is a better buffer than oxyhaemoglobin

Another factor which makes haemoglobin an important buffer is the phenomemon of isohydric
exchange. That is, the buffer system (HHbO2-HbO2-) is converted to another more effective
buffer (HHb-Hb-) exactly at the site where an increased buffering capacity is required. More
simply, this means that oxygen unloading increases the amount of deoxyhaemoglobin and this
better buffer is produced at exactly the place where additional H+ are being produced because of
bicarbonate production for CO2 transport in the red cells.

2.2.5 Link between Intracellular & Extracellular

The two major processes involved are:

 Transfer of CO2 across the cell membrane

Important points to note about CO2 are:

 It is very lipid soluble and crosses cell membranes with ease causing acid-base changes
due to formation of H+ and HCO3-. Because of this ease of movement, CO2 is not
important in causing differences in pH on the two sides of the cell membrane.
 Extracellular buffering of CO2 is limited by the inability of the major extracellular buffer
(the bicarbonate system) to buffer changes in [H+] produced from the reaction between
CO2 and water.

The result is that buffering for respiratory acid-base disorders is predominantly intracellular:
99% for respiratory acidosis and 97% for respiratory alkalosis.