2.2.
1 Definition of a Buffer
A buffer is a solution containing substances which have the ability to minimise changes in pH
when an acid or base is added to it. (Worthley, 1977).
A buffer typically consists of a solution which contains a weak acid HA mixed with the salt of
that acid & a strong base eg NaA. The principle is that the salt provides a reservoir of A- to
replenish [A-] when A- is removed by reaction with H+.
2.2.2 Buffers in the Body
Key Fact: The body has a very large buffer capacity.
This can be illustrated by considering an old experiment (see below) where dilute hydrochloric
acid was infused into a dog.
Swan & Pitts Experiment
In this experiment, dogs received an infusion of 14 mmols H+ per litre of body water. This
caused a drop in pH from 7.44 ([H+] = 36 nmoles/l) to a pH of 7.14 ([H+] = 72 nmoles/l) That is,
a rise in [H+] of only 36 nmoles/l.
SO: If you just looked at the change in [H+] then you would only notice an increase of 36
nmoles/l and you would have to wonder what had happened to the other 13,999,964 nmoles/l
that were infused.
Did they magically disappear? The answer of course is that they were buffered and so these
hydrogen ions were hidden from view.
Before we proceed, lets just make sure we appreciate what this experiment reveals. The dogs
were infused with 14,000,000 nmoles/l of H+ but the plasma [H+] only changed by bit over
0.002%. By any analysis, this is a system which powerfully resists change in [H+]. (My
personal perspective on appreciating the magnitude of this is to use the analogy of depositing
$14,000,000 in the bank, but then finding that after 'bank charges' my account only went up by
$36.)
Make no mistake: the body has:
a HUGE buffering capacity, and
this system is essentially IMMEDIATE in effect.
For these 2 reasons, physicochemical buffering provides a powerful first defence against acid-
base perturbations.
Key Fact: Buffering hides from view the real change in H+ that occurs.
This huge buffer capacity has another not immediately obvious implication for how we think
about the severity of an acid-base disorder. You would think that the magnitude of an acid-base
disturbance could be quantified merely by looking at the change in [H+] - BUT this is not so.
Because of the large buffering capacity, the actual change in [H+] is so small it can be ignored in
any quantitative assessment, and instead, the magnitude of a disorder has to be estimated
indirectly from the decrease in the total concentration of the anions involved in the buffering.
The buffer anions, represented as A-, decrease because they combine stoichiometrically with H+
to produce HA. A decrease in A- by 1 mmol/l represents a 1,000,000 nano-mol/l amount of H+
that is hidden from view and this is several orders of magnitude higher than the visible few
nanomoles/l change in [H+] that is visible.) - As noted above in the comments about the Swan &
Pitts experiment, 13,999,994 out of 14,000,000 nano-moles/l of H+ were hidden on buffers and
just to count the 36 that were on view would give a false impression of the magnitude of the
disorder.
The Major Body Buffer Systems
Site Buffer System Comment
ISF Bicarbonate For metabolic acids
Phosphate Not important because concentration too low
Protein Not important because concentration too low
Blood Bicarbonate Important for metabolic acids
Haemoglobin Important for carbon dioxide
Plasma protein Minor buffer
Phosphate Concentration too low
ICF Proteins Important buffer
Phosphates Important buffer
Urine Phosphate Responsible for most of 'Titratable Acidity'
Ammonia Important - formation of NH4+
Bone Ca carbonate In prolonged metabolic acidosis
2.2.3 The Bicarbonate Buffer System
The major buffer system in the ECF is the CO2-bicarbonate buffer system. This is responsible for
about 80% of extracellular buffering. It is the most important ECF buffer for metabolic acids but
it cannot buffer respiratory acid-base disorders.
The components are easily measured and are related to each other by the Henderson-Hasselbalch
equation.
Henderson-Hasselbalch Equation
pH = pK’a + log10 ( [HCO3] / 0.03 x pCO2)
The pK’a value is dependent on the temperature, [H,sup>+] and the ionic concentration of the
solution. It has a value of 6.099 at a temperature of 37C and a plasma pH of 7.4. At a
temperature of 30C and pH of 7.0, it has a value of 6.148. For practical purposes, a value of 6.1
is generally assumed and corrections for temperature, pH of plasma and ionic strength are not
used except in precise experimental work.
The pK'a is derived from the Ka value of the following reaction:
CO2 + H2O <=> H2CO3 <=> H+ + HCO3-
(where CO2 refers to dissolved CO2)
The concentration of carbonic acid is very low compared to the other components so the above
equation is usually simplified to:
CO2 + H2O <=> H+ + HCO3-
By the Law of Mass Action:
Ka = [H+] . [HCO3-] / [CO2] . [H20]
The concentration of H2O is so large (55.5M) compared to the other components, the small loss
of water due to this reaction changes its concentration by only an extremely small amount. This
means that [H2O] is effectively constant. This allows further simplification as the two constants
(Ka and [H2O] ) can be combined into a new constant K’a.
K’a = Ka x [H2O] = [H+] . [HCO3-] / [CO2]
