Acid-Base balance and buffer

systems in the human body

Department of General Chemistry

Water and pH relationship

In solution water shows a very low
dissociation (probability of H+ in water is 1.8 x 10-9)
H2O

H+ + OH-

H+ is actually associated with a cluster of

water molecules and exists in solution as
H3O+ or H5O2+ or H7O3+

Ionization of Water

Ionization of Water
H20 + H20

H20
Keq= [H+] [OH-]
[H2O]

H3O+ + OH-

H+ + OHKeq=1.8 X 10-16M
[H2O] = 55.5 M

[H2O] Keq = [H+] [OH-]

(1.8 X 10-16M)(55.5 M ) = [H+] [OH-]
1.0 X 10-14 M2 = [H+] [OH-] = Kw

pH Scale

Devised by Sorenson (1902) [H+] can range

from 1M and 1 X 10-14M using a log scale
simplifies notation pH = -log [H+]

Water and pH relationship

For Dissociation of water,

[H ][OH ]
K
[H 2 O]
Where [ ] denotes concentration and K is
the dissociation constant

Water and pH relationship

1 mole of water = 18g
1 L of water contains
1000 18 = 55.56 mol (ie pure water = 55.56M)
Molar concentration of H+ (or OH-) ions can be calculated
[H+] = 1.8x10-9 x 55.56 = 1.0x10-7
In order to avoide using ve numbers,
the [H+] is expressed as pH which is ve log (base 10) of [H+]
pH = - log10 [H+]
pH of pure water is 7
Acidic solutions have pH < 7
while basic solutions have pH > 7

Water and pH relationship

[H+]=1x10-6 what is the pH?
pH= - log10 [H+]

[H+]=0.24x10-4 what is the

pH?

4.6

[H+]=3.4x10 what is the pH?

2.5

-3

Calculating pH

Effect of pH Shifts in Biological Systems

Biochemical processes
& reactions are pHdependent.
Cells & organisms
maintain constant
specific pH value that
is optimal for function.
Changes in charge can
alter molecular
conformation and
activity.
pH sensitive molecules
include enzymes,
receptors, ligands, ion
channels, transporters
and structural
proteins.

Weak Acids and Bases Equilibria

Strong acids / bases disassociate completely
Weak acids / bases disassociate only partially
Enzyme activity sensitive to pH
weak acid/bases play important role in
protein structure/function

Shape of titration curve is same for all weak

acids

Acid/conjugate base pairs

HA + H2O
HA

A - + H 3O +
A- + H+

HA = acid ( donates H+)(Bronstad Acid)

A- = Conjugate base (accepts H+)(Bronstad Base)

Ka = [H+][A-]
[HA]
pKa = - log Ka

Ka & pKa value describe tendency to

loose H+
large Ka = stronger acid
small Ka = weaker acid

Henderson-Hasselbach Equation
Consider the dissociation of a general acid HA
HA

H + + A-

We can define a dissociation constant (K) where

[H ][A ]
K
[HA]

Rearranging gives

K[HA]
[H ]
[A ]

Taking logarithms on both sides and multiplying by -1 gives:

-log[H+] = -logK log [HA]/[A-] or

pH = pK + log [A-]/[HA]

Henderson-Hasselbach Equation
1) Ka = [H ][A ]
[HA]
+

HA = weak acid
A- = Conjugate base

2) [H+] = Ka [HA]
[A-]
3) -log[H+] = -log Ka -log [HA]
[A-]
4) -log[H+] = -log Ka +log [A-]
[HA]
5) pH = pKa +log [A-]
[HA]

* H-H equation
describes
the relationship
between
pH, pKa and buffer
concentration

Henderson-Hasselbalch Equation
This equation can be used to determine the pH
if the pK and ratio of the ionised and unionised
forms is known.
The pKa (a for acid) is the ve log of the
dissociation constant of the acid. It is the pH at
which the ratio of the ionised and unionised
species is equal to 1. ie the molar concentration
of the ionised and unionsed species is the same.
Similarly pKb is ve log of the dissociation
constant of the base

Regulation of H+ concentration
Concentration of hydrogen ions is regulated sequentially
by:
Chemical buffer systems act within seconds
The respiratory center in the brain stem acts within
1-3 min
Renal mechanisms require hours to days to effect pH
changes
Sources of hydrogen ions anaerobic and aerobic
respiration of glucose incomplete oxidation of fatty
acids oxidation of sulfur-containing amino acids
hydrolysis of phosphoproteins and nucleic acids

Buffers
Definition: A weak acid plus its conjugate base
that cause a solution to resist changes in pH when
an acid or base are added
Effectiveness of a buffer is determined by:
1) the pH of the solution, buffers work best
within 1 pH unit of their pKa
2) the concentration of the buffer; the more
present, the greater the buffering capacity

Buffer capacity
The buffer capacity of a system is
already defined as the amount of strong
acid or base added to one litre (l) of the
system in order to change the pH one
unit

The Major Body Buffer Systems

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

Bicarbonate

Important for metabolic acids

Haemoglobin

Important for carbon dioxide

Plasma protein

Minor buffer

Phosphate

Concentration too low

Proteins

Important buffer

Phosphates

Important buffer

Phosphate

Responsible for most of 'Titratable Acidity'

Ammonia

Important - formation of NH4+

Ca carbonate

In prolonged metabolic acidosis

Blood

ICF

Urine

Bone

 Carbonic Acid
Bicarbonate
Buffer
System
~ Most
important in
the ECF

Bicarbonate buffer system

Present in intra-and extracellular fluid
Bicarbonate ion acts as weak base, carbonic
acid acts as a weak acid
Bicarbonate ions combine with excess
hydrogen ions to form carbonic acid
Carbonic acid dissociates to release
bicarbonate ions and hydrogen ions

H+ + HCO3- H2CO3 H+ + HCO3-

The blood buffering system, simplified

Phosphate Buffer
system
~ Important in ICF & urine

Phosphate buffer system

Important in intracellular fluid and urine pH
regulation
Consists of two phosphate ions
Monohydrogenphosphate ions act as a weak base
and combine with hydrogen ions to form
dihydrogenphosphate
Dihydrogenphosphate dissociates to release
hydrogen ions

H+ + HPO4-2 H2PO4- H+ + HPO4-2

Phosphate has three ionizable H+ and

three pKas

Protein Buffer
Systems

~ Important in ECF and ICF

~ Interact with other buffer systems

Protein buffer system

Consists of Plasma Proteins (albumin,
hemoglobin)
Remember proteins are just chains of AAThe
exposed amine group of the AA (NH2) accepts
H+ ions when conditions are acidic
The exposed carboxyl group of AA can release
H+ ions when conditions are basic Proteins can
act as Acids or Bases

Hemoglobin 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+.

The acid-base buffering systems of the body.

The two buffer systems are in
dynamic equilibrium with the
same hydrogen ion concentration
(pH), so that a change induced in
the concentration of any one
factor in either buffer system
rapidly affects the other
system and a new hydrogen ion
concentration in the blood is
established.
1. The lungs assist in maintaining a
constant blood pH by removing
CO2,
2. while the kidney excretes acid in
the form of H2PO4- and NH4 and
alkali in the form of HCO3-.

Respiratory Buffer Systems

The respiratory system regulation of acidbase balance is a physiological buffering
system
There is a reversible equilibrium between:
Dissolved carbon dioxide and water
Carbonic acid and the hydrogen and bicarbonate
ions

CO2+ H2O H2CO3H++ HCO3

Respiratory Buffer Systems

CO2 is produced by
cellular respiration.
CO2 is converted to
bicarbonate by
carbonic anhydrase.
results in LOWER pH in
respiring tissues.
CO2 is exhaled in lungs.

Haemoglobin binds both CO2 and H+ and so is a powerful buffer. Deoxygenated haemoglobin has the strongest
affinity for both CO2 and H+; thus, its buffering effect is strongest in the tissues. Little CO2 is produced in red
cells and so the CO2 produced by the tissues passes easily into the cell down a concentration gradient. Carbon
dioxide then either combines directly with haemoglobin or combines with water to form carbonic acid. The CO2 that
binds directly with haemoglobin combines reversibly with terminal amine groups on the haemoglobin molecule to form
carbaminohaemoglobin. In the lungs the CO2 is released and passes down its concentration gradient into the alveoli.

Respiratory Acidosis and

Alkalosis
Result from failure of the respiratory system to
balance pH
PCO2is the single most important indicator of
respiratory inadequacy PCO2 levels
Normal PCO2 fluctuates between 35 and 45 mm
Hg
Values above 45 mm Hg signal respiratory acidosi
Values below 35 mm Hg indicate respiratory
alkalosis

Respiratory Acidosis

Respiratory acidosis is the most common cause of acid-base imbalance

Occurs when a person breathes shallowly, or gas exchange is hampered
by diseases such as pneumonia, cystic fibrosis, or emphysema

Respiratory alkalosis
A common result of hyperventilation
Excessive loss of CO2 & subsequent loss
of carbonic acid
Caused by hyperventillation:too much
CO2 lost (carbonic acid and H+ ions)
Anxiety, high altitudes (low O 2 levels),
musicians,
Symptoms:lightheadedness, agitation,
dizziness,

Metabolic Acidosis
All pH imbalances except those caused by
abnormal blood carbon dioxide levels
Metabolic acidosis is the second most common
cause of acid-base imbalance
Typical causes are ingestion of too much
alcohol and excessive loss of bicarbonate ions
Other causes include accumulation of lactic
acid, shock, ketosis in diabetic crisis,
starvation, vomiting, and kidney failure

Metabolic Alkalosis

Rising blood pH and bicarbonate levels indicate metabolic alkalosis

Typical causes are:
Vomiting of the acid contents of the stomach
Intake of excess base (e.g., from antacids)
Constipation, in which excessive bicarbonate is reabsorbed

Clinical Application
Acid-Base Imbalances
If the pH of arterial blood drops to 6.8 or rises to 8.0 for
more than a few hours, the person usually cannot survive

acidosis versus alkalosis

factors that lead to

respiratory acidosis

Clinical Application

Metabolic acidosis

Respiratory alkalosis

Metabolic alkalosis

Renal Mechanisms of Acid-Base

Balance
Chemical buffers can tie up excess acids or
bases, but they cannot eliminate them from
the body
The lungs can eliminate carbonic acid by eliminating
carbon dioxide
Only the kidneys can rid the body of metabolic
acids (phosphoric, uric, and lactic acids and
ketones) and prevent metabolic acidosis
The ultimate acid-base regulatory organs are the
kidneys

Kidney buffering power

Whatever the nature of the
disturbance, the response of the
kidney leads to the formation and
extraction from the plasma of a fluid
with an excess or a deficit of acid.
The primary result is a return of the
H ion concentration of the blood
toward the normal level.

Renal Mechanisms of Acid-Base Balance

The most important renal mechanisms for regulating acid-base balance are:
Conserving (reabsorbing) or generating new bicarbonate ions: decreases acidity
of ECF
Excreting bicarbonate ions: increases acidity of ECF
Excreting excess H+

Respiratory and Renal Compensations

Acid-base imbalance due to inadequacy
of a one system is compensated for by
the other system
The respiratory system will attempt to
correct metabolic acid-base imbalances
The kidneys will work to correct
imbalances caused by respiratory
disease

Respiratory Compensation
In metabolic acidosis:

The rate and depth of breathing are elevated Blood

pH is below 7.35 and bicarbonate level is low
As carbon dioxide is eliminated by the respiratory
system, PCO2falls below normal

In respiratory acidosis, the respiratory rate is

often depressedand is the immediate cause of
the acidosis
In metabolic alkalosis:
Compensation exhibits slow, shallow breathing,
allowing carbon dioxide to accumulate in the blood
Correction is revealed by:High pH (over 7.45) and
elevated bicarbonate ion levelsRising PCO 2

Clinical Applications
Homeostatic mechanisms slow down with
age
Elders may be unresponsive to thirst clues
and are at risk of dehydration
The very young and the very old are the
most frequent victims of fluid, acid-base,
and electrolyte imbalances

Buffering power of saliva

1.The pH of the mouth must be maintained near neutral for normal tooth
maintenance.
2.Oral pH is buffered to a small extent by saliva proteins and phosphate.The
major influence on saliva pH is bicarbonate ion which is a by-product of cell
metabolism
3.Bicarbonate concentration increases in saliva as the flow rate rises and is due to
the increased metabolic rate. This, in turn, raises the pH (more alkaline) of
saliva.
4.Bicarbonate ions diffuse into dental plaque and neutralise acid produced by
plaque bacteria when carbohydrate is fermented.
5.This reaction is driven by a unique type of carbonic anhydrase which is secreted
into saliva by serous acinar cells of the parotid and submandibular glands.
6.Bicarbonate ions maintain the pH of saliva above 6.3.

Role of Bone Buffering

Bone consists of matrix within which specialised cells are dispersed. The matrix
is composed of organic [collagen and other proteins in ground substance] and
inorganic [hydroxyapatite crystals: general formula Ca10(PO4)6(OH)2] components.

The hydroxyapatite crystals make up two-thirds of the total bone volume but
they are extremely small and consequently have a huge total surface area. The
crystals contain a large amount of carbonate (CO3-2) as this anion can be
substituted for both phosphate and hydroxyl in the apatite crystals. Bone is the
major CO2 reservoir in the body and contains carbonate and bicarbonate
equivalent to 5 moles of CO2 out of a total body CO2 store of 6 moles. (Compare
this with the basal daily CO2 production of 12 moles/day)

CO2 in bone is in two forms: bicarbonate (HCO3-) and carbonate (CO3-2). The
bicarbonate makes up a readily exchangeable pool because it is present in the
bone water which makes up the hydration shell around each of the
hydroxyapatite crystals. The carbonate is present in the crystals and its release
requires dissolution of the crystals. This is a much slower process but the
amounts of buffer involved are much larger.

Role of Bone Buffering

Bone is an important source of buffer in chronic
metabolic acidosis (ie renal tubular acidosis & uraemic
acidosis)
Bone is probably involved in providing some buffering
(mostly by ionic exchange) in most acute acid-base
disorders but this has been little studied.
Release of calcium carbonate from bone is the most
important buffering mechanism involved in chronic
metabolic acidosis.
Loss of bone crystal in uraemic acidosis is
multifactorial and acidosis is only a minor factor
BOTH the acidosis and the vitamin D3 changes are
responsible for the osteomalacia that occurs with
renal tubular acidosis.

Ammonia is produced in renal tubular

cells by the action of the enzyme
glutaminase on the amino acid glutamine.
This enzyme functions optimally at a
lower (more acidic) than normal pH.
Therefore, more ammonia is produced
during acidosis improving the buffering
capacity of the urine. Ammonia is
unionised and so rapidly crosses into the
renal tubule down its concentration
gradient. The ammonia combines with H+
to form the ammonium ion, which being
ionised does not pass back into the
tubular cell. The ammonium ion is
therefore lost in the urine, along with
the hydrogen ion it contains