ACID-BASE BALANCE(1)

Dr. HENRY EKORIKO

DEPT OF MEDICAL BIOCHEMISTRY
UNIUYO.
INTRODUCTION (1)

 The acid-base balance or pH of the body fluids is maintained by a closely regulated mechanism.

 This involves the body buffers, the respiratory system and the kidney.

 Hydrogen ion concentration decides the ionization of weak acids and thus affects their
physiological functions

 According to Bronsted, acids are substances that are capable of donating protons

 bases are those that accept protons.

 Acids are proton donors and bases are proton acceptors.

INTRODUCTION (2)
HA H+ + A-

HCl H+ + CL-

H2 CO3 H+ +HCO3 -

BASE

H+ + NH3 NH4

H+ + HCO3 - H2 CO3
STRONG AND WEAK ACIDS
 Strong acids dissociate completely in solution e.g HCL

 In a solution of HCl, almost all the molecules dissociate and exist as H+ and Cl– ions.
Hence, the concentration of H+ is very high and it is a strong acid

 weak acids ionize incompletely, for example H2CO3

 weak acid (e.g. acetic acid), it will ionize only partially. So, the number of acid molecules
existing in the ionized state is much less, may be only 50%.

 the dissociation of an acid is a freely reversible reaction

 at equilibrium the ratio between dissociated and undissociated particle is a constant.

HYDROGEN ION CONCENTRATION (pH)

 pH is defined as the negative logarithm of hydrogen ion concentration to the base 10. The
hydrogen ion concentration [H+] is expressed in moles/litre. pH = –log [H+] = log 1/[H+]

 The acidity of a solution is measured by noting the hydrogen ion concentration in the solution

 pH scale ranges from 0 to 14.pH = 7 is neutral, while pH > 7 is alkaline and pH < 7 is acidic

 Pure water or neutral aqueous solutions have [H+] = 1 × 10–7 mol/litre. Therefore their pH
according to definition can be calculated to be equal to 7.

 A rise or fall in pH by 1 signifies a tenfold fall or rise in the H+ conc. respectively. pOH is the
negative logarithm or [OH–] to the base 10
 pH value is inversely proportional to the acidity. Lower the pH, higher the acidity or
hydrogenion concentration while higher the pH, the acidity is lower

 At a pH of 1, the hydrogen ion concentration is 10 times that of a solution with a pH 2 and

100 times that of a solution with a pH of 3 and so on. The pH 7 indicates the neutral pH,
when the hydrogen ion concentration is 100 nanomoles/liter.

 Ionic product of water = [H+][ OH–] = 10–14 i.e ; pKw = pH+ pOH. pKw of water at room temp
is 14.
Henderson-Hasselbalch Equation
HA = H+ + A–

 Equilibrium constant K can be written as, k= [H+] + [A–]/ [HA]

 pH = pKa + log[A-]/[HA] ; this is Henderson-Hasselbalch Equation where Ka = acid dissociation

constance; [A-] = conjugate base, [HA] = acid

 pH = pKa + log [base]/acid When [base] = [acid]; then pH = pKa

 pKa of an acid group is that pH at which the protonated and unprotonated species are present at equal
concentrations. pKa is the pH at which the acid is half ionised; Salt : Acid= 1 : 1.

 Therefore, when the concentration of base and acid are the same, then pH is equal to pKa. When the
acid is half ionized, pH and pKa have the same values.
Applications of Henderson-Hasselbalch Equation
 The Henderson-Hasselbalch’s equation, has great practical application in clinical practice in assessing
the acid-base status, and predicting the limits of the compensation of body buffers.

 It can be used to determine pH of blood, if the concentration of salt, i.e. bicarbonate and acid (carbonic
acid) is known

 To determine the pH of a buffer solution if pKa of the buffer acid and the molar ratio of salt to acid in
the solution are known

 Example; if approx. concentration of bicarbonate in healthy individual is 0.025 M, approx.

concentration of carbonic acid is 0.00125 M and pKa of carbonic acid = 6.1. Calculate the pH of blood.
BIOLOGICAL IMPORTANCE OF pH
 Enzymes function at optimum pH. Various body fluids and cell organelles, etc. maintain a specific pH
in order to allow the activity of the enzymes located there

 Amino acids and proteins exist as Zwitterions at isoelectric pH. The magnitude of the charge depends
on the pH

 Specific tautomeric form of nucleic acid bases exist at pH 7.4. This helps in proper hydrogen bonding
between the complementary base pairs

 pH is responsible for Gibbs-Donnan membrane equilibrium

 pH and Keq : pH influences the Keq product yield and spontaneity of metabolic oxidation-reduction and
some nonenzymatic acid-base catalysis.
 BUFFERS

 Buffers are solutions which can resist changes in pH when acid or alkali is added

 Buffers consist of mixtures of weak acids and their corresponding salts, alternatively, weak bases and their salts.
The former type is the more important and common in human body

 The process by which the added H+ or OH– are removed is called as buffering action.

 The extent of resistance of change in pH by buffer is called its buffering capacity

 the buffer capacity is determined by the actual concentrations of salt and acid present, as well as by their ratio

 The pH of a buffer solution is determined by two factors:

 The value of pK: The lower the value of pK, the lower is the pH of the solution.

 The ratio of salt to acid concentrations

 A few examples are : H2 CO3 /NaHCO3 (Bicarbonate buffer) (carbonic acid and sodium

bicarbonate); CH3 COOH/CH3 COONa (Acetate buffer) (acetic acid and sodium acetate);

Na2 HPO4 /NaH2 PO4 (Phosphate buffer)

MECHANISM OF ACTION OF A BUFFER
 MECHANISM OF ACTION OF A BUFFER (2)
The sketchy diagram above shows the weak acid HA and its completely ionised salt B+A–.

 Added H+ ions, in the form of strong acid, combine with anions A– to form the weakly dissociable HA, so that pH does
not become more acidic as it would be in the absence of the buffer. The capacity to combine with added acid remains
so long as there is a supply of the buffer salt in the medium

 Added OH– ions, in the form of a strong base, combine with H+ ions derived from the acid HA and form the weakly
dissociable H2O molecules and pH does not become as alkaline as would happen in absence of the buffer.

 The buffer capacity is determined by the absolute concentration of the salt and acid. But the pH of the buffer is
dependent on the relative proportion of the salt and acid
MECHANISMS OF REGULATION OF pH

 “Front-line” defence: They are mainly:

 Buffer systems in the blood

 Respiratory mechanisms: Regulation of excretion of CO2 and hence, regulation of H2CO3

concentration in EC fluid

 “Second-line” defence: This is achieved by kidneys(Renal mechanisms).

 Dilution factor: The acids introduced into and formed in the body are distributed throughout
the ECF volume
BUFFER SYSYTEMS OF THE BODY

Extracellular Intracellular Erythrocyte

fluid fluid fluid
1 NaHCO3/H2CO3 K2 HPO4/KH2 PO4 K+Hb/H+Hb
(phosphate) (hemoglobin)
(bicarbonate)
2 Na2HPO4/NaH2PO4 K+ Protein/H+Protein K2HPO4/KH2 PO4
(phosphate) (protein buffer) (phosphate)
3 Na+ Albumin/H+ Albumin KHCO3/H2CO3 KHCO3/H2CO3
 Bicarbonate Buffer System
 The most important buffer system in the plasma is the bicarbonate-carbonic acid system
(NaHCO3/H2CO3). It accounts for 65% of buffering capacity in plasma and 40% of buffering
action in the whole body.

 Normal ratio in blood = NaHCO3/H2CO3 = 1/20, pH 7.4

 It accounts for 65% of buffering capacity in plasma and 40% of buffering action in the whole
body. The normal bicarbonate level of plasma is 24mmol/L.

 They are the chief buffers of blood and constitute the so called alkali reserve.
Neutralisation of strong and non-volatile acids entering the ECF is achieved by the
bicarbonate buffers.
 Alkali reserve : It is represented by the NaHCO3
The bicarbonate carbonic acid concentration in the blood that has not yet
buffer system is the most combined with strong and non-volatile acid.
important for the following reasons:
 Presence of bicarbonate in
relatively high concentrations.
 The components are under
physiological control, CO2 by lungs
and bicarbonate by kidneys
 Produces H2CO3, which is a weak
acid and volatile and CO2 is exhaled
out
 Phosphate Buffer System
 It is mainly an intracellular buffer. Its concentration in plasma is very low. The pKa value is 6.8. In the body,
Na2HPO4/NaH2PO4 is an effective buffer system, because its pKa value is nearest to physiological pH.

 Na2HPO4 / NaH2PO4 = [Alk PO4]/[Acid PO4]) Normal ratio in plasma is 4:1. This ratio is kept constant with the
help of the kidneys. Thus, phosphate buffer system is directly linked up with the kidneys. Below is the illustration
of the action of phosphate buffer when an acid(HCl) and when the base(NaOH) enter the bood
Protein Buffer System

 Buffering capacity of protein depends on the pKa value of ionizable side chains. The
most effective group is histidine imidazole group with a pKa value of 6.1

 In acidic medium: protein acts as a base, NH2 group takes up H+ ions from the
medium forming NH+3, Proteins become +vely charged

 In alkaline medium: Proteins act as an acid. Acidic COOH gr dissociates and gives H+,
forming COO–. H+ combines with OH– to produce a molecule of water, proteins
become –vely charged.
Hemoglobin as a Buffering Agent
 Like other proteins, the buffering action of Hb depends on the following : acidic-COOH gr,
basic-NH2 gr, Guanidino group and most important is imidazole group, which varies with the

pH of the medium.

 the buffering capacity of Hb is due to the presence of “Imidazole” nitrogen group which
remains dissociated in acidic medium and conjugate base forms
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