Acid base balance

Dr. Prathima MBBS, MD

Professor
Dept of Biochemistry
 Specific learning objectives
• Define acids and bases

• Derive Henderson Hassebalch's equation and discuss its

importance

• Define buffer, buffering capacity and its significance

• Classify buffers in the body and plasma

• Explain role and mechanism of bicarbonate buffer,

phosphate buffer and protein buffer system in maintenance
of pH.
 Specific learning objectives
• Explain the role of respiratory system in maintaining acid
balance
• Explain the isohydric transport of CO2 in blood
• Discuss the key mechanisms by which kidneys help in
maintaining acid base balance.
• Define titrable acidity
 Specific learning objectives

• Classify acid base disorders based on the metabolic/

respiratory component and pH
• Describe the causes, pathophysiology and compensatory
mechanisms – all the acid base balance disorders
• Define Anion gap and write its reference range
• Classify metabolic acidosis based on anion gap giving at least
three causes in each type
• Classify metabolic alkalosis based on urinary chloride levels
 Specific learning objectives
• Recognize the relationship between serum potassium and
acidosis / alkalosis
• Explain the application of ABG analysis in clinical practice
• Interpretation of results of Arterial Blood Gas (ABG) analysis
data.
 According to Bronsted and Lowry

Acids: substances capable of donating protons.

bases: substances capable of accepting protons

• Strong acids: eg HCl
• Weak acids : eg H2CO3

HCl H+ + Cl-
Cl- is the conjugate base.
H2CO3 H+ + HCO3
HCO3- is the conjugate base
Bases:
Eg: NH3+ H+ =NH4
HCO3+ H+ = H2CO3
 pH
• pH = -log( H+) or 1/ log (H+)
• pH is inversely proportional to the H+
concentration.
Eg : Gastric juice = 1.5 -3
Pancreatic juice= 7.5 – 8
Urine = 5- 7.5
Arterial blood pH= 7.35-7.45
* Intracellular pH : 6.8-7.3
 Ka=dissociation constant

• Dissociation of an acid is a freely reversible reaction .

At equilibrium, the ratio of the dissociated and undissociated
particle is a constant
• Ka= [H+][A-]
[HA]
• pKa= pH at which the acid is half ionised.
• Strong acids have low pKa, weak acids have high pKa
 Buffers:
Buffers are solutions which can resist changes in the pH
when an acid or an alkali is added.

Composition of a buffer
vMixtures of weak acids + their salt with a conjugate
base.
vMixtures of weak bases + their salt with a conjugate
acid.
Bicarbonate buffer made up of :

H2CO3- is the weak acid

NaHCO3 salt of the conjugate base
pH= pKa+ log (base )
(acid)

*Henderson Hasselbalch equation

How do buffers act?
Buffering capacity
Capacity of the buffer to resist any change in pH when
an acid or base is added.

It is the number of grams of strong acid or alkali that is

needed for the change in pH by 1 unit of 1 litre of
buffer.

Effective range of a buffer:

Its 1 pH unit higher or lower than the pKa.
But buffer is most effective when pH=pKa.
 Henderson Hasselbalch equation

• pH= pKa+ log (base )

(acid)

When the concentration of base by acid is the

same pH=pKa.
Derivation of HH equation
Factors affecting the pH of the buffer:
1. pKa: Lower the pKa, lower is the pH
2. Ratio of the salt (base) to the acid
concentration
 Applications of HH Equation
• To calculate the concentration of acid and
base to be added to prepare a particular
buffer.
• Clinical applications –to assess the acid base
balance status .
• To predict the limits of compensation of body
buffers.
 Buffers of the body
• Buffers of the body fluids – First line of
defence(blood buffers)
• Respiratory system-second line of defence
• Renal system-third line of defence.
• Normal arterial pH=7.35-7.45
• <7.35 acidosis >7.45 alkalosis

• During normal metabolism acids are produced:

1. Volatile acids(20,000meq/day): H2CO3
2. Fixed acids(60-80meq/day) : lactic acid, keto acids,
sulphuric, phosphoric acid.
• Volatile acids eliminated by lungs as CO2
• Fixed acids are buffered by the body fluids
and excreted later by the kidneys.
In the body :
• 58% of buffering is done by ICF buffers ( 52%
tissue cells + 6% by RBC’s)
• 42% is done by ECF buffers
 Buffers of the body fluid
ECF (42%) ICF( 52%) RBC FLUID
(6%)
Bicarbonate Phosphate Hb buffer
buffer buffer
Phosphate Protein buffer Phosphate
Buffer buffer
Protein buffer Bicarbonate Bicarbonate
buffer buffer
Buffers of the body fluids
 Bicarbonate buffer system

• Most imp buffer system in the ECF

• The normal HCO3 level 22-26 mmol/L.
Reasons for bicarbonate buffer to be the most
imp buffer

• Relatively high concentration of HCO3

• They can act at 2 levels :
Acid component(H2CO3)regulated by RS
Base component(HCO3) reg by kidneys.
• The ratio of base to the acid is 20:1
• Concentration of HCO3 is 20 times more than
H2CO3 - alkali reserve
pH= pKa+ log (base )
(acid)
pKa for carbonic acid=6.1
7.4=6.1+log 24/0.03X 40
7.4=6.1+log 24/1.2
=6.1+log 20

=6.1+1.3
=7.4
Mechanism of action of bicarbonate buffer
 Phosphate buffer system
• Important ICF buffer and also PH regulation of urine
• HPO4 / H2PO4
• pKa of 6.8 is close to the intracellular pH 6.8 – 7.3
• They are effective at a wide range of pH as they have
more than one ionizable group and the pKa value
are different for the different ionizable groups.
 Has more than 1 ionizable group with pKa
values
pKa=1.96
H3PO4----------H+ H2PO4
pka=6.8 (close to intracellular pH )
H2PO4--------H+ HPO4
pKa=12.4
HPO4--------H+ +PO4
pH = pka + log base /acid

7.4 = 6.8 + log base /acid

7.4 - 6.8 = log base /acid
.6 = log base /acid
Antilog 0.6 = 4/1
 Protein & Hb buffer
• Imp buffer in blood and ICF( after phosphate )
• Buffering action of proteins are due to amino acids with
ionizable side chains.
• Histidine residues are more common
• Histidine residues with imidazole side chain (pka= 6) causes
buffering action
• Eg 16 residues of histidine in albumin
38 residues of histidine in Hb.
• Body buffers can act quickly but NOT
permanently
• Do not help in acid elimination from the body.
• Respiratory & Renal systems are important for
the elimination .
 Respiratory regulation of pH

• Second line of defence

• Rapid action but short term and taken over by
kidneys
• When PC02 increases ,chemoreceptors of the
brain get stimulated .
• This increases the rate and depth of
respiration
• Thus eliminating the CO2 from the body.
• This cannot continue for a long time
 Action of hemoglobin

• Isohydric transport: Hb transports CO2 with

little change in pH
• Generates HCO3 or alkali reserve by the
activity of an enzyme -carbonic anhydrase.
• O2 & CO2 exchange occurs
at Pulmonary & Peripheral
tissue capillaries
• CO2 is transported from
tissue to pulmonary
capillaries by 3 mechanism
as :
Ø HCO3 (70%)
Ø CarbaminoHb( 20%)
Ø Dissolved state ( 10%)
Transport of C02 from tissue capillaries
70% in the form of HCO3
20% as carbaminohemoglobin
In the pulmonary capillaries
 Renal regulation
• One of the functions of kidney – regulation of pH of the ECF
• Normally urine pH is acidic, pH is lower than ECF , kidneys
contribute to acidification of urine .
• Normal urine pH is slightly acidic
• Normal range is 4.5 to 7.8.
• pH of the glomerular filtrate is about 7.4, falls to 6.9 in
proximal tubule, 6-6.5 in distal tubule and to 4.5- 4.7 in the
collecting duct
 Renal regulation mechanisms
1. Excretion of H+(and reabsorption of HCO3-)
2. Reabsorption of HCO3 (No excretion of H+)
3. Excretion of titrable acids
4. Excretion of ammonium ions (NH4+)
Structure of nephron
Structure of nephron
Excretion of H+ (proximal tubule )
• 85% of the H+ excreted from PCT.
• 15% of the H+ excreted from DT &CD.
• H+and Na +exchange takes place throughout
the formation of urine .
Reabsorption of HCO3 ( no excretion of H+)
• Glomerular filtrate contains the same amount of HCO3- as
plasma.
• The entire HCO3- has to be reabsorbed so normal urine is
almost HCO3 free.
• The luminal surface of the Renal tubular cells are
impermeable to HCO3, so direct absorption is not possible.
• So HCO3 converted to H2CO3 and then H20 +CO2 in lumen.
• Water moves into the cells using aquaporins water channels
Titrable acids:
• It is a measure of net acid excretion by the
kidneys.
• 10-40mmol per day
• It is the number of milliliters of N/10 NaOH
required to titrate 1 litre of urine to PH of 7.4.
• The major titrable acid present in the urine is
sodium acid phosphate
Excretion of titrable acids
Excretion of ammonium ions
• Normally 70meq/L of acid is excreted /day
• In acidosis increases to 400 meq/L .
• Glutamine is produced in liver and transported to kidneys via
circulation
• Increase in glutaminase and NH4+ excretion takes 3 to 4 days
to begin .
• NH4 +accompanies acid ions and conserves Na+ & K+.
• High capacity of acid elimination ie ½ to 2/3 of body acid load.
 Urinary buffers

• Nephrons cannot produce a urine pH <4.5

• So in order to excrete more H+, they must be buffered.
• Thus the ability of the kidneys to eliminate large amount of
H+ in a normal volume of urine depends on the buffers
present in the urine.
• H+ combines with HPO4-2 or NH3

ü Phosphate : HPO4+ H+ --àH2PO4-2

ü Ammonia : NH3+ + H+--à NH4+
 Intracellular buffers
• 58% of the total buffering capacity of the body
• Skeletal muscle and bone
• H+ in the Skeletal muscle is removed by
exchanging with Na & K ions
• In the bone , its buffered Ca/Na/Carbonate
ions

* Decalcification of bone in chronic acidosis