ACID BASE BALANCE

Review
 Our pH homeostasis mechanism
When there is decrease in pH [any cause]

⬇︎⬇︎⬇︎⬇︎⬇︎ 
Blood buffers try to bring the pH back to normal
[ the bicarbonate buffer[HCO3-/H2CO3] is the main buffer in this setup]


Bicarbonate converted to carbondioxide[HCO3] and this is exhaled through
lungs

Now there is bicarbonate is deficit

Here the kidneys come into action
[regeneration of new bicarbonate ,reclaiming the bicarbonate escaping into the urine]


When all these defense fronts are exhausted, then there result in acidosis.
 Syllabus objectives
 describe buffer systems of plasma, interstitial
fluid and cells.
 state the dissociation constant of physiological
organic phosphates, histidine side chain N-
terminal aminogroups.
 explain role of kidneys in acid base
maintenance.
 recall
 hydrogen ions are
present in all body
compartments

 Maintenance of
appropriate
concentration of H+ is
must for proper cellular
function

 Maintenance of
appropriate
Inhibition of glycolysis in acidosis
concentration of H+ is
called acid base balance
or pH homeostasis
 Lines of defense in pH alteration
pH homeostasis is maintained by
 First line of defense :buffers

 Second line defense:lungs[to excrete volatile

acid(co2)
 Third line of defense: kidneys [ to excrete fixed

acid.pyruvic acid,lactic acid]

 Sources of Hydrogen Ions in Body
 In 24 hour, a person weighing 70 kg disposes about 20 mol of
carbon dioxide (volatile) through lungs and 70 – 100 mmol of
nonvolatile acids (sulfuric and phosphoric acid) through kidney

 Still the extracellular H+ concentration is maintained to about 40

nmol/L (pH 7.4)

Arterial Blood pH: Venous Blood pH:

7.35 – 7.45 7.32 – 7.38

Intracellular fluid
pH:
6.0 – 7.4
 buffer
 a solution which resists changes in pH when
acid or alkali is added to it.
 Buffer solution is the mixture a weak acid and
its conjugate base pair
 E.g
 H2CO3 weak acid,its conjugate base [pair] is HCO3-
 Bicarbonate buffer is the mixture of H2CO3 : HCO3-
 Unbuffered water and addition of strong
acid

 When 1mmole of HCl added to 1L of

unbuffered water
pH Water:7=10-7 H+
1mmoles HCl =10-3 H+
New [H+] = 0.0000001 +0.001
=0.0010001
~ 10-3moles
pH = 3
 Bicarbonate Buffer solution[pH6.3] and
addition of strong acid

 When 1mmole of HCl added to 1L of

10mmoles bicarbonate buffer solution
 Acid component increase by 1mmoles
 Base component decrease by 1mmoles

Use of hendersen hasselbalch equation for determining final pH

pH=pK + log [HCO3-/H2CO3]
=6.1 +log[9/11]
=6.1 –o.o87
= 6.o13
Henderson-Hasselbach equation

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

pK= negative log of dissociation constant

 Significance of Henderson Hasselbach
equation
1. Prepare buffer solution of different pH range
2. pK value determines the maximum buffering
capacity range of a given buffer
3. Lower the pK value lower is the pH
4. The pH remains the same till the ratio of base to
acid remains constant
 Working of buffer:
Titration Curve

 Used to determine the amount of

weak acid in solution which also
yields pKa value
 Conjugate acid (HA) of a weak
acid is stoichiometrically
converted to its conjugate base
(A-) by addition of strong base
 pH thus obtained is plotted
against the concentration of OH-
added
 Buffering capacity
Buffers ability to maintain the pH of the given
solution depends on the
1. Pk value :
 buffers best work to maintain the pH which is equal
to its pK value
 Buffers work better to maintain the pH which is in
the range of +1 to -1 of pK value
 E.g if pk =6.8, the range of buffering is good in pH
range 5.8 to 7.8
2. Concentration of the buffer components.
 What we need in our body buffers
 Buffer system with higher ratio of anions
which can buffer the acids produced in our
body
 i.e why all our buffer system have higher anion ratio
comparison to acid component
 Buffer system whose pK is around our physiological
pH [7.35 -7.45]

 Buffer system which is available in sufficient

concentration
 Buffers of our body
 Bicarbonate buffer: NaHCO3/H2CO3
 Phosphate buffer: Na2HPO4/NaH2PO4-
 Organic phosphates:2,3 bisphosphiglycerate
 Protein buffer: Pr-/HPr
 Hemoglobin buffer: Hb/HHb
 Albumin buffer :Alb/HAlb
 Ammonia Buffer: NH3/NH4+
 Concentration of buffers
 Bicarbonate buffer:21 -28mmoles/L
 Phosphate buffer: 1.2- 2.2mmoles/L[Intracellular

phosphates ~100mmoles/L]
 Ammonia buffer : on demand [from glutamine

and glutamate]
 Protein buffer:
 Hemoglobin : Males :14-18 g/dL [ 8.7-11.2 mmol/L]
-  Female: 12-16 g/dL[ 7.4-9.9 mmol/L]
 Albumin :4-6gms/dl [60.18µmol/L - 90.28µmol/L ]
Na2HPO4/
K H2PO4
 Normal ranges
NORMAL RANGE CRITICAL LIMIT
 H + = 35 -45nmoles/L
 pH =7.35 -7.45 pH : <7.2 and >7.6
 pCO2 = 35 – 45 mmHg pCO2 : <20 and > 70
 PO 2= >80mmHg pO2 : <45
 HCO3- = 21 -28mmoles/L HCO3 : <10 and
 Base excess = -2 to +2 >40
 Oxygen saturation: >95%
Bicarbonate buffer system [recall]
 Characteristic of bicarbonate
bufferBicarbonate buffer: NaHCO3-/H2CO3

1. In our body carbonic acid mainly found in

dissolved form of carbon dioxide dCO2.
i.e H2CO3= dCO2
2. Conc of dCO2 = αPCO2 =0.031x 40
=1.24mmoles/L
[α= burns coefficient of solubility=0.031mmoles/L/mmHg]
[P= partial pressure of CO2 = 40mmHg]
 Characteristic of bicarbonate buffer

3. Concentration of bicarbonate ~ 25mmoles/L

[Bicarbonate is synthesized through the enzyme carbonic anhydrase
with very high turnover, high catalytic activity.]

4.HCO3-/H2CO3 ~ 20:1 [ 25/1.24 = 20.16:1]

5.pK=6.1 [best working range 5.1 – 7.1]

 Bicarbonate buffer in Henderson
Hasselbach equation
pH= pk +log [HCO3-]/H2CO3]
= 6.1 + Log 20/1
=6.1+ 1.3
= 7.4 which is the pH of blood
 Characteristic of bicarbonate buffer

7.Bicarbonate buffer system is said to be open

buffers system
*Bicarbonate level is regulated by kidney

* Carbondioxide level is regulated by lungs

8.Taken as an index of acid base balance in the

body
9.Main buffer for metabolic acidosis
Phosphate buffer system
Characteristics of phosphate buffer system

 Both organic and inorganic phosphates act as

buffers
 In plasma inorganic phosphate is involved

 Base /acid ratio = 4:1 [ NaHPO42-/NaH2PO4- ]

 concentration in plasma is 1.2-2.2 mmoles/L

 pK=6.8 [best working range 5.8 to 7.8]
phosphate buffer in Henderson Hasselbach
equation
pH= pk +log [HPO4-]/H2PO4-]
= 6.8 + Log 4/1
=6.8+ 0.6
= 7.4 which is the pH of blood
 Three ionizable inorganic phosphate
buffer

Phosphoric acid H3PO4 has three acid forms

 H2PO4-/H3PO4 ------------- pk=2.6

 HPO42-/H2PO4- ----------------- pk= 6.8

 PO4-/HPO4 2- ------------- pk=12.5

Characteristics of phosphate buffer system

 Major role in kidney to excrete H+ and

regeneration of bicarbonate ions

 Intracellular phosphates ~100mmoles/L;majorly

they are Organic phosphate buffer
 2,3 diphosphoglycerate in RBC
 Glucose 1P
 ATP,ADP,AMP
Ammonia Buffer System
Characteristics of ammonia buffer system

 Pk = 9.1

 NH3:NH4+ = 1:100

 In body mainly remains as NH4+

 Major role in kidney to excrete H+ and

regeneration of bicarbonate ions
pH= pk +log [NH3]/NH4+]
= 9.1 + Log 1/100
=9.1+log 10 -2
=9.1- 2
= 7.1 which is the pH of urine
 Characteristics of ammonia buffer system

 This buffer system is inducible. it is induced by

increased acid load in the renal tubule

 Ammonia is generated by following reactions

which immediately convert to ammonium
binding with H+
NH3 NH3
Glutamine ---> glutamic acid ---> alphaketoglutarate
glutaminase glutamic oxidase
Protein buffer system
 Characteristics of protein buffer
 Proteins have various dissociable groups
contributed by their constituent aminoacids
 Dissociable groups can give or take H+
depending on their different pk values
 Dissociable groups whose pK value is nearer to
physiological pH will only contribute to
buffering
Dissociable groups with their pK value
 Alpha COO --------------- 3.5 – 4
 Non alpha carboxyl in Asp & Glu ------- 4 - 4.8
 Imidazole of histidine ---------- 6.5 – 7.4
 Sufhydral of cysteine ---------- 8.5 – 9
 Hydroxyl group of tyrosine ------ 9.5 – 10.5
 Alpha aminoacid ----------- 8 – 9
 Epsilon aminoacid of lysine ------ 9.8 – 10.4
 Guanidium of arginine --------- 12
 Ionization of Histidine

 The R group (side chain) of histidine has an

imidazole functional group that undergoes
reversible protonation.
 Three ionizable groups:
 Acid (-COOH)
 Protonated Imidazole
 - (NH3)+
 At pH 7.3, carboxyl group of histidine is
entirely deprotonated (-COO-) and α-amino
group is fully protonated (- NH3+),
imidazole is partially dissociated
 pK1=1.8, pK2=6.0, pK3=9.2
 After calculations 4% histidine protonated
at pH 7.3
 Characteristics of protein buffer
 Hb ,avg pK = 7.3

 Plasma proteins ---- pK = 6.5 --- 7

 Hemoglobin has 36 histidine aminoacids

 Albumin has 16 aminoacids

 Characteristics of protein buffer
 Concentration of Hb = 16g/dL and Concentration of
albumin = 4g/dL

 They transport CO2 with buffering of proton

component

 Deoxy Hb is better buffer [pK= 7.9]than oxy Hb [pk

= 6.7]

 For each mmoles of O2 liberated Hb removes

0.7mmoles of H+
 Intracellular buffering
 Physicochemical buffers
 Intracellular proteins~50%
 Organic phosphates ~48%
 Bicarbonates~2%
 aminoacids
 Biochemical buffers
 Contributes 50% that of physicochemical buffers
 Defined as change in cellular metabolism with change in
pH[increase glycolysiswith increase in pH]
 Organelles as buffers
 Sequestering of H+ into lysosomes,mitochondria,golgi
apparatus
Role of kidney in Regulation
of Acid-Base Balance
 Excretion of acid
 Reclamation /regeneration of bicarbonate
 Excretion of bicarbonate [alkalosis]
 Renal Mechanism
 Average pH of glomerular filtrate = 7.4
 Average urinary pH = 6.0
 Indicates renal excretion of non-volatile acids
 Acidosis: excretion of acids is increased and
base is conserved and regenerated
 Alkalosis: excretion of bicarbonate
 Renal Mechanism of excretion of H+

 NaH exchanger[mainly]
 H+ ATPase [A intercalated cell]
 H+K+ATPase
[during K depletion in Alpha intercalated cell]
 Na+ - H+ Exchange/bicarbonate
reclammation
 Nearly all mammalian cells
contain a plasma membrane
ATP-hydrolyzing protein
capable of exchanging
sodium ions for protons :
Na+ - H+ exchangers

 In the renal tubules, NHE-1

and NHE-3 appear to be the
predominant isoforms that
extrude H+ ions into the
tubular fluid in exchange for Bicarbonate filtered through glomerulus is
Na+ ions reclaimed into circulation
For each mmol of H+ secreted into tubular fluid,
1 mmol Na+ and 1 mmol (HCO3)- enter
the tubular cell and return to general circulation
 Reclamation of Filtered Bicarbonate

 Unmodified glomerular filtrate and plasma have

same concentration of bicarbonate ion
 Increasing acidification of proximal tubular
urine causes decreased bicarbonate
concentration
 Excreted H+ reacts with bicarbonate to form
carbonic acid and subsequently carbon dioxide
and water
 CO2 diffused into cell and thus via cytoplasmic
anhydrase there is reclamation of bicarbonate
 Normally 90% of the filtered bicarbonate ion
(4500 mmol/d) is reclaimed in PCT For each mmol of H+ secreted
 Threshold: 28 mmol/l into tubular fluid, 1 mmol Na+
 Type II RTA: decrease in blood pH due to and 1 mmol (HCO3)- enter the
decreased ability of PCT to reabsorb bicarbonate tubular cell and return to general
ion circulation
 Secretion of H+ with Regeneration of
bicarbonate

1.H+ secreted is buffered

by dibasic phosphate
Under normal physiologic
condition, 30 mmol of H+
is excreted per day as
(H2PO4)-

2. H+ secreted is buffered
by ammonia released from
deamination of glutamine
and glutamate
 Renal Production of Ammonia and
Excretion of Ammonium Ions
 In normal individuals,
(NH4)+ production in
tubular lumen accounts to
excretion of 60% (30 to 60
mmol) of hydrogen ions
 At normal blood pH, ratio of
(NH4)+ to NH3 is about 100
to 1

At acidic pH of urine, the equilibrium between (NH4)+ and NH3 shifts

markedly to the left (10000 to 1) strongly favoring formation of (NH4)+
Role of Potassium in H+ Exchange
 Potassium ions compete with H+ in renal
tubular NHE
 As intracellular K+ levels of renal tubular cells
is high, more K+ and less H+ are exchanged for
Na+ (Urine becomes less acidic)  Increased
acidity of body fluids Hyperkalemia
Acidosis
 As K+ is depleted, more H+ ions are exchanged
for Na+, urine becomes more acidic 
alkalinity of body fluid increased Hypokalemia
Alkalosis
Bicarbonate chloride exchanger for bicarbonate
excretion

When bicarbonate concentration> 26mmoles/l

 Which is a better urinary buffer:
Phosphate or Ammonia?
 In normal conditions: Phosphate is the
primary urinary buffer.
 During acidosis: The role of phosphate is
limited as it’s secretion is not regulated by
kidney but is dietary dependent. In such
conditions, ammonia is the important buffer
 Acidosis associated with kidneys
 Uremic acidosis
 Renal tubular acidosis
 URAEMIC ACIDOSIS
 Uraemic acidosis results from the loss of functional nephrons
 involves injury to glomeruli and tubules
 decreased glomerular filtration rate (GFR) (e.g. <20 mL/min)
 failure to excrete acid anions
 accumulation of acidic anions such as phosphate and sulfate
occurs
 causes high anion gap metabolic acidosis (HAGMA)
 low plasma HCO3
 patients manifest as renal failure, often have prolonged
survival and develop chronic complications such as bone
demineralisation
RENAL TUBULAR ACIDOSIS (RTA)
 Renal tubular acidosis (RTA) involves defects isolated to the
renal tubules only
 GFR may be normal or only minimally affected[occurs despite a
normal or only mildly reduced glomerular filtration rate (GFR)]
 primary problem is impaired ability to acidify the urine and
excrete acid [defective renal acid-base regulation]
 results in net acid retention and hyperchloremic normal anion
gap metabolic acidosis (NAGMA)
 may be incomplete and only develop in the presence of an acid
load
 RTA is often detected incidentally through an abnormal blood
workup, but some patients present with clinical features such as
poor growth, dehydration, or altered mental state
 Type 1 distal Type 2 proximal Type 4
causes Hereditary ,autoim Hereditary,part of Primary/secondary
mune, Fanconi syndrome, aldosterone
nephrotoxins, lead nephropathy, deficiency
nephrocalcinosis amylodoisis
 Type 1 distal Type 2 proximal Type 4
Defect reduced H+ impaired HCO3 impaired cation
excretion in distal reabsorption in exchange in distal
tubule proximal tubule tubule
Hyperchloremic yes yes yes
NAGMA
Minimum urine >5.5 <5.5 (but usually <5.5
pH >5.5 before
acidosis becomes
established)
plasma HCO3 <15 usually >15 usually >15
Plasma K low-normal low-normal high
Renal stones yes no no