Learning objectives for all three lecture hours:

23. Define acids, bases, buffers and buffering capacity

24. Describe the interrelationships of pH, PCO2 and bicarbonate
concentration, and state the Henderson-Hasselbalch equation
25. List criteria for evaluation of buffering capacity
26. List the buffer systems available in the human body.
27. List the specific properties of CO2 - HCO3- buffer system
28. Describe production of acids and bases in the body and their excretio
29. State the normal ranges of arterial pH, PCO2, HCO3-,base excess
and anion gap concentration
30. Define alkalosis and acidosis and differentiate between their
respiratory or metabolic origin
31. Discuss the respiratory and renal mechanisms that help
compensate for acidosis and alkalosis.
32. Evaluate blood gas data to determine a subject's acid-base status
 Acid - base balance in the body
CO2
Tissue
Blood

CO2

CO2 Lung

HCO3-

OH- +

H+ H+ + B- HB
Kidney

H+ oder HCO3-

Production Buffering Excretion

and transport
 Pairs of acid - base
( BRØNSTED -Definition )
HA A- + H +
Conjugated acid Conjugated base
Examples: NH4+ NH3 + H+
H3PO4 H2PO4- + H+
H2PO4- HPO4-- + H+

Acid : dissociates H+

Base : binds H+
Buffer: a pair of acid and base
 Law of Mass action

Qualitative : HA A- + H+

[ H+ ] · [ A- ]
Quantitative : = K'
[ HA ]

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

Henderson-Hasselbalch- equation
= logarithmic form of the law of mass action
 Buffer curve
100
[A- ]
pH = pK' + log
99 [HA]
Acid
75
Base
Base, A- (%)

Buffer capacity
50 D[A- ]
b= 91
DpH

25
50

1 9
0
pK'-2 pK'-1 = pK' pK'+1 pK'+2
PH
Buffer capacity depends on: pH – pK’ difference & Total concentration of buffer
 Buffer curves of three biological
acid-base pairs

pK'
3,9 6,8 9,4
Addition of OH-  Increase of [A- ] (mmol/l)

100 0

Addition of H+  Increase of HA (mmol/l)

HLa H2PO4- NH4+

50 50

La- HPO4-- NH3

0 100
3 5 7 9 11
pH
 Examples of biological Buffers
pH range of the most body fluids: 6,8 - 7,4

1. Phosphate Buffer : pK' = 6,8

H2PO4- H+ + HPO4--
High intracellular concentration: (exeption erythrocytes)
(also phosphorylated substances)

2. Bicarbonate / CO2 -Buffer : pK' = 6,1

CO2+ H2O H++ HCO3-

3. Proteins :
a) Imidazolring of Histidine
b) Terminal amino group
c) Sulfhydril group of Cysteine
 Properties of protein buffer:

NH2 H+ H
H+ C H+
C=NH HN N
NH2 NH C=CH
Lysine Arginine Histidine
H+ NH2 COO- H+
Cysteine Tyrosine Asparagine acid
Glutamine acid
S- COO- H+

H+ O- H+

His Arg
Asp, Glu Cys Lys
-COOH
-NH2 Tyr

3 5 7 pH 9 11 13

: indicates pK’range
 7,4 24 mmol/l

[ HCO3- ]
pH = 6,1 + log
[ CO2 ]

1,2 mmol/l

In arterial blood plasma: [ HCO3- ] = 24 mmol/l

Graphic ilustration of the Henderson- [ HCO3- ]
pH = 6,1 + log
Hasselbalch equation [ CO2 ]
60

PCO2 = 60
50

45
[HCO3-] mmol/L

40

35
PCO2 = 40
30

25

20

15 PCO2 = 20

10

0
7.1 7.2 7.3 7.4 7.5 7.6 7.7 7.8
pH
 Increases in PCO2 in a solution
Containing only HCO3-, and not other buffers

CO2 exhalation is reduced

[ H+ ] · [ HCO3- ]
H+ + HCO3
- H2O + CO2 = K’
[ CO2 ]

By increases in CO2 , only a not measurable small amount of HCO3- is produced,

because a new equilibrium is soon established through H+ ions that are also
Produced from the reaction of CO2 and watter.
 Increases in PCO2 in a solution containing only HCO3-
CO2 + H2O HCO3- + H+

PCO2 (mm Hg)

120 80 60 40 30 20

40
Plasma [HCO3-] (mmol/L)

30

Hemoglobin
20 0 g/dl

10

6.8 7.0 7.2 7.4 7.6 7.8 8.0

pH
Increases in PCO2 in a solution containing HCO3- and
other buffers (proteins, phosphate)

[ H+ ] · [ HCO3- ]
= K’
[ CO2 ]
CO2 exhalation reduced

H+ + HCO3- H2O + CO2

+
B-

HB
[HCO3- ] = [B- ] = [HB]
 Increases in PCO2 in a solution containing HCO3- and other buffers
(proteins, phosphate)
PCO2 (mm Hg)
120 80 60 40 30 20
Buffer
40 lines
Plasma [HCO3-] (mmol/L)

30 Hemoglobin
g/dL
0

20 5 Anemia

15 Normal
10
20 Polycythemia

6.8 7.0 7.2 7.4 7.6 7.8 8.0

pH
 key notes
Criteria for evaluation of buffering capacity:
a) pH - pK´ difference: The smaller this difference, the greater buffering capacity
b) Total concentration of a buffer: The higher the concentration, the greater
buffering capacity
Based on these criteria, Phosphate and Protein buffers are effective biological buffers

Specific properties of CO2/HCO3- buffer system:

 Because of a large pH - pK´difference, HCO3- buffer can not effectively
buffer in a closed system. Despite this, CO2/HCO3- system is a
powerful buffer in the body, because in the body buffering occurs
like in an open system e.g. CO2 is rapidly regulated to a constant level
by lung ventilation.
 HCO3- buffer system can not participate to buffering during respiratory
acid/base disturbances
 Increase of CO2 in a solution containing only HCO3- buffer causes no
measurable increase in HCO3- concentration, whereas the same CO2
increase in a solution containing other buffers in addition to HCO3-
produces a significant increase in HCO3- concentration
 CO2 production in metabolism

Example:

Glucose : C6H12O6 + 6 O2 6 CO2 + 6 H2O + Energy

Per minute (during rest) : 280 ml CO2 = 13 mmol CO2

Per day 20 000 mmol CO2

CO2 transport from tissue CO2
to lung
CO2

Venous
CO2 Arterial

CO2

CO2 transport in Blood causes

"Respiratory acidosis" of venous blood
compared to arterial blood :
PCO2 , [ HCO3- ] , pH
 Synthesis of H+ and OH- in metabolism

1. Carbohydrate : Glucose + 6 O2 6 CO2 + 6 H2O

neither H+ nor OH- is produced

2. Fat : Fatty acid + O2 CO2+ H2O

neither H+ nor OH- is produced

H+
3. Amino acid : partially R produced
OH-
 Formation of H+ and OH- from amino acids
1. Sulfur containing amino acids ( Methionine, Cysteine, Cystine )
Example : Cys° Glucose° + Urea° + SO4-- + 2 H+

2. Cationic amino acids ( Lysine, Arginine, Histidine )

Example : Arg+ Cl- Glucose° + Urea° + H+ + Cl-

3. Anionic amino acids ( Glutamate, Aspartate )

Example : Glutamate Glucose° + Urea° + OH-

Formation of H+and OH- from salt of organic acids

(Acetate, Lactate, Malate, Citrate, Gluconate ....... )

Example : 2K+ + COO- -CHOH-CH2 -COO- + 3O2

4CO2 + H2O + 2K++ 2OH-
 Daily H+ balance :
( Mixed food, Nord America )
From S- containing amino acids 70 mmol H+
From cationic amino acids 140 mmol H+
210 mmol H+

From anionic amino acids 70 mmol OH-

From organic acids 60 mmol OH-
130 mmol OH-

Nett balance: 210 mmol H+ being formed

130 mmol OH- being formed
80 mmol H+
should be excreted.
This is done by kidney.
 Blood
H+ excretion
in the kidney
Tubule cell
HPO4-- HCO3-
+ +
H+ H+ H+ + HCO3-

Tubular CA
lumen
H2PO4- CO2 CO2 + H2O
+ Glutamine H+ secretion is increased by increased reaborption of
sodium through apical sodium channel
H 2O
2 NH4+
aKetoglutarate
2 NH3 Glucose
+
2 H++ 2 HCO3-
2 H+
CA
2 NH4+ 2 CO2+ 2 H2O

Urine: Titration NH +
acidity 4
(40 mmol/day), but can increase to 500 mmol/day
(40 mmol/day)
 Blood Buffers

a) CO2 / HCO3- in plasma and in erythrocytes

b) Hemoglobin in erythrocytes

c) Plasma proteins in plasma

 Parameters for analysing acid/base status
1. pH : Concentration of free [ H+]
Indicates
2. PCO2
respiratory influences
3. Actual [ HCO3- ]
4. Standard [ HCO3- ] Indicates
5. Buffer bases non respiratory (metabolic)
6. Base excess influences

Standard [ HCO3- ]
[ HCO3- ] in Plasma of a voll oxygenized Blood sample
By a normal PCO2( 40 mmHg ) and at 37°C.
doese not depend on respiratory changes
Buffer bases (BB): Total concentration of all bases (HCO3- + B-)
CO2 + H2O HCO3- + H+
+ Not dependent on PCO2, because
B- increases in HCO - equals
3

decreases in B- and vice versa

HB
Base excess = BE
Difference of actual BB from normal value (48 mmol ·l-1).
BE = BBactual – BB normal; Most used parameter in the clinic.
 Average values and normal range of important acid/base
Parameters in arterial blood plasma

Parameter Symbol Unit Average value Normal range

CO2 Partial pressure PaCO2 mmHg 40 32 - 45

Plasma pH pHPl
_ 7,40 7,37 - 7,45

Plasma bicarbonate [HCO3- ]Pl mmol·l-1 24 20 - 27

concentration
Standard bicarbonate [HCO3- ]St mmol·l-1 24 21 - 26
concentration
Base excess BE mmol·l-1 0 -3 - +3

Buffer bases BB mmol·l-1 48 42 - 54

Definitions:

pH lower than normal Acidosis

pH higher than normal Alkalosis

 Acidosis

Caused by: Changes in PCO2 Changes in [ H+ ] or [ OH- ]

Respiratory Non-respiratory
acidosis acidosis
(= Metabolic acidosis)

The same for Alkalosis

 Respiratory acidosis
CO2 + H2O HCO3- + H+
+
B-

HB
Primarily: PCO2 Compensation:

[ HCO3- ]
Resulting in: pH ( Acidosis ) pH = 6,1 + log
[ CO2 ]
[HCO3-]
HCO3- (by kidney)
[B-]
BB > normal
Because: HCO3-  B-
BE = + X mmol/L
Thus: BB = normal
compensation: metabolic alkalosis
BE = 0 mmol/L
 Respiratory acidosis and its compensation

PCO2 (mm Hg)

Plasma [HCO3-] (mEq/L)

120 80 60 40 30 20

40
2
30
1

20
Buffer
line
10
6.8 7.0 7.2 7.4 7.6 7.8 8.0

pH
1: Respiratory acidosis 2: Renal compensation
 Metabolic acidosis

CO2 + H2O HCO3- + H+

Lung keeps
+
arterial PCO2 B-
constant

Primarily: [ H+ ] , pH HB
Resulting in: [ HCO3- ] Compensation:
[ B- ] pH = 6,1 + log
[ HCO3- ]

BB < normal [ CO2 ]

BE (-)
but PaCO2 normal PCO2 (By Ventilation)
 Metabolic acidosis and its compensation

PCO2 (mm Hg)

Plasma [HCO3-] (mmol/L)

120 80 60 40 30 20

40

30

20 1
Buffer
line
2
10
6.8 7.0 7.2 7.4 7.6 7.8 8.0

pH
1: Metabolic acidosis 2: Respiratory compensation
 PCO2 (mm Hg)
Plasma [HCO3-] (mmol/L) 120 80 60 40 30 20

40 2

30 1

20 3
4
Buffer
10 line

6.8 7.0 7.2 7.4 7.6 7.8 8.0

pH
1. Metabolic alkalosis 3. Respiratory alkalosis
2. Respiratory compensation of 4. Renal compensation of respiratory
metabolic alkalosis alkalosis
 PCO2 (mm Hg)
120 80 60 40 30 20

Plasma [HCO3-] (mmol/L) 40

30

20
Buffer
line
10
6.8 7.0 7.2 7.4 7.6 7.8 8.0

pH
Any point on buffer line (except at pH 7.4) indicates a primary respiratory
change without compensation:
Acidosis by pH lower than 7.4
Alkalosis by pH higher than 7.4
 PCO2 (mm Hg)
120 80 60 40 30 20

Plasma [HCO3-] (mmol/L) 40

30

20
Buffer
line
10
6.8 7.0 7.2 7.4 7.6 7.8 8.0

pH
Any point on isobar 40 (except at pH 7.4) indicates a primary metabolic
change without compensation:
Acidosis by pH lower than 7.4
Alkalosis by pH higher than 7.4
 PCO2 (mm Hg)
120 80 60 40 30 20

Partially
40
Plasma [HCO3-] (mmol/L)

compensated
respiratory acidosis

30

20 Combined
respiratory and
metabolic acidosis
Buffer
10 line

6.8 7.0 7.2 7.4 7.6 7.8 8.0

pH
 PCO2 (mm Hg)
120 80 60 40 30 20

40
Plasma [HCO3-] (mmol/L)

30

20

Buffer
10 line

6.8 7.0 7.2 7.4 7.6 7.8 8.0

pH
 PCO2 (mm Hg)
120 80 60 40 30 20
Partially comp.
Metabolic
40
Plasma [HCO3-] (mmol/L)

Partially compensated Alkalosis

Respiratory Acidosis

30
Combined Respiratory
and Metabolic Alkalosis

Combined Respiratory
and Metabolic Acidosis

20

Partially compensated
Buffer
10 Partially compensated Respiratory Alkalosis line
Metabolic Acidosis

6.8 7.0 7.2 7.4 7.6 7.8 8.0

pH
pH PCO2 BE (+ ≈ HCO3- )
( - ≈ HCO3- )
7.4 40 0

7.2 Acidosis 60 Res. Acid 0 Normal

disturbance uncompensated

7.2 Acidosis 60 Res. Acid + 10 Meta. Alka

disturbance compensation

7.2 Acidosis 60 Res. Acid - 10 Meta. Acid

disturbance disturbance

7.2 Acidosis 30 Res. Alka + 10 Meta. Alka

Because both PCO2 and BE do not match with
pH, it is a wrong measurement
7.2 Acidosis 30 Res. Alka - 10 Meta. Acid
compensation disturbance
Anion Gap if calculated with K+

-
AG = (Na + +
K - ( Cl
+
+ -
HCO 3 ))

14 - 18

144

14
Anion Gap if calculated without K+
 Anion Gap Acidoses
• High AG (new • Normal AG (HCO3-
unmeasured anion; loss)
exogenous acid) • High Cl-, replaces lost
HCO3- (elec. Neutrality)
• nl Cl-, low HCO3-

•Lactic acidosis •Diarhhea

•Ketoacidosis •RTA
•Chronic renal failure •HCl ingestion
•Methanol, Ethy. Gly. •CA inhibitors
•Rhabdomyolysis
 Correction of a metabolic Acidosis

Base required = negative BE · 0,3 · body weight

( mmol ) ( mmol · l-1) ( kg )

1 molar NaHCO3 solution ( 1 ml = 1 mmol HCO3- )

Example : BE = - 10 mmol · l-1 BW = 70 kg

Base needed = 210 mmol

Infusion of 210 ml 1M NaHCO3

 Correction of a metabolic alkalosis

Acid required = positive BE · 0,2 · body weight

( mmol ) ( mmol · l-1 ) ( kg )

1 molar Arginine+ Cl-

Lysine+ Cl

( 0,1 molar HCl only in difficult cases)