BUFFER SOLUTION IN HUMAN PHYSIOLOGY

(2022-2026)
SEM - 3
Under the guidance of
Professor – N.N CHOUDHRY (HOD)
- ANURAG MEHTA (GUIDE)
Department of Chemistry
 ACKNOWLEDEGMENT
• We deem it a pleasure to acknowledge our sense of gratitude to our project
guide prof .( Anurag sir ) under whom we have carried out the project work . His
incise and objective guidance and timely advice encourage us with constant flow
energy to continue the work .
• We wish to reciprocate in full measure the kindness shown by Mr . N.N.Choudhry
(HOD ,department of chemistry) who inspired us with his valuable suggetions in
successfully completing the project work .
• We shall remain grateful to him ,for providing us a strong academic atmosphere
by enforcing strict discipline to do the project work with most concentration and
dedication .
• NAME- SARWESH ANAND
• BSC – CHE004
INTRODUCTION
• Blood, like many solutions, is a buffer. Now you can define Buffers as solutions consisting of a weak acid and
its conjugate base—these solutions resist changes in pH when either acid or base is added to it. Let us
consider a buffered solution that contains acetic acid as its weak acid and sodium acetate as its conjugate
base. If some hydrochloric acid was added to this solution, the sodium acetate would react with it by the
following process: The quantity of acid or base that a buffer can neutralize before the pH changes greatly is
called the "buffering capacity." Buffered solutions with high concentrations of weak acid and conjugate base
have higher buffering capacities than those with low concentrations. In the above reaction, the very strong
HCl that has been added to the solution has been converted to acetic acid, which is a weak acid. Because
weak acids cause a much smaller disruption in pH than strong acids, the pH of the solution will decrease
much less than if it contained no sodium acetate.
• Likewise, if sodium hydroxide is added to this solution, the acetic acid would react to it by the following
process: Because the strong base NaOH has been converted to the weak base sodium acetate, the pH of the
solution does not rise nearly as much as if the acetic acid was not present in the first place
CONTENT
NO TOPICS PAGES

1 OBJECTIVE / AIMS 5

2 PROJEPLAN / METHODOLOGY 6 – 11

3 PROJECT STRATEGY / EXECULATION 11-16

4 RESULTS / YOUR FINDINGS 17 – 20

5 CONCLUSION 21

6 REFERENCE 22
OBJECTIVE
• This module aims to 1. Define acidity and buffers 2. Describe the
different physiological buffers and their role 3. Comprehend the
significance of these buffers This module describes the role of
haemoglobin and other buffers in the body. It will enable the learner
to define and comprehend the terms associated with the role of
buffers.
Carbonic Acids
Metabolism of fats and carbohydrates result in the production of 15-20 mol of CO2 per day. Before elimination by
the lungs, most of the CO2 is taken up by red blood cells, reacting with H2O to form carbonic acid as shown
below:
Intracellularly, carbonic acid dissociates to form hydrogen and bicarbonate ions. The latter is pumped out of the
cell into plasma. At the alveoli, bicarbonate ions re-enter the RBC and the above equation is driven to the left, re-
producing CO2 which is then eliminated by the lung. Under normal circumstances, CO2 produced via metabolism
is primarily eliminated via alveoli ventilation. Any increases in CO2 production is matched by an increase in
alveolar ventilation leading to a stable PCO2 level .
. Noncarbonic acids on the other hand are primarily derived from the metabolism of proteins and dietary intake
of foods containing sulphates (H2SO4) and phosphates (H2PO4 - ) and are primarily responsible for the daily acid
load that requires excretion by the kidneys. A normal diet results in the generation of 50 -100 meq of H+ per day.
Most of this comes from the metabolism of sulphur containing amino acids such as cysteine and methionine
which yield sulphuric acid and from metabolism of lysine, arginine and histidine which yield hydrochloric acid.
Ingestion of alkali containing foods (e.g. citrate), and metabolism of amino acids such as aspartate and glutamate
that lead to alkali production offset the daily acid load but the net effect is daily acid addition.
Acid buffering
• One of the major ways in which large changes in H+ concentration are
prevented is by buffering. The body buffers which are primarily weak
acids are able to take up or release H+ so that changes in the free H+
concentration are minimized. Buffers are located in the extracellular
fluid (ECF), intracellular fluid and bone.
• Chloride shift
• Chloride Shift, Hamburger shift or Hamburger phenomenon, (named
after Hartog Jakob Hamburger) is a process which occurs in a
cardiovascular system and refers to the exchange of bicarbonate
(HCO3 − ) and chloride (Cl− ) across the membrane of red blood cells
(RBCs)
Extracellular buffers
• The most important buffer in the ECF and the body is HCO3- (bicarbonate)
which combines with excess H+ ions to form carbonic acid.
• Take for instance an acid load of H2SO4 produced via metabolism of
methionine: Note in this buffering reaction that bicarbonate reacts with a
strong acid to form a weaker acid (H2CO3) which then dissociates into CO2
and H2O. The CO2 produced here does not reform H2CO3 because it is
then excreted by the lungs. Note that HCO3- is used up in this reaction. So
that pH is not affected, the HCO3- used up in this process must be
regenerated and the Na2SO4 must be excreted by the kidneys. The
CO2/HCO3- buffer system is considered very effective because of the vast
quantity of bicarbonate in the body and the ability to excrete the CO2
formed via ventilation. Other less important buffers in the ECF are plasma
proteins and inorganic phosphates.
 Intracellular buffers
• The primary intracellular buffers are proteins, organic and inorganic
phosphates and in the RBC, hemoglobin (Hb-). Whereas buffering by plasma
HCO3- occur almost immediately, approximately 2-4 hours is required for
buffering by cell buffers due to slow cell entry.
• Hemoglobin is a very important buffer in RBCs, particularly in the role of
carbonic acid buffering.
It should also be noted that transcellular uptake of hydrogen ions by cells
result in the passage of Na+ and K+ ions out of cells to maintain
electroneutrality.
These are the only blood buffer systems capable of buffering respiratory acid-
base disturbances as the bicarbonate system is ineffective in buffering changes
in H+ produced by itself.
Homoglobin is an important blood buffer particularly for
buffering CO2
• Deoxygenation of the blood increases its ability to carry carbon dioxide; this
property is the Haldane effect. Conversely, oxygenated blood has a reduced
capacity for carbon dioxide. The Haldane effect is a property of hemoglobin first
described by John Scott Haldane
Carbamino hemoglobin
Carbon dioxide can bind to amino groups, creating carbamino compounds. Amino
groups are available for binding at the N-terminals and at side-chains of arginine
and lysine residues in haemoglobin. This forms carbaminohaemoglobin.
Carbaminohaemoglobin is the major contributor to the Haldane effect .
Buffeing
• Histidine residues in haemoglobin can accept protons and act as
buffers. Deoxygenated hemoglobin is a better proton acceptor than
the oxygenated form. In red blood cells, the enzyme carbonic
anhydrase catalyzes the conversion of dissolved carbon dioxide to
carbonic acid, which rapidly dissociates to bicarbonate and a free
proton:
• CO2 + H2O → H2CO3 → H+ + HCO3−
• By Le Chatelier's principle, anything that stabilizes the proton produced will
cause the reaction to shift to the right, thus the enhanced affinity of
deoxyhemoglobin for protons enhances synthesis of bicarbonate and
accordingly increases capacity of deoxygenated blood for carbon dioxide.
• The majority of carbon dioxide in the blood is in the form of bicarbonate.
Only a very small amount is actually dissolved as carbon dioxide, and the
remaining amount of carbon dioxide is bound to hemoglobin.
• In addition to enhancing removal of carbon dioxide from oxygen-
consuming tissues, the Haldane effect promotes dissociation of carbon
dioxide from hemoglobin in the presence of oxygen.
• In the oxygen-rich capillaries of the lung, this property causes the
displacement of carbon dioxide to plasma as low-oxygen blood enters the
alveolus and is vital for alveolar gas exchange. Protein buffers in blood
include haemoglobin (150g/l) (Figure 3) 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 than 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+ . Figure 3 depicts the role of haemoglobin as a
buffer.
• Deoxyhaemoglobin is a more effective buffer than oxyhaemoglobin and
this change in buffer capacity contributes about 30% of the Haldane effect.
The major factor accounting for the Haldane effect in CO2 transport is the
much greater ability of deoxyhaemoglobin to form carbamino compounds.
Isohydric Principle
• All buffer systems which participate in defence of acid-base changes are in
equilibrium with each other. There is after all only one value for [H+ ] at any
moment. This is known as the Isohydric Principle .
• Principle
• It means that an assessment of the concentrations of any one acid-base
pair can be utilised to provide a picture of overall acid-base balance in the
body. This is fortunate as the measurement of the concentrations of all the
buffer pairs in the solution would be difficult. Conventionally, the
components of the bicarbonate system (ie [HCO3] and pCO2) alone are
measured. They are accessible and easy to determine. Blood gas machines
measure pH and pCO2 directly and the [HCO3] is then calculated using the
Henderson-Hasselbach equation.
Buffering in different sites :
• Respiratory disorders are predominantly buffered in the intracellular
compartment. Metabolic disorders have a larger buffering
contribution from the extracellular fluid (eg ECF buffering of 40% for a
metabolic acidosis and 70% for a metabolic alkalosis). Various buffer
systems exist in body fluids to minimise the effects of the addition or
removal of acid from them. In ECF, the bicarbonate system is
quantitatively the most important for buffering metabolic acids. Its
effectiveness is greatly increased by ventilatory changes which
attempt to maintain a constant pCO2 and by renal mechanisms which
result in changes in plasma bicarbonate. In blood, haemoglobin is the
most important buffer for CO2 because of its high concentration and
its large number of histidine residues.
Deoxyhomoglobin is a better than oxyhemoglobin
• Another factor which makes haemoglobin an important buffer is the
phenomemon of isohydric exchange. That is, the buffer system
(HHbO2-HbO2-) is converted to another more effective buffer (HHb-
Hb-) exactly at the site where an increased buffering capacity is
required. More simply, this means that oxygen unloading increases
the amount of deoxyhaemoglobin and this better buffer is produced
at exactly the place where additional H+ are being produced because
of bicarbonate production for CO2 transport in the red cells.
Link between intracellular & extracellular compartment

• The two major processes involved are:

• Transfer of CO2 across the cell membrane
• Ionic shifts (ie proton-cation exchange mechanisms)
Important points to note about CO2 are:
• It is very lipid soluble and crosses cell membranes with ease causing acid-
base changes due to formation of H+ and HCO3 - . Because of this ease of
movement, CO2 is not important in causing differences in pH on the two
sides of the cell membrane.
• Extracellular buffering of CO2 is limited by the inability of the major
extracellular buffer (the bicarbonate system) to buffer changes in [H+ ]
produced from the reaction between CO2 and water.
• The result is that buffering for respiratory acid-base disorders is
predominantly intracellular: 99% for respiratory acidosis and 97% for
respiratory alkalosis. The second major process which allows transfer of H+
ions intracellularly is entry of H+ in exchange for either K+ or Na+ .
Exchange is necessary to maintain electroneutrality. This cation exchange is
the mechanism which delivers H+ intracellularly for buffering of a
metabolic disorder. In the cell, the protein and phosphates (organic and
inorganic) buffer the H+ delivered by this ion exchange mechanism.
Experiments in metabolic acidosis have shown that 57% of buffering occurs
intracellularly and 43% occurs extracellularly. The processes involved in this
buffering are given .
• Thirty-two percent (32%) of the buffering of a metabolic alkalosis occurs
intracellularly and Na+ - H + exchange is responsible for most of the
transfer of H+ as shown in Figure
Renal buffering
• The process of renal buffering and acid excretion is complex. In order
to conceptualize this process, let us consider the follow equation: The
above equation represents the process of buffering of the daily
nonvolatile acid load. Buffering minimizes the effect that strong acids
such as HCl would have on the pH. Nonetheless the pH will be
affected if the bicarbonate lost in this process is not regenerated,
because as we will learn; loss of bicarbonate from the ECF lowers the
extracellular pH, leading to acidosis. One way to regenerate the lost
bicarbonate would be for the kidney to reverse the above equation,
and excrete HCl in the urine as free H+ ions. Unfortunately this would
require a urine pH of 1.0, an impossible task since the minimum
attainable urine pH is 4.0 to 4.5 .
In order to maintain acid base balance , the kidney must accompolish
two tasks :
• 1) Reabsorption of all filtered bicarbonate
• 2) Excrete the daily acid load
The kidney achieves these three tasks effectively via the processes of
hydrogen secretion, bicarbonate reabsorption and
excretion of hydrogen ions with urinary buffers (titratable
acids and ammonium).
Conclusion
• The human body has a series of physiological buffers which maintain
the intricate mileu interior of the body. The different buffers prevent
both respiratory and metabolic acidosis and protect the system from
collapse and death.
References :
• 1. Worthley LI. Hydrogen ion metabolism. Anaesth Intensive Care
1977 Nov; 5(4) 347-60. pmid:23014. PubMed
• 2. Pitts RF. Mechanisms for stabilizing the alkaline reserves of the
body. Harvey Lect 1952- 1953; 48 172-209. PubMed
• 3. Bernards WC Interpretation of Clinical Acid-Base Data. Regional
Refresher Courses in Anesthesiology. 1973; 1: 17-26
• 4. Bushinsky DA. Acidosis and bone. Miner Electrolyte Metab 1994;
20(1-2) 40-52.
• Wekipedia and internet sources