Calculating The PH of A Buffer

Calculating the pH of a buffer
Buffer capacity
pH of blood
Buffers
Buffers are chemicals that resist a change in the pH of a solution
when either acids or bases are added to the solution.
The buffers in the body fluids contain salts of either weak acids or
weak bases that combine with H+ when H+ increases in those
fluids, or release H+ when H+ decreases in those fluids.
Buffers tend to keep the H+ concentration, and thus the pH, within
a narrow range of values because of these characteristics.
What is a buffer in chemistry and how does it work?
• A buffer is simply a mixture of a weak acid and its conjugate base or a

weak base and its conjugate acid. Buffers work by reacting with any
added acid or base to control the pH.
Example 1 An acetate buffer is made by mixing 250 mL of 0.5 molar
acetic acid and 250 mL of 1 molar sodium acetate. What
will be the pH value of the prepared buffer? The pKa of
acetic acid is 4.7.
or
What is the pH value of a buffer prepared by adding 70 mL of 0.2 molar CH3COOH
Example 2
to 60 mL of a 0.5 molar CH3COONa solution? pKa = 4.7.
• What is the pH value of this buffer after adding 20 mL of 1 molar HCl
• What is the pH value of this buffer after adding 2 mL of 2 molar NaOH
• What is the pH value of this buffer after adding 20 mL of 1 molar HCl
CH3COONa + HCl  CH3COOH + NaCl

n=C×v=0.5×0.06=0.03 n=C×v=0.2×0.07=0.014 at the beginning
- 0,02 n=1×0.02=0,02 + 0.02 for the reaction
0,01 0.034 after the reaction
• What is the pH value of this buffer after adding 2 mL of 2 molar NaOH
CH3COOH + NaOH  CH3COONa + H 2O

n=C×v= 0.2×0.07=0.014 n=C×v=0.5×0.06=0.03 at the beginning
- 0.004 n=2×0.002=0,004 + 0.004 for the reaction
0.01 0.034 after the reaction
Buffering Capacity
How to calculate buffer capacity?
Example 3
For 100 mL of buffer was added 1 mL of 0.9 M HCl, calculate the buffer capacity:
Step 1: Calculate pH of the buffer after adding 1 mL 0.9M HCl 0.9 mol ------ 1000 mL
0.0009 mol ------ 1 mL
Step 2: Calculate ∆pH of the buffer: ∆pH = 4.13-3.99 = 0.14
Step 3: Calculate 𝛽 0.0009 mol ------ 101 mL

0.0089 mol ------ 1000 mL
Mechanisms of pH
regulation in the body
Organ regulation:
• pulmonary regulation (respiratory
system)
• kidney regulation
• liver regulation
• bone regulation
Buffer regulation:
• Protein buffer (including hemoglobin
buffer): Hemoglobin and Protein of
plasma
• Phosphate buffer: H2PO4-/HPO42-
• Bicarbonate buffer: H2CO3-/H2CO3
Buffers in Biological Systems
Biological functions: The equation of buffer system in blood is,

H2CO3 + H2O  H3O+ + HCO3-
If any acid (H+) is added, the reaction will be,
H+ (acid) + HCO3-  H2CO3
H2CO3  H2O + CO2
If any base (OH-) is added, the reaction will be,
OH- + H2CO3  HCO3- + H2O
As such, the buffer of blood neutralizes the addition of acids

and bases and keeps the pH constant.
Why is pH so important?
In body: [H+] ~ nmol/L, while [K+], [Cl-], [HCO3-] ~ mmol/L
However, [H+] is crucial:

 pH affects function of proteins (hydrogen bonds  3-D structure = function
 All the know low molecular weight and water soluble biosynthetic
intermediates possess, groups that are almost completely ionised at neutral pH
 pH-dependent ionisation (i.e. charge) serves to an efficient intracellular
trapping of ionised compounds within the cell and its otganelles
Respiratory system
The respiratory system responds rapidly to a change in pH and helps bring the pH of body fluids
back toward normal. Increasing CO2 levels and decreasing body fluid pH stimulate neurons in the
respiratory center of the brain and cause elevated rate and depth of ventilation. As a result, CO2 is
eliminated from the body through the lungs at a greater rate, and the concentration of CO2 in the
body fluids decreases. As CO2 levels decline, the concentration of H+ also declines. The pH therefore
rises back toward its normal range
If CO2 levels become too low or the pH of the body fluids is elevated, the rate and depth
of respiration decline. As a con-sequence, the rate at which CO2 is eliminated from the
body is reduced. Carbon dioxide then accumulates in the body fluids because it is
continually produced as a by-product of metabolism. As CO2 accumulates in the body fluids,
so does H+, resulting in a decreased pH.
Kidney regulation
The nephrons of the kidneys secrete H+ into the urine and therefore can directly regulate the
pH of the body fluids. The kidney is a powerful regulator of pH, but it responds more slowly
than does the respiratory system. Cells in the walls of the distal convoluted tubule are
primarily responsible for the secretion of H+. As the pH of the body fluids drops below normal,
the rate at which the distal convoluted tubules secrete H+ increases.
At the same time, reabsorption of HCO3− increases. The increased rate of H+ secretion
and the increased rate of HCO3− reabsorption both cause the blood pH to rise toward its
normal value. On the other hand, as the body fluid pH elevates above normal, the rate
of H+ secretion by the distal con-voluted tubules declines, and the amount of HCO3− lost
in the urine increases. Consequently, the blood pH drops toward its normal value.
Liver and Bone regulation
Metabolic regulation of liver: Bone inorganic matrix consists of
 Metabolism of organic acids anions such as hydroxyapatite crystals (Ca10(PO4)6(OH)2
lactate, ketones and mino acids Bone can take up H+ in exchange for Ca2+,
 Metabolism of ammonium Na+ and K+ (ionic exchange)
 Conversion of NH4+ to urea in the liver Release of HCO3-, CO3- or HPO42-
consumes HCO3-
 Production of glutamate  urine
buffering
 Production of plasma proteins e.g. albumin
contributing to the anion gap
Bicarbonate buffer
The most important buffer system

 Buffer capacity is strong, its content
account for 53% of total buffer
system
 A open system:
• Regulated by respiratory
regulation
• Regulated by renal regulation
Phosphate buffer
Kidneys help regulate blood pH by excreting H+ and reabsorbing HCO3-
• Most of the H+ secretion occurs across the walls of the proximal convoluted
tubule in exchange for Na+ (antiport mechanism  moves Na+ and H+ in opposite
directions)
• Normal urine normally is slightly acidic becease the kidneys reabsorb almost all
HCO3- and excrete H+ (returns blood pH back to normal range)
H2PO4- HPO42- + H+
KH2PO4- = 6,2×10-8, pK2=6.8
• In blood: the ratio HPO42- : H2PO4- = 4:1

• In urine: the ratio HPO42- : H2PO4- = 1:4
Protein buffer
• Behaves as a buffer in both plasma and cells
• Hemoglobin is by far the most important protein buffer.
• Most important intracellular buffer (ICF)
• The most plentiful buffer of the body
• Protein are excellent buffer because they contain both acid and base groups that
can give up ot take up H+
• Proteins are extremely abundant in the cell
• The more limited number of proteins in the plasma reinforce the bicarbonate
system in the ECF
Regulation of Acid-Base Balance in Human Body
The concentration of H+ in the body fluids is reported as the pH. The body fluid
pH is maintained between 7.35 and 7.45; any deviation from that range is life-
threatening. Consequently, the mechanisms that regulate body fluid pH are
critical for survival. The pH of body fluids is mainly controlled by three factors:
buffers, the respiratory system, and the kidneys. When the pH of body fluids is
not properly maintained, the result is acidosis or alkalosis.
Different buffer systems assume dominant roles in different parts of the body
Extracellular Intracellular
Blood Urine
Fluid Fluid
Major Buffer
Major Buffer
● Bicarbonate
● Bicarbonate buffer system
buffer system
● Hemoglobin Major Buffer Major Buffer
Minor Buffer Minor Buffer ● Proteins ● Ammonia
● Intracellular ● Plasma ● Phosphate ● Phosphate
proteins proteins
● Phosphate ● Phosphate
buffer system buffer system
Blood Gas Testing blood gas testing
Blood collection under anaerobic conditions

 From the artery
 Capillary blood
 Indications: to draw a small amount of blood
 For infants and young children
Normal Arterial and Venous values
pH of human blood 7.35-7.45 (or 7.38-7.42)
The physiological levels of the main gas parameters of the acid-base balance are:
pO2: 70-95 mmHg
pCO2: 35—45 mmHg
HCO3- concentration: 22-26 mmmol/L
Hb saturation by oxygen: 95-98%
𝜶- solubility of CO2
in water at 37 ℃
Acidosis
Acidosis occurs when the blood pH falls below 7.35.

The central nervous system malfunctions, and the
individual becomes disoriented and, as the condition worsens,
may become comatose. Acidosis is separated into two categories.
Respiratory acidosis results when the respiratory system

is unable to eliminate adequate amounts of CO2.
Carbon dioxide accumulates in the circulatory system,
causing the pH of the body fluids to decline.
Metabolic acidosis results from excess production of acidic substances,

Such as lactic acid and ketone bodies, because of increased metabolism
or decreased ability of the kidneys to eliminate H+ in the urine.
Alkalosis
Alkalosis occurs when the blood pH increases above7.45.

A major effect of alkalosis is hyperexcitability of the nervous system.
Peripheral nerves are affected first, resulting in spontane-ous nervous
stimulation of muscles. Spasms and tetanic contrac-tions result, as can
extreme nervousness or convulsions.
Tetany of respiratory muscles can cause death.
Respiratory alkalosis results from hyperventilation, as can occur in response to stress.
Metabolic alkalosis usually results from the rapid elimination of H+ from thebody,
as occurs during severe vomiting or when excess aldosterone is secreted by
the adrenal cortex.
This presentation has been prepared for educational purposes as part of the Medicinal Chemistry
course for Students of Faculty Medicine Wroclaw Medicine University.

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Calculating the pH of a buffer

• A buffer is simply a mixture of a weak acid and its conjugate base or a

CH3COONa + HCl  CH3COOH + NaCl

CH3COOH + NaOH  CH3COONa + H 2O

Step 2: Calculate ∆pH of the buffer: ∆pH = 4.13-3.99 = 0.14

Step 3: Calculate 𝛽 0.0009 mol ------ 101 mL

Biological functions: The equation of buffer system in blood is,

As such, the buffer of blood neutralizes the addition of acids

In body: [H+] ~ nmol/L, while [K+], [Cl-], [HCO3-] ~ mmol/L

However, [H+] is crucial:

The most important buffer system

• In blood: the ratio HPO42- : H2PO4- = 4:1

Blood collection under anaerobic conditions

Acidosis occurs when the blood pH falls below 7.35.

Respiratory acidosis results when the respiratory system

Metabolic acidosis results from excess production of acidic substances,

Alkalosis occurs when the blood pH increases above7.45.

Respiratory alkalosis results from hyperventilation, as can occur in response to stress.

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