Acids, Bases and Buffers

Lecture 14&15
Noor Ullah
PhD Scholar (Biochemistry)
Lecturer MLT, KMU IPMS
noor.ipms@kmu.edu.pk
Acids and Bases

• The concentration of the hydrogen ion, one of the most important ions in
biological systems, affects most cellular and organismal processes

• For example, the structure and function of proteins and the rates of most
biochemical reactions are strongly affected by hydrogen ion concentration

• Additionally, hydrogen ions play a major role in processes such as energy

generation and endocytosis

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Acids and Bases

• Many biomolecules have acidic and/or basic properties

• Large polymers and macromolecular complexes usually have amphoteric

surfaces; that is, they possess both acidic and basic groups

• A side group of a molecule is said to be an acid if it is a proton donor and a

base if it is a proton acceptor

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Acids and Bases

• Strong acids (e.g., HCl) and bases (e.g., NaOH) ionize almost completely in
water:

• HCL

• Organic acids (compounds with carboxyl groups) are referred to as weak acids
because of their partial dissociation in water

• Organic bases have a small but measurable capacity to combine with hydrogen
ions

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Acids and Bases

• Many common weak bases contain amino groups

• The dissociation of an organic acid is described by the following reaction

• HA (weak acid) (A- conjugate base of HA)

• The deprotonated product of the dissociation reaction is referred to as a

conjugate base

• For example, acetic acid (CH3COOH) dissociates to form the conjugate base
acetate (CH3COO−)

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Strength of acids

• The strength of a weak acid (its capacity to release hydrogen ions) may be
determined using the following expression:

• The larger the value of Ka, the stronger the acid is

• Because Ka values vary over a wide range, they are expressed using a logarithmic
scale:

• The lower the pKa, the stronger the acid

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The pH scale

The pH scale can be used to

determine hydrogen ion
concentration [H+]

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pH

• pH= Power of hydrogen (amount of hydrogen)

• It could also be the amount of hydronium or just free hydrogen ion inside the
water

• pH 7 means that the concentration of hydrogen ions is very small in pure water
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 Production of acids by the body- sources of H + ions

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Regulation of body pH (H+)

• Concentration of hydrogen ions(changes in blood pH) is regulated sequentially by:

1. Chemical buffer systems – act within seconds

2. Physiological buffer systems

• The respiratory mechanisms – hyperventilation or hypoventilation- acts
within 1-3 minutes

• Renal mechanisms – secretion of H+ and reabsorption of HCO3- - require

hours to days to affect pH changes

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Regulation of body pH (H+) and Buffers

• The regulation of pH is a universal and essential activity of living organisms

Hydrogen ion concentration must typically be kept within narrow limits

• For example, normal human blood has a pH of 7.4. It may vary between 7.35 and
7.45, depending on the concentrations of acidic and basic waste products and
metabolites

• Certain disease processes cause pH changes that, if not corrected, can be

disastrous

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Regulation of body pH (H+) and Buffers

• Acidosis- when human blood pH falls below 7.35, results from excessive production of acid
in the tissues, loss of base from body fluids, or failure of the kidneys to excrete acidic
metabolites

• Acidosis occurs in certain diseases (e.g., diabetes mellitus) and during starvation If blood
pH drops below 7, the central nervous system becomes depressed, resulting in coma and
eventually death

• Alkalosis-When blood pH rises above 7.45

• This condition, brought on by prolonged vomiting or by ingestion of excessive amounts of

alkaline drugs, overexcites the central nervous system

• If this situation is uncorrected, convulsions and respiratory arrest develop.

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Buffers

• A buffer is a solution of a weak acid (HA) and its salt (BA) with a strong base.

• A buffer cannot remove H+ ions from the body

• It temporarily acts as a shock absorbent to reduce the free H+ ions and later to be
removed by the renal mechanism

• Buffers help maintain a relatively constant H+ ions concentration

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Buffers

• Consider a solution containing acetate buffer- acetic acid and sodium acetate

• If hydrogen ions are added, the equilibrium shifts toward the formation of acetic
acid with the [H+] changing little

• If hydroxide ions are added, they react with the free hydrogen ions to form water,
the equilibrium shifts to the acetate ion, and the pH changes little

• ++OH= H20

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Buffering capacity

• The capacity of a buffer to maintain a specific pH depends on two factors

1. The molar concentration of the acid–conjugate base pair

2. The ratio of their concentrations

• Buffering capacity is directly proportional to the concentration of the buffer

components- the more molecules of buffer present, the more H+ and OH− ions
can be absorbed without significantly changing the pH

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Buffer concentration
• The concentration of the buffer is defined as the sum of the concentration of
the weak acid and its conjugate base

• For example, a 0.2 M acetate buffer may contain 0.1 mol of acetic acid and 0.1
mol of sodium acetate in 1 L of H2O

• Such a buffer may also consist of 0.05 mol of acetic acid and 0.15 mol of sodium
acetate in 1 L of H2O.

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Buffer concentration

• The most effective buffers are usually those that contain equal concentrations
of both components or the pH is equal to the pKa.

• Biological systems generate acids during metabolism, and buffer capacity for
acid neutralization must be maximized

• Consequently, biological buffers often contain a higher concentration of the

conjugate base

• Bicarbonate buffer is an example of such a buffering system

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Henderson–Hasselbalch equation

• In choosing or making a buffer, the pH and pKa concepts are useful

• The relationship between the pH and pKa values is expressed in the Henderson–
Hasselbalch equation

• A-/HA

• pH= acidity of the buffer solution

• pKa= Negative log of Ka

• Ka= Acid dissociation constant

• HA= concentration of acid

• A= concentration of conjugate base

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Blood/ Physiological buffers

• The 3 most important buffers in the blood

1. Bicarbonate buffer

2. Phosphate buffer

3. Protein buffer

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Bicarbonate buffer

• One of the most important buffers in blood, has three components

• The first of these, carbon dioxide, reacts with water to form carbonic acid:

• Carbonic acid then rapidly dissociates to form H+ and HCO3− ions

• Because the concentration of H2CO3 is very low in blood, the preceding

equations may be simplified to

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Bicarbonate buffer

• Recall that buffering capacity is greatest at or near the pKa of the acid–conjugate
base pair

• Carbonic acid is a diprotic acid (it can donate two hydrogen ions) with a pK1 of
6.3

• In blood, there is a critical need to maintain the pH at the high end of the
buffering range of this acid and to maximize buffering capacity for acid

• Therefore, it is optimal for the concentration of the conjugate base,

bicarbonate, to be high compared to H2CO3 (or CO2), typically 20 to 1.

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Bicarbonate buffer

• This ratio, which differs from the ideal weak acid:conjugate base ratio of 1:1,
indicates that the bicarbonate buffer is operating in blood at the limit of its
buffering capacity

• Nevertheless, the bicarbonate buffer is effective for two reasons: there is a

high bicarbonate concentration in blood, and the components are under
physiological control

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Bicarbonate buffer

• The uncatalyzed conversion of CO2 to HCO3− and H+ is a slow process:

• In blood, this reaction is catalyzed by carbonic anhydrase

• With rates as high as 106 molecules of CO2 converted to bicarbonate per second
per enzyme molecule, carbonic anhydrase is one of the most efficient enzymes
known

• The CO2 level is kept low and is regulated through changes in the respiratory rate

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Bicarbonate buffer

• The bicarbonate level stays high because the kidneys excrete H+

• When excessive amounts of HCO3− are produced, the kidney excretes bicarbonate

• As acid, a metabolic waste product, is added to the body’s bicarbonate system,

the concentration of HCO3− decreases and CO2 is formed

• Because the excess CO2 is exhaled, the ratio of HCO3− to CO2 remains essentially
unchanged

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Phosphate buffer system

• (NaH2PO4 – Na2HPO4)

• It is mostly an intracellular buffer and is of less importance in plasma due to its

low concentration

• With pKa 7.2, it would appear that phosphate buffer is an excellent choice for
buffering the blood

• Although the blood pH of 7.4 is well within this buffer system’s capability, the
concentrations of H2PO4− and HPO4 − in blood are too low to have a major effect

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Phosphate buffer system

• Instead, the phosphate system is an important buffer in intracellular fluids

where its concentration is approximately 75 milliequivalents (mEq) per liter

• Phosphate concentration in extracellular fluids such as blood is about 4 mEq/L

• Because the normal pH of cell fluids is approximately 7.2 (the range is from 6.9 to
7.4), an equimolar mixture of H2PO4− and HPO4 − is typically present

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Protein buffers

• The plasma proteins and hemoglobin together constitute the protein buffer
system of the blood

• Composed of amino acids linked together by peptide bonds, proteins contain

several types of ionizable groups in side chains that can donate or accept
protons

• Protein molecules are powerful buffers because they are present in significant
concentration in living organisms

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Protein buffers
• For example, hemoglobin is the most abundant biomolecule in red blood cells

• Hemoglobin plays a major role in maintaining blood pH because of its structure

and high cellular concentration

• It mainly buffers the fixed acids, besides being involved in the transport of gases
(O2 and CO2)

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