Water, pH and Buffers

MBC226
DR AYYUB PATEL

HARPER’S 29TH EDITION : PAGES 14-26

 Outline
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• Homeostasis

• Medical relevance and importance of water

• The structure and function of water

• Dissociation of water, acids and bases

• Medical relevance and importance of body pH

• pH calculations and the Henderson-Hasselbalch equation

• Buffers, biological/physiological examples

• Medical relevance and importance of buffers

HOMEOSTASIS
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●The dynamic that defines the distribution of water and

the maintenance of pH and electrolyte concentrations

●Water distribution is maintained by the kidneys,

antidiuretic hormone, hypothalamic thirst response,

respiration and perspiration

 HOMEOSTASIS
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• Clinically, need to be aware of water

depletion caused by decreased intake
(coma, wandering the desert) or increased
loss (diarrhea, renal malfunction, over-
exercise), and excess body water due to
increased intake (too much I.V.) or decreased
excretion (renal failure)
 WATER
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● Comprises approx 60-70% of body weight

(45-60% intracellular, 25% extracellular/blood
plasma). Under normal conditions, a person need
1,5 - 2,5 litres per day. Our blood contains 90% of
water.

● dipolar: partial negative charge on oxygen, partial

positive charge on hydrogens

● dipolar nature leads to formation of many low

energy hydrogen bonds
Structure of H20
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From Lehninger, 2nd ed., Ch 4

Biological
Hydrogen
Bonds
 Functions of Water in body
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• Cell life Water is a carrier, distributing essential nutrients to cells

• Chemical and metabolic reactions Water removes waste via urines and faeces.
• Transport of nutrients Water participates in the biochemical break-down of food
• Body temperature regulation helps limit changes in body temperature

• Elimination of water Water is an effective lubricant around joints. It also acts as a shock
absorber for eyes, brain, spinal cord and even for the foetus through amniotic fluid.
 Properties of water

• Very polar

• Oxygen is highly electronegative

• H-bond donor and acceptor
• High b.p., m.p., heat of vaporization,
surface tension
Water dissolves polar compounds(salts)as it forms
hydrogen bond with positive and negatively charged ions

solvation shell
or
hydration shell
Non-polar substances are insoluble in water

Many lipids are amphipathic

 How detergents work?

Micelle formation is essential for the absorption of fat-soluble

vitamins and complicated lipids within the human body. Bile salts
formed in the liver and secreted by the gall bladder allow micelles of
fatty acids to form. This allows the absorption of complicated lipids
(e.g., lecithin) and lipid-soluble vitamins (A, D, E, and K) within the
micelle by the small intestine.
 Hydrogen Bonding of Water

One H2O molecule can

associate with 4
other H20 molecules

•Ice: 4 H-bonds per water

molecule
•Water: 2.3 H-bonds per water
molecule
Crystal lattice of ice
 Relative Bond Strengths

Bond type kJ/mole

H3C-CH3 88
H-H 104
Ionic 40 to 200
H-bond 2 - 20
Hydrophobic interaction 3 -10
van der Waals 0.4 - 4
Molecules in which the atoms are held together by
_Covalent bonds have the strongest chemical linkages
 Water and pH relationship
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● In solution water shows a very low

dissociation (probability of H+ in water
is 1.8 x 10-9)
● H2O H+ + OH-
● H+ is actually associated with a cluster
of water molecules and exists in
solution as H3O+ or H5O2+ or H7O3+
 Ionization of Water 0

H20 + H20
H3O+ + OH-H20 H+ +
Keq= [H+] OH- Keq=1.8 X 10-
[OH-] 16M
[H2O] = 55.5
[H2O] M
[H2O] Keq = [H+]
[OH-]
(1.8 X 10-16M)(55.5 M ) = [H+]
[OH-]
1.0 X 10-14 M2 = [H+] [OH-] = Kw= ionic product of
water

If [H+]=[OH-] then [H+] = 1.0 X 10-

7
 pH Scale
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● Devised by Sorenson (1902)
● [H+] can range from 1M and
1 X 10-14M
● using a log scale simplifies
notation
● pH = -log [H+]
●Neutral pH = 7.0
●pH of a solution is the relative
concentration of H+ donors and acceptors
 Medical Importance of pH
If your urinary pH fluctuates between 6.0 to 6.5 in the morning and between 6.5 and 7.00 in
the evening, your body is functioning within a healthy range. If your saliva stays between
6.5 and 7.5 all day, your body is functioning within a healthy range.
Most people who suffer from unbalanced pH are acidic
•Mild acidosis can cause such problems as:
•Cardiovascular damage, including the constriction of blood vessels and the reduction of
oxygen.
•Weight gain, obesity and diabetes.
•Bladder and kidney conditions, including kidney stones.
•Immune deficiency.
•Acceleration of free radical damage, possibly contributing to cancerous mutations.
•Hormone concerns.
•Premature aging.
•Osteoporosis; weak, brittle bones, hip fractures and bone spurs.
•Joint pain, aching muscles and lactic acid buildup.
•Low energy and chronic fatigue.
•Slow digestion and elimination.
•Yeast/fungal overgrowth.
 Water and pH relationship
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● [H+]=1x10-6 what is the pH?

● pH= - log10 [H+]
6

● [H+]=0.24x10-4 what is the pH? 4.6

● [H+]=3.4x10-3 what is the pH?
2.5
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 Henderson-Hasselbach Equation
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Consider the dissociation of a general/weak acid HA

HA H+ + A-

We can define a dissociation constant (K) where

Rearranging gives

Taking logarithms on both sides and multiplying by -1 gives

-log[H+] = -logK – log [HA]/[A-] or
pH = pK + log [A-]/[HA]
Dissociation Constant and pH
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From Marks, Marks, Smith, Ch

 Henderson-Hasselbalch Equation
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• This equation can be used to determine the pH if the pK

and ratio of the ionised and unionised forms is known.

• The pKa (a for acid) is the –ve log of the dissociation

constant of the acid. It is the pH at which the ratio of the
ionised and unionised species is equal to 1. ie the molar
concentration of the ionised and unionsed species is the
same.

• Similarly pKb is –ve log of the dissociation constant of

the base
 Henderson-Hasselbach Equation
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HA = weak acid
1) Ka = [H+]
[A-] A- = Conjugate base
[HA]
2) [H+] = Ka
[HA]
[A-]= -log Ka -log
3) -log[H+]
[HA] * H-H equation describes
the relationship between
[A-] pH, pKa and buffer
4) -log[H+] = -log Ka +log [A-
concentration
]
[HA]
5) pH = pKa +log [A-]
[HA]
Acid/conjugate base pairs 0

HA + H2O A- + H3O+
HA A- + H+
HA = acid ( donates H+)(Bronstad Acid)
A- = Conjugate base (accepts H+)(Bronstad Base)

Ka & pKa value describe tendency to

Ka = [H+] loose H+
[A-]
[HA] large Ka = stronger acid
small Ka = weaker acid
pKa = - log
Ka
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From Devlin, 3rd ed., Ch 1

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 Weak Acids and Bases Equilibria
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•Strong acids / bases – disassociate completely
•Weak acids / bases – disassociate only partially
•Enzyme activity sensitive to pH
• weak acid/bases play important role in
protein structure/function
pKa values determined by titration
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Phosphate has three ionizable H+ and three pKas:polyprotic acid
act as acid-base buffer at pH values around any of theirˇpKa's0
Buffers
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• Buffers are aqueous systems that resist changes in pH

when small amounts of a strong acid or base are
added.
• A buffered system consist of a weak acid and its
conjugate base.
• The most effective buffering occurs at the region of
minimum slope on a titration curve
(i.e. around the pKa).
• Buffers are effective at pHs that are within +/-1 pH
unit of the pKa, the concentration of the buffer; the
more present, the greater the buffering capacity
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Sample pH problems
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Sample pH Problem (cont)
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 Physiological Buffers 0

• Carbon Dioxide-Bicarbonate System; a major

regulator of blood pH

• Phosphate System; major regulator of cytosolic pH

• Proteins as buffers

• [CO2] and [HCO3] are much higher than [PO4] in

blood; the reverse is true in the cytosol, [PO4]
>>> [HCO3]
Examples - Physiological Buffers
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From Marks, Marks, Smith, Ch

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Lungs and kidney are the organs that control blood pH.
From Marks, Marks, Smith, Ch 4
Blood Bicarbonate and Metabolic Acidosis
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The bicarbonate blood buffer in a normal adult
maintains the blood pH at about 7.40. pH of blood
range(7.35 - 7.45) average = 7.4. However the pH of
blood varies from 7.3 to 7.5 without disturbing the
normal functions of the body.
If the blood pH drops below 7.35, the condition is
referred to as an ACIDOSIS. A prolonged blood
pH below 7.0 can lead to death. Clinically for an
acidosis, the acid-base parameters (pH, [HCO3- ],
[CO2] )of the patients blood should be monitored.
The normal values for these are pH = 7.40;
[HCO3- ] = 24 mM; [CO2] = 1.2 mM.
Sample Problem – Metabolic 0

Acidosis
• The blood values of a patient were pH
= 7.03 and [CO2] = 1.1 mM. What is
the patient’s blood [HCO3-] and how
much of the normal [HCO3-] has been
used in buffering the acid causing the
condition?

• The pK’ for [HCO3-]/[CO2] = 6.10

 Solution 0

● Substitute into Henderson-Hasselbalch equation:

● 7.03 = 6.10 + log [HCO3-]/1.1 mM, or
● 0.93 = log [HCO3-]/1.1 mM
● The anti-log of 0.93 = 8.5, thus:

● 8.5 = [HCO3-]/1.1 mM, or [HCO3-] = 9.4 mM

● Since normal [HCO3-] equals 24 mM, there was a

decrease of 14.6 mmol of [HCO3- per liter of blood in
this patient. This would be approaching the point
where, if left untreated, the HCO3- buffering capacity
would be no longer effective in this patient.