Medical Chemistry

Chapter 4 Buffer Solution

 Study Goals
1. Master the composition and the mechanism of buffer
solution

2. Master the concepts such as buffer capacity, buffer

range, the factors influencing buffer capacity

3. Master how to prepare a buffer

4. Understand the application of buffer action in medicine

 Two experiments
Experiment 1

Add 2 drops of 1 mol/L HCl to 1L of water

pH change from 7 to 2
The solution used
in Experiment 2 is
Experiment 2
a kind of buffer
Add 2 drops of 1 mol/L HCl to a specific solution
solution.

pH unchanged (slightly changed)

 4.1 Composition and Action
of Buffer Solutions
4.1.1 The Essential Feature of a Buffer

Buffer Solution:

Definition of buffer solution：solutions that resist

change in hydronium ion, H+, and the hydroxide ion,OH-,
concentration (and consequently pH) upon addition of
small amounts of acid or base, or upon dilution.
4.1.2 Composition of buffers solutions

weak acid and its conjugate base

a weak base and its conjugate acid

Notice:
Weak acid and its conjugate base must be both
present and in large amount compared to the
acid/base added
ECNU
• Buffers are mixtures containing a common
ion: either weak acids and their conjugate
bases or weak bases and their conjugate
acid.
• Two common buffers:
ammonium-ammonia, carbonate-bicarbonate
NH4+(aq)  H+(aq) + NH3 (aq)
CO3-2(aq) + H2O (l)  H CO3-1 (aq) + OH -1 (aq)
Buffer systems that are useful at various pH values
Conjugate Conjugate
Buffer system pKa( at 25℃)
acid base

HAc－NaAc HAc Ac- 4.76

H2CO3 － NaHCO3 H2CO3 HCO3- 6.35

H3PO4 － NaH2PO4 H3PO4 H2PO4- 2.16

Tris·HCl － Tris Tris·H+ Tris 7.85

H2C8H4O4 － KHC8H4O4 H2C8H4O4 HC8H4O4- 2.89

NH4Cl － NH3 NH4+ NH3 9.25

CH3NH3+Cl- － CH3NH2 CH3NH3+ CH3NH2 10.63

NaH2PO4 － Na2HPO4 H2PO4- HPO42- 7.21

Tris: Tris(Hydroxymethy)methanamin
三羟甲基氨基甲烷

NH2
HOH2C C CH2OH
CH2OH
 Which are buffers
A. mix acetic acid and sodium acetate solutions.
B. mix NH4Cl and HCl solutions.
C. mix NH4Cl and NH3 solutions.
D. mix NH3 and NaOH solutions.
 A Buffered Solution
 resists change in its pH when either H+ or
OH− are added.
1.0 L of 0.50 M CH3COOH
+ 0.50 M CH3COONa
pH = 4.74
 Adding 0.010 mol solid NaOH raises the
pH of the solution to 4.76, a very minor
change.
The effect of addition of acid or base to …

acid added base added

an unbuffered solution

acid added base added

a buffered solution
 4.1.3 How a Buffer solutions Works

Buffer with equal

concentrations of
Buffer after conjugate base Buffer after
addition of H3O+ and acid addition of OH-
H3O+ OH-

H2O + CH3COOH H3O+ + CH3COO- CH3COOH + OH- H2O + CH3COO-

 How buffer solutions work(A)

Buffer solution composed of CH3COOH/CH3COONa

CH3COONa(aq) CH3COO-(aq)+H+ (aq)

CH3COOH CH3COO-(aq)+H+ (aq)

 Little H+
Balance left
+ -
HAc + H2O H3O + Ac

+ -
NaAc Na + Ac
anti-acid component

Acid Equilibrium Acid added was

pH unchanged
added Shift to the left consumed

base
added
 Little OH-
Shift to the right
+ -
HAc + H2O H3O + Ac
anti-base component

+ -
NaAc Na + Ac

Acid Equilibrium Acid added was

pH unchanged
added Shift to the left consumed

base Added OH -react base added was

pH unchanged
added With CH3COOH consumed
 How buffer solutions work(B)
Buffer solution composed of NH3/NH4Cl
NH4Cl(aq) Cl-(aq)+NH4+ (aq)
Produce NH4+ ions

NH 3(aq) + H2O NH 4+(aq)+OH- (aq)

Added H+ react
Acid Acid added was
With NH 3 to pH unchanged
added consumed
Form NH 4+

Base added
Base Equilibrium
was pH unchanged
added Shift to the left
consumed
 anti-acid mechanism
+
H
Shift left
+
+ –
HAc + H2O H3O + Ac
Shift right + Anti-acid
Anti-base –
OH

anti-base mechanism
H2O
The amounts of weak acid and weak base in the buffer
must be significantly larger than the amounts of H3O+ or
OH- that will be added, otherwise the pH cannot remain
approximately constant. Thus addition of limited
amounts of a strong acid or base is counteracted by the
species present in the buffer solution, and the pH
changes very little. No solution can keep the pH
approximately constant if you add larger amounts of
either acid or base that are present in the original buffer.
① either weak acids and their conjugate bases or

weak bases and their conjugate acid.

② they must be in approximately equal amounts

③ in substantial amounts
 4.1.4 pH of Buffer Solution
The Henderson-Hasselbalch Equation
For a HB-NaB buffer system,

HB + H2O H3O+ + B–
NaB Na+ + B–
[H+][B–] + [HB]
Ka = [H ] = Ka [B–]
[HB]
Apply –log on both sides of above equation,
[B–] The Henderson-
pH = pKa + lg Hasselbalch Equation
[HB]
 The Henderson-Hasselbalch Equation
[B–]
pH = pKa + lg [conjugate base]
= pKa + lg
[HB] [conjugate acid]

pKa ：the –log of Ka of the conjugate acid

[B–]、[HB]：equilibrium concentration

[B–] / [HB]：buffer ratio

[B–]+[HB]： total concentration
[HB] = cHB − cHB(dissociated)  cHB
[B–] = cNaB + cHB(dissociated)  cNaB
 Sample Problem 1 Calculating the pH of a
Buffer Solution-1
PROBLEM

A buffer is prepared by mixing equal volumes of 0.2 mol·L-

1 NaAc and 0.2 mol·L-1 HAc. What is the pH of the final
solution? The pKa of HAc is 4.74.
SOLUTION

[ A− ] 0.1
pH = pK a + lg = 4.74 + lg
log 1/1= =4.4.74
44
[ HA] 0.2
 Buffer capacity
Buffer capacity is the amount of acid or base
that can be added to a buffer solution before a
major pH change (more than +- 0.5 units)
occurs.
The buffer’s capacity is determined by the
amount of acid and conjugate base.
A buffer with a large capacity has a large
concentration of HA and of A-.
 How to choose a buffer
For a buffer with pH 3, we choose an
acid/conjugate base combination that has a pKa
close to 3.

For a buffer with a pH of 9, we choose an

acid/conjugate base combination that has a pKa
close to 9.
 4.2 Preparation of Buffer
General steps for a buffer preparation:
1. Choose the conjugate acid-base pair.

2. Determine the buffer concentration.

3. Calculate the ratio of buffer component

concentrations.
4. Mix the solution and adjust the pH.
pH=pKa±1

For example:
pH=5.0,
Choose pKa=4~6

pH=7.0,
Choose pKa=6~8

pH=9.0,
Choose pKa=???
 4.2 Preparation of Buffer
General steps for a buffer preparation:
1. Choose the conjugate acid-base pair.

2. Determine the buffer concentration.

As we all know, buffer capacity depends on the concentrations of
buffer components. The higher concentrations of buffer
components, the greater the buffer capacity. Usually the
reasonable concentration of a buffer is in the range from 0.05
mol/L to 0.2 mol/L
 4.2 Preparation of Buffer
General steps for a buffer preparation:
1. Choose the conjugate acid-base pair.

2. Determine the buffer concentration.

3. Calculate the ratio of buffer component

concentrations.
4. Mix the solution and adjust the pH.
 The Henderson-Hasselbalch Equation
[B–]
pH = pKa + lg [conjugate base]
= pKa + lg
[HB] [conjugate acid]

pKa ：the –log of Ka of the conjugate acid

[B–]、[HB]：equilibrium concentration

[B–] / [HB]：buffer ratio

[HB] = cHB − cHB(dissociated)  cHB

[B–] = cNaB + cHB(dissociated)  cNaB

 4.2 Preparation of Buffer
General steps for a buffer preparation:
1. Choose the conjugate acid-base pair.

2. Determine the buffer concentration.

3. Calculate the ratio of buffer component

concentrations.
4. Mix the solution and adjust the pH.
Preparation of NaAc solution Preparation of HAc solution

Buffer of the desired pH

4.3 Buffering Action in Medical

A pH value of bodily fluids in a certain range of constant

B The application of buffer action in medicine

C Medical experiments are performed at precise pH

4.3.1 pH value of bodily fluids in a certain
range of constant

体液 pH 体液 pH
large intestinal
blood 7.35~7.45 8.3~8.4
secretion
Adult gastric
0.9~1.5 Milkis 6.0~6.9
juice
Baby's gastric
~5.0 tear ~7.4
juice

saliva 6.35~6.85 urine 4.8~7.5

cerebrospinal
pancreatic juice 7.5~8.0 7.35~7.45
fluid
intestinal fluid ~7.6
 4.3.2 Buffering Action in Human Blood

The pH of the blood in a healthy individual remains

remarkably constant at 7.35 to 7.45. This is because the
blood contains a number of buffers that protect against
pH change due to the presence of acidic or basic
metabolites. From a physiological viewpoint, a change of
±0.3 pH unit is extreme. Acid metabolites are ordinarily
produced in greater quantities than basic metabolites,
and carbon dioxide is the principal one.
 The blood pH must maintain ranging from 7.35~7.45.

alkalescence 弱碱性

acid-base balance
酸碱平衡

Research has found that the metabolite of fruits

and vegetables are alkaline matters.
If everyone will fall ill from eating acidic food?

The acid metabolite is more than alkalines

in the human body.
人体内酸性代谢产物比碱性代谢产物更多。

acid-base unbalance
酸碱失调
It shows that the blood has the seltf-regulation ability for pH values.
事实证明人体血液具有自身调节pH值的能力。

Buffer Action 缓冲作用

strong acid The ability which can resist any change

or base when the strong acid or base is added to,
the pH is still in the normal range,it is
defined buffer action.
 What information can you get from this experiment?

It tells us buffer solution has buffer action.

缓冲溶液具有缓冲作用。

Why the buffer solution has the buffer action?

Buffer Mechanism缓冲机制

Essence: conjugate acid-base pair(共轭酸碱对)

conjugate acid—antalkaline（抗碱成分）

conjugate base—antiacid（抗酸成分）
Π The buffer system of human blood

Why the human blood can maintain the pH in such

a narrow scope?
Main buffer pairs（缓冲对） of human blood
H2CO3——NaHCO3
NaH2PO4——Na2HPO4

7.35～7.45
If the pH of the blood drops below 7.35, a potentially fatal
condition called acidosis results. If the pH of the blood
increases to over 7.45, another serious condition known as
alkalosis results.

The HCO3-/H2CO3 buffer system is the most important

one in buffering blood in the lung (alveolar blood). As
oxygen from inhaled air combines with hemoglobin, the
oxygenated hemoglobin ionizes, releasing a proton. This
excess acid is removed by reacting with HCO3-:
 B-

CO2 + H2O H2CO3 H++ + HCO3-

H
acid base

Anti-base component Anti-acid component

 H 2CO3 H + + HCO3−
Which direction of the balance will shift to?

acidic materials H+ increase shift to the left

How does the body to adjust the products?

H2CO3 increase CO2 breath out
HCO3- decrease pH normal
HCO3- is replenished
H2CO3 = CO2  +H 2O

Does the buffer solution can resist the foreign without limit?
 OH-
H2CO3 HCO3- Buffer system
H3O+

Lungs CO2 + H2O Kidneys

Buffer action of any buffer solution is limited.
任何缓冲溶液的缓冲能力都是有限的。 alkalosis
buffer capacity 缓冲容量
碱中毒
If the foreign is outside of the buffer capacity,
the buffer solution will lose buffer action.

HCO3− lose too much H 2CO3 H + + HCO3−

fail to adjust
酸中毒 pH decreases
(acidosis)
An important diagnostic analysis is the CO2/HCO3- balance in blood.
This ratio is related to the pH of the blood by the Henderson-
Hasselbalch equation:
− −
[ HCO ] [ HCO 3]
pH = pK a + lg
' 3
= 6.10 + lg
[CO 2 ]dissolved [CO 2 ]dissolved

where H2CO3 can be considered equal to the concentration of

dissolved CO2 in the blood; 6.10 is pKa1' of carbonic acid in blood at
body temperature (37℃). Normally, the bicarbonate concentration
in blood is about 26.0 mmol·L-1, while the concentration of carbon
dioxide is 1.3 mmol·L-1. Accordingly, for the blood,

[HCO 3− ] 26mmol  L−1

pH = 6.10 + lg = 6.10 + lg −1
= 7.40
[CO 2 ]dissolved 1.3mmol  L
You know that carbonic acid also breaks down into water and
gaseous carbon dioxide. The extra carbon dioxide is released in the
lungs and exhaled. When extra OH- finds its way into the blood
system, it reacts with the H2CO3. The extra bicarbonate ions formed
are eventually expelled by the kidneys.
But note that the [HCO3-]/[H2CO3] ratio at pH 7.4 is 26 mmol·L-1/1.3
mmol·L-1 = 20:1. This is not a very effective buffer ratio; and as
significant amounts of HCO3- are converted to H2CO3, the pH would
have to decrease to maintain the new ratio. But, fortunately, the
H2CO3 produced is rapidly decomposed to CO2 and H2O by the
enzyme decarboxylase(脱羧酶), and the CO2 is exhaled by lungs.
Hence, the ratio of HCO3-/ H2CO3 remains constant at 20: 1.
The buffer capacity of blood for handling CO2 is estimated to be
distributed among various buffer systems as follows: hemoglobin
and oxyhemoglobin, 62%; H2PO4-/HPO42-, 22%; plasma protein,
11%; bicarbonate, 5%. Proteins contain carboxylic(羧基) and amino
groups, which are weak acids and bases, respectively. They are,
therefore, effective buffer agents. The combined buffer capacity of
blood to neutralize acids is designated by clinicians as the "alkali
reserve," and this is frequently determined in the clinical laboratory.
Certain diseases cause disturbances in the acid balance of the body.

For example, diabetes may give rise to: "acidosis," which can be
fatal.
 碱中毒
(alkalosis)

Ⅳ Sum up

1. Buffer solution

2. Buffer mechanism

3. Buffer pairs in blood

Questions
In addition to the human blood, the pH value of the other
body fluid should control in limited scope. For example,
gastric juice（胃液） is a typical buffer solution as well.
Please look into some information after class to see the
buffer mechanism of it.
 Summarize
After this class we should know:

1) The definition of a buffer solution

2) The composition of a buffer solution

3) explain the role of each component in a buffer

solution in the control of pH

4) how to choose a buffer

5) Buffering Action in Medical

 Homework
1. What is a buffer solution? Illustrate H2CO3-HCO3−1 in human
blood to explain how a buffer works and how important buffer
solutions are in medicine.
2. Which of the following sets of chemicals can be used to prepare
buffer solutions?
(1) HCl+NH3·H2O; (2) HCl+Tris; (3) HCl+NaOH;
(4) Na2HPO4+Na3PO4; (5) H3PO4+NaOH; (6) NaCl+NaAc
3. Caculate the pH of a buffer prepared by mixing 0.01 mol/L acetic
acid(CH3COOH, HAc) and 0.01 mol/L sodium acetate(CH3COONa,
NaAc) solutions with the volume ratio of 1:1.
 Homework
4. How to the anti-acid component and anti-base component of a
buffer work? Why are they typically a conjugate acid-base pair?

5. Choose the factors that determine the capacity of a buffer from

among the following and explain your choices.

(a)Conjugate acid-base pair; (b) pH of the buffer; (c) Buffer range;

(d)Concentration of buffer components; (e) Buffer-component ratio.