Buffers

Chem 600 Lab

 Buffers
 Buffers are of main importance in regulating the pH of
the blood, cells and tissues.
 Many biochemical reactions including those catalyzed by
enzymes require pH control which is provided by buffers.
 Mammalian tissues in the resting state have a pH of
around 7.4

2H2O + CO2  H2CO3 + H2O ↔ HCO3- + H3O+

Buffer Solutions
-solutions that resist pH changes

COMPONENTS:
• Weak acid + its conjugate base (salt)

• Weak base + its conjugate acid (salt)

NOTE:

A buffer solution which contains a weak acid and its salt is

considered to be acidic. One which contain a weak base and its
salt is considered to be a basic buffer.
Buffer Solutions

CH3COOH and CH3COO-

Weak Acid + Conjugate Base =Buffer

Note: Conjugate Bases are sometimes

given as salt (conjugate base and cation)
i.e. CH3COONa, CH3 COOK and
(CH3COO)2Ca
Buffer Solutions

NH3 + H2O NH4+ + OH-

NH3 and NH4+

Weak Base + Conjugate Acid =Buffer

Note: Conjugate Acids are sometimes

given as salt (conjugate acid and anion)
i.e. NH4Cl, NH4Br and etc
Buffers
 The buffering power is greatest when pH=pKa , i.e.
when the acid and the salt are at the same
concentration

A buffer works most effectively at pH values

that are + 1 pH unit from the pKa (the buffer
range)
pH = pKa ± 1
➢ Concentration of the buffer components The
greater the concentration of the weak acid and
its conjugate base, the greater the buffering
capacity
THE MOST COMMONLY USED BUFFERS IN
THE LABORATORY:
1. Acetate buffer: CH3COONa/CH3COOH (pKa =4.75)

2. Phosphate buffer: Phosphate buffer

Na2HPO4/KH2PO4 (pKa =7.20)

3. Citrate buffer: Sodium Citrate/Citric acid

Which of the following solutions can be classified as buffer
systems?

(a) KH2PO4/H3PO4

(b) NaClO4/HClO4

(c) C5H5N/C5H5NHCl (C5H5N is pyridine; its Kb is 1.7 x 10-9)

(d) NH3/(NH4)2SO4

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9

Consider mixture of salt NaA and weak acid HA.

NaA (s) Na+ (aq) + A- (aq)

[H+][A-]
Ka =
HA (aq) H+ (aq) + A- (aq) [HA]

Ka [HA]
[H+] =
[A-] Henderson-Hasselbalch
equation
[HA]
-log [H+] = -log Ka - log
[A-] [conjugate base]
-] pH = pKa + log
[A [acid]
-log [H+] = -log Ka + log
[HA]

[A-]
pH = pKa + log
[HA]
WEAK BASES: BOH B+ + OH-
Kb = [B+] [OH-]
[BOH]

Solving for [OH-]: [OH-] = Kb [BOH]

[B+]

Taking negative logarithms of both sides:

-log [OH-] = -log Kb - log [BOH]

[B+]

pOH = pKb + log [B+]

[BOH]
(a) Calculate the pH of a 0.20 M CH3COOH solution.
The Ka of CH3COOH is 1.8 x 10-5.

(b) What is the pH of a solution containing both 0.20 M

CH3COOH and 0.30 M CH3COONa? The Ka of CH3COOH
is 1.8 x 10-5.

Answers:
a) pH= -log (1.9 x 10-3 ) = 2.72
b) pH = 4.92

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Acetic acid (50.0 mL, 0.100M) and sodium acetate
(20.0mL, 0.200 M) were mixed to form a buffer. Calculate
the pH of the solution given that the Ka of acetic acid is
1.75 x10-5

pH= pKa + log [CH3COO-]

[CH3COOH]

pH= 4.757 + log (0.200M x 0.020 L)

(0.100M x 0.050L)

pH= 4.66
 What will be the pH of the buffer system, which consists of acetic
acid (50.0 mL, 0.100M) and sodium acetate (20.0mL, 0.200 M),
after the addition of 30.0 mL 0f 0.010 M HCl? pKa = 4.757
CH3COOH (aq)  H+ (aq) + CH3COO- (aq)

The addition of HCl shifts the equilibrium to the left!

pH= 4.757 + log (0.200M x 0.020 L)- (0.010M x 0.030L)

(0.100M x 0.050L) + (0.010M x 0.030L)

pH= 4.757 + (-0.156)

pH= 4.60
What will be the pH of the buffer system, which
consists of acetic acid (50.0 mL, 0.100M) and sodium
acetate (20.0mL, 0.200 M), after the addition of 15.0
mL 0f 0.025 M NaOH?
CH3COOH (aq)  H+ (aq) + CH3COO- (aq)

NaOH will react with H+ and shifts the equilibrium to the right!

pH= 4.757 + log (0.200M x 0.020 L)+ (0.025M x 0.015L)

(0.100M x 0.050L) - (0.025M x 0.015L)

pH= 4.757 + (-0.0241)

pH= 4.73
An important buffer stabilizing the blood plasma is the
bicarbonate-carbonic acid system. The conc. of the
H2CO3 in blood is 0.00125M. Calculate the [NaHCO3]
in the plasma if the pH = 7.4.

H2CO3 + H2O = H3O+ +HCO3- ;Ka = 4.46 x10 -6; pKa =6.35
HCO3- + H2O = H3O+ + CO32-;Ka= 4.67X10-11; pKa =10.33

pH= pKa + log [conj. base]

[acid]

7.4= 6.35 + log [HCO3-]

[0.00125M]
7.4= 6.35 + log [HCO3-] - log[0.00125M]
7.4= 6.35 + log [HCO3-] – (-2.90)
7.4-9.25= log [HCO3-] Antilog(-1.85)= [HCO3-]
-1.85= log [HCO3-] [HCO3-] = 0.0141M =[NaHCO3]
Exercises
(a) Calculate the pH of a buffer system containing 1.0 M
CH3COOH and 1.0 M CH3COONa. The Ka of CH3COOH is
1.8 x 10-5
pH = 4.74
(b) What is the pH of the buffer system after the addition of 0.10
mole of gaseous HCl to 1.0 L of the solution? Assume that
the volume of the solution does not change when the HCl is
added.
pH = -log (2.2 x 10-5 ) = 4.66

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Exercises

(c) What is the pH of a solution that is 0.400 M in formic acid and

1.00 M in sodium formate? The Ka of HCOOH is 1.8 x 10-4
pH = 4.14

(d) Calculate the pH of a solution that is 0.200 M in NH3 and

0.300 M in NH4Cl. Ka for NH4+ = 5.70 x 10-10

pH = 9.06

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Exercises

(e) Calculate the pH of a mixture made up of 100.0 mL of 0.25M

HAc and 100 mL of 0.10 M NaAc. The pKa of HAc is 4.76

pH = 4.36

(f) Calculate the pH of a mixture made up of 100.0 mL of 0.25M

HAc and 200.0 mL of 0.10M NaAc. The pKa of HAc is 4.76
pH = 4.67

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Exercises

(g) Determine the ratio of lactic acid to lactate required in a

buffer system which has a pH of 4.0. The pKa of lactic acid is
3.86
1:1.38

(h) Determine the volume of 0.1M lactate and 0.1 M lactic acid
that must be mixed to prepare 500.0 mL of 0.1M buffer with a
pH of 4.0.
Lactate: 1.38/2.38 x 500 = 289.9 mL
Lactic acid: 500 mL – 289.9 mL = 210.10 mL

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Exercises

(i) What is the pH of a buffer solution made by dissolving 2.0g

of benzoic acid and 5.0g sodium benzoate in 250.0 mL of
water? Ka of benzoic acid is 6.3 x 10-5
Molar masses: Benzoic = 122 g/mol
sodium benzoate = 144 g/mol
pH = 4.53

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Decide on the Buffer Properties
Before making a buffer you must know the
following;

1. concentration

2. volume

3. desired pH

• Most buffers work best at concentrations

between 0.1 M and 10 M.

• The pH should be within 1 pH unit of the

acid/ conjugate base pKa.
Describe the preparation of 1L 0.1M acetate buffer at
pH = 5.22 from NaAc( MW= 87 g/mole) and glacial
acetic acid (density= 1.0498 g/L), pKa= 4.73. MW of
Hac = 60 g/mol. Hac = H+ +Ac- Nac = H+ +Ac-
Moles of 0.1M buffer soln = M x L = 0.1 x 1 = 0.1 mole
pH= pKa + log [conj. base]
Portion of Ac and HAc needed to prepare the buffer
[acid]
Ac- = 0.7555 x 0.1mole = 0.07555 mole
5.22= 4.73 + log [Ac-] HAc = 0.2445 x 0.1mole = 0.02445 mole

[HAc] Convert moles into measurable qty

log [Ac-] = 5.22-4.73 Ac- = source solid NaAc, find the mass needed
[HAc] 0.07555 mole x 87 g/mole = 6.57 g
[Ac-] =3.09 HAc = source liquid acetic acid, volume needed
[HAc] 1 0.02445 mole x 60 g/mole x 1 mL/1.0498 = 1.40 mL
ntotal = 3.09 + 1 = 4.09
Dissolve 6.57 g NaAc in 1.40 mL glacial HAc,
XAc-= 3.09/4.09 = 0.7555
then dilute to 1L with distilled water in a
XHAc= 1/4.09 = 0.2445 volumetric flask
Describe the preparation of 500.0 mL if 0.10M acetate buffer at
pH 5.22 from sodium acetate (MW: 87.0 g/mol) and glacial
acetic acid (MW: 60 g/mol). Density of glacial acetic acid is
1.0498 g/mL. pKa = 4.73

Mix 3.29 g of NaAc and 0.70 mL of glacial acetic acid in

water and dilute with distilled water to the 500 mL mark
of the volumetric flask

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Describe how you would prepare a 1.0L of a 1.0M “phosphate
buffer” with a pH of about 7.40.

3 stages of ionization of phosphoric acid

H3 PO 4 (aq) ƒ H + (aq ) + H 2 PO -4 (aq) K a1 = 7.5 × 10-3 ; pK a1 = 2.12

H 2 PO-4 (aq) ƒ H + (aq) + HPO 42- (aq) K a 2 = 6.2 × 10-8 ; pK a 2 = 7.21
HPO 2-
4 (aq ) ƒ H +
( aq ) + PO 3-
4 (aq ) K a3 = 4.8 × 10-13 ; pK a 3 = 12.32

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Strategy

For a buffer to function effectively, the concentrations of the acid

component must be roughly equal to the conjugate base
component. The desired pH is close to the pKa of the acid, that
is, when pH ≈ pKa,

[conjugate base]
log  0
[acid]
or

[conjugate base]
 1
acid 25
 2- -
The most suitable of the three buffer systems is HPO 4 /H 2 PO 4,
-
because the pKa of the acid H 2 PO 4 is closest to the desired pH.
From the Henderson-Hasselbalch equation we write

[conjugate base]
pH = pK a + log
[acid]
[HPO 2-4 ]
7.40 = 7.21 + log
[H 2 PO -4 ]
[HPO 2-4 ]
log -
= 0.19
[H 2 PO 4 ]

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Taking the antilog, we obtain

[HPO 2-4 ] 0.19

-
= 10 = 1.5
[H 2 PO 4 ]

Thus, one way to prepare a phosphate buffer with a pH of 7.40

is to dissolve disodium hydrogen phosphate (Na2HPO4) and
sodium dihydrogen phosphate (NaH2PO4) in a mole ratio of
1.5:1.0 in water.

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 Na2HPO4 = 1.5 (1.0 mol) = 0.6 mol
2.5

NaH2PO4 = 1.0 (1.0 mol) = 0.4 mol

2.5

Given the molar masses:

Na2HPO4 = 141.98 g/mol; NaH2PO4 = 119.98 g/mol

 Weigh 85.19 g of Na2HPO4 and 47.99 g of NaH2PO4 ,

dissolve in water till the 1-L mark.
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