Preparation of Buffer Solution: Prepared By: Dr. Wafa Farooq

Lab-6
Preparation of Buffer Solution
Prepared by :
Dr. Wafa Farooq
Objectives:
▪ Definition of buffer solution.
▪ Henderson Hasselbach Equation.
▪ Buffer Capacity.
▪ Preparation of buffer solution.
2
➢ A buffer is a solution characterized by the ability to resist
changes in pH when limited amounts of acid or base are
added to it.
➢ Importance : pharmaceutical solutions are buffered as the
buffer solutions are capable of maintaining pH at some fairly
constant value when even small amounts of acid or base are
added.
Biological fluids, such as blood, are usually buffer solutions; the
control of pH is vital to proper functioning of these fluids.
3
▪ Buffers contain either a weak acid and its conjugate base or a
weak base and its conjugate acid.
▪ For example: Mix acetic acid & sodium acetate, Mix ammonia
& ammonium chloride.
▪ Blood, as a buffer solution, contains H2CO3 ,HCO-3 , CO32-and

CO2as well as other conjugate acid–base pairs.
4
Henderson Hasselbach Equation
[𝐴− ] For acidic
pH= pKa +log
[𝐻𝐴 ] buffer
[𝐵] For basic

pH= pKa +log
[𝐵𝐻 + ] buffer
5
The buffer capacity is the number of moles of strong acid or
strong base that 1 L of the buffer can absorb without changing
pH by more than 1.
OR represents the amount of H+ or OH− the buffer can absorb

without a significant change in pH.
𝑑𝑐𝑏 𝑑𝑐𝑎
𝛽= =-
𝑑𝑝𝐻 𝑑𝑝𝐻
where dcb is the number of moles per liter of strong base, and dca is the number of
moles per liter of strong acid added to the buffer. Since adding strong acid to a buffer
causes the pH to decrease, dca/dpH is negative, and buffer capacity is always positive.
6
Buffer capacity
• Another equation
Ka • [H3O+]
β = 2.303 • C •
(Ka + [H3O+])2
c : total buffer conc.(sum of the molar conc. of the acid & the salt)
7
Maximum Buffer capacity
• βmax occurs where pH = pKa ([H3O+] = Ka)

+ 2
[H3O ] 2.303
βmax = 2.303 • C • = •C
(2 [H3O+])2 4
βmax = 0.576 • C
( pH = pKa )
8
▪ An acid buffer is most effective when the concentration of the
base component equals the concentration of the acid
component.
i.e when pH= pka
▪ A buffer solution works best when the acid/base to salt ratio

is range from 1 :10 to 10:1 and when pH of buffer is equal
pka ± 1.
▪ Two important characteristics of a buffer are:

its pH and its buffer capacity.
9
Three practical methods to prepare a buffer:
1- First Method : By the Titration, in the presence of
one of the two buffer forms with strong base or acid:
 Prepare a buffer composed of an acid and its salt by

adding a strong base(e.g. NaOH) to a weak acid (e.g.
Acetic acid) until the required pH is obtained.
 If the other form of buffer is available (in this case
sodium acetate), a strong acid is added (e.g. HCl)
until the required pH is obtained.
CH3COONa+HCl→CH3COOH+NaCl
 So acetate buffer is formed(CH3COOH/CH3COONa)
10
 Advantages:
1. Easy to understand.
2. Useful when only one form of the buffer is available (in this
case acetic acid)
 Disadvantages:
1. Slow.
2. May require lots of base (or acid).
11
12
2- Second Method: Using the buffer pKa , calculate the
amounts (in moles) of acid/salt or base/salt present in the
buffer at the desired pH.
 If both forms (i.e., the acid and the salt) are available,
convert the amount required from moles to grams ,using the
molecular weight of that component, and then weigh out
the correct amounts of both forms. Or convert moles to
volume if the stock is available in the liquid form.
13
 Advantages:
1. Fast.
2. Easy to prepare.
3. Additional pH adjustment is rarely necessary, and when
necessary, the adjustment is small.
 Disadvantages:
1. Requires the buffer pKa
2. and solving two equations.
14
15
16
17
3- The Third Method: Using table.
 Find a table of the correct amounts of acid/salt or base/salt
required for different pH's
 Dissolve the components in slightly less water than is required for
the final solution volume.
 Check that the pH and correct if necessary.
 Add water to the final volume.
18
19
 Advantages:
1. Easy to do (with appropriate table).
2. Convenient for frequently prepared buffers.
 Disadvantages:
1. May be impossible to find table.
2. Table may be incorrect.
3. Requires both forms of buffer.
4. Component amounts from table will need to be adjusted to give
the buffer concentration and volume in your solution.
20
preparation of buffer solution
1- Choose the proper acid and it salt to get high capacity buffer solution.
For example if you want to prepare buffer solution of pH =5 of 0.025 M
(500mL), look for acid pKa which is ± 1, the best acid is acetic acid with
pKa= 4.76 and its salt sodium acetate.
−
[𝐴 ]
pH= pKa +log
[𝐻𝐴]
−
[𝐴 ]
5 – 4.76= log [𝐻𝐴]
−
[𝐴 ]
0.24 = log 𝑏𝑦 𝑡𝑎𝑘𝑖𝑛𝑔 𝑎𝑛𝑡𝑖 log 𝑜𝑓 0.24
[𝐻𝐴]
−
[𝑠𝑎𝑙𝑡 ]
1.7378 = [𝑎𝑐𝑖𝑑]
salt= 1.7378acid
21
…. Cont.
Salt + acid = 0.025 M
By Substituting salt into 1.7378 acid
1.7378acid + acid =0.025M
2.7378 Acid =0.025 M Divide both side by 2.7378
Acid= 9.13x10-3 M salt = 1.7378 acid
Salt= 9.13x10-3 x1.7378= 0.01586 M

22
…. Cont.
For Salt
𝑚𝑜𝑙𝑒
Molarity = 𝐿 mole = MxL = 0.01586 MX 0.5 L = 7.93x10-3 mole
𝑤𝑒𝑖𝑔ℎ𝑡
mole = 𝑀.𝑤𝑡
weight= Mole x molecular weight= 7.93x10-3 x 82.03= 0.651 g

of salt
23
…. Cont.
For Acid
𝑚𝑜𝑙𝑒
Molarity = mole = M x L = 9.13x10-3 MX 0.5 L = 4.565X10-3 mole
𝐿
𝑤𝑒𝑖𝑔ℎ𝑡
mole =
𝑀.𝑤𝑡
weight= Mole x molecular weight= 4.565x10-3 x 60.05= 0.274 g
Take 0.274 g of acid
24
…. Cont.
To change grams to mL you have to use density of acid which

equal to 1.05 g/mL
0.274
mL= 1.05
= 0.261 𝑚𝐿
Take 0.651 g of salt in a little water in 500 mL volumetric flask,

then add 0.261 mL of acid and complete with water up to 500 mL.
25
Calibration of pH-meter
1-Preparing for Calibration.

2- Clean your electrode
3- Prepare your buffers 4, 7, 10
4- Place your electrode in the buffer with a pH value of 4 and begin reading.
5- clean your electrode and immerse it in second buffer solution =7 then
solution 10.
26
Set to calibration mode Three buffer solution 4,7,10 Rinse the electrode with water
Dry the electrode with tissue Start measuring the Buffer solution 27
28

Preparation of Buffer Solution: Prepared By: Dr. Wafa Farooq

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Lab-6

Preparation of Buffer Solution

▪ Blood, as a buffer solution, contains H2CO3 ,HCO-3 , CO32-and

[𝐵] For basic

OR represents the amount of H+ or OH− the buffer can absorb

• βmax occurs where pH = pKa ([H3O+] = Ka)

▪ A buffer solution works best when the acid/base to salt ratio

▪ Two important characteristics of a buffer are:

 Prepare a buffer composed of an acid and its salt by

By Substituting salt into 1.7378 acid

1.7378acid + acid =0.025M

2.7378 Acid =0.025 M Divide both side by 2.7378

Acid= 9.13x10-3 M salt = 1.7378 acid

Salt= 9.13x10-3 x1.7378= 0.01586 M

weight= Mole x molecular weight= 7.93x10-3 x 82.03= 0.651 g

weight= Mole x molecular weight= 4.565x10-3 x 60.05= 0.274 g

Take 0.274 g of acid

To change grams to mL you have to use density of acid which

Take 0.651 g of salt in a little water in 500 mL volumetric flask,

1-Preparing for Calibration.

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