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PH and Buffer System - Notes

This document provides information about pH, buffers, and buffer preparation. It discusses how buffers resist changes in pH when acids or bases are added. Common buffer systems include phosphate and Tris buffers. The Henderson-Hasselbalch equation describes buffer behavior. Several examples are provided for calculating pH values of buffer solutions and determining amounts of chemicals needed to prepare buffers at specific pH values. The document also discusses characteristics of effective biological buffers and structures of synthetic zwitterionic buffers.

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katherine moreno
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426 views29 pages

PH and Buffer System - Notes

This document provides information about pH, buffers, and buffer preparation. It discusses how buffers resist changes in pH when acids or bases are added. Common buffer systems include phosphate and Tris buffers. The Henderson-Hasselbalch equation describes buffer behavior. Several examples are provided for calculating pH values of buffer solutions and determining amounts of chemicals needed to prepare buffers at specific pH values. The document also discusses characteristics of effective biological buffers and structures of synthetic zwitterionic buffers.

Uploaded by

katherine moreno
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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NOTES

pH and Buffer system


Preparing buffer solution

A solution containing a conjugate weak acid/weak base pair that is


resistant to a change in pH when a strong acid or strong base is
added.
Many metabolic reactions are accompanied
by the release or uptake of protons.

• Oxidative metabolism produces CO , the anhydride of


2

carbonic acid, which if not buffered would produce severe


acidosis.

• Biologic maintenance of a constant pH involves buffering by


phosphate, bicarbonate, and proteins, which accept or
release protons to resist a change in pH.
Virtually all biochemical investigations must be carried
out in buffered aqueous solutions


The natural environment of biomolecules and cellular
organelles is under strict pH control, with normal pH range
usually 6 to 8.

An artificial buffer system is found to be the best substitute
for the natural cell milieu.
The Henderson-Hasselbalch equation describes
the behavior of weak acids & buffers
Derivation of the Henderson-Hasselbalch equation
Derivation of the Henderson-Hasselbalch equation
The Henderson-Hasselbalch equation has great
predictive value in protonic equilibria
Buffer solutions can be made to maintain almost
any pH, depending on the acid-base pair used.

If a strong acid (H3O+) is added, the basic component of
the buffer (A–) can react with it.


If a strong base (OH–) is added, the acidic component of
the buffer (HA) will react with it.
•Buffer solutions are most effective when both components,
the conjugate acid and the conjugate base, are present in
reasonably large concentrations (high buffer capacity)


In order for the solution to be considered a buffer, this ratio must be between
a 1/10 and a 10/1 ratio of [X-]/[HX].

A buffer is most effective when there are approximately equal concentrations of the
two buffer components (a ratio of [X–]/[HX] close to 1/1).

The solution will guard against large pH changes equally well whether acid is added
or base is added.
1/10 and a 10/1 ratio of [X ]/[HX] -

• This restriction means that the [H3O+] in the buffer will


be within a factor of 10 of the Ka value of the conjugate
acid.

• It also means that the pH of the buffer will be within one


pH unit of the value of the pKa of the conjugate acid.
Several characteristics essential in a
biological buffer system
1) pKa between 6 and 8.
2) Highly soluble in aqueous systems.
3) Exclusion or minimal transport by biological membranes.
4) Minimal salt effects.
5) Minimal effects on dissociation due to ionic composition, concentration, and temperature.
6) Buffer–metal ion complexes nonexistent or soluble and well defined.
7) Chemically stable.
8) Insignificant light absorption in the ultraviolet and visible regions.
9) Readily available in purified form.
Effective buffering ranges of
several common buffers.

Although most biochemical


solutions require buffer systems
effective in the pH range 6 to 8,
there is occasionally a need for
buffering over the pH range 2 to
12.
Structures and Properties of Several Synthetic Zwitterionic Buffers
Structures and Properties of Several Synthetic Zwitterionic Buffers
Preparation of Common Laboratory Buffers
0.2 M Acetate Buffer (pKa 4.86)

Prepare 0.2 M acetic acid and 0.2 M sodium acetate solutions separately. Add two
solutions in different proportions to obtain various pH solutions.

Sample calculation:

Since 1000 mL of 0.2 M acetic acid contain = 0.2 moles,

so 46 mL of 0.2 M acetic acid will contain = 0.2 _x005F_x0002_ x 46/1000


= 9200 μmoles
Preparation of Common Laboratory Buffers
0.2 M Sodium Phosphate Buffer (pKa 6.86)

Prepare 0.2 M solutions of sodium dihydrogen phosphate (NaH2PO4 ) and disodium
hydrogen phosphate (Na2HPO4) salts separately and mix dibasic solution in monobasic
solution

Sample calculation:
Since 1000 ml of 0.2 M NaH2PO4 contain= 0.2 moles,
so 46 ml of 0.2 M NaH2PO4 will contain- 0.2 x 46/1000 = 9200 μmoles
Preparation of Common Laboratory Buffers
0.2 M Tris-HCl Buffer (pKa 8.1)

Tris buffer is prepared by mixing appropriate volumes of Tris and HCl
Describe how you would prepare a “phosphate
buffer” with a pH of about 7.40
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.

For example, we could dissolve 1.5 moles of Na2HPO4 and 1.0


mole of NaH2PO4 in enough water to make up a 1-L solution.
Calculate the pH of a buffer that is 0.020 M in NH 3 and 0.030 M in
NH4Cl. What is the pH after adding 1.00 mL of 0.10 M NaOH to 0.10 L of
this buffer?

ka pka
What is the pH after adding 1.00 mL of 0.10 M NaOH to 0.10 L of this buffer?
Calculate the pH of a buffer system containing
1.0 M CH3COOH and 1.0 M CH3COONa
Sodium hypochlorite (NaOCl, the active ingredient of almost all
bleaches) was dissolved in a solution buffered to pH 6.20. Find
the ratio [OCl ]/[HOCl] in this solution.
Find the pH of a 1.00-L aqueous solution prepared with 12.43 g of
tris (121.135) plus 4.67 g of tris hydrochloride (157.596).

Tris= 0.1026 M
Tris hydrochloride = 0.0296M
If we add 12.0 mL of 1.00 M HCl to the solution in the
previous example, what will be the new pH?
12. 0 mL x 1.00 mol/L HCl= 0.0120 mol H+
How many milliliters of 0.500 M NaOH should be added to
10.0g of tris hydrochloride to give a pH of 7.60 in a final volume
of 250 mL?
Moles tris hydrochloride= 10.0 g x 157.596 g/mol =0.0635 mol

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