CHAPTER 2

Solutions
 Objective
At the end of the chapter, the student will be
able to
• Define solution, types of solutions and expressing
concentration of solutions
• Discuss purpose and how to prepare dilutions
• Explain standard, primer, and secondary solutions
• Discuss concept, preparation, and calculations of pH, Strong
and weak acid solutions, and Strong and weak basic solutes.
• Discuss concept, and calculations of dissociation constants.
 Outline of solution lecture
• Definition of solution
• Types of solutions
• Expressing concentration of solutions
• Chemical units: molarities, normality
• Dilution: Definition and preparation
• Standard, primary, and secondary solutions
• Concepts and characteristics of acids and bases
• pKa, dissociation constant of strong acids & bases,
dissociation constant of weak acids and bases
• pH- concept and calculations
• Henderson-Hasselbach equation
• Buffer solutions: Characteristics and classifications.
 2-Solution
2.1. Definition
• A solution is a homogeneous mixture of one
or more substances dispersed molecularly in a
sufficient quantity of dissolving medium
• A solution is made up of solute and solvent
• Solute + solvent = Solution
2.2 Types of solution

• Solid solution:
• Liquid solution:
• Gaseous solution:

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2.3. Expressing concentration of solutions
2.3.1 Relative expression
-Dilute solution:
-Concentrated solution
-Saturated solution
-Super saturated solution

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2.3.2 Quantitative expression of the
concentration

A. Physical units
– Parts per hundred (Percentage)
%W/W,
%W/V
, % V/V
– Parts per unit
– Parts per million
7
 B. Chemical units

• Mole is the amount of a given substance in grams.

• terms like normality, molarity, or molality are used.
• If the quantity of the solute is very small millimole,
micromole or nanomoles may be used.
o 1 mole = 1,000 milimoles
o 1mole = 1,000,000 micromoles
o 1mole = 1,000,000,000 nanomoles

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 a. Molar solution (Molarity, M)
• number of moles of solute per liter of solution or the
gram molecular weight of a compound per liter of
solution.

E.g.. Prepare an aqueous glucose solution with two

moles of glucose per liter

• Solution: 1 mole= its molecular weight in grams

o mole of glucose = 180g
o 2 moles = 2X180g = 360g

• 360g glucose is dissolved in 1 liter of water to yield 2

moles/l of glucose. 9
• Formula:

Molarity = Actual weight in grams

Molecular Weight x volume of solution in
liter.
• E.g.. What is the molarity of NaOH if 20g NaOH is
dissolved in 200ml of solution?

• Solution: actual mass = 20g, molecular weight = 40

Volume = 0.2 liters
Molarity = 20g = 2.5 M
NaOH
40g X0.2 liter
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a. Normal solution (Normality, N)
• is the number of grams equivalents of solute by
weights per liter of solution (not solvent).

• An equivalent weight is equal to the molecular weight

of a substance divided by its valance.

• The valance is the number of unit that can combine

with or replace one mole of hydrogen ion

• Normality (N) = actual weight in grams

Equivalent weight x Vol. in liter

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Eg. What is the normality of KOH if 5.6g KOH is
dissolved in 1 liter of solution?

• Solution: actual mass = 5.6g

Volume of solution= 1 liter
KOH equivalent weight = (56/1) = 56

• Therefore 5.6g = 0.1 N KOH

56 X 1 liter

• Exercise. How do you make 0.4 N solution of

KOH.

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2.4.Making dilution
2.4.1.Introduction:
• Dilution is defined as a process by which the
concentration of a given solution is decreased by
the addition of solvents to make a weaker solution

• It represents the ratio of concentrated material to

the total volume of a solution

• Total volume consists of the volume or weight of

the concentrated plus the volume of the diluents .

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 Dilution, continued…
• This ratio of concentrated or stock solution to the
total volume equals the dilution factor

Dilution factor (df) = volume of stock

Total volume of solution

• The df is inversely related to the concentration

thus, the dilution factor increases as the
concentration decreases.
14
 Reasons for dilution
• To prepare a weaker solution from stronger one

• If the specimen at hand is less than a procedure

calls for

• If the concentration of substances (analyte) is

too high to be accurately measured.

• To remove undesirable substances from the

specimen
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Methods of dilution
o Dilution can be made singly or in series.

o It usually expressed as a ratio rather than

with units of concentration.

o X/Y where X= sample volume

oY= is the sum of the sample
volume and the diluents
oE.g. 1:2, 1:5, 1:10 or fraction ½,
1/5, 1/10.
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 2.4.2.Simple/single dilution

• Is defined as one part of the original solution to

the total part of final solution, which include
both solute & solvent.

• In making a simple dilution, the laboratory

technician must decide on the total volume
desired & amount of stock solution to use

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• In the performance of single dilution, the most
commonly used equation is:
a. Using Proportion
• It is used when reagents are prepared by adding
a specific amount of one solution to a specific
amount of an other solution.
C
V= A+B

Where: C – total volume of final reagent

A – total parts of solution A
B – total parts of solution B
V – volume of each part
• Example 1: a buffer made by adding two parts of
‘solution A’ to five parts of solution B would be required
to make 70 mL of the buffer.

• Formula: C V = 70mL required

2 parts of A + 5 part of B
A+B
= 70 mL
7 part
= 10 volume of one part
A = 2 x 10 = 20 mL
B = 5 x 10 = 50 mL

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• Example 2: a 100mg/mL N2 standard is diluted
1:10. then the concentration of the resulting
solution is
100mg/mL x 1/10 = 10 mg/mL

b. Using C1V1 = C2V2

• This formula is useful only if the units for
concentration & volume are the same & if three
of the four variables are known.

• Example what volume is needed to make 500ml

of 0.1M solution of tris-buffer from a solution of 2
M tris-buffer?
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2.4.3. Serial dilution
• Large dilutions may be difficult to make because
of the amount of diluent that needs to be added.

• For example, a 1/1000 dilution may be difficult to

create accurately even with 0.1mL of serum &
99.9 mL of diluent.

• A series of dilutions, also called serial dilutions,

may be a better way to make the dilutions

• Generally, it is a multiple progressive dilution

process in which the original solution is further
diluted.

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 Serial dilution, continued…
• It is required for certain quantitative tests

• Serial dilution is extremely useful when the

volume of the concentrate &/or diluents is in
short supply

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 1 mL 1 mL 1 mL
1 mL -discard

1ml 1ml 1ml 1ml

Sample: serum tube 1 tube 2 tube 3

Dilution: 1:2 1:4 1:8
Concentration: 1 0.5 0.25

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• The dilution fold of a system can be determined
by the formula:
1 = volume transferred
dilution fold total volume
• Volume transferred = is equal to the constant
volume transferred to each successive tubes in
the serial dilution system.

• Total volume = is equal to the volume being

transferred plus the volume of diluents already in
the tube.

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• E. g. 1. What is the dilution fold of the following
serial dilution system consisting of five tubes?
The following amount of diluents have been
added to the tubes; 0.5 mL to tube 1 & 0.5 mL to
tube 2 to 5. Next, 0.5 mL of patient serum is
added to tube 1 and 0.5 mL is serialy transferred
through tube 5. finally, 0.5 mL is discarded from
tube 5.

o 1/Y = 0.5/1.0
o Y x 0.5 = 1
o Y = 1/0.5 = 2

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E. g. 2. it is often desirable to determine the
dilution of a given tube (Y) in a serial dilution
system. This dilution can be calculated by
Solution of tube 1 = dilution of Y x [
1/dilution fold]
• What is the dilution of tube 3 in the preceding
example?
Y= ½ x (½) (Y-1)
= ½ x (½)2
=
½ x ½ x ½
=
1/8 ;
The dilution of serum in the tube 3 is 1/8.

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 2.5.Standard Solution
is a solution whose concentration is exactly known.

• There are two types of standard solutions:

– primary standard

– secondary standard solution.

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 2.5.1.Primary standard solution

• a highly purified chemical that can be

measured directly to produce a substance of
known concentration in a given solution.

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• The solution must satisfy the following criteria:

o Must be essentially free of impurities

o Should be stable in both solid & solution form

o The concentration should be accurate.

o Should not be hygroscopic or vaporize at 20oc

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• Example
o Sodium carbonate(NaCO3)

o Sodium oxalate

o Sodium chloride,

o Potassium dichromate

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 2.5.2 Secondary standard solution

• is defined as a substance of lower purity &

whose concentration is determined by
comparison to a primary standard.

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• To prepare secondary standard
o Weigh some amount of the substance using
analytical balance.

o Dissolve in a given volume of solvent.

o Using primary standard solution, determine

the exact concentration of the substance.

o If necessary dilute to some volume to get the

concentration required.

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• Example secondary standard
o Oxalic acid

o Nitric acid

o Hydrochloric acid

o Sulfuric acid.

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2.6 Concept of acids and bases
A. Acids & Bases
• According to Bronsted Lowry theory and
Arrhenus theory respectively

Acids – are proton donors

- dissociate to furnish hydrogen ions
Bases – are proton acceptors
-dissociate to furnish hydroxyl ions in queous

solution
 Characteristics of acids
 Characteristics of bases (alkaline solution)
 Dissociation constant
HA + H2O H3O+ + A-

K = [H3O+ ] [A-]
[HA ] [H2O]

Ka = K [H2O] = [H3O+ ] [A-]

[HA ]
• For weak acids Ka < 1
• For strong acids Ka > 1
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pKa of acids or bases
• Strength of an acid or base is determined by its
pKa which is defined as the negative logarithm of
the ionization constant.

o HA H + + A-

o K = [H+] [A-]
[HA]
o pKa = -log Ka

• Strong acids has a low pka value where as a

weak acids has a high pKa value.
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Dissociation of strong acids or bases
o Strong acids are those which dissociate
completely into their respective cations (H+)
&anions in aqueous (A-) solution.
 HA H+ + A-

 HNO3 H+ + NO3-

 H2SO4 2H+ + SO4=

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• Strong bases are those which dissociate
completely into their respective hydroxyl ions
(OH+) & respective cations (B-) in aqueous
solution.

o BOH B+ + OH-

o NaOH Na+ + OH-

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Dissociation of weak acids or bases
o Weak acids (HA) are those which partially
dissociate when they are in aqueous solution

o HA H + + A-

o Examples: Acetic acid, Carbonic acid,

phosphoric acids, lactic acids, benzoic acids etc.

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• Weak bases are those which dissociate partially
when dissolved in aqueous solution.

 BOH B+ + OH-

 Eg NH3, Ca (OH)2, Al2(OH)3

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 Hydrogen ion concentration (pH)

To express hydrogen ion concentration, the

symbol pH was introduced by Sorensen in 1909.

• pH is defined as the negative logarithm of the

Hydrogen ion concentration.
pH = - log [H+] in mol/L
pH of pure water = 7
pH of 1 M solution of strong acid = 0
pH of 1 M solution of strong base = 14
12/16/22 41
1. What is the pH of a solution whose hydrogen
ion concentration is 3.2 x 10-4 mol/L

2. What is the pH of a solution whose hydroxide

ion concentration is 4 x 10-4

3. Calculate the hydrogen ion concentration of a

solution of HNO3 with pH 3.4.

42
Handerson-Hasselbalch equation
o HA H + + A-

o K = [H+] [A-]
[HA]
o [H+] = Ka x [HA]/ [A-]

o Log [H+] = log Ka + log [HA]/ [A-]

o - Log [H+] = -log Ka - log [HA]/ [A-]

o PH = PKa + log [A-]/ HA]

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• The pH of the system is determined by the pKa
of the acid & the ratio of [A-] to [HA].

o pH = pKa + log101

o pH = pKa + 0

o pH = pKa

• The buffer has greatest buffer capacity at its

pKa, i.e., that at which the [A-] = [HA].

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• The capacity of the buffer decreases as the ratio
deviates from 1.

• In general, a buffer should not be used at a PH

> 1 from its PKa.

o What is the pH of a solution whose hydrogen

ion concentration is 3.2 × 10-4 mol/l?

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 Buffer solution & its action

• Buffers are chemical system that prevents change in the

concentration of another chemical substance.

• For example, a proton donor & acceptor system of

solutions are used as buffers to prevent change in
hydrogen ion concentration.

• It consists of a mixture of weak acid (or base) & its

conjugate base (or acid).

• E.g. CH3COOH(ethanoic acid) & CH3COONa (its salt)

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• Buffers resists changes in pH when small
quantities of an acid or a base is added to it.

• A buffer act as like an base if acid is added &

like an acid if base is added.

• The useful buffer range is at PKa + 1

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• A good buffer is prepared by mixing equal
amount of the acid (base) & its salt.

• The action of the buffer & their role in the

maintenance of solutions PH is expalined with
the aid of Henderson-Hasselbatch equation
o HA H + + A-
o BOH B+ + OH-

o HA - is a weak acid, indicate that H is

combined with an other chemical element or
compound A.
o A – is a conjugate base since it can accept H+
& act as a base.

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• Maximum buffering capacity is achieved when
[A-] = HA, such that the ratio of [A-]/ HA] is 1 & pH
= pKa.

• A good buffer maintains its pH change within the

limit of + 1 PH unit.
• Similarly a buffer solution is effective if the pH of
prepared buffer is adjusted between + 1 pKa or
pKb.
• E.g. the pKa acetic acid at 25 0C is 4.76. to
prepare effective acetate bufer, the pH should be
between 3.76 & 5.76.

12/16/22 49
Types of buffers
1.Neutral buffer – phosphate buffer

2. Acidic buffer – citrate buffer

3. Alkaline buffer – borate buffer

12/16/22 50
 Neutral buffers

• It is a buffer solution that has pH value of

around 7.
• Used for maintaining reaction around
neutral pH
1. Acidic buffer solutions
• It is a buffer solution which has a pH less than 7.
• They are commonly made from a weak acid &
one of its salts - often a sodium salt.
• For example, a mixture of ethanoic acid & sodium
ethanoate in solution.
• In this case, if the solution contained equal molar
concentrations of both the acid & the salt, it would
have a pH of 4.76 (its PKa).
• You can change the pH of the buffer solution by
changing the ratio of acid to salt, or by choosing a
different acid & one of its salts.

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2. Alkaline buffer solutions
• It is a buffer solution which has a pH greater
than 7.

• they are commonly made from a weak base &

one of its salts.

• For example, a mixture of ammonia &

ammonium chloride solutions.

• If they are mixed in equal molar proportions, the

solution would have a pH of 9.25.

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How do buffer solutions work?

o A buffer solution has to contain things which

will remove any H+ or OH- ions that you might
add to it - otherwise the pH will change.

o Acidic & alkaline buffer solutions achieve this

in different ways.

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A. Acidic buffer solutions preparation

o We'll take a mixture of ethanoic acid & sodium

ethanoate.
o Ethanoic acid is a weak acid, & the position of this
equilibrium will be well to the left:

o Adding sodium ethanoate to this adds lots of extra

ethanoate ions.

o According to Le Chatelier's Principle, that will tip the

position of the equilibrium even further to the left.
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o The solution will therefore contain these
important things:

o lots of un-ionised ethanoic acid;

o lots of ethanoate ions from the sodium

ethanoate;

o enough hydrogen ions to make the solution

acidic.

12/16/22 56
12/16/22 57
 B. Alkaline buffer solutions
preparation
o We'll take a mixture of ammonia and ammonium
chloride solutions as typical.

o Ammonia is a weak base, and the position of

this equilibrium will be well to the left:

o Adding ammonium chloride to this adds lots of

extra ammonium ions. According to Le
Chatelier's Principle, that will tip the position of
the equilibrium even further to the left.

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o The solution will therefore contain these
important things:
lots of unreacted ammonia;

lots of ammonium ions from the ammonium

chloride;

enough hydroxide ions to make the solution

alkaline.

12/16/22 59
12/16/22 60
1. Calculate the pH of a solution containing 0.08
M acetic acid & 0.02 M sodium acetate. [pKa =
4.7]

2. Calculate the pH of 500 ml of 0.1 M weak acid

after addition of 100 ml of 0.1 M KOH. [pKa =
5]

12/16/22 61
 Summary
 Solution are homogeneous mixture of one or
more substances( solutes) dispersed
molecularly in a sufficient quantity of dissolving
medium (solvents)
 Solutions amount may expressed as dilutions,
concentrations, saturated, and super saturated
solution.
 Molarity of solution (Molarity, M) number of
moles of solute per liter of solution or the gram
molecular weight of a compound per liter of
solution, where as a Normality of a solution
(Normality, N) is the number of grams
equivalents of solute by weights per liter of
solution (not solvent
 Summary….
 Standard Solution is a solution whose
concentration is exactly known. Primary
standard solutiona highly purified chemical that
can be measured directly to produce a
substance of known concentration in a given
solution. Secondary standard solutionis
defined as a substance of lower purity & whose
concentration is determined by comparison to a
primary standard.
• Acids – are proton donors, or dissociate to
furnish hydrogen ions; where as Bases – are
proton acceptors or dissociate to furnish
hydroxyl ions in aqueous solution
 pH is defined as the negative logarithm of the
Hydrogen ion concentration. pH = - log [H+] in
mol/L
 Buffers are chemical system that prevents
change in the concentration of another chemical
substance. There are three types of buffers:
Neutral buffer eg. phosphate buffer; Acidic buffer
eg. citrate buffer; and Alkaline buffer eg.– borate
buffer
 Reference
1. Burtis, Carl A., and Ashwood, Edward R.. Tietz:
Fundamentals of Clinical Chemistry. Philadelphia, 2001.
2. Arneson, W and J Brickell: Clinical Chemistry: A
Laboratory Perspective 1st ed. 2007 FA Davis
3. Burtis, Carl A., and Ashwood, Edward R.. Tietz:
textbook of Clinical Chemistry. Philadelphia, 1999.
 Next Chapter

CHAPTER 3.

Introduction to radiant energy