Solutions

*Concentration Expressions
*Serial Dilution
 Units of Concentration
 A solution is a homogeneous
mixture of one substance (the
solute) dissolved in another
substance (the solvent).
 Concentration is a ratio of the
amount of solute to the amount
of solvent.
 Units of Concentration
 Percent volume
% volume = volume solute (ml) x 100
volume solution (ml)
 Percent mass
% mass = mass solute (g) x 100
mass solution (g)

Solution = solvent + solute

 Units of Concentration

Example 1:
What is the percent by volume concentration of a
solution in which 75.0 ml of ethanol is diluted to a
volume of 250.0 ml?

75.0 ml x 100 = 30.0%

250.0 ml
 Units of Concentration

Example 2:
What volume of acetic acid is present in a bottle containing
350.0 ml of a solution which measures 5.00%
concentration?

x = 0.05
350.0 ml

x = 17.5 ml
 Units of Concentration

Example 3:
Find the percent by mass in which 41.0 g of NaCl is
dissolved in 331 grams of water.

41 g x 100 = 11.0%
372 g
 Units of Concentration

 Molarity (M) is the most common unit of

concentration
 Molarity is an expression of moles/Liter of
the solution.
 Units of Concentration
 A mole is the SI unit of number of particles and
can be used as an expression of the molecular
weight of a substance.

The formula weight of an

element is expressed as
grams/mole
 Units of Concentration
 The molar mass of a compound can be
calculated by adding the molar mass of the
individual elements.

22.99 + 35.45 = 58.44 g/mol

 Making Solutions
 You just calculated the molar mass of sodium
chloride to be 58.44 g/mol.
 To determine how to make a stock solution of
sodium chloride, use the formula:

g = M x L x molar mass
 Making Solutions
 How many grams of NaCl would you need to prepare
200.0 mL of a 5 M solution?
g = M x L x molar mass
g = (5mol/L) (0.2L) (58.44g/mol)
g = 58.44 g
 Diluting Solutions

 Often once you have made a stock

solution, you need to dilute it to a working
concentration.
 To determine how to dilute the stock
solution, use the formula:
C1 – concentration of stock
C1V1 = C2V2 C2 - concentration of diluted solution
V1 – volume needed of stock
V2 – final volume of dilution
 Diluting Solutions

Example 5:
How many milliliters of a 5 M stock solution of NaCl are
needed to prepare 100 ml of a 0.4 M solution?

C1 V 1 = C 2 V 2
(5) V1 = (0.4)(100)
V1= 8 ml
 Diluting Solutions
 Serial Dilutions are dilutions made in series (for
example, if you needed to make solutions that
were 2M, 1M, 0.5M, and 0.25 M)
 The formula for serial dilutions is:

Dilution Factor = (V1 + V2) V1 – volume of solution being diluted

V1 V2 – volume of solvent
 Units of Concentration

Example 6:
Propose a method to prepare 100 ml of a 0.5 M glucose
solution from a 5 M glucose solution.
Serial Dilutions
Introduction
 Many of the laboratory procedures involve the
use of dilutions.
 It is important to understand the concept of
dilutions, since they are a handy tool used
throughout all areas of the clinical laboratory.
 These dilutions have to be considered as they
make a quantitative difference in what is going
on.
Serial Dilutions

 A serial dilution is any dilution where the

concentration decreases by the same
quantity in each successive step.
 Serial dilutions are mutiplicative.
What Does This Mean??
 If a solution has a 1/10 dilution the
number represents 1 part of the solute
added to 9 parts of diluent.
 So the volumes used would be 10-1= 9.
 This represents 1 part soluteadded to 9
parts of diluent.
Dilutions

 If a 1/8 dilution of the stock solution is

made followed by a 1/6 dilution what is
the final dilution.
 The final dilution is: 1/8 x 1/6 = 1/48
 These type of dilutions are trickier and not
used very frequently in the clinical lab.
Doubling Dilutions

 “Doubling dilutions” are very popular.

 This is a series of ½ dilutions. Each
successive tube will ½ the amount of the
original concentrated solution.
 If this is done 6 times this is what you
would end up with:
Doubling Dilution 6 Times
 1st dilution = 1 /2
 2nd dilution = 1 /2 x 1 /2 = 1/4
 3rd dilution = 1/4 x 1 /2 = 1/8
 4th dilution = 1/8 x 1 /2 = 1/16
 5th dilution = 1/16 x 1 /2 - 1/32
 6th dilution = 1/32 x 1 /2 = 1/64
 This results in a series of dilutions, each a
doubling dilution of the previous one
Dilution Factor
 The dilution factor is the final uses the formula
volume/aliquot volume.
 EXAMPLE: What is the dilution factor if you add
0.1 mL aliquot of a specimen to 9.9 mL of
diluent?
 The final volume is equal to the aliquot volume PLUS
the diluent volume:
0.1 mL + 9.9 mL = 10 mL
 The dilution factor is equal to the final volume
divided by the aliquot volume:
10 mL/0.1 mL = 1:100 dilution
Practice

 Problem: What is the dilution factor when

0.2 mL is added to 3.8 mL diluent?
Set Up The Problem

 dilution factor = final volume/aliquot

volume
 0.2 +3.8 = 4.0 total volume
 4.0/0.2 = 1:20 dilution
Problem Continued

 Remember that serial dilutions are always

made by taking a set quantity of the initial
dilution and adding it successively to
tubes with the same volume.
 So each successive dilution would be
multiplied by the dilution factor.
Problem Continued

 So in the above problem all successive

tubes would have 3.8 mLs of diluent.
 You would then transfer 0.2 of the initial
diluted sample into the next tube, mix
transfer 0.2, mix and so on.
 If you had 4 tubes what would be the
final dilution of tube 4?
Solving the Problem -
*Calculate DF of tube 1
Tube 1 2 3 4

Aliquot 0.2 0.2 0.2 0.2

Diluent 3.8 3.8 3.8 3.8

Math *4/0.2 1/20x1/20 1/400x1/20 1/8000x1/20

Dilution 1:20 1:400 1:8000 1:160,000

Solving the Problem

 Or if you simply wanted to know the

dilution of the final tube you could just
multiply them together:
 1/20 x 1/20 x 1/20 x 1/20 = 1:160,000
Next

 What if we added the indicator system to the

test system?
 You must create additional rows, if you are
showing your work, to take into consideration
the dilutional effect that will have on the
dilution.
 When an indicator (X) is added ; it is CRITICAL
to remove an aliquot from the last tube.
Adding Indicator Cells
Tube 1 2 3 4

Dilution 1:20 1:400 1:8000 1:160,000

X 0.2 0.2 0.2 0.2

Volume 3.8 3.8 3.8 3.8

DF 4/1

Math 4x20 4x400 4x8000 4x160,000

Dilution 1:80 1:1600 1:32,000 1:640,000

Comparing from the
original dilution
Tube 1 2 3 4

Aliquot 0.2 0.2 0.2 0.2

Diluent 3.8 3.8 3.8 3.8

Math *4/0.2 1/20x1/20 1/400x1/20 1/8000x1/20

Dilution 1:20 1:400 1:8000 1:160,000

Serial Dilutions
 When performing serial dilutions in the clinical
laboratory one must know the dilution of each
tube so a chart is always created to indicate the
dilution of each tube.
 If you know the dilution factor you multiply each
successive tube by it and can easily determine
the concentration of each tube to create your
chart.