LABORATORY CALCULATIONS/ DILUTIONS

USE OF DILUTIONS:
The ability to make dilutions accurately is

critical as incorrect reagent /sample preparation will..

Dilutions
Dilutions are performed to obtain a

desired concentration .
It will always_________ the concentration

of the sample/solution
They are ratios hence rarely expressed as

whole numbers

 In general dilution involves adding more

solvent to a solution so that the concentration of the solute becomes lower

Total number of solutes in the solution

remains the same but the volume of the

solution becomes greater , resulting in a

____________________

Identifying the volumes

1:2 1 to 1

1 in 2
1/2 50% v/v

DILUTION RATIO
It is defined as the volume of sample per

total volume
Total volume is equal to the volume of the

sample plus the volume of the buffer used to make the dilution

Useful definitions
Aliquot- A measured sub-volume of the original
sample Diluent- Material with which the sample is diluted Dilution factor(DF)- Ratio of final volume/aliquot volume Concentration factor(CF)- Ratio of aliquot volume divided by final volume (inverse to DF)

Final volume= aliquot+diluent

Sample volume: it is the volume of sample

/solution required to prepare a given

dilution.

Diluent volume: Solvent required to

perform the dilution. Type of diluent varies from test to test.

Types of dilutions
PART DILUTIONS SERIAL DILUTIONS

Dilution of solutions and body fluids

Dilutions are expressed as 1 unit of the

original solution to the total number of units of the final solution.

 and 1/10 dilution

Units of concentration
Rather than particles/ml ,concentration are

expressed as follows:
mg/mL

moles/mL
cells/mL

Any other unit that is used to express

concentration

how to make dilution

Dilutions can be made in infinite number of

ways
When numerator and denominator of a

dilution are multiplied by the same number we have the same dilution with a different volume
Volume of sample-decides the dilution

Work out different ways for a 1/10 dilution

Part dilutions
When X part of sample is added with Y

parts of diluent.
Example:

Difference between and 1:2 dilution.

or 1in 2 dilution:

Means 1 part of aliquot added to 1 part of

diluent
Total volume will be 2 part. 1 part of aliquot in 2 parts

NOTE: What will be the dilution if 1 part of sample was mixed with 2 parts diluent?

 1:2 dilution

1 part of sample is mixed with 2 parts of diluent.

Formular for calculation:

1. Dilution = sample volume/total volume 2. Part of sample/Part of total volume= parts

sample volume/parts total volume

Example: To prepare 100L of 1in 6 dilution ? What is the volume required?

Mixing parts or volumes

To make 1:3 acetic:ethanol solution
One part acetic acid Three part ethanol

To make 1/3 dilution of acetic acid in ethanol

solution
One part acetic acid Two part ethanol

Dilutions of solutions from a higher concentration to a lower concentration C1 X V1 = C2 X V2

Example.. To prepare 100 ml of 25 grams/liter sucrose solution from a stock sucrose solution of 50 g/liter
C1 =50g/L C2= 25g/L V2=100ml
=

V1 = C2 xV2 C1

25 x 100 = 50 ml
50

Calculating new concentration after dilution

Example : What would be the concentration of a solution made by diluting 45.0 mL of 4.2 M KOH to 250 mL ?
C1 = 4.2 , V1 =45.0 mL ,V2= 250 mL ,C2=? C2 =4.2 x 45 = 0.76M 250

Dilution factor
Dilution factor is the final volume / aliquot

volume Example: 1. What is the dilution factor and the concentration factor if you add 0.1 mL aliquot of a specimen to 9.9mL of diluent?

Dilution of body fluids

METHOD:
Calculate the volume of the aliquot
Calculate the volume of the diluent Measure the correct volume of aliquot and

diluent and mix

Example:
To make 10ml of a 1/20 dilution of blood
Volume blood required = Final volume needed

X dilution required
10 X 1/20= 0.5ml

To prepare 10ml of a 1/20 dilution , add

0.5ml of blood to 9.5 ml of diluting fluid.

Calculating the proper dilution

TOTAL VOLUME /SAMPLE VOLUME

and taking the reciprocal of that

number.
Calculate the dilution of blood when using

50l of blood and 950l of diluting fluid?

en

SERIAL DILUTIONS:
Used to prepare larger dilutions with just a

small amount of sample.

They are a sequential set of dilutions in

which the stock for each dilution in the series is the working solution from previous dilution.

Example:
Take 8 test tubes

Measure 500l of diluent into each tube

500l sample is placed in the 1st tube and

mixed
To make the total volume of the 1st tube to Dilution in the tube will

be_______________

 Remove 500l from the first tube and add

to the diluent in the second tube and mix.

What will be the dilution in the second

tube?

 This procedure is repeated serially with

the six remaining tubes

Calculate the dilution of the successive

tubes?

Serial dilution

THANK YOU!