BCH312 [Practical] 1

¨ Understanding how to prepare solutions and make dilutions is an essential skill for biochemists which
is necessary knowledge needed for doing any experiment.

¨ What is SOLUTIONS ?
A simple solution is basically two substances that are evenly mixed together.
è One of them is called the solute and the other is the solvent.
è Solution can be composed from one or more solute dissolved in a solvent forming
a homogenous mixture.
¨ Example:

solution Solute è is the substance to be dissolved (sugar)

Solvent è is the one doing the dissolving (water)

2
3
¨ Dilution of solution: means to add more solvent without the addition of more
solute è To make it less concentrated.

1. Volume to volume dilutions (ratio).

2. Preparing dilutions by using the V1XC1=V2XC2 formula.
3. Serial Dilutions.

4
¨ This type of dilutions describes the ratio of the solute to the final volume of the dilute solution.
¨ For example: to make 1:10 dilution of 1M NaCl solution, one part of the 1M NaCl solution, should
be mixed with nine parts of water, for a total of ten parts.
¨ Therefore 1:10 dilution means è1 part of 1M NaCl + 9 parts of water.
¨ Thus:
è if 10 ml of the 1:10 dilution was needed, then 1ml of 1M NaCl should be mixed with 9 ml of water.
è if 100 ml of 1:10 dilution was needed, then 10 ml of the 1M NaCl should be mixed with 90 ml of
water. [The final concentration of NaCl in both cases will be 0.1 M (1/10) = 0.1]

¨ Example:
1:4 dilution
volume of solute 1 ml from solute + 3 ml from solvent = Total volume 4
total volume 5
 how many ml of 7M solution A we need
to make 20 ml of 2:10 A solution?
2 ml à 10 ml

? ml à 20 ml

= (2 X 20) / 10 = 4 ml
So,
So, 4 ml from solution (A) of 7 M is needed and complete volume up to 20 ml (adding 16 ml water).
Note: [16 ml water= 20 ml -4 ml].
How to Know the concentration of solution A after dilution?

First we will find the DILUTION FACTOR by the following :

Dilution factor (D.F) = final volume / aliquot volume
=10/2 = 5
Then we will divide the stock concentration (before dilution) by the D.F:
7/5 = 1.4M

Note: To find out the stock concentration you will multiply the diluted concentration by the D.F
¨ Sometimes it is necessary to use one solution to make a specific amount of a more
dilute solution .

¨ To do this the following formula can be used:

V1 X C1=V2 X C2
¨ Where:
Ø V1= Volume of starting solution needed to make the new solution (volume of stock solution).
Ø C1= Concentration of starting solution (stock solution).
Ø V2= Final volume of new solution.
Ø C2= Final concentration of new solution.
7
 how many ml of 1M solution we need to
make 5 ml of 0.25M solution?

è V1XC1=V2XC2
Where: V1 = ? , C1= 1M ,V2= 5ml ,C2 = 0.25M

So: ( V1) x (1M) = (5ml) x (0.25M)

è V1 = (5 x 0.25)/1 = 1.25 ml

So 1.25ml of the 1M solution is needed (starting solution) then complete the volume up to 5 ml by diluent (generally water).
¨ It is a stepwise dilution of a solution, where the dilution factor is constant at each step.
¨ The source of dilution material for each step comes from the diluted material of the previous step.

1:10
Dilution factor (D.F) = final volume / aliquot volume
= 10 /1 = 10 (for each step)

100 /10 = 10 10 /1 = 10 1 /0.1 = 10 0.1 /0.01 = 10 9

Dilution factor (D.F) = final volume / aliquot volume = 10 /1 = 10 (for each step)

From the lower concentrated solution to the higher one

X10 X10 X10 X10

÷10 ÷ 10 ÷ 10 ÷ 10
From the more concentrated solution to the lower one
10
 è Dilution factor (D.F) = final volume / aliquot volume
= 2/1 = 2 è 1:2 how to calculate the concentration of the
diluted solutions if they unknown ?
-To prepare standard solution 1:
1 ml of the stock 2.0M solution is needed and volume made up to 2 ml è First: find the D.F:
with distilled water (never forget to mix properly). Dilution factor (D.F) = final volume / aliquot volume
-To prepare standard solutions 2-4: = 2/1 = 2
1 ml of the previously diluted solution is taken and volume is made
up to a final volume of 2 ml by the addition of distilled water. è Second: divide the previous solution concentration
1 1 1 1 by the D.F:
1 ml of 2 M 1 ml of 1 M 1 ml of 0.5M 1 ml of 0.25
solution solution solution M solution
-concentration of solution 1 = 2.0 M stock solution /2 =1 M
-concentration of solution 2 = 1M/2 = 0.5 M
-concentration of solution 3 = 0.5M/2 =0.25 M
-concentration of solution 4 = 0.25/2 =0.125 M

2M 1M 0.5 M 0.25 M 0.125 M

12
¨ To get familiar with solution dilutions by different methods.

13
(1)……………………

¨ Prepare 50ml with 1:20 dilution using the 0.08M NaOH solution you previously prepared.

¨ Calculation:
……………………………………………………………………………………………………………………………………
……………………………………………………………………………………………………………………………………

èTo prepare the 1:20 dilution ……....ml of the starting solution (0.08M NaOH) is needed and volume
made up to a final volume of ………..ml.

14
(2)……………………

¨ Prepare 100ml of 0.2M HCl from the previously 0.4M HCl solution you previously prepared.

¨ Calculation:
……………………………………………………………………………………………………………………………………
……………………………………………………………………………………………………………………………………

è To prepare the 0.2M HCl ……..ml of the starting solution (0.4M HCl) is needed and volume made up
to a total volume of ……..ml by adding water.

15
(3)……………………

¨ Starting with a 3 M Copper Sulfate stock solution, prepare 8ml of four standard solutions (1 to 4)
of the following Molarity respectively (dilution 2:8) :
(1) …………. M (2) …………. M (3) …………. M (4) …………. M .

¨ Calculation:
……………………………………………………………………………………………………………………………………
……………………………………………………………………………………………………………………………………
è To prepare standard solution 1: ……… ml of the stock 2.0M solution is needed and volume made up to
…….. ml with distilled water.
è To prepare standard solution 2-4: ……… ml of the previously diluted solution (8.00×10-2 M) is taken
and volume is made up to a final volume of …… ml by the addition of distilled water.

16
1. A solution was prepared by taking 6ml of a 0.22M solution and then the volume was made up to a
final volume of 30ml .What is the concentration of the final solution.?
2. How would you prepare 80ml of a 1:25 dilution of a 2.1M KCl solution? And what is the
concentration of the diluted solution?

17