SIJ1005 : PRINCIPLES OF

BIOCHEMICAL
TECHNIQUES
Basic Laboratory
 USING CHEMICALS
• SAFETY ASPECTS
Risk assessment & relevant safety information
PREPARING SOLUTIONS
Molar concentration (mmol-1, molm-3) or mass concentration (gl-1, kgm-3)
concentration = amount/volume
refer slide steps to make up a solution
use distilled/deionized water to make up aqueous solution and stir to dissolve
• STOCK SOLUTION
Valuable - range of solution with different concentrations of a reagent/common
ingredients
More concentrated than the final requirement and diluted when final solutions are
made up
HOW to make aqueous solution of known concentration

• Decide the concentration and volume • Weigh out the required mass of chemical
required
• Add chemical to a beaker/conical flask and stir to
• Relative molecular mass of the chemical (Mr) dissolve

• Check and adjust pH of solution

• Work out the mass of chemical that will give the
concentration desired and volume required
• Make up the solution to the desired volume
Express all volumes in same units
Calculate the number of moles required
• Transfer the solution to a reagent bottle and label
Convert from mol to g by multiplying by relative
molecular mass (Mr for NaCl=58.44)
 PREPARATION OF DILUTIONS
• Single dilution
procedure
1. Transfer an accurate volume of the stock solution to a volumetric flask
2. Make up to the calibration mark with appropriate solvent
3. Mix thoroughly

Dilution factor – ratio of the initial concentration of stock and final concentration
eg. You want to prepare 100 ml solution of NaCl at 0.2 moll-1 using stock of 4.0 moll-1
The dilution factor is 0.2/4.0 =0.05 = 1/20 ( 20 fold dilution)
• Dilution Series
use – prepare standard curve for calibration
Preparation of linear dilution series - the concentrations are
separated by an equal amount
Eg. Series containing protein at 0, 0.2, 0.4, 0.6, 0.8, 1.0 -1
use [C1]V1= [C2]V2 to determine the amount of stock solution required for
each member of the series
https://www.youtube.com/watch?v=MCrNjHcfcpY
Study exercise
1. What mass of substance would be required to prepare each of the following;
a) 100 ml of NaCl at 50 mmoll-1 (Mr = of NaCl = 58.44)
b) 250 ml of mannitol at 0.10moll-1 (Mr = of mannitol= 182.17)
c) 200 ml of BSA at 800 -1

2. If you added 1.0 ml of an aqueous of NaCl at 0.4 moll-1 to 9.0 ml of water what would be the final concentration of NaCl
in mmoll-1?

3.How would you prepare 250 ml of KCl at a final concentration of a 20.0 mmol-1 from a solution containing KCl at 2.0 moll-
1
?

4. Suppose you had the following stock solutions : NaCl 100.0 mmol-1 , KCl 200.0 mmol-1 , glucose 5.0 mmol-1 . Calculate the
volume of water required to prepare each of the following
d) 1.0 ml of a solution containing only KCl at 10.0 mmol-1
e) 50 ml of a solution containing NaCl at 2.5 mmol-1 and glucose at 0.5 mmol-1
f) 100 ml solution containing NaCl at 5.o mmol-1 , KCl at 2.5 mmol-1 , and glucose 5.0 (0.25) mmol-1

(answer in each case to three significant figure)

END