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Basic Laboratory

This document provides information on basic biochemical techniques for preparing solutions of known concentrations from stock solutions. It discusses calculating molar concentrations and mass concentrations, making stock solutions, and preparing single dilutions and dilution series. Specific steps are outlined for making aqueous solutions of a given concentration by determining the mass of chemical needed based on its molar mass and the desired volume and concentration. Methods are described for performing single dilutions by transferring an accurate volume of stock solution and diluting to the final volume, as well as preparing dilution series using the equation that concentration 1, volume 1 equals concentration 2, volume 2. Practice problems are included on calculating amounts needed and volumes for various dilution scenarios.

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harminder singh
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32 views7 pages

Basic Laboratory

This document provides information on basic biochemical techniques for preparing solutions of known concentrations from stock solutions. It discusses calculating molar concentrations and mass concentrations, making stock solutions, and preparing single dilutions and dilution series. Specific steps are outlined for making aqueous solutions of a given concentration by determining the mass of chemical needed based on its molar mass and the desired volume and concentration. Methods are described for performing single dilutions by transferring an accurate volume of stock solution and diluting to the final volume, as well as preparing dilution series using the equation that concentration 1, volume 1 equals concentration 2, volume 2. Practice problems are included on calculating amounts needed and volumes for various dilution scenarios.

Uploaded by

harminder singh
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© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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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

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