5 Steps to Teach Solution Prep

• Biotech 1 SLOP
• Text - Chapter 3
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Ellyn Daugherty
SM Biotech Career Pathway
www.BiotechEd.com
www.emcp.com/biotech
www.sargentwelch.com/biotech
aeedaugher@aol.com
Five Steps to Teach Solution Prep

1. What is a solution?
2. Solvent Measurement
3. Measure the Solute
4. Mix the Solute and Solvent
5. Check the Solution Preparation
 Step 1. What is a Solution ?
• Solutes (usually measured in g)
• Solvents (usually measured in mL)
• Mixed together until solute
dissolves in solvent
solute 8 mg/mL 4 mg/mL 2 mg/mL
concentration =
solution
• Described by the portion of solute
to solvent usually in one of 3 ways:
•5 g/L protease
- mass/volume
•175 mg/mL rennin
•25 !g/!L chymosin

- % mass/volume • 2% glucose solution

• 10% sodium hydroxide solution

• 1M NaOH
- molarity • 50 mM TRIS
• 5 !M CaCl2
 Step #2 Solvent Measurement
Volume of solvent is usually measured using one of 3
instruments. For each, ask:
• What is the total volume that can be measured?
• What is the value of each type of graduation?
Review: Metric Measurements and Conversions

1000 1000
L mL !L

92 ml = _________ !L 357 !L = _________ mL

0.5 L = _________ mL 0.557 mL = _________ !L

 Step #3 Solute Measurement
Solutes are usually dry chemicals
and their mass is measured on a
scale/balance and reported in
grams (g).
For a specified concentration of a
specified volume of solution, the
technician must determine the
amount of mass of solute need for
the volume of solution at some final
concentration.
3 different equations, one for each
concentration units group
- mass/volume
- % mass/volume
- molarity
Determine the Mass of Solute (g)
3 different equations, one for each concentration units group

___________ _____________ __________________

• mass/volume concentration desired X volume desired = mass of solute needed

__________ ____________ _________________

• % mass/volume conc desired (decimal) X volume desired = mass of solute needed

_________ ____________ __________ __________

• molarity concentration X volume desired X molecular weight = mass of solute needed

desired (mol/L) (L) (g/mol) (g)
 Measuring for Mass/Volume Solutions

To make 100 mL of a 5 mg/mL hemoglobin solution

___________ _____________ _________________

concentration desired X volume desired = mass of solute to be weighed

5 mg/mL 100 mL 500 mg = 0.5 g

concentration desired X volume desired = mass of solute to be weighed
Measuring for % Mass/Volume Solutions
To make 100 mL of a 3% CuSO4 solution
3 % = 0.03

__________ ____________ _________________

conc desired (decimal) X volume desired = mass of solute needed

0.03 g/mL 100 mL 3 g CuSO4

conc desired X volume desired = mass of solute needed
 Measuring for Molar Solutions
To make 300 mL of a 0.5M NaOH solution.
Convert 300 mL = 0.3 L

_________ ____________ __________ _______

concentration X volume desired X molecular weight = mass of solute
desired (mol/L) (L) (g/mol) (g)

Molecular Weight ?

Moles are used to count molecules and atoms since they are too small to
count individually. 1 mole equals the amount of molecules that gives a
mass, in grams, equal to that substance’s molecular or formula weight.

The formula weight can be determined using a periodic table or by looking

at a chemical reagent bottle.
For Molar Solutions you need the MW
Determine the molecular weight of the solute

The formula weight can be determined using a periodic table or by looking

at a chemical reagent bottle
 Measuring for Molar Solutions
To make 300 mL of a 0.5M NaOH solution.
Convert 300 mL = 0.3 L

0.5 mol/L 0.3 L 40 g/mol 6g

concentration X volume desired X molecular weight = mass of solute
desired (mol/L) (L) (g/mol) (g)

6 g of NaOH is measured on a balance and put into an appropriate vessel.

Then, solvent is added up to the 300 mL graduation.
 4. Mixing the Solute and Solvent

Mass/Volume solutions

100 mL of a 5 mg/mL hemoglobin solution.

5 mg/mL 100 mL 500 mg = 0.5 g

concentration desired X volume desired = mass of solute to be weighed

250 mL beaker

100 mL
solvent (dH20 or buffer)
0.5 g hemoglobin
Mass/Volume Demo
 Mixing % Mass/Volume Solutions

100 mL of a 3% CuSO4 solution

0.03 g/mL 100 mL 3 g CuSO4

conc desired (decimal) X volume desired = mass of solute needed

250 mL beaker

100 mL
solvent (buffer or dH20)
3 g CuSO4
% Mass/Volume Demo
 Mixing Molar Solutions
300 mL of a 0.5M NaOH solution.
Convert 300 mL = 0.3 L

0.5 mol/L 0.3 L 40 g/mol 6g

concentration X volume desired X molecular weight = mass of solute
desired (mol/L) (L) (g/mol) (g)

600 mL beaker
300 mL
solvent (dH20 or buffer)
6 g NaOH
Molar Solution Demo
Step 5 Checking the Solution Preparation

I like to teach solution prep with copper sulfate since it is blue

and can be visualized (relative number of molecules compared)
in the Spec 20D+ at 600 nm)

I also check the volume and labels on all tubes.

Other methods of checking solution prep?

 Now YOU try it.
Make the following solutions but before you do:
• Do all the calculations showing the equations with all the units of measurement shown.
• Diagram how you are going to prepare the solution.

After you prepare the solution, check it in the spec. Demonstrate how to take up and dispense a sample.

• 5 mL of 0.2 g/mL cupric sulfate pentahydrate

• 7 mL of 50 mg/mL cupric sulfate pentahydrate
• 4 mL of 2% cupric sulfate pentahydrate
• 6 mL of 1% cupric sulfate pentahydrate
• 5 mL of 0.5 M cupric sulfate pentahydrate
• 4 mL of 250 mM cupric sulfate pentahydrate

Check each solution in the spec at 600 nm.

Use 3 mL of the solution to be checked, in a 13x100 mm cuvette.