solution preparation steps
DESCRIPTION
Learn how to prepare solutionsTRANSCRIPT
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Ellyn DaughertySM Biotech Career Pathway
www.BiotechEd.com
www.emcp.com/biotech
www.sargentwelch.com/biotech
• Biotech 1 SLOP
• Text - Chapter 3
• Lab Manual - Chapter 3
• Student CD Tutorials
• Course Planner
5 Steps to Teach Solution Prep
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1.What is a solution?
2.Solvent Measurement
3.Measure the Solute
4.Mix the Solute and Solvent
5.Check the Solution Preparation
Five Steps to Teach Solution Prep
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• Solutes (usually measured in g)
• Solvents (usually measured in mL)
• Mixed together until solute
dissolves in solvent
• Described by the portion of solute
to solvent usually in one of 3 ways:
- mass/volume
- % mass/volume
- molarity
Step 1. What is a Solution ?
8 mg/mL 4 mg/mL 2 mg/mL
concentration =solute
solution
• 2% glucose solution
• 10% sodium hydroxide solution
•5 g/L protease
•175 mg/mL rennin
•25 !g/!L chymosin
• 1M NaOH
• 50 mM TRIS
• 5 !M CaCl2
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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?
Step #2 Solvent Measurement
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L mL !L
Review: Metric Measurements and Conversions
1000 1000
92 ml = _________ !L 357 !L = _________ mL
0.5 L = _________ mL 0.557 mL = _________ !L
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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
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Determine the Mass of Solute (g)3 different equations, one for each concentration units group
• mass/volume
• % mass/volume
• molarity
___________ _____________ __________________
concentration desired X volume desired = mass of solute needed
__________ ____________ _________________
conc desired (decimal) X volume desired = mass of solute needed
_________ ____________ __________ __________
concentration X volume desired X molecular weight = mass of solute needed
desired (mol/L) (L) (g/mol) (g)
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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
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Measuring for % Mass/Volume Solutions
__________ ____________ _________________
conc desired (decimal) X volume desired = mass of solute needed
To make 100 mL of a 3% CuSO4 solution
3 % = 0.03
0.03 g/mL 100 mL 3 g CuSO4
conc desired X volume desired = mass of solute needed
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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 tocount individually. 1 mole equals the amount of molecules that gives amass, in grams, equal to that substance’s molecular or formula weight.
The formula weight can be determined using a periodic table or by lookingat a chemical reagent bottle.
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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 lookingat a chemical reagent bottle
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Measuring for Molar Solutions
To make 300 mL of a 0.5M NaOH solution.
Convert 300 mL = 0.3 L
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.
0.5 mol/L 0.3 L 40 g/mol 6 g
concentration X volume desired X molecular weight = mass of solute
desired (mol/L) (L) (g/mol) (g)
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4. Mixing the Solute and Solvent
Mass/Volume solutions
100 mL of a 5 mg/mL hemoglobin solution.
solvent (dH20 or buffer)
0.5 g hemoglobin
100 mL
5 mg/mL 100 mL 500 mg = 0.5 g
concentration desired X volume desired = mass of solute to be weighed
250 mL beaker
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Mass/Volume Demo
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Mixing % Mass/Volume Solutions
100 mL of a 3% CuSO4 solution
3 g CuSO4
solvent (buffer or dH20)
100 mL
0.03 g/mL 100 mL 3 g CuSO4
conc desired (decimal) X volume desired = mass of solute needed
250 mL beaker
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% Mass/Volume Demo
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Mixing Molar Solutions
300 mL of a 0.5M NaOH solution.
Convert 300 mL = 0.3 L
6 g NaOH
300 mL
solvent (dH20 or buffer)
0.5 mol/L 0.3 L 40 g/mol 6 g
concentration X volume desired X molecular weight = mass of solute
desired (mol/L) (L) (g/mol) (g)
600 mL beaker
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Molar Solution Demo
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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?
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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.