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Solution Preparation and Dilutions

Part A. Making Solutions using Mass/Volume


Concentrations
Many solutions are prepared with a certain mass of solute in a certain
volume of solvent. Any metric mass in any metric volume is possible,
but the most common units of mass/volume concentrations are as
follows:
g/mL
g/L
mg/mL
g/mL
g/L
ng/L
ng/L

grams per milliliter


grams per liter
milligrams per milliliter
micrograms per milliliter
micrograms per microliter
nanograms per liter
nanograms per microliter

To determine how to prepare a certain volume of a solution at a certain


mass/volume concentration, use the equation below. Convert units as
necessary to make sure units that are used can be cancelled out.
Mass/Volume Concentration Equation
concentration desired x total volume desired = mass of solute in the
total volume desired
(ex. g/mL)
(ex. mL)
(ex.
g)
Ex. A technician needs 50 mL of 15 mg/mL pepsin solution for an
experiment. Using the Mass/Volume Equation, the calculation would be
as follows:
15 mg/mL x 50 mL = 750 mg = 0.75 g pepsin
The technician would add 0.75 g pepsin to a container and fill up to the
50 mL mark with solvent (usually deionized water). Most balances
weigh in grams, so the conversion from mg to g was necessary.
Determine the calculations for the solutions in the Practice Problems
section for Part A. Making Solutions Using Mass/Volume Concentrations
at the end of this handout.

Part B. Making SolutionsUsing Molarity


Concentrations
Solution Preparation 1

The concentration of many solutions is reported as moles/liter (mol/L or


M; the M is spoken molar) or some function of those units. This
concentration measurement is called molarity. Molarity is sometimes a
challenging concept to understand. However, with your recently
acquired solution preparation skills, you will see that making molar
solutions requires only one extra calculation.
To understand how to make a solution of a given molarity, you must
know what a mole is. A mole of a compound is equal to 6.02 x 1023
molecules, but that is not really a very useful number. So, in
biotechnology, it is easier to use this definition: The unit 1 mole is
the mass, in grams, equal to the molecular weight (MW), also
called formula weight (FW), of the substance. The FW can be
determined by using a Periodic Table or by adding the atomic weights
of the atoms in the molecule. An easy way, though, is to just read the
label of a chemical reagent bottle, which lists the MW or FW. The
molecular weight of NaCl is 58.5 atomic mass units (amu) since the Na
atom weighs 23 amu, and a Cl atom weighs 35.5 amu.
Molarity concentrations are reported as the number of moles per liter
(mol/L or M). If the concentration is very low, then the concentration
could be reported in millimoles/liter (mmol/L or mM). If you wanted a
1-M NaCl solution, you would measure out 1 mole of NaCl (58.5 g) and
dissolve it in water to a total volume of 1 L. This gives you 1 mole of
NaCl per liter of solution, 1 M NaCl.
A liter of solution is a large volume for most research and development
purposes. In research and development labs, mL or L quantities are
usually used. To determine how to mix up a smaller volume of a
solution of some molarity, follow the example below.
Multiply the volume desired (L) by the concentration (molarity) desired
(mol/L), as you did in the mass volume calculations. Then, multiply the
result by the compounds molecular weight (g/mol) to account for
measuring in moles, as in the following equation:
Molarity Concentration Equation
volume x
molarity x
molecular weight =
grams of solute to be dissolved in
wanted
desired
of the solute solvent to the final desired
volume
(L)
(mol/L)
(g/mol)
Convert smaller or larger units to these as necessary. The L units
cancel out and the mol units cancel out, leaving the mass in grams
of the solutes needed to make the solution.

Solution Preparation 2

Ex. A technician needs 50 mL of 0.5 M NaCl solution for an experiment.


Using the Molarity Concentration Equation, the calculation would be as
follows:
0.05 L x 0.5 mol/L x 58.5 g/mol = 1.46 g NaCl
The technician would add 1.46 g NaCl to a container and fill up to the
50 mL mark with solvent.
Determine the calculations for the solutions in the Practice Problems
section for Part B. Making Solutions Using Molarity Concentrations at
the end of this handout.

Part C. Making Dilutions of Concentrated Solutions

Making dilutions of concentrated solutions is a common practice in a


biotechnology lab. A concentrated solution is generally called a stock
solution, and the diluted solution is called the working solution.
Preparing a concentrated stock solution saves a lot of time and is
easier to store than large volumes of diluted working solutions. Making
a working solution simply requires diluting some volume of stock
solution to the concentration needed.
When a number of dilutions must be made, and each is proportionally
the same dilution as the one before, it is called a serial dilution. Doing
a serial dilution makes sense for many experiments when many
samples of varying concentrations are needed. A serial dilution is also
useful for preparing very dilute solutions that are hard to make from
scratch, because the solute masses can be too small to measure on a
balance.
Each succeeding sample is made with the
same ratio of sample and diluent as the one before
500 mL
1M NaCl

1 M NaCl

500 mL
0.5M NaCl

0.5 M NaCl

500 mL
0.25M NaCl

0.25 M NaCl

0.125 M NaCl

Each of the above dilutions is one part previous sample and one part
solvent. This is called a 1:2 dilution, or one part sample to two total
parts.
To figure out how to prepare a working solution from a stock solution,
we use the process bulleted below:
Restate the problem:
o What do you have, i.e., what is the concentration of the stock
solution and how much of it do you have?

Solution Preparation 3

o What is needed, i.e., what is the concentration of the working


solution and how much of it do you need?
Convert all concentrations and volumes to the same units.
Calculate the Dilution Factor: Concentration of
stock/Concentration of working solution
Calculate the volume of stock needed: Volume needed/Dilution
factor
Calculate the volume of solvent (usually water) needed: Volume
of working solution needed Volume of stock needed

For example, a technician needs to prepare 150 mL of 0.1 M TRIS (the


working solution), from 100 ml of a stock soltuion of 1 M TRIS.
What you have: 100 ml of 1.0 M Tris
What you need: 150 ml of 0.1M Tris
All volumes and concentrations are already in the same units
The dilution factor = 1M Tris/0.1M Tris = 10
The volume of stock needed = 100/10 = 10ml
The volume of solvent needed = 100 ml 10 ml = 90 ml of
water.
Determine the calculations for the solutions in the Practice Problems
section for Part C. Making Dilutions of Concentrated Solutions at the
end of this handout.

Solution Preparation 4

Practice Problems
Part A. Making Solutions Using Mass/Volume
Concentrations

1. Describe how you would prepare 25 mL of a NaCl solution at a


concentration of 2.5 g/mL.

2. Describe how you would prepare 2 L of a 0.5 g/mL dextrose solution.

Part B. Making Solutions Using Molarity


Concentrations

1. Describe how you would prepare 125 mL of a 10 M NaOH solution.

2. Describe how you would prepare 75 mL of a 0.1 M NaCl solution.

Solution Preparation 5

Part C. Making Dilutions of Concentrated Solutions


1.

Describe how you would prepare 950 mL of a 1M CuSO45H2O


solution from a 25M CuSO45H2O stock.

2.

Describe how you would prepare 50 mL of a 5 mM NaCl solution


from a 1 M NaCl stock.

Solution Preparation 6