Examples:
Making Solutions
Two simple examples are presented here. A third
example is of a complex solution for which the
description lists the concentrations of components
using different expressions.
Weight in volume: Prepare 2 liters 0.85% sodium
chloride
With 1% defined as 1 gram per 100 ml, 0.85% is
0.85 grams per 100 ml. Since two liters is
20x the volume of 100 ml, we need 20 x 0.85
grams which is 17 grams NaCl. For this quantity
we can use a top loading balance or even a
trip balance.
A typical electronic balance is accurate to one
hundredth of a gram, which is sufficiently accurate
for weighing out 17 grams. First we "tare" the
instrument by placing a weigh boat onto the pan
and setting it to “zero.” We don't
want to contaminate our chemical stocks, so we
either clean the spatula or spoon before dipping
it into the container or we simply shake the chemical
out onto the boat.
Suppose that we tap out 16.97 grams of NaCl. Should
we go to the trouble to get that last 0.03 gram?
Nope! Consider that if it was
necessary to be more accurate, we would describe
the formula as something like 0.846% NaCl, or maybe
0.8495%. If there is some advantage to being precise
then we should exercise precision, otherwise trying
to be too precise just wastes time.
Remember how to use significant digits? Seventeen
grams means greater than 16.5 grams and less than
17.5 grams. If we wanted to be more accurate we
would write “17.0” grams, meaning greater
than or equal to 16.95 grams and less than or equal
to 17.05 grams.
Molarity: Prepare 200 ml of 70 mM sucrose
Suppose that you need 200 milliters of a 70 mM
solution of sucrose. Two hundred milliliters
is 0.2L and 70 mM is 0.07M. The molecular weight
of sucrose can be determined from its chemical
formula, namely C12H22O11 and the atomic weights
of carbon, hydrogen, and oxygen. The formula
weight for sucrose is identical to its molecular
weight, namely 342.3 grams per mole. A 1M solution
would consist of 342.3 grams sucrose in one
liter final volume.
A concentration of 70 mM is the same as 0.07 moles
per liter. Take 0.07 moles/liter times 342.3 grams
per mole and you have 23.96 grams needed per liter.
To make 200 milliliters of your solution multiply
grams/liter by liters needed. Since 200 milliliters
is 0.2L, multiply 23.96 grams by 0.2L to get 4.792
grams needed. Since a typical top loading electronic
balance displays mass to the nearest 0.01 gram,
the amount to be weighed should be rounded to 4.79
grams, although it is perfectly acceptable and
perhaps even preferable to round to 4.8 grams.
Complex solution: Prepare a sample buffer for
SDS-PAGE
The following formula describes the composition
of the 2x concentrated buffer that we use to denature
proteins for electrophoresis. The formula descriptions
v/v or w/w would not be listed in a methods section
since it is obvious which components are liquids
or solids.
50% (v/v) concentrated SDS-PAGE stacking gel buffer,
pH 6.8
4.6% (w/v) sodium dodecyl sulfate
20% (v/v) glycerol
160 mM dithiothreitol (Cleland's reagent)
0.01% bromphenol blue dye
For 100 ml of sample buffer a 125 ml erlenmeyer
flask is an appropriate mixing vessel. The liquid
components will take up 70% of the total volume
so we start by placing 50 ml stacking gel buffer
and 20 ml glycerol in the flask. Glycerol is very
viscous, so to be accurate you might use a syringe
to deliver the stuff. We need 4.6 grams (4.6%)
of sodium dodecyl sulfate (also called lauryl sulfate).
To mix it evenly it should be added while stirring
the solution. The concentration of dye is also
given as weight-in-volume. One hundredth of 1%
is only 0.1 mg/ml. You'll need just 10 mg of it.
Because the bromphenol blue serves as a tracking
dye and its concentration is not critical, you
can weigh out something close to 10 mg or just
use a "pinch" by adding the amount on the narrow
end of a spatula.
The formula weight for Cleland's reagent is 154
grams/mole. The amount to weigh is given by (0.16
mole/L)(0.1 L)(154 gms/mole) = 2.46 gms (rounds
off to 2.5 gms).
Most of the time it is not necessary to heat a
solution to mix it, but in this case the detergent
does not go into solution completely until it is
heated. After popping the flask in a microwave
oven for a minute or so on a low setting and stirring
a bit the solution should be ready to pour into
a 100 ml graduated cylinder for topping off with
distilled water.
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