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Dilutions

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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 liter

milligrams per milliliter

micrograms per milliliter

micrograms per microliter

nanograms per liter

nanograms per microliter

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.

Concentrations

Solution Preparation 1

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

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.

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

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

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

concentration of 2.5 g/mL.

Concentrations

Solution Preparation 5

1.

solution from a 25M CuSO45H2O stock.

2.

from a 1 M NaCl stock.

Solution Preparation 6

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