A.

Units, Amounts and Concentrations

Units and abbreviations

The common units, and their respective abbreviations, used in biomedical sciences
are based on the International System of Measurements (SI units): grams (g), metres
(m) etc. Often it is necessary to deal with large quantities e.g. 1500 g or, more
commonly, very small quantities, e.g. 0.0000015 g. In such cases, such numbers are
either expressed by use of powers of 10 (positive or negative) or by use of the
appropriate prefix.

Thus: 1500 g = 1.5 x 103 g = 1.5 kg (kilograms)

and 0.0000015 g = 1.5 x 10 -6 g = 1.5 g (micrograms)

Laboratory scientists most commonly use the prefix notation, particularly those that
differ by a factor of 1000 in magnitude, so you should try to become familiar with them.
Use of the prefixes can greatly simplify calculations, especially mental ones.

Note also that biomedical scientists normally express volumes and concentrations in
terms of litres rather than in cubic measurements:

e.g. 1 litre, rather than 1 dm3

1 millilitre (1 ml) rather than 1 cm3

A concentration of 1 milligram per litre is also still commonly expressed in the form 1
mg/l rather than 1 mg.l-1 or 1 mg.dm-3.

The commonly used prefixes are:

Prefix Name Which modifies an amount by Examples

m milli 1/1000th, i.e. by 10-3 mmol, mg, ml

micro 1/1000 000th, i.e. by 10-6 mol, g, l

n nano by 10-9 nmol, ng

p pico by 10-12 pmol, pg

f femto by 10-15 fmol, fg

k kilo by 1000 times, i.e. by103 kg

M mega by 106 MPa

The prefixes centi (10-2) and deci (10-1) are only commonly used in specific cases e.g.
cm

Concentrations
The determination of the concentration of a substance in a biological fluid is central to
many areas of medical and dental practice (e.g. electrolytes in serum or glucose in
urine). It is important, therefore, that concentrations are expressed in clear
unambiguous terms. There are several ways of doing this. The simplest is to
express the concentration as the weight or mass of the substance per unit volume:

e.g. 10 g/l or 20 mg/ml or 2 g/ml

Another way is to express the concentration of a solution or mixture in terms of per

cent (%). This is a somewhat outdated method but you may still come across it,
particularly if you read the older medical literature, so you should know what it means.
Note that there are different types of % concentration:

% (v/v) (volume by volume)

% (w/v) (weight by volume)
% (w/w) (weight by weight)

A 1% (v/v) concentration is obtained by diluting 1 volume of a substance into 100

volumes (total) of solution, e.g. 1 ml ethanol diluted with water to a final volume of
100 ml gives a 1% (v/v) ethanol solution.

A 1% (w/v) concentration is obtained by dissolving 1 g of substance in a final

volume of 100 ml solution, e.g. 1 g glucose dissolved in water to a final volume of
100 ml solution gives a 1% (w/v) glucose solution.

A 1% (w/w) concentration is obtained by mixing 1 g of substance with something

else in a total weight of 100 g, e.g. 1% (w/w) salt in sand.

Another old method of expressing concentration that you may still see (just look at the
side of a tube of toothpaste) is "parts per million (ppm)". One ppm is one part of
anything in one million parts of total material, e.g. 1 g of compound X in a million g
total, or 1 litre of Y in a million litres total.

Moles and molarity

By far the most important unit defining an amount of a biological substance is the
mole, with the corresponding concentration being the molarity. As far as possible,
the concentrations of specific compounds in serum, urine etc, are now expressed as
molarities in clinical laboratories.

One mole of a substance is the molecular weight of that substance expressed in

grams. Thus, the molecular weight of glucose is 180, so:

1 mole of glucose = 180 g

1 mmol glucose = 180 mg
1 mol glucose = 180 g
100 mol glucose = 18,000 g = 18 mg

Note the abbreviations: 1 mmol = 1 millimole; 2 mmol = 2 millimoles; 5 mol

= 5 micromoles.

Concentrations in molarities are given by expressing the number of moles of the

substance present in a defined volume of solution:

A 1 molar (1 M) solution contains 1 mole per litre (1 mol/l)

a 1 millimolar (1 mM) solution contains 1 millimole per litre (1 mmol/l)

Note: mol and moles mean the same thing (an amount) and moles/litre (long
winded but correct) is a concentration and can be expressed as mol/l, or mol.l-1, or
(best and simplest of all) M. Ensure you know the distinction between
concentrations and amounts.

So, if you dissolve 0.5 mol of a compound in one litre of solvent the concentration of
the compound is 0.5 mol/l, or 0.5 M. 100ml of the solution contains 0.05 mol.

Prefix notation
The value of the prefix notation can now be seen as it allows rapid mental calculations
to be performed (after much practice!). The following concentrations are all the same:

0.5 M, 0.5 mol/l, 0.5 mmol/ml, 0.5 mol/l, 0.5 pmol/pl

You see that by scaling both the units (amount and volume) in the concentration up or
down by a factor of 1000, the value of the concentration remains the same. If you only
scale one of the terms (amount or volume), then you can express the same
concentration in yet more ways:

0.5 mol/l = 0.5 mmol/ml = 500 mol/ml = 500,000 pmol/l

Now, lets say you wanted to determine the amount of cholesterol in the blood of a
newborn infant. You know it will be around 5 mM. The detection limit of the method
you are going to use is about 20 nmol and you can only take 20 l of blood. Will this
be enough?

5 mM cholesterol contains 5 mmol/l, or 5 mol/ml, or 5

nmol/l.

It is easy to see now that 20l contains 100 nmol of

cholesterol - ENOUGH.

Examples
How many mol are dissolved in 2 l of a 20 mM solution?

20 mM = 20 mmol/l, so 2 l contain 40 mmol. 40 mmol = 40,000 mol.

The molecular weight of NaCl is 58. How many mg are in 50 mol of NaCl?

1 mol of NaCl is 58 g, so 1 mol is 58 g, so 50 mol is 2,900 g = 2.9 mg.

What is the molarity of a 1% (w/v) solution of glucose? (molecular weight = 180)

1% (w/v) contains 1 g in 100 ml and, therefore, 10 g in 1 litre.

A 1 M solution of glucose contains 180 g/l, so 10 g/l represents a molarity of
10/180 = 0.056 M (or 56 mM).

B. Dilutions
This is another source of great confusion. Most experiments require you to make
dilutions of reagents, either before use or as a consequence of the actual assay.
When you are asked to make a ten-fold dilution of a reagent, the objective is to
produce a solution that has a reagent concentration one-tenth of the original. It
follows that the molecules of reagent that occupied a volume of "x" before must now
occupy a volume of "10x". This is obtained by adding to one volume of reagent to nine
volumes of diluent (sometimes referred to as a 1 plus 9 dilution or, more commonly,
a "1 in 10" dilution).

When a solution of known concentration is diluted, it is obvious that the concentration

will fall. Less obvious is the amount by which it falls! Take a typical example:
A solution of a compound is maintained as a stock solution at 5 mM. What is the final
concentration of the compond in which 0.15 ml of stock solution is mixed with 0.4 ml of
buffer and 0.2 ml of water?

There are many ways to derive this answer. Here are a few different approaches
all equally acceptable. Take the one that seems most logical to you!

i. The final volume is 0.75 ml, therefore the concentration of the substrate
will be 0.15/ 0.75 x 5 mM = 1 mM

ii. 5 mM is 5 mmol/l, or 5 mol/ml. Therefore 0.15 ml of stock solution

contains 0.15 x 5 = 0.75 mol. This is expanded into a volume of 0.75 ml
and therefore there are now 0.75 mol/0.75 ml, or 1 mol/ml, or 1 mmol/l,
or 1 mM

iii. The formula for a dilution is v1 x c1 = v2 x c2 (where c1 and c2 are the

concentrations before and after dilution, and v1 and v2 are the volumes
before and after dilution). Therefore, c2 = v1/v2 x c1, which works out to be
c2 = 0.15 / 0.75 x 5 mM , i.e. 1 mM

Note that the last and first are identical, except that the latter defines the problem as a
formula. It may be valuable for dilutions when a solution of known concentration must
be diluted to a new concentration. The formula will give the new volume, but
remember that part of that volume will come from the sample before dilution! Method
2 seems tortuous with this example, but it suits some to work this way.

Lastly, remember to think about the answer its not unusual to see erroneous
calculations giving serum concentrations of metabolites in the tens or hundreds of
molar - try to imagine 10 M glucose in the blood? (the molecular weight of glucose is
180 imagine a cross between a Mars bar and black pudding!). You should always
do rough calculations to ensure that your answers make biological (and logical!)
sense.