DILUTIONS

It is often necessary to make dilutions of specimens being ana- lyzed or to make

weaker solutions from stronger solutions in various laboratory procedures.
Clinicians must be able to work with various dilution problems and dilution
factors. They often need to determine the concentration of antibody in each solu-
tion, the actual amount of material in each solution, and the total volume of each
solution. All dilutions are a form of ratio. Dilution is an indication of relative
concentration.
Diluting Specimens
In most laboratory determinations, a small sample is taken for analysis and the
final result is expressed as concentration per some convenient standard volume. In
a certain procedure, 0.5 mL of blood is diluted to a total of 10 mL with various
and 1 mL of this dilution is then analyzed for a particular chemical
constituent.The final result is to be expressed in terms of the concentration of
that substance per 100 mL of blood.
Dilution Factor
A dilution factor is used to correct for having used a diluted sample in a
determination rather than the undiluted sample. The result (answer) using the
dilution must be multiplied by the reciprocal of the dilution made. For example, a
dilution fac- tor by which all determination answers are multiplied to give the
concentration per 100 mL of sample (blood) may be calcu- lated as follows.
First, determine the volume of blood that is actually ana- lyzed in the procedure.
Using a simple proportion, it is evident that 0.5 mL of blood diluted to 10 mL is
equivalent to 1 mL of blood diluted to 20 mL:
0.5 mL blood 1 mL blood 10 mL solution = x mL solution
x= 1 mL blood×10 mL =20 mL 0.5 mL
The concentration of specimen (blood) in each milliliter of solution may be
determined by the use of another simple pro- portion to be 0.05 mL of blood per
milliliter of solution:
1 mL blood x mL blood 20 mL solution = 1 mL solution
x= 1 mL×1 mL =0.05 mL 20 mL
Because 1 mL of the 1:20 dilution of blood is analyzed in the remaining steps of
the procedure, 0.05 mL of blood is actually analyzed (1 mL of the dilution used ×
0.05 mL/mL = 0.05 mL of blood analyzed).
To relate the concentration of the substance measured in the procedure to the
concentration in 100 mL of blood (the units in which the result is to be
expressed), another proportion may be used:
the desired terms of measurement. Thus, in the previous exam- ple, the following
equations may be used:
0.5 mL
(volume of blood used) =
10 mL
(volume of total dilution)
x mL
(volume of blood analyzed)
1 mL
(volume of dilution used)
x = 0.05 mL (volume of blood actually analyzed) 100 mL (volume of blood
required for expression of result)
0.05 mL (volume of blood actually analyzed)
Single Dilutions
= 2000 (dilution factor)
100 mL
(volume of blood desired)
0.05 mL (volume of blood used)
The concentration of the substance being measured in the vol- ume of blood actually
tested (0.05 mL) must be multiplied by 2000 to report the concentration per 100 mL
of blood.
The preceding material may be summarized by the follow- ing statement and
equations. In reporting results obtained from laboratory determinations, one must
first determine the amount of specimen actually analyzed in the procedure and then
calculate the factor that will express the concentration in
When the concentration of a particular substance in a speci- men is too great to be
determined accurately, or when there is less specimen available for analysis than
the procedure requires, it may be necessary to dilute the original specimen or
further dilute the initial dilution (or filtrate). These single dilutions are
usually expressed as a ratio, such as 1:2, 1:5, or 1:10, or as a fraction, 1⁄2,
1⁄5, or 1⁄10. These ratios or fractions refer to 1 unit of the original specimen
diluted to a final volume of 2, 5, or 10 units, respectively. A dilution,
therefore, refers to the volume or number of parts of the substance to be diluted
in the total volume, or parts, of the final solution. A dilution is an expression
of concentration, not volume; it indicates the relative amount of substance in
solution. Dilutions can be made singly or in series.
To calculate the concentration of a single dilution, multi- ply the original
concentration by the dilution expressed as a fraction.
Example of Calculation of Concentration of a Single Dilution
A specimen contains 500 mg of substance per deciliter of blood. A 1:5 dilution of
this specimen is prepared by volumet- rically measuring 1 mL of the specimen and
adding 4 mL of diluent. The concentration (C) of substance in the dilution is
calculated as follows:
C=500 mg/dL×1/ =100 mg/dL 5
Note that the concentration of the final solution (or dilution) is expressed in the
same units as that of the original solution.
To obtain a dilution factor that can be applied to the determi- nation answer and
express it as a concentration per standard vol- ume, proceed as follows. Rather
than multiply by the dilution expressed as a fraction, multiply the determination
value by the reciprocal of the dilution fraction. In the case of a 1:5 dilution,
the dilution factor applied to values obtained in the procedure would be 5, because
the original specimen was five times more concentrated than the diluted specimen
tested in the procedure.