Calibrations,

Standardizations, and
Blank Corrections
 An analytical method is
standardized by determining its
sensitivity
 In a quantitative analysis, we
measure a signal and calculate the
amount of analyte using one of
the following equations.
Smeas = knA + Sreag
or Smeas = kCA + Sreag
 Where
Smea measured signal
K the method’s sensitivity
Sreag. any signal due to the reagents,
nA is the moles of analyte
CA is the analyte's concentration
 To obtain accurate results
we must eliminate
determinate errors
affecting the measured signal
This is accomplished by combination
of
Calibrations,
Standardizations,
Reagent blanks
Calibrating Signals
Signals are measured using
equipment or instruments that
must be properly calibrated
 To ensure that Smeas is determined
accurately, we calibrate the equipment
or instrument used to obtain the signal.
Calibration is accomplished against a
standard, adjusting Smeas until it
agrees with the standard’s known
signal.
When the signal is a measurement of
mass, Smeas is determined with an
analytical balance. Before a balance can
be used, it must be calibrated against a
reference weight meeting standards
established by either the National
Institute for Standards and Technology
or the American Society for Testing and
Materials
 Balances are calibrated using
standard weights. When necessary,
we can also correct for the
buoyancy of air
With an electronic balance the sample’s
mass is determined by the current
required to generate an upward
electromagnetic force counteracting the
sample’s downward gravitational force.
The balance’s calibration procedure
invokes an internally programmed
calibration routine specifying the
reference weight to be used.
The reference weight is placed on the
balance’s weighing pan, and the
relationship between the displacement
of the weighing pan and the
counteracting current is automatically
adjusted.
An object’s true weight in vacuo, Wv, is
related to its weight in air, Wa, by the
equation

where Do is the object’s density,

Dw is the density of the calibration weight,
0.0012 is the density of air under normal laboratory conditions
(all densities are in units of g/cm 3).
Clearly the greater the difference between Do and Dw
the more serious the error in the object’s measured weight.
Volumetric glassware can be calibrated by
measuring the mass of water
contained or delivered and
using the density of water
to calculate the true volume
 Example:
A 10-mL volumetric pipette was calibrated by following the procedure just
outlined, using a balance calibrated with brass weights having a density
of 8.40 g/cm3. At 25 °C the pipette was found to dispense 9.9736 g of
water. What is the actual volume dispensed by the pipette?
SOLUTION
At 25 °C the density of water is 0.99705 g/cm3. The water’s true weight,
therefore, is

and the actual volume of water dispensed by the pipette is

If the buoyancy correction is ignored, the pipette's volume is reported as

Introducing a negative determinate error of –0.11%.

Most instruments have calibration
standards suggested by the
manufacturer.
3.2. Standardizing Methods
The American Chemical Society’s
Committee on Environmental
Improvement defines standardization as
(the process of determining the
relationship between the measured
signal and the amount of analyte).
A method is considered
standardized when the value of k
in equation 1 or 2 is known.
. Reagents Used as Standards
Primary reagent
Reagents used as standards are divided into
primary reagents and secondary reagents. A
primary reagent (A reagent of known purity
that can be used to make a solution of known
concentration) can be used to prepare a
standard containing an accurately known
amount of analyte.
A primary reagent must:
1-have a known stoichiometry,
2-have a known purity (or assay)
3-stable during long-term storage both
in solid and solution form.
Because of the difficulty in establishing the degree
of hydration, even after drying, hydrated
materials usually are not considered primary
reagents. Reagents not meeting these criteria are
called secondary reagents. (A reagent whose
purity must be established relative to a primary
reagent.) The purity of a secondary reagent in
solid form or the concentration of a standard
prepared from a secondary reagent must be
determined relative to a primary reagent.
Other Reagents Preparing a standard often requires
additional substances that are not primary or
secondary reagents. When a standard is prepared
in solution, for example, a suitable solvent and
solution matrix must be used. Each of these
solvents and reagents is a potential source of
additional analyte that, if unaccounted for, leads to
a determinate error. If available, reagent grade
chemicals conforming to standards set by the
American Chemical Society should be used.
Types of Water
 There are several approaches to
standardization, including
the use of external standards,
the method of standard addition,
and the use of an internal standard.
Preparing Standard Solutions

Solutions of primary standards generally are

prepared in class A volumetric glassware to
minimize determinate errors.
3.4.Single-point standardization

Any standardization using a single standard

containing a known amount of analyte.
The simplest way to determine the value of k in
equation 2 is by single point standardization.
A single standard containing a known
concentration of analyte, CS, is prepared and
its signal, Sstand, is measured. The value of k
is calculated as 3
Multiple-point standardization

 The preferred approach to standardizing a method is to

prepare a series of standards, each containing the analyte
at a different concentration. Standards are chosen such
that they bracket the expected range for the analyte’s
concentration. Thus, multiple-point standardization
(Any standardization using two or more standards
containing known amounts of analyte.) should use at
least three standards, although more are preferable. A
plot of Sstand versus CS is known as a calibration curve.
The exact standardization, or calibration relationship, is
determined by an appropriate curve-fitting algorithm
The most desirable standardization
strategy is an external
standardization.
External Standards
The most commonly employed standardization method
uses one or more external standards (A standard
solution containing a known amount of analyte,
prepared separately from samples containing the
analyte) containing known concentrations of analyte.
These are identified as external standards because they
are prepared and analyzed separately from the
samples. A quantitative determination using a single
external standard was described at the beginning of
this section, with k given by equation 4. Once
standardized, the concentration of analyte, CA, is
given as External Standards
A multiple-point external
standardization is accomplished
by constructing a calibration
curve
The method of standard additions, in
which known amounts of analyte
are added to the sample, is used
when the sample’s matrix
complicates the analysis.
An internal standard, which is a
species (not analyte) added to all
samples and standards, is used
when the procedure does not
allow for the reproducible
handling of samples and
standards.
Standardizations using a single
standard are common, but also
are subject to greater
uncertainty. Whenever possible,
multiple point standardization is
preferred
internal standard (A standard, whose
identity is different from the analyte’s,
that is added to all samples and standards
containing the analyte)
The results of multiple-point
standardization are graphed as a
calibration curve.
A linear regression analysis can provide
an equation for the standardization.
 A reagent blank corrects the
measured signal for signals due to
reagents other than the sample that
are used in an analysis.
The most common reagent blank is
prepared by omitting the sample.
When a simple reagent blank does
not compensate for all constant
sources of determinate error,
other types of blanks, such as the
total Youden blank, can be used