Download 2.2.21. Fluorimetry

2.2.22. Atomic emission spectrometry
EUROPEAN PHARMACOPOEIA 5.0
electrode or 2 indicator electrodes) immersed in the solution
to be examined and maintained at a constant potential
difference as a function of the quantity of titrant added.
The potential of the measuring electrode is sufficient to
ensure a diffusion current for the electroactive substance.
Apparatus. The apparatus comprises an adjustable voltage
source and a sensitive microammeter ; the detection system
generally consists of an indicator electrode (for example,
a platinum electrode, a dropping-mercury electrode, a
rotating-disc electrode or a carbon electrode) and a reference
electrode (for example, a calomel electrode or a silver-silver
chloride electrode).
A three-electrode apparatus is sometimes used, consisting of
an indicator electrode, a reference electrode and a polarised
auxiliary electrode.
Method. Set the potential of the indicator electrode as
prescribed and plot a graph of the initial current and the
values obtained during the titration as functions of the
quantity of titrant added. Add the titrant in not fewer than
3 successive quantities equal to a total of about 80 per cent
of the theoretical volume corresponding to the presumed
equivalence point. The 3 values must fall on a straight
line. Continue adding the titrant beyond the presumed
equivalence point in not fewer than 3 successive quantities.
The values obtained must fall on a straight line. The point
of intersection of the 2 lines represents the end-point of the
titration.
For amperometric titration with 2 indicator electrodes, the
whole titration curve is recorded and used to determine the
end-point.
Method. Dissolve the substance to be examined in the
solvent or mixture of solvents prescribed in the monograph,
transfer the solution to the cell or the tube of the fluorimeter
and illuminate it with an excitant light beam of the
wavelength prescribed in the monograph and as near as
possible monochromatic.
Measure the intensity of the emitted light at an angle of
90° to the excitant beam, after passing it through a filter
which transmits predominantly light of the wavelength of
the fluorescence. Other types of apparatus may be used
provided that the results obtained are identical.
For quantitative determinations, first introduce into the
apparatus the solvent or mixture of solvents used to dissolve
the substance to be examined and set the instrument to zero.
Introduce the standard solution and adjust the sensitivity
of the instrument so that the reading is greater than 50. If
the second adjustment is made by altering the width of the
slits, a new zero setting must be made and the intensity of
the standard must be measured again. Finally introduce
the solution of unknown concentration and read the result
on the instrument. Calculate the concentration cx of the
substance in the solution to be examined, using the formula :
cx
=
concentration of the solution to be examined,
cs
=
concentration of the standard solution,
Ix
=
intensity of the light emitted by the solution to
be examined,
01/2005:20220 Is
=
intensity of the light emitted by the standard
solution.
2.2.20. POTENTIOMETRIC TITRATION
In a potentiometric titration the end-point of the titration
is determined by following the variation of the potential
difference between 2 electrodes (either one indicator
electrode and one reference electrode or 2 indicator
electrodes) immersed in the solution to be examined as a
function of the quantity of titrant added.
The potential is usually measured at zero or practically zero
current.
Apparatus. The apparatus used (a simple potentiometer or
electronic device) comprises a voltmeter allowing readings
to the nearest millivolt.
The indicator electrode to be used depends on the substance
to be determined and may be a glass or metal electrode (for
example, platinum, gold, silver or mercury). The reference
electrode is generally a calomel or a silver-silver chloride
electrode.
For acid-base titrations and unless otherwise prescribed,
a glass-calomel or glass-silver-silver chloride electrode
combination is used.
Method. Plot a graph of the variation of potential difference
as a function of the quantity of the titrant added, continuing
the addition of the titrant beyond the presumed equivalence
point. The end-point corresponds to a sharp variation of
potential difference.
If the intensity of the fluorescence is not strictly proportional
to the concentration, the measurement may be effected using
a calibration curve.
In some cases, measurement can be made with reference
to a fixed standard (for example a fluorescent glass or a
solution of another fluorescent substance). In such cases,
the concentration of the substance to be examined must
be determined using a previously drawn calibration curve
under the same conditions.
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2.2.22. ATOMIC EMISSION
SPECTROMETRY
Atomic emission spectrometry is a method for determining
the concentration of an element in a substance by measuring
the intensity of one of the emission lines of the atomic
vapour of the element generated from the substance. The
determination is carried out at the wavelength corresponding
to this emission line.
Apparatus. This consists essentially of an atomic generator
of the element to be determined (flame, plasma, arc, etc.),
a monochromator and a detector. If the generator is a
flame, water R is the solvent of choice for preparing test and
reference solutions, although organic solvents may also be
01/2005:20221 used if precautions are taken to ensure that the solvent does
not interfere with the stability of the flame.
Method. Operate an atomic emission spectrometer in
2.2.21. FLUORIMETRY
accordance with the manufacturer’s instructions at the
prescribed wavelength setting. Introduce a blank solution
Fluorimetry is a procedure which uses the measurement
into the atomic generator and adjust the instrument reading
of the intensity of the fluorescent light emitted by the
to zero. Introduce the most concentrated reference solution
substance to be examined in relation to that emitted by a
and adjust the sensitivity to obtain a suitable reading.
given standard.
General Notices (1) apply to all monographs and other texts
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