EUROPEAN PHARMACOPOEIA 10.0 2.2.34.

Thermal analysis

01/2016:20234 The technique is used to determine the difference in heat

flow (with reference to the temperature) evolved or absorbed
by the test sample compared with the reference cell, as a
function of the temperature. Two types of DSC instruments
are available, those using power compensation to maintain
2.2.34. THERMAL ANALYSIS(3) a null temperature difference between sample and reference
and those that apply a constant rate of heating and detect
Thermal analysis is a group of techniques in which the temperature differential as a difference in heat flow between
variation of a physical property of a substance is measured sample and reference.
as a function of temperature. The most commonly used Instrument. The instrument for power compensation DSC
techniques are those which measure changes in the mass or consists of a furnace containing a sample holder with a
energy of a sample of a substance. reference cell and a test cell. The instrument for heat flow DSC
These techniques have different applications : consists of a furnace containing a single cell with a sample
– determination of phase changes ; holder for the reference crucible and the test crucible.
– determination of changes in chemical composition ; A temperature-programming device, thermal detector(s) and
– determination of purity. a recording system which can be connected to a computer
are attached. The measurements are carried out under a
THERMOGRAVIMETRY controlled atmosphere.
Thermogravimetry (TG) or thermogravimetric analysis Calibration of the instrument. Calibrate the instrument
(TGA) is a technique in which the mass of a sample of a for temperature and enthalpy change, using suitable certified
substance is recorded as a function of temperature according materials or reference standards.
to a controlled temperature programme.
Temperature calibration. It can be performed using certified
Instrument. The essential components of a thermobalance reference materials having an intrinsic thermal property, such
are a device for heating or cooling the substance according as the melting point of pure metals or organic substances,
to a given temperature programme, a sample holder in a or the phase transition point of crystalline inorganic salts or
controlled atmosphere, an electrobalance and a means of oxides. Melting points of indium, tin and/or zinc are usually
electronic signal output to a recorder or a computer. employed for calibration.
Temperature calibration. The temperature sensor close to
or in contact with the sample is calibrated using the Curie Heat-quantity calibration. For accurate estimation of
temperature of a ferromagnetic substance such as nickel. the quantity of heat change (enthalpic change) of a test
In the case of an instrument capable of simultaneously sample, caused by a certain physical change accompanying a
conducting TG/TGA and differential thermal analysis (DTA) temperature change, it is necessary to calibrate the instrument
or differential scanning calorimetry (DSC), the same certified using suitable certified reference materials. Similar to
reference materials as those for DTA and DSC may be used, temperature calibration, heat-quantity calibration may be
for example, indium, tin and/or zinc. performed using suitable certified reference materials showing
a known definite enthalpic change caused by physical changes,
Calibration of the electrobalance. Place an appropriate such as the melting of pure metals and/or organic substances,
quantity of a suitable certified reference material (e.g. calcium or the phase transition of crystalline inorganic salts. The heats
oxalate monohydrate CRS) in the sample holder and record of fusion of indium, tin and/or zinc are usually employed for
the mass. Set the heating rate according to the manufacturer’s calibration.
instructions (e.g. 5 °C/min) and start the temperature
increase. Record the thermogravimetric curve as a graph Operating procedure. Weigh in a suitable crucible an
with temperature or time on the abscissa, increasing from left appropriate quantity of the substance to be examined and
to right, and mass on the ordinate, decreasing downwards. place it in the sample holder. Place an empty crucible in the
Stop the temperature increase at about 250 °C. Measure reference holder. Set the initial and final temperatures, and the
the difference on the graph between the initial and final heating rate according to the operating conditions prescribed
mass-temperature or mass-time plateaux, which corresponds in the monograph.
to the loss of mass. The declared loss of mass for the certified Begin the analysis and record the DSC curve with the
reference material is stated on the label. temperature or time on the abscissa (values increasing from
Method. Apply the same procedure to the substance to be left to right) and the energy change on the ordinate (specify
examined using the conditions prescribed in the monograph. whether the change is endothermic or exothermic).
Calculate the loss of mass of the substance to be examined The temperature at which the phenomenon occurs (the
from the difference measured in the graph obtained. Express onset temperature) corresponds to the intersection (A)
the loss of mass as Δm/m (per cent). of the extension of the baseline with the tangent at the
If the instrument is in frequent use, carry out temperature point of greatest slope (inflexion point) of the curve (see
calibration regularly. Otherwise, calibration is carried out Figure 2.2.34.-1). The end of the thermal phenomenon is
before each measurement. indicated by the peak of the curve.
As the operating conditions are critical, the following The enthalpy of the phenomenon is proportional to the area
parameters are noted for each measurement : pressure or under the curve limited by the baseline ; the proportionality
flow rate, gas composition, mass of the sample, heating rate, factor is determined from the measurement of the heat of
temperature range and sample pre-treatment including any fusion of a known substance (e.g. indium) under the same
isothermal period. operating conditions.
DIFFERENTIAL SCANNING CALORIMETRY Each thermogram may be accompanied by the following
Differential scanning calorimetry (DSC) is a technique that data : conditions employed, record of last calibration, mass
can be used to demonstrate the energy phenomena produced of the sample and identification (including thermal history),
during heating (or cooling) of a substance (or a mixture of container, atmosphere (identity, flow rate, pressure), direction
substances) and to determine the changes in enthalpy and and rate of temperature change, instrument and recorder
specific heat and the temperatures at which these occur. sensitivity.
(3) This chapter has undergone pharmacopoeial harmonisation. See chapter 5.8 Pharmacopoeial harmonisation.

General Notices (1) apply to all monographs and other texts 61

2.2.34. Thermal analysis EUROPEAN PHARMACOPOEIA 10.0

Figure 2.2.34.-1. – Thermogram

Applications Determination of purity. The measurements of the fraction

Phase changes. Determination of the temperature, heat of substance melted at a given temperature and the heat of
capacity change and enthalpy of phase changes undergone by fusion by DSC enable the impurity content of a substance
a substance as a function of temperature. The transitions that to be determined from a single thermal diagram, requiring
may be observed include those shown in Table 2.2.34.-1. the use of only a few milligrams of sample with no need for
repeated accurate measurements of the true temperature.
Table 2.2.34.-1.
In theory, the melting of an entirely crystalline, pure substance
solid - solid transition : allotropy - polymorphism at constant pressure is characterised by a heat of fusion ΔHf
desolvation in an infinitely narrow range, corresponding to the melting
amorphous-crystalline point T0. A broadening of this range is a sensitive indicator
of impurities. Therefore, samples of the same substance,
solid - liquid transition : melting whose impurity contents vary by a few tenths of a per
glass-transition cent, give thermal diagrams that are visually distinct (see
Figure 2.2.34.-2).
solid - gas transition: sublimation
The determination of molar purity by DSC is based on the
liquid - solid transition : freezing use of a mathematical approximation of the integrated form
recrystallisation of the van’t Hoff equation applied to the concentrations
glass-transition (not the activities) in a binary system [ ln(1 − x2) » -x 2 and
T  ´ T0 » T02 ]. For low amounts of impurities ( x 2  1) and
liquid - gas transition: evaporation for temperatures close to the melting point T0 the equation
can be written as follows, in which T and x 2 are variables :
Changes in chemical composition. Measurement of heat
and temperatures of reaction under given experimental RT02
conditions, so that, for example, the kinetics of decomposition T = T0 - ´ x2 (1)
ΔH f
or desolvation can be determined.
Application to phase diagrams. Establishment of phase
diagrams for solid mixtures. The establishment of a phase
diagram may be an important step in the preformulation and
optimisation of the freeze-drying process.

Figure 2.2.34.-2. – Thermal diagrams according to purity

62 See the information section on general monographs (cover pages)

EUROPEAN PHARMACOPOEIA 10.0 2.2.35. Osmolality

T = temperature of the sample, in kelvins ; is dependent on the molal concentration of the solute(s) in
the solution, on their dissociation and on the deviation of the
T0 = melting point of the chemically pure substance, in solution from ideal behaviour (Raoult’s law).
kelvins ;
The unit of osmolality is the osmole per kilogram (osmol/kg),
R = gas constant for ideal gases, in but the submultiple milliosmole per kilogram (mosmol/kg) is
joules·kelvin− 1·mole− 1 ; more commonly used.
ΔHf = molar heat of fusion of the pure substance, in The osmolality (ξm) of a solution containing i solutes is given
joules·mole− 1 ; by the expression :
x2 = mole fraction of the impurity, i.e., the number
of molecules of the impurity divided by the total ξm = å νimiΦm , i
number of molecules in the liquid phase (or molten vi = number of entities formed by the dissociation
phase) at temperature T (expressed in kelvins). of one molecule of the ith solute ; if the solute is
Hence, the determination of purity by DSC is limited to the non-ionic (non-dissociating), νi equals 1,
detection of impurities forming a eutectic mixture with the mi = molality of the ith solute in the solution, in moles
principal compound and present at a mole fraction of typically per kilogram of solvent,
less than 2 per cent in the substance to be examined. Φm,i = molal osmotic coefficient, a dimensionless factor.
This method cannot be applied to :
– amorphous substances ; The molal osmotic coefficient is a measure of the deviation
of the solution from ideal behaviour. For an ideal solution,
– solvates or polymorphic compounds that are unstable osmolality equals molality (Φ=1).
within the experimental temperature range ;
In the case of a real, non-ideal solution, the molal osmotic
– impurities forming solid solutions with the principal coefficient is influenced by the interactions occurring amongst
substance ; the components (i.e. molecules, ions, solvent) of the solution.
– impurities that are insoluble in the liquid phase or in the The more complex the composition of the solution, the harder
melt of the principal substance. it becomes to determine Φ.
During the heating of the substance to be examined, the For this reason, the measurement of a colligative property such
impurity melts completely at the eutectic temperature. Above as the freezing-point depression is used as a practical means
this temperature, the solid phase contains only the pure of determining osmolality by obtaining an overall measure of
substance. As the temperature increases progressively from the contribution of the various solutes present in a solution.
the eutectic temperature to the melting point of the pure
substance, the mole fraction of the impurity in the liquid PRINCIPLE OF MEASUREMENT
phase decreases, since the quantity of liquefied pure substance Unless otherwise prescribed, osmolality is determined by
increases. measuring the freezing-point depression (ΔTf) of a solution.
The relationship between osmolality and freezing-point
For all temperatures above the eutectic point : depression is given by the expression :
1
x2 = ´ x 2* (2) ΔT f = k f ξm
F
where kf is the molal cryoscopic constant, which is
F = molten fraction of the analysed sample ; solvent-dependent. For water, the value of kf is 1.86 K/osmol
x 2* = mole fraction of the impurity in the analysed (i.e. adding 1 mol of a non-dissociating solute to 1 kg of water
sample. results in a decrease in freezing-point of 1.86 K).
When the entire sample has melted, F = 1 and x 2 = x 2* . EQUIPMENT
If equation (2) is combined with equation (1), the following An osmometer for freezing-point depression measurement
equation is obtained : typically consists of :
RT02 1 – an appropriate sample container ;
T = T0 - ´ ´ x 2* – a means of cooling the sample ;
ΔH f F
– a temperature-sensitive resistor (thermistor), with an
The value of the heat of fusion of the pure substance is appropriate current or potential difference measurement
obtained by integrating the melting peak. device that can indicate a temperature depression or give
The melting point T0 of the pure substance is extrapolated osmolality values directly ;
from the plot of temperature T (expressed in kelvins) versus – a means of mixing the sample and/or inducing solidification
1/F. The slope α of the curve (obtained after linearisation, when supercooling occurs.
x*
if necessary) corresponding to RT02 ΔH2 , allows x 2* to be
f PROCEDURE
evaluated.
CALIBRATION
The fraction x 2* multiplied by 100 gives the mole fraction
in per cent for total eutectic impurities. Prepare reference solutions as specified in Table 2.2.35.-1,
as necessary, using dried sodium chloride R. Commercially
available certified solutions for osmometer calibration, with
07/2019:20235 osmolalities equal or similar to those listed in Table 2.2.35.-1,
may be used. Calibrate the equipment according to the
manufacturer’s instructions using water R to determine the
zero value and at least 2 of the reference solutions listed in
Table 2.2.35.-1. Confirm the calibration using at least one
2.2.35. OSMOLALITY additional reference solution with a known osmolality (see
Table 2.2.35.-1). Select a reference solution preferably with
PRINCIPLE an osmolality within ± 50 mosmol/kg of the expected value
GENERAL for the solution to be examined or close to the centre of the
Osmolality is a measure of the total number of chemical expected osmolality range of the solutions to be examined. It
entities per kilogram of solvent, and thus provides an is recommended that the reading is within ± 4 mosmol/kg of
indication of the osmotic pressure of the solution. Osmolality the osmolality of the chosen reference solution.

General Notices (1) apply to all monographs and other texts 63