Differential Scanning

Calorimetry
“Cooking with Chemicals”
Clare Rawlinson
School of Pharmacy
University of Bradford

EYP 2006
 Outline
„ Brief history of thermal analysis

„ Theory of thermal analysis techniques

– Thermal Gravimetric Analysis (TGA)
– Differential Scanning Calorimetry (DSC)

„ Generating valid data

– Calibration
– Sample preparation

„ Interpreting data and Applications

– Real events
– Artefacts

„ Recent advances

EYP 2006
 Calorimetry
„ Calorimetry
– The study of heat transfer during
physical and chemical processes

„ Calorimeter
– A device for measuring the heat
transferred

Lavoisier and Laplace (1782-1784):

„ oil was burned in a lamp (Fig 9) held in
a bucket (Fig. 8) held in a wire mesh
cage (f)
„ surrounded by ice in spaces b and a of
the double walled container a foot in
diameter
„ lid (F) was topped with ice, as was a
mesh lid (not shown) beneath it that
covered the inner volume b
EYP 2006
 Oil lamps to Guinea Pigs…
„ Measured heat production of
the metabolic processes in
the ice bath calorimeter
„ Outer jacket prevented
conduction of heat from the
external environment which
would have also melted the
ice
„ From latent heat of fusion for
ice (334 J/gram ice at 0 ºC)
Lavoisier converted the rate
of water formation to heat
production
„ In 10 hours 370 grams of ice
melted

Guinea pig produced 12,358 J per hour of heat

(12.4 kJ/hr)
EYP 2006
 Basic Principles of Thermal Analysis

Modern instrumentation used for thermal analysis usually

consists of four parts:

ƒ sample/sample holder

ƒ sensors to detect/measure a property of the sample and the

temperature

ƒ an enclosure within which the experimental parameters may be

controlled

ƒ a computer to control data collection and processing

EYP 2006
 TGA and DSC
„ Thermogravimetric Analysis (TGA)
– mass change of a substance measured as function of
temperature whilst the substance is subjected to a controlled
temperature programme1
– mass is lost if the substance contains a volatile fraction

„ Differential Scanning Calorimetry (DSC)

– provides information about thermal changes that do not involve a
change in sample mass1
– more commonly used technique than TGA
– Two basic types of DSC instruments: heat-flux and power
compensation

1Haines, P. J. (2002) The Royal Society of Chemistry, Cambridge.

EYP 2006
 Heat Flux DSC
Sample holder :
ƒ sample and reference are connected by a low-resistance heat flow
path
ƒ Aluminium, stainless, platinum sample pans heating
coil

Sensors:
„ temperature sensors sample reference
„ usually thermocouples pan pan

Furnace:
ƒ one block for both sample
and reference cells inert gas
vacuum thermocouples

Temperature controller:
• temperature difference between the sample and reference is
measured
EYP 2006
 Power Compensated DSC
Sample holder :
ƒ Aluminium, platinum, stainless steel pans
individual
heaters
Sensors:
ƒ Pt resistance sample reference
thermocouples. pan pan

ƒ Separate sensors
and heaters for the
sample and reference inert gas
vacuum
inert gas
vacuum
thermocouple ΔT = 0
Furnace:
ƒ separate blocks for sample and reference cells

Temperature controller:
ƒ differential thermal power is supplied to the heaters to maintain the
temperature of the sample and reference at the program value
EYP 2006
 Outline
„ Brief history of thermal analysis

„ Theory of thermal analysis techniques

– Thermal Gravimetric Analysis (TGA)
– Differential Scanning Calorimetry (DSC)

„ Generating valid data

– Calibration
– Sample preparation

„ Interpreting data and Applications

– Real events
– Artefacts

„ Recent advances

EYP 2006
 DSC Calibration

Baseline
Calibration

ƒ evaluation of the
thermal resistance of
the sample and
reference sensors

ƒ measurements
over the temperature
range of interest

EYP 2006
 DSC Calibration
ƒ Temperature
• match melting onset temperatures to the known melting points of
standards analyzed by DSC
• should be calibrated as close to desired temperature range as possible
ƒ Heat flow
• use calibration standards of known heat capacity, slow accurate heating
rates (0.5–2.0 °C/min), and similar sample and reference pan weights

metals
• Indium 156.6 °C; 28.45 J/g
calibrants • Zinc 419.47°C, 108.17 J/g
• high purity inorganics
• accurately known enthalpies • KNO3 128.7 °C
• thermally stable • KClO4 299.4 °C
• light stable organics
• not hygroscopic • polystyrene 105 °C
• do not react (pan, atmosphere) • benzoic acid 122.3 °C; 147.3 J/g
EYP 2006
 Sample Preparation
ƒ accurately-weighed samples (~3-20 mg, usually 3-5 mg for simple
powders)
ƒ small sample pans (0.1 mL) of inert or treated metals (Al, Pt, stainless)
ƒ several pan configurations, e.g., open , pinhole, or hermetically-sealed
pans
ƒ same material and configuration should be used for the sample and the
reference
ƒ material should completely cover the bottom of the pan to ensure good
thermal contact
ƒ avoid overfilling the pan to minimize thermal lag from the bulk of the
material to the sensor

* small sample masses and

low heating rates
increase resolution, but
at the expense of Al Pt alumina Ni Cu quartz
sensitivity
EYP 2006
 Purge Gases
„ Sample may react with air - oxidising or burning
„ Control moisture content of atmosphere
„ Use inert gas e.g. nitrogen or argon
„ Flowing purge gas
„ In some cases deliberately choose reactive gas, e.g.
– hydrogen to reduce an oxide to metal
– carbon dioxide which affects decomposition of metal carbonate

„ Removes waste products from sublimation or

decomposition

EYP 2006
 Outline
„ Brief history of thermal analysis

„ Theory of thermal analysis techniques

– Thermal Gravimetric Analysis (TGA)
– Differential Scanning Calorimetry (DSC)

„ Generating valid data

– Calibration
– Sample preparation

„ Interpreting data and Applications

– Real events
– Artefacts

„ Recent advances

EYP 2006
 Typical Features of a DSC Trace
(Polymorphic System)
endothermic events
Exo melting
sublimation
solid-solid transitions
desolvation
chemical reactions

exothermic events
crystallization
sulphapyridine solid-solid transitions
decomposition
chemical reactions

baseline shifts
glass transition
EYP 2006
 Melting Processes by DSC

Pure substances Impure

substances
• linear melting curve
• Broad,
• melting point eutectic
melt asymmetric
defined by onset melting peak
temperature
• melting
characterized at
Melting with
peak maxima
decomposition
• eutectic impurities
• exothermic
may produce a
second peak
• endothermic

EYP 2006
 Definition of Transition Temperature
0.5

156.50°C
28.87J/g
0.0
Exo
extrapolated
-0.5
onset temperature
Heat Flow (W/g)

-1.0

-1.5

peak melting
-2.0
temperature
157.81°C

-2.5
140 145 150 155 160 165 170 175
Exo Up Temperature (°C) Universal V3.3B TA Instruments

EYP 2006
 Enthalpy of Fusion
0.5

156.50°C
28.87J/g
0.0
Exo

-0.5
Heat Flow (W/g)

-1.0

-1.5

-2.0

157.81°C

-2.5
140 145 150 155 160 165 170 175
Exo Up Temperature (°C) Universal V3.3B TA Instruments

EYP 2006
 Enthalpy of Fusion by DSC

For a single (well-defined)

melting endotherm
ƒ area under peak
ƒ minimal
decomposition/sublimation
ƒ readily measured for high
melting polymorph
ƒ can be measured for low
melting polymorph Endo

More difficult where multiple thermal events leading to stable melt

ƒ e.g. solid-solid transitions (A to B) before melt, or where melt /
recrystallisation before melt
ƒ Estimate from sum all areas

EYP 2006
 Purity by DSC
ƒ 1-3 mg samples in hermetically-
sealed pans are recommended
ƒ Peak width a valuable measure Exo
of purity:
ƒ impurities lower the melting
97%
point
ƒ Less pure (non-perfect)
crystals melt first followed by 99%
purer larger crystals
ƒ polymorphism interferes with benzoic acid 99.9%
purity determination, especially
when a transition occurs in the
middle of the melting peak
ƒAccurate measurement of
Plato, C.; Glasgow, Jr., A.R. Anal.
ΔHf needs pure samples of
Chem., 1969, 41(2), 330-336.
polymorphs
EYP 2006
 Glass Transitions
ƒ characterized by change in heat capacity (no heat absorbed or
evolved)
ƒ transition from a disordered solid to a
liquid
ƒ appears as a step (endothermic
direction) in the DSC curve
ƒ gradual enthalpy change may occur,
producing an endothermic peak
superimposed on the glass transition
Exo

EYP 2006
 Effect of Heating Rate
ƒ many transitions (evaporation,
crystallization, decomposition, etc.) are
kinetic so shift to higher temp. when
heated at a higher rate
ƒ increasing the scanning rate increases
sensitivity, while decreasing the
scanning rate increases resolution
ƒ to obtain thermal event temperatures
close to the true thermodynamic value,
slow scanning rates (e.g., 1–5 K/min)
should be used
ƒ Rapid scanning can obscure thermal
events
ƒAdvantageous in fast scan DSC,
e.g. 500K/min
EYP 2006
 Recognizing Artefacts
sample mechanical
Sample Pan cool air entry
pan shock / knock
movement in moves in into cell
distortion bench
pan furnace

sensor
contamination

atmosphere Closing /
electrical effects, changes opening pan
power spikes, etc. hole, e.g.
burst of
pan lid sublimation
EYP 2006
Ensuring correct interpretation of DSC

„ You can’t
„ Can minimise misinterpretation
„ Essential to have valid data to interpret
– Calibration, reproducible data, appropriate sampling etc

„ Kinetics / thermodynamics at elevated temps

– High temp can speed kinetics – event would happen at room
temperature but slowly
– Effect activated by increased temp (overcome activation energy)
- event would not happen at room temperature
„ DSC shows excipients interact at 120ºC
– Does not necessarily show interaction at room temp

EYP 2006
Polymorph Screening and Indentification
–––––– Form
Form II 1.0

–––––– Form IIII

Form
–––––– Variable
Form III Hydrate 0.5
–––––– Dihydrate
–––––– Acetic acid solvate 0.0

Heat Flow (W/g)

-0.5
„ thermal stability
– melting -1.0

– crystallization -1.5

– solid-state transformations
––––––– Form II
––––––– Form IIII
––––––– Variable
Form III Hydrate
––––––– Dihydrate
-2.0

– desolvation
––––––– Acetic acid solvate

– glass transition -2.5

0 50 100 150 200 250 300 350

– sublimation Exo Exo Up Temperature (°C)

– decomposition „ mixture analysis

„ heat flow
– physical purity (crystal
forms, crystallinity)
– heat of fusion
– chemical purity
– heat of transition
– heat capacity „ phase diagrams / interactions

EYP 2006
 Effect of Phase Impurities
ƒ lots A & B of polymorph (identical by XRD) are different by DSC:

2046742
Lot A - pure
FILE# 022511DSC.1

-1
Heat Flow (W/g)

-2

2046742
FILE# 022458 DSC.1 Form II ?
-3

Lot B - seeds
-4

-5
80 130 180 230 280
Exo Up Temperature (°C) Universal V3.3B TA Instruments

ƒ Lot A: pure low melting polymorph – melting observed

ƒ Lot B: seeds of high melting polymorph induce solid-state transition
below the melting temperature of the low melting polymorph
EYP 2006
 Outline
„ Brief history of thermal analysis

„ Theory of thermal analysis techniques

– Thermal Gravimetric Analysis (TGA)
– Differential Scanning Calorimetry (DSC)

„ Generating valid data

– Calibration
– Sample preparation

„ Interpreting data and Applications

– Real events
– Artefacts

„ Recent advances

EYP 2006
 Microcalorimetry
„ High sensitivity DSC
„ Solutions
„ Scan range typically
0-120 °C
„ Scanning rate of 0-120
°C/hr
„ Reverse scan rate 0-45
°C/hr
(depending on efficiency
of cooling tank)
„ Useful for looking at low
energy modifications
„ e.g. protein relaxation and trehlose
refolding, polymer
characterisation
EYP 2006
 Modulated DSC (MDSC)
ƒ introduced in 1993; “heat flux” Modulated DSC Heating Profile
design
ƒ sinusoidal (or square-wave or
sawtooth) modulation is
superimposed on the underlying
heating ramp
ƒ total heat flow signal contains all
of the thermal transitions of
standard DSC

ƒ Fourier Transformation analysis is used to separate the total heat flow

into its two components: reversing and non-reversing heat flow
ƒ increased sensitivity, resolution and the ability to separate multiple
thermal events

EYP 2006
 MDSC for Polymorph Characterization
Heat capacity Kinetic
(reversing heat flow) (non-reversing heat flow)
glass transition crystallization
melting decomposition
evaporation

0.00 0.2
Reversing (heat flow component) Non-reversing (heat flow component)

-0.05 reversing heat flow non-reversing heat flow

-0.10 0.0

-0.15 Lot A Lot A

Nonrev Heat Flow (W/g)

Rev Heat Flow (W/g)

-0.20 -0.2

-0.25

-0.30
Lot B -0.4
Lot B
-0.35

-0.40 -0.6

-0.45 DSC010622b.1 483518 HCL (POLYMORPH 1) DSC010622b.1 483518 HCL (POLYMORPH 1)

DSC010622d.1 483518 HCL DSC010622d.1 483518 HCL

-0.50 -0.8
110 120 130 140 150 160 170 180 110 120 130 140 150 160 170 180
Exo Up Temperature (°C) Universal V3.3B TA Exo Up Temperature (°C) Universal V3.3B TA

EYP 2006
 ‘Hyper’ DSC
„ Fast scanning DSC
„ Only possible with power compensated
„ Normal equipment ≈ 100 ºC/min
„ Specialised up to 500 ºC/min
„ Increased sensitivity, loss of resolution
„ e.g. amorphous content in mainly crystalline sample
– change of specific heat at Tg is linear relationship to the
amorphous content
– Conventional DSC 10% amorphous limit of detection
– Hyper DSC <1% amorphous easily detected

Lappalainen, M., I. Pitkanen, et al. (2006). International Journal of

Pharmaceutics 307(2): 150-155.

EYP 2006
 Best Practices for Thermal Analysis
ƒ proper instrument calibration
ƒ use purge gas (N2 or He) to remove corrosive off-gases
ƒ small sample size
ƒ good thermal contact between the sample and the
temperature-sensing device
ƒ proper sample encapsulation
ƒ start temperature well below expected transition temperature
ƒ slow scanning speeds
(Unless aiming to obscure thermal transitions, e.g fast scan DSC)
ƒ avoid decomposition in the DSC
(Run TGA first – its easier to clean up!)
EYP 2006
 Caution…
„ It is a bulk tool
– Analysing the gross average of events in a sample
– Conversely, small powder sample in DSC may not
represent packing of powder bulk in
decomposition studies

„ Instrument error in DSC typically ± 0.5 - 1ºC

„ In Scanning modes, thermal events may be

“smeared” by a thermal lag
– Sample temperature not keeping up with
instrument
– Bigger effect at higher heating rates
– Typically 1ºC at 10ºC/min

EYP 2006
 And more caution!
„ Thermal analysis tells you what is happening at
the temperature it happens at!
– Care when extrapolating to room temperature
stability / interaction

„ Don’t over-interpret data

„ Care when using thermal analysis in isolation

ƒ Artefacts / heating rate effects etc
ƒ Couple with other analytical tools
– Heated X-ray, heated vibrational
spectroscopy, hot stage microscope

EYP 2006
 Acknowledgements

Professor Adrian Williams, University of Reading

Dr Ian Grimsey, University of Bradford
Dr Peter Timmins, Bristol Myers Squibb

Dr Wendy Hulse, University of Bradford

Luciana DeMatos, University of Sheffield

EYP 2006
Questions

EYP 2006
 Reversing and Non-Reversing
Contributions
to Total DSC Heat Flow

total heat flow

resulting from
dQ/dt = Cp . dT/dt + f(t,T)
average heating
rate
reversing signal non-reversing
heat flow resulting from signal
sinusoidal temperature (kinetic
modulation component)
(heat capacity component)

e.g. see Pharmaceutical Research: 17 (6): 696-700, June 2000 Craig, DQM et al.

EYP 2006
 Some Common Thermal Analysis Techniques
There are various techniques in which a physical property is measured as a
function of temperature, while the sample is subjected to a predefined
heating or cooling program.

Differential Thermal Analysis (DTA)

• the temperature difference between a sample and an inert reference material, ΔT = TS -
TR, is measured as both are subjected to identical heat treatments

Differential Scanning Calorimetry (DSC)

• the sample and reference are maintained at the same temperature, even during a
thermal event (in the sample)

• the energy required to maintain zero temperature differential between the sample and
the reference, dΔq/dt, is measured

Isothermal titration calorimetry (ITC)

• The temperature of a “reaction” is kept constant whilst the energy change is measured

Thermogravimetric Analysis (TGA)

• the change in mass of a sample on heating is measured

EYP 2006
 Thermogravimetric Analysis (TGA)
• thermobalance to monitor
sample weight as a function
of temperature

• weight calibration using

known weights

• temperature calibration
based on ferromagnetic
120

transition of Curie point

standards (e.g., Ni)
100
12.15%

80 19.32%
Weight (%)

• larger sample masses, lower

60

temperature gradients, and 29.99%

higher purge rates minimize

40

undesirable 20
buoyancy
effects
0 20 40 60 80 100 120 140 160
Time (min) Universal V3.7A TA Instruments

EYP 2006
 Differential Thermal Analysis
Sample holder: Sample and reference cells

Sensors: Thermocouples, one for the sample

and one for the reference

Furnace: Block containing sample and

reference cells

Temperature controller: Controls temperature

program

Advantages:
Disadvantages:
„ instruments can be used at very high
temperatures „ uncertainty of heats of
„ instruments are highly sensitive
„ flexibility in sample volume/form
fusion and transition
„ characteristic transition or reaction temperatures
temperatures can be determined

EYP 2006
 “Hyphenated” Techniques
• thermal techniques alone are insufficient to prove the existence of polymorphs
and solvates
• other techniques should be used, e.g., microscopy, diffraction, and spectroscopy

• development of “hyphenated” techniques for simultaneous analysis

TG-DTA 120
4.2
15.55%
(0.9513mg)

TG-DSC 24.80°C
100.0%
3.2

80

Temperature Difference (µV/mg)

179.95°C
84.45%
2.2
TG-FTIR
Weight (%) 40 1.2

TG-MS
0.2

-0.8

-40 -1.8
20 70 120 170 220 270
Exo Up Temperature (°C) Universal V3.3B TA Instruments

TG-DTA trace of sodium tartrate

EYP 2006