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
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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 C) 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)

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Basic Principles of Thermal Analysis

Modern instrumentation used for thermal analysis usually consists of four parts:
sample/sample holder sensors to detect/measure a property of the sample and the temperature an enclosure within which the experimental parameters may be controlled a computer to control data collection and processing

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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.

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Heat Flux DSC

Sample holder :
sample and reference are connected by a low-resistance heat flow path Aluminium, stainless, platinum sample pans heating
coil

Sensors:
temperature sensors usually thermocouples
sample pan reference pan

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

Temperature controller:
temperature difference between the sample and reference is measured
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Power Compensated DSC

Sample holder :
Aluminium, platinum, stainless steel pans
individual heaters
sample pan reference pan

Sensors:
Pt resistance thermocouples. Separate sensors and heaters for the sample and reference

inert gas vacuum thermocouple T = 0

inert gas vacuum

Furnace: Temperature controller:

separate blocks for sample and reference cells

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
evaluation of the thermal resistance of the sample and reference sensors measurements over the temperature range of interest

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DSC Calibration
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

Heat flow
use calibration standards of known heat capacity, slow accurate heating rates (0.52.0 C/min), and similar sample and reference pan weights metals Indium 156.6 C; 28.45 J/g Zinc 419.47C, 108.17 J/g inorganics KNO3 128.7 C KClO4 299.4 C organics polystyrene 105 C benzoic acid 122.3 C; 147.3 J/g
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calibrants
high purity accurately known enthalpies thermally stable light stable not hygroscopic do not react (pan, atmosphere)

Sample Preparation
accurately-weighed samples (~3-20 mg, usually 3-5 mg for simple powders) small sample pans (0.1 mL) of inert or treated metals (Al, Pt, stainless) several pan configurations, e.g., open , pinhole, or hermetically-sealed pans same material and configuration should be used for the sample and the reference material should completely cover the bottom of the pan to ensure good thermal contact 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 sensitivity

Al
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Pt

alumina

Ni

Cu

quartz

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)

Exo endothermic events melting sublimation solid-solid transitions desolvation chemical reactions exothermic events crystallization solid-solid transitions decomposition chemical reactions baseline shifts glass transition

sulphapyridine

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Melting Processes by DSC

Pure substances linear melting curve melting defined by temperature point onset
eutectic melt

Impure substances Broad, asymmetric melting peak melting characterized peak maxima at

Melting with decomposition exothermic endothermic

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eutectic impurities may produce a second peak

Definition of Transition Temperature

0.5
156.50C 28.87J/g

0.0

Exo
-0.5

extrapolated onset temperature

Heat Flow (W/g)

-1.0

-1.5

-2.0

peak melting temperature

157.81C

-2.5 140
Exo Up

145

150

155

160

165

170

175
Universal V3.3B TA Instruments

Temperature (C)

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Enthalpy of Fusion
0.5
156.50C 28.87J/g

0.0

Exo

-0.5

Heat Flow (W/g)

-1.0

-1.5

-2.0

157.81C

-2.5 140
Exo Up

145

150

155

160

165

170

175
Universal V3.3B TA Instruments

Temperature (C)

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Enthalpy of Fusion by DSC

For a single (well-defined) melting endotherm area under peak minimal decomposition/sublimation readily measured for high melting polymorph can be measured for low melting polymorph More difficult where multiple thermal events leading to stable melt
e.g. solid-solid transitions (A to B) before melt, or where melt / recrystallisation before melt Estimate from sum all areas
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Endo

Purity by DSC
1-3 mg samples in hermeticallysealed pans are recommended Peak width a valuable measure of purity: impurities lower the melting point Less pure (non-perfect) crystals melt first followed by purer larger crystals polymorphism interferes with purity determination, especially when a transition occurs in the middle of the melting peak Accurate measurement of Hf needs pure samples of polymorphs
EYP 2006 benzoic acid

Exo
97%

99%

99.9%

Plato, C.; Glasgow, Jr., A.R. Anal. Chem., 1969, 41(2), 330-336.

Glass Transitions
characterized by change in heat capacity (no heat absorbed or evolved) transition from a disordered solid to a liquid appears as a step (endothermic direction) in the DSC curve gradual enthalpy change may occur, producing an endothermic peak superimposed on the glass transition Exo

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Effect of Heating Rate

many transitions (evaporation, crystallization, decomposition, etc.) are kinetic so shift to higher temp. when heated at a higher rate increasing the scanning rate increases sensitivity, while decreasing the scanning rate increases resolution to obtain thermal event temperatures close to the true thermodynamic value, slow scanning rates (e.g., 15 K/min) should be used Rapid scanning can obscure thermal events Advantageous in fast scan DSC, e.g. 500K/min
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Recognizing Artefacts
Sample movement in pan sample pan distortion mechanical Pan shock / knock moves in bench furnace cool air entry into cell

sensor contamination

atmosphere electrical effects, changes power spikes, etc.

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burst of pan lid

Closing / opening pan hole, e.g. sublimation

Ensuring correct interpretation of DSC

You cant 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 120C

Does not necessarily show interaction at room temp
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Polymorph Screening and Indentification

Form II Form Form II Form II Variable Form III Hydrate Dihydrate Acetic acid solvate
Heat Flow (W/g)
1.0 0.5 0.0

thermal stability melting crystallization solid-state transformations desolvation glass transition Exo sublimation decomposition heat flow heat of fusion heat of transition heat capacity

-0.5

-1.0

-1.5

Form II II Form II Variable Form III Hydrate Dihydrate Acetic acid solvate

-2.0

-2.5

50

100

150

200

250

300

350

Exo Up

Temperature (C)

mixture analysis physical purity (crystal forms, crystallinity) chemical purity

phase diagrams / interactions

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Effect of Phase Impurities

lots A & B of polymorph (identical by XRD) are different by DSC:
0
2046742 FILE# 022511DSC.1

Lot A - pure

-1

Heat Flow (W/g)

-2

2046742 FILE# 022458 DSC.1 Form II ?

-3

Lot B - seeds
-4

-5
Exo Up

80

130

180

230

280
Universal V3.3B TA Instruments

Temperature (C)

Lot A: pure low melting polymorph melting observed Lot B: seeds of high melting polymorph induce solid-state transition below the melting temperature of the low melting polymorph
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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 refolding, polymer characterisation
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trehlose

Modulated DSC (MDSC)

introduced in 1993; heat flux design sinusoidal (or square-wave or sawtooth) modulation is superimposed on the underlying heating ramp total heat flow signal contains all of the thermal transitions of standard DSC Fourier Transformation analysis is used to separate the total heat flow into its two components: reversing and non-reversing heat flow increased sensitivity, resolution and the ability to separate multiple thermal events
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Modulated DSC Heating Profile

MDSC for Polymorph Characterization

Heat capacity (reversing heat flow) glass transition melting Kinetic (non-reversing heat flow) crystallization decomposition evaporation
0.2
Reversing (heat flow component)

0.00 -0.05 -0.10 -0.15

reversing heat flow

0.0

non-reversing heat flow

Nonrev Heat Flow (W/g)

Non-reversing (heat flow component)

Lot A

Lot A
-0.2

Rev Heat Flow (W/g)

-0.20 -0.25 -0.30 -0.35 -0.40 -0.45

DSC010622b.1 483518 HCL (POLYMORPH 1) DSC010622d.1 483518 HCL

Lot B

-0.4

Lot B

-0.6
DSC010622b.1 483518 HCL (POLYMORPH 1) DSC010622d.1 483518 HCL

-0.50 110
Exo Up

120

130

140

150

160

170

180
Universal V3.3B TA

-0.8 110
Exo Up

120

130

140

150

160

170

180
Universal V3.3B TA

Temperature (C)

Temperature (C)

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Hyper DSC
Fast scanning DSC Only possible with power compensated Normal equipment 100 C/min Specialised up to 500 C/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.

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Best Practices for Thermal Analysis

proper instrument calibration use purge gas (N2 or He) to remove corrosive off-gases small sample size good thermal contact between the sample and the temperature-sensing device proper sample encapsulation start temperature well below expected transition temperature slow scanning speeds
(Unless aiming to obscure thermal transitions, e.g fast scan DSC)

avoid decomposition in the DSC

(Run TGA first its easier to clean up!)
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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 - 1C 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 1C at 10C/min
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And more caution!

Thermal analysis tells you what is happening at the temperature it happens at! Care when extrapolating to room temperature stability / interaction Dont over-interpret data Care when using thermal analysis in isolation Artefacts / heating rate effects etc Couple with other analytical tools Heated X-ray, heated vibrational spectroscopy, hot stage microscope
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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

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Questions

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Reversing and Non-Reversing Contributions to Total DSC Heat Flow

total heat flow resulting from average heating rate

dQ/dt = Cp . dT/dt + f(t,T)

reversing signal heat flow resulting from sinusoidal temperature modulation (heat capacity component) non-reversing signal (kinetic component)

e.g. see Pharmaceutical Research: 17 (6): 696-700, June 2000 Craig, DQM et al.
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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, T = 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, dq/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
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Thermogravimetric Analysis (TGA)

thermobalance to monitor sample weight as a function of temperature weight calibration known weights using

temperature calibration based on ferromagnetic transition of Curie point standards (e.g., Ni)
120 100
12.15%

80

19.32%

larger sample masses, lower temperature gradients, and higher purge rates minimize undesirable buoyancy effects
60
29.99%

Weight (%)

40

20

20

40

60

80

100

120

140

160

Time (min)

Universal V3.7A TA Instruments

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

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 TG-DSC
80 120
15.55% (0.9513mg) 24.80C 100.0% 179.95C 84.45%

4.2

3.2

Weight (%)

TG-FTIR TG-MS

2.2

40

1.2

0.2 0 -0.8

-40
Exo Up

20

70

120

170

220

270

-1.8

Temperature (C)

Universal V3.3B TA Instruments

TG-DTA trace of sodium tartrate

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Temperature Difference (V/mg)