Thermal Analysis

Applications
Booklet

Thermal Analysis of Food

Application Examples
2 METTLER TOLEDO Thermal Analysis
Introductory Handbook

Thermal Analysis of Food

Application Examples
Preface

Thermal analysis was first used as a standard method for the investigation of polymers. However, it is well proven
that thermal analysis also benefits the testing of food and food ingredients. Techniques and methods dedicated to
testing of food products such as proteins, carbohydrates, fats, oils and packaging materials have been well devel-
oped over recent years.

Proteins, together with fats and carbohydrates, are among the most important constituents of food due to their sig-
nificance in nutrition and physiology and their role in the development of functional properties. They are important
for processing food (e.g. emulsification) and for qualifying the consumable product (sensory properties). Differential
Scanning Calorimetry (DSC) and Thermogravimetric Analysis (TGA) are regarded as being the main techniques
applied to improving the functional properties and process conditions.

Heat and time are two of the main parameters in food processing. DSC allows cooking and freezing processes to
be investigated in respect to heat and time. It can also be used to determine other properties of food products and
ingredients.

Plastic films make up approximately 10% by weight of packaged food products. The variety of film composites
designed for particular applications is enormous. Analysis of films is necessary to check their barrier properties and
composition. Individual film components as well as auxiliary materials (e.g. adhesives) can be identified by DSC.
Thermomechanical Analysis (TMA) and Dynamic Mechanical Analysis (DMA) can also provide additional informa-
tion regarding the mechanical properties of packaging materials.

We hope that the applications described here will be of interest and

• make you aware of the potential of thermal analysis methods for the investigation of food and related materials.
• provide you with some hints to assess the quality of the results.
• encourage you to explore these topics further by reading the recommended application handbooks,
or by attending respective webinars.

Disclaimer

This application handbook presents selected application examples. The experiments were conducted with the utmost
care using the instruments specified in the description of each application. The results were evaluated according to
the current state of our knowledge.
This does not however absolve you from personally testing the suitability of the examples for your own methods,
instruments and purposes. Since the transfer and use of an application is beyond our control, we cannot of course
accept any responsibility.

When chemicals, solvents and gases are used, general safety rules and the instructions
given by the manufacturer or supplier must be observed.
® TM All names of commercial products can be registered trademarks, even if they are not denoted as such.

METTLER TOLEDO Thermal Analysis 3

4 METTLER TOLEDO Thermal Analysis
Contents

Preface 3

1 What Thermal Analysis Can Do for Food  6

2 Applications and Their Thermal Analysis Techniques 8

3 Examples 9
3.1 Protein denaturation of four seeds 9
3.2 Influence of the starch content on swelling in water 11
3.3 Influence of moisture on milk powder 13
3.4 Crystallization of vegetable oils 14
3.5 Identification of plastic films by DSC 16

4 Food Regulatory Compliance 18

5 Method Development 20

6 Excellent Crucibles, Accessories and Reference Materials 22

7 For More Information 23

METTLER TOLEDO Thermal Analysis 5

 1. What Thermal Analysis Can Do for Food

The most important effects or properties for food and food ingredients that can be investigated by thermal
analysis are discussed briefly in the following sections.

1.1 Proteins
Proteins are long chain molecules consisting of one or more polypeptides typically folded into a globular or a
fibrous form. A polypeptide is a single linear polymer of amino acids linked together by so-called peptide bonds.
They differ not only in their molecular composition but also in their spatial structure, a factor that is decisive in
the development of the techno-functional properties.

Denaturation of Proteins
When a protein is denatured, the secondary and tertiary structures are altered but the peptide bonds of the
primary structure between the amino acids are left intact. Since all structural levels of the protein determine
its function, the protein can no longer perform its function once it has been denatured. Protein denaturation is
the reaction that occurs most frequently in food processing. It is a collective term that describes a multitude of
chemical reactions. Denaturation can be induced thermally as well as by treatment with detergents, acids and
bases or through mechanical stress. Once initiated, the reactions run in parallel or consecutively rapidly, and
hence only the states “native” and “denatured” can be distinguished. In general, denaturation occurs in foods
as a complex, usually irreversible system change. DSC detects denaturation as an endothermic1 signal, usually
in the range 40 to 100°C. The DSC signal also yields information on the fraction of denatured protein in the
sample.
1
endothermic: consumes heat

Influence of Process Parameters on Protein Quality

The development of techno-functional properties of the proteins such as the ability to bond with water,
emulsifying power, foam stabilization or aroma complexation depends to a large extent on their “intactness”. The
degree of denaturation can be directly correlated with a number of techno-functional properties.
DSC is an ideal tool for optimizing protein processing methods. In addition, it can be applied to intermediate
products in a multi-stage process. It can also easily measure samples of different consistency (solid/liquid and
liquid/solid dispersions as well as emulsions).
DSC is further applied to evaluate the influence of thermal treatment (freezing, heating, drying), pH, water
content, the addition of salts, sugars, minerals or detergents on the quality of the protein fractions of fish
muscle, milk and whey, hen’s eggs, bovine muscle, grain, oil seeds, beans. It measures quality criteria such as
solubility, texture, emulsifying power, or surface activity.

1.2 Carbohydrates and starch

In the field of carbohydrates, DSC is mainly used to characterize the gelatinization and the retrogradation of
starches. Processes such as the baking of bread, the extrusion of grain products or the thickening of sauces are
based on the gelatinization of starches. Many techniques are used to investigate the properties of starches (e.g.
viscosity measurement, X-ray diffraction, microscopic investigations, etc.). DSC however has a wide application
range with respect to both temperature and possible water/starch ratios.
The thermal degradation of carbohydrates is most easily investigated by thermogravimetric analysis (TGA).

The Gelatinization of Starch

The gelatinization of starch in water on heating is the result of a number of conformational changes. In the
course of the reaction, the crystallinity of the starch granules is reduced, heat is absorbed and hydration of
the starch occurs with swelling of the starch granules. Further uptake of water leads to an increase in volume
through to the complete breakup of the oriented structure of the granules. This process occurs when water is
available in excess of 60%. The gelatinization process of native starches is expected to produce an endothermic
peak between 50 and 80 °C. DSC is the preferred measurement technique.

6 METTLER TOLEDO Thermal Analysis

Additional applications
Thermal analysis is applied in food development by characterizing modified starches (e.g. acetylation, hydrolysis, etc.). These
techniques can also be used to evaluate the swelling properties and behavior, determine the thermo-mechanical properties (e.g. of
pasta or candies) and to follow the thermal decomposition process.

1.3 Fats, oils and lipids

Lipids are a broad group of naturally occurring molecules which includes fats, waxes, sterols, fat-soluble vitamins (such as
vitamins A, D, E and K), monoglycerides, diglycerides, phospholipids, and others. Fats (solids) or oils (liquid) are a subgroup of
lipids called triglycerides. This category of molecules is important for many forms of life, serving both structural and metabolic
functions. The investigation of fats and oils is one of the most frequently performed routine applications of DSC in food processing.
The solidification properties of fats are of particular technological interest for the processing of such foodstuffs. For instance, the
crystallization of cocoa butter is a process that is crucial for the quality of chocolate. This is the step in which chocolate products
obtain their desired properties such as glaze, breaking strength and melting behavior (e.g. “softness”).

Polymorphism
Particular attention must be paid to polymorphism of the fats, i.e. triglycerides of the same composition can differ in their
arrangement in the crystal lattice and show hence different melting points. Since natural fats such as cocoa butter contain
numerous different triglycerides, these mixed systems do not have a narrow melting point, but melt over a temperature range.

Other effects
The autooxidation of edible oils and fats can have a negative influence on the storage and processing of products that contain fats.
The oxidation rate is relatively low at normal room temperatures, but increases rapidly at higher temperatures in the presence of
oxygen. Above 150°C, for example in roasting or frying, autooxidation begins followed by thermal decomposition.
The stability of a fat (characterized by the temperature at which the oxidation begins) and the kinetics of the oxidation reaction can
both be determined by DSC measurements. This allows, as an example, used fats to be distinguished from fresh fat.

Additional applications
Additional thermal analysis applications include the behavioral study of cooking fat under different conditions, the influence of
emulsifying agents on fat crystal structures and the determination of the solid fat index (liquid reaction), to name a few.

1.4 Water
The water content of food and ingredients has great influence on many food properties. Two examples include:
• Too high water content (activity) can lead to microorganism growth, which consecutively turns the food inedible.
• Too low a water content can make make the food dry and hard, which impacts on the texture and taste.
• Water acts as a plasticizer and my shift the glass transition temperature which in many cases determines the texture of food.
For the analysis of water and moisture content, Karl Fischer instruments and halogen moisture analyzers are often used. However,
to investigate adsorption and desorption properties of water on food products, (TGA) is the ‘method of choice’. TGA can measure, as
a function of temperature or time, how much water is adsorbed by the food.

1.5 Packaging materials

Packaging increasingly contributes to food quality, storage life and marketing. Hence, the evaluation of the most suitable packaging
materials and stringent quality control are prerequisiteo food products being consumed. Thermal analysis techniques are
ideally suited to identifying components of packaging materials such as films, laminates and adhesives. DSC provides valuable
information such as glass transition temperatures, crystallinity formation and melting points. This information helps identify the
polymer type and chemical or physical attributes. In addition, oxidation induction times (OIT) can be routinely measured by DSC to
measure the material quality for longer term stability testing.

Quality control
DSC measurements allow different compositions and conformations of polymer fractions to be determined within minutes.

More details
A comprehensive presentation is given in the application handbook
G. Widmann, T. Oberholzer: Food– Collected applications, Mettler-Toledo AG, 51725004B, © 06/2014

METTLER TOLEDO Thermal Analysis 7

 2. Food Applications and Their Thermal Analysis Techniques

The table shows the effects and properties of the main components of food products that can be investigated by
thermal analysis.

DSC Differential scanning calorimetry

TGA Thermal gravimetric analysis
TMA Thermomechanical analysis
DMA Dynamic mechanical analysis

Main components of food products Suitable TA technique

Effects Protein Lipid Carbo- Water Packaging DSC TGA TMA DMA
hydrate
Compositional analysis • • • • •
Chemical reaction • • • • • • •
Crystallinity • • • •
Denaturation • •
Desorption/drying/evaporation • • •
Enthalpy changes • •
Freezing • • •
Gelatinization • •
Glass transition • • • • •
Identification • • • •
Melting point/melting range • • • •
Oxidative stability • • • •
Polymorphism • • •
Purity determination • •
Solid Fat Index (SFI) • •
Specific heat capacity • •
Swelling • • • •
Thermal stability • • • • • •
Table 1: Overview

8 METTLER TOLEDO Thermal Analysis

3. Examples

The following examples provide a short selection of thermal analysis techniques and methods to food samples.
All examples provide measurement parameters, interpretation remarks and a conclusion.

3.1 Protein denaturation of four seeds

Sample Commercial seeds from health food store

Conditions Measuring cells: DSC
Crucibles: Aluminum standard 40 µL, hermetically sealed
Reference: approximately the same amount of water in a sealed pan
Sample preparation: Mill seeds, aqueous extraction at pH 4.5, centrifuge, discard super-
natant, re-extract at pH 8.5, supernatant evaporated to desired dry
substance content under vacuum.
DSC measurement: Heating from 30 to 110 °C at 5 K/min

Interpretation
Four different untreated vegetable proteins were investigated. Each curve shows an endothermic peak in the
range between 72 °C and 100 °C that is characteristic for protein denaturation. The proteins differ in their thermic
stability and enthalpy of denaturation.
With comparable sample weights and protein concentrations, the integrals of the peaks are a direct measure of
the specific reaction enthalpies.
Spelt contains the proteins that are most temperature sensitive and that have the highest enthalpy of denatur-
ation. Whereas the wheat proteins are already extensively denaturated at 86 °C, the main fraction of the soybean
protein is still native. In this case, denaturation does not set in until 88 °C.
The enthalpies of denaturation for grain and oil seed proteins usually lie in the range 3–10 J/g protein.
The enthalpy changes depend on the phase of the proteins, so that the enthalpy of denaturation of seed proteins
in the original conditions is less than in the dissolved state in aqueous solutions.

METTLER TOLEDO Thermal Analysis 9

 Evaluation
Sample DS content in % Sample weight in mg DS Reaction enthalpy in J/g DS
Spelt 20.0 5.40 17.2
Soybean 13.0 4.78 5.2
Wheat 12.5 3.44 3.5
Lupines 23.0 6.80 4.2
DS: dry substance

The higher the enthalpy of denaturation of the native protein, the better the measurement of the fractions
of denatured protein in processed protein samples by DSC.
The free selection of the heating rate with DSC can be of use to help improve the measurement and detection of
reactions. The higher the heating rate, the larger the detected signal, i.e. the larger the measured peak.
The calculated peak area should of course be the same if the reaction process is the same. However, it should
be noted that at high heating rates the reaction is shifted to higher temperatures.
Identification of vegetable proteins by DSC is an application that is less likely to be used in practice and would
also be complicated since the reaction enthalpy can vary depending on the variety and growth conditions.
A native protein must, therefore, always be measured as a reference when estimating the degree of denaturation.
The DSC curve is a ‘fingerprint’ of the measured sample. Such fingerprints of protein fractions in foods are
characteristic of the condition of the protein.

Conclusion
The measurements show that DSC can be used to investigate food products containing proteinaceous material.
This applies also particularly to animal proteins (e.g. egg, muscle protein).
Further, DSC can be used to follow the extent of the thermal treatment (to verify the treatment) and to determine
the influence of the process parameters (temperature, time) on protein quality for process optimization purposes.

10 METTLER TOLEDO Thermal Analysis

3.2 Influence of the starch content on swelling in water

Measurement by DSC
If warm water is added to starch granules, the granules swell and, at a certain point, burst. Some of the starch
molecules – mainly linear amylose molecules – are released into the aqueous phase. The interaction of water
with amylose outside the granules thickens the aqueous phase and leads to the formation of a gel. However, if
cold water is added to starch granules, they do not change much.

Sample Potato starch

Measurement Measuring cell: DSC
Crucible: Aluminum standard 40 µL, hermetically sealed
Sample preparation: Preparation of the starch suspensions by the addition of varying
amounts of stilled water; stirring (Ultra-Turrax for 1 min);
homogenized suspension weighed into the crucibles.
TGA measurement: Heating from 20 °C to 110 °C at 5 K/min

Interpretation
The diagram shows the DSC heating curves of potato starch in water at different concentrations.

One of the most decisive factors in the swelling of starch is the starch/water ratio. If an excess of water (more
than 4 molecules of water per anhydroglucose unit), the endothermic peak is only about 10 K wide, (see also
previous example with 20% starch in water). At starch concentrations greater than 40%, the reaction range
broadens or a second endothermic peak is formed. Whereas the first peak (between 60 °C and 80 °C) appears
at constant temperature, a reaction at higher temperatures is evident, which initially leads only to the formation
of a shoulder on the first peak and the blurring of the temperature range of the reaction but then, at higher ratios,
to the appearance second peak. The area of the first peak decreases, but the total enthalpy remains the same.
At high water contents, the system is destabilized through hydration (water uptake). Increased motion of the
polymer chains (heat uptake) and melting of the starch granules begins (first endotherm). In more concentrated
starch solutions, this effect is reduced as a result of the limited water content. Direct thermic melting of the starch
granules occurs at higher temperatures.

METTLER TOLEDO Thermal Analysis 11

 Evaluation
Starch content in % Sample weight of starch in mg Reaction enthalpy in J/g starch Peak temperature in °C
49 26.12 7.0 62.4
58 26.37 7.4 62.2
70 22.58 7.8 62.7

Conclusions
The curves show that the water/starch ratio is important for obtaining reproducible results. To know exactly
how much starch is in the crucible we recommend weighing starch and water directly into the pan (no mixing
required, diffusion adjusts local gradients).

12 METTLER TOLEDO Thermal Analysis

3.3 Influence of Moisture on Milk Powder

Purpose
Amorphous lactose is a one of the main constituents of milk powder and is highly hygroscopic. If milk powder
is stored in an open container, it becomes lumpy due to the uptake of moisture. The lactose present in the milk
powder can also crystallize as a result of increased water content. This can lead to changes in the flavor and
taste of products containing milk powder. Here we show how milk powder behaves in a humid atmosphere.

Sample Skimmed milk powder (nonfat milk powder)

Measurement Measuring cells: TGA, coupled to a moisture generator
Crucible: Aluminum 40 µL, open
Sample preparation: As received, no preparation
TGA measurement: The test specimens have first been dried at 30 °C (relative humidity
0%) during 10 hours and afterwards exposed to 60% relative humid-
ity. This cycle was then repeated.
Atmosphere: Humidified nitrogen gas flow rate 100 mL/min, protective dry gas
(nitrogen) flow rate 5 mL/min

Interpretation
The curves show, that the milkpowder is still not completely dry after 10 hours drying at 0% relative humidity.
We estimate an initial moisture content in the milkpowder of about 3.5%. The peak in the first sorption cycle is due
to crystallization of the lactose in the milkpowder: the initially glassy lactose is highly hygroscopic and incorporates
rapidly significant amounts of moisture. The increasing moisture content leads to a reduction of the glass transition
to below the measuring temperature (here 30 °C). Thus, the mobility of the lactose molecules increases and enables
the lactose to crystallize. Since crystalline lactose is considerably less hygroscopic than amorphous lactose, excess
water is released during crystallization. This explains the peak observed during the first cycle. Since the crystallization
of the lactose is an irreversible process, this peak only shows up during the first cycle.

Conclusion
In a humid atmosphere, milkpowder adsorbs within a short time significant amounts of moisture: from
the sorption experiment shown here, we conclude that at 30 °C and in air with 60% RH about 10% water
is incorporated in the milkpowder within 150 min.

METTLER TOLEDO Thermal Analysis 13

 3.4 Crystallization of vegetable oils

Measurement by DSC
Vegetable oils and fats are mainly esters of glycerol with fatty acids, so-called triglycerides. Oils are free-flowing
at room temperature; fats are solid. Vegetable oils often contain a higher content of unsaturated fatty acids than
animal fats and are therefore considered to be healthier. Furthermore, vegetable oils do not contain cholesterol.
Olive oil consists predominantly of triglycerides; approximately 80% of the fatty acids are unsaturated (oleic acid
and linoleic acid). Palm oil is one of the few highly saturated vegetable fats. Soybean oil is similar to olive oil
regarding the ratio of saturated to unsaturated fatty acids. Rape seed oil contains practically only unsaturated
fatty acids.

Sample Rape seed oil, pressed

Soybean oil, Homa brand
Olive oil, Dante brand, Italian first pressing
Palmoil
Measurement Measuring cell: DSC with IntraCooler
Crucible: Aluminum 40 µL, hermetically sealed
Sample preparation: As received, no special preparation
DSC measurement: Cooling from 50 °C to -100 °C at 10 K/min
Atmosphere: Nitrogen, 20 mL/min

Interpretation
Olive oil crystallizes below -10 °C. The triglyceride fraction, with in part saturated fatty acids, crystallizes between
-10 °C and -35 °C. The main fraction of olive oil, the triglyceride with 3 oleic acid units (70%), crystallizes at lower
temperatures and is recognizable as a crystallization peak at -45 °C (a temperature of -35 °C is given in the literature
for the stable α-form.
Commercial palm oil crystallizes below +15 °C. It has a high percentage of saturated fatty acids (50% C12, 18%
C14) and hence a high melting point; 8% unsaturated oleic acid lowers the melting point to values below room
temperature.
Soybean oil contains a significant fraction of saturated fatty acids (10% C16) and also a high percentage of mono-
basic and dibasic unsaturated fatty acids (nutritionally valuable). Triglyceride specimens analyzed by HPLC show
fractions with predomi-nantly saturated fatty acids, with some unsaturated fatty acids and also with highly unsatu-
rated fatty acids. These 3 triglyceride fractions are visible in the DSC curve at different crystallization temperatures.
The high fraction of linolenic acid in soybean oil (50%) leads to a high fraction of glycerides with three linolenic acid
units (melting point -45 °C), which is shown as a peak at -42 °C.

14 METTLER TOLEDO Thermal Analysis

Rape seed oil contains virtually only unsaturated fatty acids (60% C18 monobasic, 25% C18 dibasic, 10% C18
tribasic) and crystallizes at a very low temperature. Only 5% of the fatty acids are saturated and this is indicated by
‘small peaks’ between -20 °C and -40 °C.
Even though DSC cannot clearly identify fatty acids or triglyceride fractions, rapid characterization of the oils/fats is
possible.

Evaluation
Sample sample weight in mg Heat of crystallization in J/g Onset of crystallization in °C
Olive oil 24.63 61.5 -10.3
Palm oil 7.23 91.3 +10.2
Soybean oil 27.67 30.9 -10.2
Rape seed oil 11.76 46.7 -17.1

Conclusion
DSC is a rapid means of investigating the crystallization behavior of edible oils. For comparison purposes
approximately identical sample sizes should be taken, e.g. 20 ± 5 mg. The sample size may influence the
supercooling: the smaller the sample size the greater the degree of supercooling.

METTLER TOLEDO Thermal Analysis 15

 3.5 Identification of Plastic Films by DSC

Samples PE-HD/PA 6 and PE-LD/PA 6 composite films

Application Wrapping film for food packaging

Measurement Measuring cell: DSC
Crucible: Aluminum standard 40 µL, pierced lid
Sample preparation: A disk was punched out of the film and placed flat in the standard
aluminum crucible.
DSC measurement: Heating from 30 to 290 °C at 10 K/min
Atmosphere: Nitrogen, 50 mL/min

Interpretation
The diagram displays the DSC heating curves of two different polymer film laminates. The upper curve shows the
melting curve of a laminate of PE-HD (high-density polyethylene) and PA 6 (polyamide 6).This material is normally
used for packaging finished products (cooking bags). PE-HD melts at about 124 °C and PA 6 at 220 °C.
In the lower curve, the PE melting peak is about 12 K lower. This film is also a PA/PE laminate, but uses PE-LD (low-
density polyethylene), which melts at about 111 °C. This film is commonly used for packaging sausages and cheese.
Another film made of polypropylene and polyester (PET), which can also be used in microwave ovens, was easily
distinguished from the other two films: the melting point of PET (DSC peak temperature) is 256 °C with a glass tran-
sition at 69 °C while the melting point of polypropylene is 165 °C (the results are not shown).

Evaluation
Sample Weight in mg Peak temperature in °C
PE PA
PE-HD/PA 6 2.38 123.7 220.5
PE-LD/PA 6 2.64 111.4 220.5

16 METTLER TOLEDO Thermal Analysis

Interpretation
The second diagram displays the DSC heating curve of a three-layer polymer film laminate consisting of an EVAL film
(polyethylene-vinyl alcohol copolymer) between the PE-LD and the PA 6 films. EVAL is impermeable to gas and has a melting point
of about 180 °C. The melting point decreases with increasing ethylene content (32% to 44%). The composition varies depending
on the requirements. Improved barrier properties are offset by poorer processability.
The curve also shows a smaller peak at about 98 °C immediately before the melting of the PE. This peak is due to the melting of the
adhesive used to bond the films together.
Films of this type are used in the food sector for cheese and semi-finished products that require high barrier properties with regard
to oxygen (evacuated packages or those with a protective atmosphere).

Evaluation
Component Adhesive PE-LD EVAL PA 6
Peak temperature in °C 97.7 113.5 181.6 221.0

Conclusion
The identification of plastic materials is very important for the quality control of raw materials and finished goods as well as for the inves-
tigation of competitive products. DSC can identify the main components of plastic film laminates and minor components such as adhe-
sives if their melting behavior differs from that of the polymers.

METTLER TOLEDO Thermal Analysis 17

4. Food Regulatory Compliance

4.1 Why are regulations required?

The guarantee of food safety and quality along the increasingly complex global Some food safety topics and trends
food supply chain is a principal demand of the citizen and of society. Such • Pathogens and their potential risk
food safety trends and requirements increase the demand for testing, error-free • Toxic impurities
documentation and traceability. The increasingly regulated food industry is • Food allergy
faced with more work, more administration and more costs. Hence, it is neces- • Food fraud issues
sary to ensure that efficient analytical methodologies are possessed by the • Tracing an ingredient back to its source
food industry. At the same time it is a good opportunity to streamline processes • Rising demand for convenience food
to ensure the right people and machines are in the right place and make the
company lean and fit for the future.

4.2 The Global Food Safety Initiative (GFSI)

GFSI was set up in 2000 as a non-profit foundation at the request of around 30 CEOs of international retailers. Today, it is managed
by The Consumer Goods Forum. The initiative was set up against the backdrop of various food safety crisis, including BSE, with the
intention of ensuring worldwide consumer confidence in food safety. GFSI benchmarks existing food standards against food safety
criteria with a goal of standardizing certifications and eliminating multiple audits. GFSI accepted standards:
• BRC Global Standard • Global Red Meat Standard
• Dutch HACCP • IFS International Featured Standard
• FSSC 22000 • SQF 2000
• Global Aquaculture Alliance BAP • Synergy 22000

4.3 Where METTLER TOLEDO can help meet regulations

In all the standards listed above monitoring or measuring devices are mentioned regularly. They are referred to when recognized
tests, performance verification or calibration are required. This is where METTLER TOLEDO can help.

Performance verification
Documented verification that a system is capable of performing the activities it is required to perform, according to written and pre-
approved specifications, while operating in its specified operating environment. A performance qualification usually consists of a
collection of tests. Each step of the qualification should include an instruction, an expected result and the actual result.

Equipment qualification
Installation and setup of new equipment are the most critical moments to set the stage for worry free use. Steps taken at the
beginning of use can ensure peak performance and compliance with quality systems and regulatory requirements. Four crucial
steps are required during an installation, for confident release to routine operation:
• Professional installation and setup
• Calibration and performance verification
• Operator familiarization training
• Qualification testing and documentation
METTLER TOLEDO’s deliverables include professional installation by authorized service technicians, proof of proper equipment
performance at the point of installation, operational training, maintenance and care, together with supporting documentation.

Maintenance and calibration services

Preventive maintenance gives you confidence that your equipment will continue to meet operational requirements. It covers the
following activities:
• equipment inspection and cleaning
• equipment testing and adjusting if required
• recording and replacing worn out and deteriorated parts
• documenting the proper working condition of the equipment
The type of calibration required depends on a variety of factors: equipment type, country specific Government laws and regulations,
as well as application and usage of the instrument. From basic applications to highly regulated environments, METTLER TOLEDO
offers calibration certificates to meet every need. Reference materials are available for maintenance and calibration services.

18 METTLER TOLEDO Thermal Analysis

User training
In order to achieve:
• Operators skilled in equipment usage
• Personnel who is trained in workplace safety
• Personnel who can perform routine maintenance
• Process owners to ensure productivity
• Quality professionals to help maintain compliance
METTLER TOLEDO offer a wide range of thermal analysis training opportunities supporting the needs of improving skills, workplace
safety, maintenance, productivity and compliance.

Traceability of laboratory data

LIMS and ELN* primarily aggregate result data from an array of analytical tests. However, workflows behind analytical instruments
and the associated results, instrument information, user information and methods used are essential to traceability of lab data.
Regulatory standards such as FDA (21 CFR Part 11), EU (Annex 11), GMP, and ISO (ISO 17025) have increasingly established
further controls to manage both the advantages and limits of electronic data systems.
METTLER TOLEDO’s STARe Excellence thermal analysis offers several options to achieve full data traceability following
the customer’s needs.

4.4 How thermal analysis supports food regulatory compliance

Besides general rules for sampling, for thermal analysis, sample size and shape are two additional aspects that need to be consid-
ered to obtain trustworthy and reliable data.
• Size: Thermal analysis uses sample sizes that are typically regarded as being small ie. 5–20 mg.
Hence, the selection of the right part of the sample is crucial for the analysis.
• Shape: Flat surface to allow for good thermal contact.
Good thermal analysis measurements depend also on the correct choice of crucibles and lids. Crucibles and lids material are se-
lected according to the maximum temperature and to avoid or minimize reactions with the sample.
For TGA measurements, it is important to consider the sample's minimum weight. The minimum weight depends on the repeatability
of the balance, probability from the mean and precision requirements. Also, the upper sample size limit can be defined.

The use of the METTLER TOLEDO STARe software in regulated environments forms the foundation of a setup that ensures integrity,
trustworthiness and reliability. Centralized method SOP's are driven by the STARe software for the respective instrument to guide
measurements and data capture. The captured STARe original raw data cannot be changed. Evaluated data sets are stored sepa-
rately with assigned links to the raw data. The modular concept of the STARe software, as shown in Fig 4.1, enables regulatory
compliance according to the needs of the users.

Fig 4.1: The modular concept of STARe Excellence software. User Rights
and 21 CFR 11 options are part of the specification needs when regulatory
compliance is a requirement.

More details
For substantial explanations of regulatory compliance, measurement methods and validation topics please refer to the following
guides and handbooks:
Food Regulations in the Laboratory, Mettler-Toledo AG, ©02/2016
M. Schubnell: Validation in Thermal Analysis, Mettler-Toledo AG, 51725141, ©12/2008
M. Wagner: Thermal Analysis in Practice, Mettler-Toledo AG, 51725244, ©12/2009

(*) LIMS = Laboratory information and management system, ELN = Electronic laboratory notebook,
LES = Laboratory execution system, ERP = Enterprise resource planning

METTLER TOLEDO Thermal Analysis 19

5. Measurement Errors and Uncertainty of Measurement

Measurement results should reflect reality and hence serve as a basis for drawing conclusions and making decisions. In addition,
one should be able to trust the result of a single measurement without having to perform replicate (i.e. repeat) measurements.
However, this requires a thorough understanding of the possible causes of measurement errors and how these errors can be
quantified.
The results of measurements performed under identical conditions are never free of error, but are scattered around a central value.
This deviates more or less from a recognized value that is considered to be the "true" value. Deviations from the "true" value are
due to systematic errors that cause all measurement results to be too high or too low. A systematic error is often referred to as bias.
Systematic errors occur with every measurement to the same extent with the same sign and are usually difficult to eliminate. The
scatter of results around the mean value is due to random errors of measurement – some measurements are too high while others
are too low.

Systematic measurement errors

A systematic measurement error cannot be identified as such or eliminated merely by repeating the measurement. Systematic
measurement errors can only be detected if a reference value for the expected measurement quantity is available. The reference
value must be determined using the same measurement technique. Otherwise, systematic errors can occur due to the use of
different techniques.
If systematic measurement errors are detected, their origins must be investigated and measures taken to eliminate them.
Sometimes, the origin of the systematic measurement error cannot be identified. In such cases, a correction procedure is used that
describes how the measurement quantity should be corrected in order to obtain the right value.

Example: Buoyancy correction

A typical example of a systematic measurement error in thermogravimetric analysis is buoyancy. If a test specimen is heated at
ambient pressure, e.g. in air, the density of the air in the furnace decreases with increasing temperature. As a result, the buoyancy
experienced by the test specimen and the sample holder also decreases. This effect leads to an apparent increase in mass of
the test specimen. This systematic error in the measured mass of the test specimen can be corrected by performing a blank
measurement in which an empty crucible is heated under otherwise identical conditions.

Random measurement errors

Random measurement errors refer to the scatter of individual measurement results around the mean value. Random errors occur
to different extents in every measurement and determine the precision of a measurement. They cannot be eliminated. They can
however be reduced through precise control of the experimental conditions. Random measurement errors can be described by
statistical parameters such as standard deviation, distribution laws, etc.

Uncertainty of measurement
The uncertainty of measurement is a quantitative measure of the quality of the particular measurement results. It allows the user to
estimate the reliability of the measurement results. In simple terms, the uncertainty of measurement is the value range within which
the value of the quantity being measured (the measurand) is expected to lie with a stated level of confidence. It is not the same as
error because to estimate an error the "true" value must be known.
The concept of the uncertainty of measurement concerns all sources of uncertainty in connection with the sample, sample prepara-
tion, environmental influences, experimental parameters, the analyst, the evaluation of the measured data as well as the measure-
ment procedure itself.
The cause-and-effect diagram (Ishikawa diagram, fishbone diagram) is a simple analysis tool for systematically determining the
factors that contribute to a particular problem. It subdivides the possible causes leading to an overall effect into main and second-
ary factors. The resulting factors are presented in a clearly structured diagram that resembles a fishbone. The clear overview allows
you to identify all factors that contribute to a problem and to assess their importance and dependencies. The fishbone diagram is
often applied to estimate the uncertainty of measurement, for example, the uncertainty of the heat of fusion in DSC measurements.
The following main influence factors were considered: sample preparation, method development, instrument (measurement) and
evaluation. Main and secondary factors have been put into a fishbone diagram (Fig. 5.1).
The uncertainty of measurement for this example is estimated to ±3.4%.

20 METTLER TOLEDO Thermal Analysis

Fig 5.1: Cause-and-effect diagram to identify sources of uncertainty on the determination of the specific enthalpy of fusion
in DSC measurements

More details
For substantial explanations of measurement errors and uncertainty please refer to the validation handbook:
M. Schubnell: Validation in Thermal Analysis, Mettler-Toledo AG, 51725141, ©12/2008

METTLER TOLEDO Thermal Analysis 21

 6. Excellent Crucibles, Accessories and Reference Materials

Crucibles for thermal analysis

Crucibles serve as containers for samples during thermoanalytical measurements. They guarantee that the sen-
sor is not contaminated by the measurement. The type of crucible used for a measurement can have a large
effect on the quality of the results obtained, and in addition, also influences important characteristics of the DSC
measuring cell. Considering the relevant factors before the measurement can often help to save time later on
when interpreting the curve.

METTLER TOLEDO offers an extremely wide range of crucibles.

• Aluminum crucibles – various types and sizes
• Crucibles made from copper, platinum or gold
• Medium pressure crucible – stainless steel
• High pressure crucibles – stainless steel, gold plated, nimonic
• Alumina crucibles (aluminum oxide)
• Polycrystalline alumina oxide – a sapphire-like material
• Glass crucibles

Select the right crucible type – and improve your measurement quality.

Crucible sealing press

The press allows the pan to be sealed very easily. Under the pressure of the plunger the pan
is cold welded hermetically with the lid. With 3 exchangeable assemblies you adjust the sealing
press to the various crucibles. Also available: Sealing tools for high pressure crucibles.

Sample robot
All DSC and TGA models from METTLER TOLEDO can be automated. The sample robot can
process up to 34 samples even if every sample requires a different method and a different
crucible. The sample robot is very robust and operates reliably 24 hours a day and through-
out the whole year.

Reference materials
METTLER TOLEDO markets the reference substances needed for calibration and adjustment of thermal analysis
instruments. The following reference substances are traceable to the manufacturer.

Substance Indium Tin Lead Zinc Aluminum Gold Palladium

Symbol In Sn Pb Zn Al Au Pd
Tf [°C] 156.6 231.9 327.5 419.6 660.3 1064.2 1554.0
ΔHf [J/g] 28.5 60.1 23.0 107.5 397.0 63.7 162.0
Order no 00 119 442 51 140 621 00 650 013 00 119 441 51 119 701 51 140 816 51 140 817

22 METTLER TOLEDO Thermal Analysis

7. For More Information

Outstanding Services
METTLER TOLEDO offers you valuable support and services to keep you informed about new developments and
help you expand your knowledge and expertise, including:

News on Thermal Analysis

Informs you about new products, applications and events.
www.mt.com/ta-news
www.mt.com/ta-app

Handbooks
Written for thermal analysis users with background information, theory and practice, useful tables of material
properties and many interesting applications.
www.mt.com/ta-handbooks

Tutorial
The Tutorial Kit handbook with twenty-two well-chosen application examples and the corresponding test
substances provides an excellent introduction to thermal analysis techniques and is ideal for self-study.

Title Order Number

Tutorial Kit (handbook only) 30281946
Tutorial Kit (handbook and samples) 30249170

www.mt.com/ta-handbooks

Videos
Our technical videos explain complex issues concerning thermal analysis instrumentation and the STARe
software – whether it’s sample preparation, installation, creating experiments or evaluating measurement results.
www.mt.com/ta-videos

UserCom
Our popular, biannual technical customer magazine, where users and specialists publish applications from
different fields.
www.mt.com/ta-usercoms

Applications
If you have a specific application question, you may find the answer in the application database.
www.mt.com/ta-applications

Webinars
We offer web-based seminars (webinars) on different topics. After the presentation, you will have the opportunity
to discuss any points of interest with specialists or with other participants.
www.mt.com/ta-webinars (Live Webinars)
www.mt.com/ta-ondemand (On Demand Webinars)

Training
Classroom training is still one of the most effective ways to learn.
Our User Training Courses will help you get the most out of your equipment. We offer a variety of one-day theory
and hands-on courses aimed at familiarizing you with our thermal analysis systems and their applications.
www.mt.com/ta-training (Classroom)
www.mt.com/ta-etraining (Web-based)

METTLER TOLEDO Thermal Analysis 23

Overview of METTLER TOLEDO
Thermal Analysis Application Handbooks
The following application handbooks are available can be purchased:
www.mt.com/ta-handbooks

Introductory handbooks Language Order number Details

Thermal Analysis in Practice English 51725244
Volume 1
Fundamental Aspects (327 pages)
Thermal Analysis in Practice English 30306885
Volume 2 German 30315767
Tips and Hints (48 pages) French 30315768
Italian 30353357
Russian 30315769
Japanese 30315900
Korean 30315901
Thermal Analysis in Practice English 30281946 Handbook and
Volume 3 Tutorial samples
Tutorial Examples (86 pages) 30249170
Validation in Thermal Analysis English 51725141
A Guide (232 pages)

Applications handbooks Language Order number Details

Thermal Analysis of Elastomers English 51725061 Volumes 1 and 2
Volumes 1 and 2 51725057 Volume 1
Collected Applications (275 pages) 51725058 Volume 2
Thermal Analysis of Thermoplastics English 51725002
Collected Applications (150 pages)
Thermal Analysis of Thermosets English 51725069 Volumes 1 and 2
Volumes 1 and 2 51725067 Volume 1
Collected Applications (315 pages) 51725068 Volume 2
Thermal Analysis of Pharmaceuticals English 51725006
Collected Applications (100 pages)
Thermal Analysis of Food English 51725004
Collected Applications (65 pages)
Evolved Gas Analysis English 51725056
Selected Applications (65 pages)
Thermal Analysis of Polymers English 30076210
Selected Applications (40 pages) German 30076211
French 30076212
Russian 30076213
Japanese 30076214
Korean 30076215

www.mt.com/ta-news
For more information

Mettler-Toledo GmbH, Analytical

Sonnenbergstrasse 74
CH-8603 Schwerzenbach, Switzerland
Phone +41-44 806 77 11
Fax +41-44 806 72 60
Internet www.mt.com

Subject to technical changes

© 12/2016 Mettler-Toledo GmbH, 30389952
Marketing MatChar / MarCom Analytical