Spotlight on Analytical Applications e-Zine - Volume 2

CONTENTS
TABLE OF
SPOTLIGHT
ON APPLICATIONS.
FOR A BETTER
TOMORROW.

VOLUME 2

CONTENTS
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INTRODUCTION
PerkinElmer Spotlight on Applications e-Zine – Volume 2
PerkinElmer knows that the right training, methods, applications, reporting and
support are as integral to getting answers as the instrumentation. That’s why
PerkinElmer has developed a novel approach to meet the challenges that today’s
labs face – that approach is called EcoAnalytix™, delivering to you complete
solutions for your applications challenges.

In this effort, we are pleased to share with you our Spotlight on Applications
e-zine, delivering a variety of topics that address the pressing issues and analysis
challenges you may face in your application areas today.

Our Spotlight on Applications e-zine consists of a broad range of applications
you’ll be able to access at your convenience. Each application in the table of contents
includes an embedded link that will bring you directly to the appropriate page
within the e-zine.

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CONTENTS

Energy
• Biodiesel Blend Analysis by FT-IR (ASTM D7371 and EN 14078)
• Diamond ATR and Calibration Transfer for Biodiesel-Blend Analysis by ASTM D7371
• imple Method of Measuring the Band Gap Energy Value of TiO2 in the Powder Form Using
S
a UV/Vis/NIR Spectrometer
• Photovoltaic Silicon Impurity Analysis by ELAN DRC ICP-MS

Environmental
• Analysis of Micronutrients in Soil Using AAnalyst 800 Atomic Absorption Spectrophotometer
• Determination of Oil and Grease in Water with a Mid-Infrared Spectrometer
• Characterization of Soil Pollution by TG-IR Analysis
• Ozone Precursor Analysis Using a TurboMatrix Thermal Desorption-GC System

Food Beverage
• nalysis of Organic Fertilizers for Nutrients with AAnalyst 800 Atomic Absorption Spectrophotometer
A
• race Elemental Characterization of Edible Oils with Graphite Furnace Atomic Absorption
T
Spectrophotometer
• Determination of Nickel in Fats and Oils
• nalysis of Fish and Seafood with AAnalyst 800 Atomic Absorption Spectrophotometer for Trace
A
Metals Contamination, in Accordance with AOAC Methods 999.10 and 999.11

Materials Characterization
• Determining Protein Secondary Structure with Spectrum 100
• Study Rigid Amorphous Fraction in Polymer Nano-Composites by StepScan and HyperDSC

Pharmaceuticals
• volved Gas Analysis: Residual Solvent Contamination Measured by Thermogravimetric
E
Analysis-Mass Spectrometry
• Evolved Gas Analysis: High Sensitivity Study of a Solvent of Recrystallization in a Pharmaceutical

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a p p l i c at i o n n o t e

FT-IR Spectroscopy

Authors
Ben Perston
Nick Harris
PerkinElmer, Inc.
Seer Green, UK

Biodiesel Blend Analysis Introduction – FAME and FAME Blends
The increasing importance of sustainability in energy
by FT-IR (ASTM D7371 production has led to a global commitment to the use
of fuels derived from renewable biological sources,
and EN 14078) such as biodiesel produced from plant crops. Biodiesel
consists of fatty acid methyl esters (FAME) and is
produced in a transesterification reaction as depicted
in Figure 1. A range of feedstocks are used globally,
including rapeseed, soy, sunflower, palm and jatropha.

Methanol
Base Catalyst

Triglycerides from biological source
R = C14–C18, 0–3 double bonds

Figure 1. Reaction scheme for the production of biodiesel.

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Infrared Spectroscopy

Authors
Ben Perston
Nick Harris
PerkinElmer, Inc.
Seer Green, UK

Diamond ATR and Introduction

Calibration Transfer for Biodiesel and Biodiesel-Blend Analysis
The need for sustainable fuel sources has led to an increasing global
Biodiesel-Blend Analysis emphasis on fuels produced from renewable, biological sources.
Biodiesel is one such fuel, and consists of fatty acid methyl esters
by ASTM D7371 (FAMEs) produced from vegetable oils or animal fats via a trans-
esterification reaction. Biodiesel is seldom used neat (B100), typically
being blended with fossil diesel at ratios from 5% v/v (B5) to 30%
v/v (B30). Verifying the FAME content of diesel-fuel blends is an important aspect of quality control and
auditing of blending and distribution operations. Because FAME has a strong infrared absorption at 1745 cm-1
due to the ester carbonyl group, infrared spectroscopy is an excellent technique for this analysis, and there
are EN and ASTM® standard test methods describing the procedure.1,2

The PerkinElmer® EcoAnalytix™ Biodiesel IR FAME Analyzer3 consists of a Spectrum™ 100 FT-IR spectrometer
with an attenuated total reflection (ATR) accessory featuring either a diamond or zinc selenide (ZnSe) crystal.
In addition to the Spectrum Express™ or Spectrum 10 software, which includes a dedicated biodiesel-analysis
module and detailed standard operating procedures (SOPs), the system is provided with a “starter calibration”,
covering the full range from 0% to 100% FAME. This application note describes how the supplied calibration
can be optimized to give peak performance in a particular installation, without resorting to a full recalibration.


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UV/Vis/NIR Spectrometer

Author
Jayant Dharma
PerkinElmer Technical Center
Aniruddha Pisal
Global Application Laboratory
PerkinElmer, Inc.
Shelton, CT USA

Simple Method of
Measuring the Band
Gap Energy Value of
TiO2 in the Powder
Form Using a UV/Vis/ Introduction

NIR Spectrometer The measurement of the
band gap of materials
is important in the semi-
conductor, nanomaterial
and solar industries. This note demonstrates how the band gap
of a material can be determined from its UV absorption spectrum.
The term “band gap” refers to the energy difference between
the top of the valence band to the bottom of the conduction
band (See Figure 1); electrons are able to jump from one band
to another. In order for an electron to jump from a valence
band to a conduction band, it requires a specific minimum
amount of energy for the transition, the band gap energy1,2.
A diagram illustrating the bandgap is shown in Figure 1.
Measuring the band gap is important in the semiconductor and
nanomaterial industries. The band gap energy of insulators is
large ( 4eV), but lower for semiconductors ( 3eV). The band
gap properties of a semiconductor can be controlled by using
different semiconductor alloys such as GaAlAs, InGaAs, and
InAlAs. A table of materials and bandgaps is given in Reference 1. Figure 1. Explanation of band gap.


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Case
ICP-Mass Spectrometry
study

Jiabin Du, Huanhuan Pan
Technology Center, LDK Solar Co., Ltd.
Economic Development Zone
Xinyu, Jiangxi 338032, China

Jianmin Chen and Wilson You
Global Application Laboratory
PerkinElmer, Inc.
710 Bridgeport Avenue,
Shelton, CT USA

Photovoltaic Silicon Rising energy and oil prices in light of the economic
Impurity Analysis by slowdown, and a heightened awareness of the
environment, have led to an increase in global
ELAN DRC ICP-MS incentives for the diversification of energy sources
and greater utilization of renewable energy segments.
With the increased interest in renewable energy, there are growing opportunities
for photovoltaics (PV). According to market research, PV is expected to account
for over 50% of the world’s total electricity generated by renewable energy
sources by 2070. The PV market overall has grown to $19 billion globally and
is predicted to continue over the next decade at over 30% per year. Wafers are
the principal raw material used to produce solar cells, which are devices capable
of converting sunlight into electricity. Today approximately 90% of the world-
wide PV installations use mono- or multi-crystalline silicon wafers in the solar
cells and silicon wafers represent half or more of the cost of silicon solar cells.

For more information about this story,
please visit LDK Solar Limited at:
http://www.ldksolar.com/index.html

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Atomic Absorption

Author
Praveen Sarojam, Ph.D.
PerkinElmer
Global Application Center
Mumbai, India

Analysis of Micronutrients Introduction
Soil is used in agriculture, where it serves as the primary
in Soil by Using AA 800 nutrient base for plants. Soil material is a critical
component in the mining and construction industries.
Atomic Absorption Soil serves as a foundation for most construction projects.

Spectrophotometer Soil resources are critical to the environment, as well
as to food and fiber production. Waste management
often has a soil component. Land degradation is a
human-induced or natural process which impairs the
capacity of land to function. Soils are the critical
component in land degradation when it involves acidification, contamination etc. Soil contamination
at low levels is often within soil capacity to treat and assimilate. Many waste treatment processes rely
on this treatment capacity. Exceeding treatment capacity can damage soil biota and limit soil func-
tion. Derelict soils occur where industrial contamination or other development activity damages the
soil to such a degree that the land cannot be used safely or productively. The analysis of soils is an
excellent measure of soil fertility. It is a very inexpensive way of maintaining good plant health and


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Author
Aniruddha Pisal
PerkinElmer, Inc.
Shelton, CT 06484 USA

Determination of Oil Introduction
The concentration of dispersed oil and
and Grease in Water grease (OG) is an important parameter for
water quality and safety. OG in water can
with a Mid-Infrared cause surface films and shoreline deposits
leading to environmental degradation,
Spectrometer and can induce human health risks when
discharged in surface or ground waters.
Additionally, OG may interfere with aerobic
and anaerobic biological processes and lead to decreased wastewater treatment
efficiency. Regulatory bodies worldwide set limits in order to control the
amount of OG entering natural bodies of water or reservoirs through industrial
discharges, and also to limit the amount present in drinking water.

OG in water is commonly determined by extraction into a non-polar, hydrocarbon-
free solvent followed by measurement of the infrared absorption spectrum of
the extract. The absorption between 3000 and 2900 cm-1 by C-H groups in
the OG is correlated to the concentration of OG. There are several standard
test protocols based around this methodology,1–4 most commonly using
1,1,2-trichloro-1,2,2-trifluoroethane (CFC-113) or tetrachloromethane.
However, these solvents are known ozone-depleting compounds, and under
the Montreal Protocol, the use of CFC-113 was phased out by 1996 and the
use of tetrachloromethane will become illegal in 2010.


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Thermogravimetric Analysis –

Characterization There are many scenarios in which soil can become contaminated
by hydrocarbon products. Leakage from fuel storage tanks or

of Soil Pollution transfer lines as well as storm water runoff from vehicle washing
areas are just two examples. In environmental monitoring

by TG-IR Analysis or land reclamation, therefore, it is important to test soil for
contamination. Total petroleum hydrocarbon (TPH) testing
by solvent extraction and infrared spectroscopy is a sensitive
method, but has a considerable burden of sample preparation.
A gas chromatographic analysis of the extract can provide even
greater sensitivity and more detailed compositional information,
but further increases the time required for the analysis.

Thermogravimetric analysis coupled to infrared spectroscopy (TG-IR) can provide detailed information
about the amount and nature of the pollution, while requiring no sample preparation at all. This
application note illustrates the kind of data that can be obtained with a modern TG-IR system.

Experimental
A soil sample was obtained and mixed with diesel fuel at a concentration of about 10% m/m. 17 mg
of the soil was transferred to the crucible of a PerkinElmer® TGA 4000, coupled to a PerkinElmer
Spectrum™ 100 infrared spectrometer by the TL 8000 transfer line with a 10-cm gas cell. The transfer
line and gas cell were heated to 280 °C to avoid any risk of condensation of heavier organic compounds.
The purge gas through the TGA was nitrogen at flow rate of 20 mL/min with a balance purge of 40 mL/min.
This combined rate of 60 mL/min was kept constant through the transfer line and cell. The temperature
was increased from 30 to 800 °C at a constant rate of 20 °C/min. Infrared spectra over the range
4000–600 cm-1 were collected every 12 s at 8 cm-1 resolution (co-adding
four interferometer scans for each spectrum). Pyris™ software was used to
control the TGA, while TimeBase™ was used for collection and analysis of
the time-resolved IR data.


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White
paper Gas Chromatography

Authors
Graham Broadway
Andrew Tipler
PerkinElmer, Inc.
Shelton, CT USA

Ozone Precursor In the United States, the Clean Air Act of 1970 gave the
U.S. Environmental Protection Agency (EPA) responsibility

Analysis Using a Thermal for maintaining clean air for health and welfare. Six
parameters are measured routinely in ambient air: SOx,

Desorption-GC System NOx, PM10 (particulate matter less than 10 microns), Pb,
CO and ozone. In the 1990 Clean Air Act Amendments,
Title 1 expanded the measurements in air to include volatile
organic compounds (VOCs) that contribute to the formation
of ground-level ozone. These parameters are measured
in urban areas that do not meet the attainment goals for
ozone, as shown in Figure 1. These measurements are
implemented through a program known as Photochemical
Assessment Monitoring Stations (PAMS).

This program has been in place in the U.S. for a number
of years, and in 2008 the National Ambient Air Quality
Standards (NAAQS) for Ground-Level Ozone was reduced
to 0.075 ppm for an 8-hour period.1 The U.S. EPA predicts
that a large number of counties will violate the 2008 stan-
dard2 (Figure 1). Similar recommendations have also been
made in Europe. Following the 1992 Ozone Directive and
United Nations Economic Commission for Europe’s protocol
on controlling VOC emissions, a European ozone precursor
priority list was established by Kotzias et al.3 and subse-
quently modified by the EC 2002/3/CE directive.

Figure 1. Areas expected to violate the 2008 Ozone Standard.

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Atomic Absorption

Author
PerkinElmer
Mumbai, India

Analysis of Organic Introduction
The fertilizer industry helps to ensure that farmers have
Fertilizers for Nutrients the nutrients they need to grow enough crops to meet
the world's requirements for food, feed, fiber and
with AAnalyst 800 energy. Nutrients in manufactured fertilizers are in the
form that can be absorbed readily by the plants. All of
Atomic Absorption these nutrients exist in nature, but the quantities are not

Spectrophotometer sufficient to meet the needs of our growing, urbanized
population. Soils may be naturally low in nutrients, or
they may become deficient due to nutrient removal by
crops over the years without replenishment – or when
high-yielding varieties are grown that have higher nutrient requirements than do local varieties. All of
the essential nutrients are important but in varying quantities. Macronutrients (N, P, K, Ca, Mg, etc.)
are needed by plants in large quantities. The “primary nutrients” are nitrogen, phosphorus and potas-
sium. Today, sulphur is also considered a key macronutrient. Macronutrients include both primary and
secondary nutrients. Micronutrients (or “trace elements”) (Fe, Mn, Zn, Cu, Ni, etc.) are required in very
small amounts for correct plant growth. They need to be added in small quantities when they are not
provided by the soil. Every plant nutrient, whether required in large or small amounts, has a specific
role in plant growth and food production. One nutrient cannot be substituted for another. For example,
potassium activates more than 60 enzymes (the chemical substances that govern life and play a vital
part in carbohydrate and protein synthesis). It improves a plant's water regime and increases tolerance
to drought, frost and salinity. Plants that are well supplied with potassium are less affected by disease.
Magnesium is the central constituent of chlorophyll, the green pigment in leaves that functions as an
acceptor of the energy supplied by the sun: 15-20% of the magnesium in a plant is found in the green


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Atomic Absorption

Author
Praveen Sarojam Ph.D.
PerkinElmer, Inc.
Mumbai

Trace Elemental Introduction
The determination of the inorganic profile of oils is
Characterization of important because of the metabolic role of some
elements in the human organism. On the one hand,
Edible Oils with Graphite there is knowledge of the food's nutritional value,
which refers to major and minor elements. On the
Furnace Atomic Absorption other hand, there is the concern to verify that the
food does not contain some minerals in quantities
Spectrophotometer toxic for the health of the consumers, regardless
whether this presence of minerals is naturally
occurring or is due to contamination during the
production processes. Oil characterization is the basis for further nutritional and food technological
investigations such as adulteration detection1. The most common adulteration is an addition of a
cheaper vegetable oil to expensive oil. Authenticity is a very important quality criterion for edible
oils and fats, because there is a big difference in prices of different types of oil and fat products.
Adulteration detection is possible by determining the ratio of the contents of some chemical
constituents and assuming these ratios as constant for particular oil. In regard to adulteration
detection, approaches based on atomic spectroscopy can be attractive2. The quality of edible oils
with regard to freshness, storability and toxicity can be evaluated by the determination of metals.
Trace levels of metals like Fe, Cu, Ca, Mg, Co, Ni and Mn are known to increase the rate of oil
oxidation. Metals like As, Cd, Cr, Se etc. are known for their toxicities. The development of rapid
and accurate analytical methods for trace elements determination in edible oil has been a challenge
in quality control and food analysis. However, sample pretreatment procedures are required in order
to eliminate the organic matrix. These include wet, dry or microwave digestion, dilution with organic
solvent and extraction methods3. The content of metals and their species (chemical forms) in edible
seed oils depends on several factors. The metals can be incorporated into the oil from the soil or be
introduced during the production process. Hydrogenation of edible seed oils and fats has been


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a p p l i c at i o n B R i E F

Atomic Absorption

Determination of Nickel Scope
Triglyceride-based vegetable fats and oils can be trans-
in Fats and Oils formed through partial or complete hydrogenation to
fats and oils of greater molecular weight. The hydro-
genation process involves sparging the oil at high
temperature and pressure with hydrogen in the
presence of a catalyst, typically a powdered nickel
compound. Atomic Absorption Spectrometry is
commonly used to estimate the amount of nickel
left in the vegetable oils.

Typical Analytical Procedure

Materials and Methods
The following reagents and equipment are used for the measurement:

• Atomic absorption spectrometer
• Nickel metal
• Conc. nitric acid
• Conc. hydrochloric acid
• Double distilled water

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Atomic Absorption

Author
PerkinElmer, Inc.
Shelton, CT 06484 USA

Analysis of Fish and Seafoods Introduction
Increased knowledge about the nutrient content
with AAnalyst 800 Atomic of biological organisms is essential for a thorough
understanding of ecological stoichiometry and nutrient
Absorption Spectrophotometer transport in and among ecosystems. As a result of
water pollution in coastal area, many problems in food
for Trace Metal Contamination, safety like heavy metal accumulation have been recog-

in Accordance with AOAC nized in farmed fish, which is one of the important fish-
ery food resources. The heavy metals accumulated in

Methods 999.10 and 999.11 fish not only have a bad influence on fish but they also
affect the health of human beings through the food
chain. It is pointed out that remarkable heavy metals
were contained in fish meals that are used as major
raw materials for aquaculture feeds. The Itai-itai
disease of the Toyama Jintsu River area in Japan
was the documented case of mass cadmium poisoning.
Itai-itai disease is known as one of the Four Big
Pollution Diseases of Japan.


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FT-IR Spectroscopy

Author
Richard Spragg
PerkinElmer, Inc.
Chalfont Road
Seer Green
Beaconsfield
Buckinghamshire, UK

Determining Protein
Secondary Structure
with Spectrum™ 100 Although crystallography and NMR are the primary
tools for determining geometrical protein structures
there are significant limitations in the range of
proteins that these techniques can address. FT-IR gives less detailed information
but it has some important advantages. It can be applied to any protein, it requires
a relatively small sample, and the measurements can be made in solution. The
applications for FT-IR include determining secondary structure of novel proteins,
identifying conformations in formulations, measuring the kinetics of conformational
changes with temperature or other perturbations, and investigating protein-ligand
interactions.
The information about secondary structure is contained in the shape of the amide-1
band in the IR spectrum. This requires careful measurement as it overlaps with the
water band at 1640cm-1 and water vapor absorptions. Both transmission and ATR
techniques can be used, but adsorption of the protein on to the crystal surface is a
potential problem with ATR. For transmission measurements pathlengths between
6 and 10μm are generally used.
The secondary structure is analyzed as a mixture of different amounts of various
sub-structures such as α-helix and β-sheet, each of which has characteristic
absorptions. In curve-fitting and deconvolution approaches the amide-1 band is
analyzed explicitly as the superposition of such bands. Chemometric approaches
use a library of spectra from proteins of known secondary structure.

Brief

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Differential Scanning Calorimetry

Author
Christoph Schick
University of Rostock
Inst. of Physics
Universitätsplatz 3, 18051
Rostock, Germany

Study Rigid Amorphous
Fraction in Polymer
Introduction
Nano-Composites It is known that there is a rigid amorphous fraction (RAF) in semicrystalline polymers.
by StepScan and The RAF exists at the interface of crystal and amorphous phase as a result of the immo-
bilization of a polymer chain due to the crystal. There is debate on whether the crystal
HyperDSC melts first and then RAF devitrifies or the RAF devitrifies before the crystal melts. It can
not be answered easily because these two things often happen in the same temperature range. Also,
the RAF fraction sometimes exists at the surface of silica nanoparticles in the polymer silica nanocom-
posites material. However, unlike semi-crystalline polymers, the silica nanoparticle does not undergo
any transition at the temperature when RAF devitrifies. So polymer silica nanocomposites offer a good
opportunity to study the devitrification of RAF1.

Some studies have indicated a second glass transition from RAF in dynamic measurements. To the
author’s best knowledge, there is no evidence of a second Tg in polymer nanocomposites from DSC experi-
ments. In order to identify RAF in DSC, absolute heat capacity measurement is very important. A formula
has been well established for the determination of RAF in semicrystalline polymers based on accurate
heat capacity measurement as described by Wunderlich, see2 for a review.

Here, heat capacity measurement has been performed in order to detect a possible second Tg on nanocom-
posites of polymethyl methacrylate (PMMA) with silicon oxide nanoparticles of different shape. StepScan™
DSC was used for determination of precise heat capacity and HyperDSC® to prevent degradation and identify
devitrification of the RAF at elevated temperatures.


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Mass Spectrometry

Evolved Gas Analysis: Introduction
Thermogravimetric analysis (TGA) of mate-
Residual Solvent rials is commonly used to measure weight
loss from a sample as it is heated or held
Contamination isothermally. In the pharmaceutical industry,

Measured by many materials show weight losses
associated with the loss of solvent/

Thermogravimetric water, desolvation or decomposition
of the sample. This information is then

Analysis-Mass used to assess the purity and stability
of the material and its suitability for use.

Spectrometry The TGA gives a quantitative measure of
mass lost from the sample, but it does not
provide information on the nature of the
products that are lost from the sample,
and this information is often required
for complete characterization.

Coupling a mass spectrometer (MS) to a
TGA allows evolved gases to be analyzed
and identified, delivering this additional
valuable information.

Brief

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a p p l i c at i o n b r i e f

Mass Spectrometry

Evolved Gas Analysis:
Introduction
a High Sensitivity Thermogravimetric analysis (TGA) of materials is commonly used to measure

Study of a Solvent weight loss as a sample is heated or held isothermally. In the pharmaceutical
industry, materials often show weight losses associated with the loss of

of Recrystallization solvent/water, desolvation or decomposition of the sample. This information is
then used to assess the purity and stability of the material and its suitability for

in a Pharmaceutical use. The TGA gives a quantitative measure of mass lost from the sample,
but it does not provide information on the nature of the products that are
lost from the sample, and this information is often required for complete
characterization.
Typical applications of TG-MS include:
Coupling a mass spectrometer (MS) to a TGA allows evolved gases to be
• Detection of moisture/solvent loss from a
analyzed and identified giving this additional valuable information.
sample (e.g. loss on drying or dehydration
of a pharmaceutical)
Instrumental Setup
• Thermal stability (degradation) processes
All of the TGA systems supplied by PerkinElmer (Pyris™ 1 TGA, STA 6000
• Study reactions (e.g. polymerizations) and TGA 4000) can be easily interfaced to MS systems. PerkinElmer can
• Analysis of trace volatiles in a sample (e.g. supply systems with either the PerkinElmer Clarus® MS or with a Hiden
volatile organic compound (VOC) testing) Analytical MS. In this study, a Pyris 1 TGA interfaced to a Hiden Analytical
HPR-20 MS was used.

Figure 1. Pyris 1 TGA is shown interfaced to a HPR-20 MS (left) and a Clarus 600 MS (right).

Brief

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USEFUL LINKS
View previous issues of our Spotlight on Applications e-Zine
• Volume 1
• Archives

Access our application archives
By Industry:
• Consumer Product Safety
• Environmental
• Food, Flavors and Agricultural Analysis
• Forensic Analysis
• Petrochemical Analysis
• Pharmaceutical Development Manufacturing
• Polymers
• Renewable Energy
• Semiconductor Electronics

By Technology:
• Atomic Absorption (AA)
• Elemental Analysis
• Gas Chromatography (GC)
• GC Mass Spectrometry (GC/MS)
• Hyphenated Technology
• ICP Mass Spectrometry (ICP-MS)
• Inductively Coupled Plasma (ICP-OES ICP-AES)
• Infrared Spectroscopy (FT-IR IR)
• LIMS Data Handling
• Liquid Chromatography (HPLC UHPLC)
• Mass Spectrometry
• Raman Spectroscopy
• Thermal Analysis
• UV/Vis UV/Vis/NIR

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