ISO-FOOD

ERA Chair for isotope techniques

Isotopes in
in food quality, safety and traceability food research
The ISO-FOOD ERA Chair for isotope techniques in food quality,
safety and traceability develops advanced analytical methods
of chemical analysis and complementary isotope techniques,
Welcome
which can uncover information that conventional techniques
cannot provide, i.e. that of geographical origin, authenticity, or
new and emerging contaminants in foodstuffs.

Isotope techniques are used for determining isotopic ratios of

elements, which cannot be obtained by conventional chemical
analysis. Because of specific behaviour of isotopes in physical
and chemical processes, isotope ratios of individual elements in
soil, water, plant and animal tissues or molecules provide exclu-
sive information on their origin or formation pathways.

Isotope analysis is thus providing complementary information

to that obtained by conventional analytical methods for deter-
mining elemental composition, toxic ionic species, organic com-
pounds or radionuclides in the food we eat.

Isotopes, on their own or combined with chemical composition,

carry unique information supporting food safety and traceabil-
ity and authentcity.
Isotopes are atoms of the same element that contain equal
numbers of protons but different numbers of neutrons in their
nuclei, and hence differ in atomic mass. Most elements have
Isotopes:
more than one natural isotope. All of them have the same
chemical properties, regardless of whether they are stable or
radioactive (decaying with time). Differences in their masses
tiny, yet big
result in different reaction rates between heavy and light iso-
topes of the same element. differences

1
1 H 2
1 H 3
1 H
 Changing The distribution of isotopes varies in nature, depending on the
physical and (bio)chemical reactions in which they are involved
- a process known as isotope fractionation. For instance during

isotope ratios evaporation and condensation, water is always enriched with

heavy isotopes (2H, 18O) compared to water vapour. The further
from the source (ocean), the higher in the mountains, and the
lower the temperature, the more depleted in the heavier iso-
tope of oxygen and hydrogen is the rain or snow. This gives the
precipitation in each region a distinct isotopic composition.

Similar fractionation occurs during photosynthesis: plants

preferentially absorb light 12C and further fractionate it during it i te

La
In
al p io r pr io

n
recipitat ecipitat
photosynthesis. Different photosynthetic pathways are re-
flected in different C isotopic ratios in plants and crops. The
so-called C3 photosynthesis produces plants with significantly
less of the heavy carbon isotope (13C) than C4 photosynthesis.

n
o ra ti o
ap
ev
C3 C4
Authentic Food fraud refers to the addition, tampering or misrepresenta-
tion of food, food ingredients or food packaging, or false or any
other misleading statements made about a product for eco-

or fraud? nomic gain. Although it might not necessarily be dangerous for

the consumer‘s health, it is illegal and results in reduced con-
sumer confidence, economic loss, and damage to brand iden-
tity. Certificates of geographical origin or production practice
(PDO – protected designation of origin; PGI – protected geo-
graphical indication) represent a considerable added value to
the food product, therefore robust methods for fraud detec-
tion are needed.

Fraud detection is a major analytical challenge: it means

searching for region-specific indicators such as isotopic com-
positions that can differentiate between botanical origin (e.g.
soy oil in olive oil), species (e.g. cow milk in goat cheese), fraud-
ulent practices (e.g. sugar syrup in honey) or additions of illic-
it substances (e.g. melamine in milk). Statistical evaluation of
isotopic, elemental and chemical profiles of food then reveals
the accordance or deviation of analysed samples from authen-
tic food products. Obviously, this approach depends on hav-
ing the ability to compare unknown samples with a database
of authentic products of known origin and year of production.
Databases need to have a sufficient number of samples to
have good geographic coverage and list as many parameters
as possible so as to capture natural variations.
Geographical Commonly used indicators of geographical origin are the isoto-
pic ratios of oxygen (18O/16O) and hydrogen (2H/1H) in water and
organic molecules in plants and animals (e.g. cellulose, pro-
origin of food teins, lipids). The isotopic composition of water in milk or fruit
juice varies from region to region even within a small country
such as Slovenia. Combined with other geographical indicators,
such as the elemental fingerprint (derived from bedrock and
water), isotopes are the most powerful tool for determining
the geographical origin of food.
To assure optimal yields, fertilisation with nitrogen is inevitable
in both conventional and organic agriculture. Isotopes can help
differentiate between the two production regimes: synthetic
Organic or
nitrate and ammonia have different isotopic compositions than
sludge or organic fertilisers, and this difference can be traced
to plants. In a similar way, the type of feed fed to cattle and
conventional?
poultry can also be traced: maize is enriched in 13C compared to
grass, and this difference is reflected in the isotopic composi-
tion of carbon in animal products, such as meat and milk.

NPK

> -3

-4.5 — -3

Synthetic fertilisers have more

-13
light 14N than organic fertilisers.
Plants fertilised with organic
fertilisers are enriched in 15N
-6.5 < -4.5 compared with those fertilised
δ O in rain water
18
δ O in cow milk
18
with synthetic nitrate.
Non-traditional While stable isotopes of water and nutrients have been anal-
ysed for decades, the analyses of isotopic ratios of other ele-
ments (‘non-traditional isotopes‘) are relatively new since they
The European Commission has recently adopted the regulation
of permissible levels of the most common natural (polonium,
radium, uranium-238, thorium-232) and man-made radionu-
Radionuclides
stable isotopes require expensive state-of-the-art analytical equipment and
exhaustive extraction procedures.
clides (plutonium, americium, and by-products of nuclear fis-
sion, such as cesium-137 and strontium-90) in water, food and
feed. To assure consumer safety, rapid methods for determin-
The relative abundances of stable and radioactive isotopes of
ing low levels of radionuclides, and dose assessments due to
certain food contaminants, such as lead, zinc or mercury, vary
intake of various foodstuffs and water for infants, children and
between different geological sources, therefore isotopic fin-
adults have been developed. Since most foodstuffs are highly
gerprinting of contaminants can provide information on their
complex matrices and the concentrations of radionuclides are
source. Based on the isotopic ratios of an element, for instance
usually extremely low, new opportunities are being explored by
lead, it is possible to estimate whether the lead in the soil and
combining radiochemical and mass-spectrometric methods.
crops is of natural origin (e.g. weathering of bedrock), or if it
derives from anthropogenic sources.

Pb
206
Pb
204

Energy spectrum of polonium

Po
209
Po
210
radioisotopes in mussels
 Isotopes in Isotope dilution and isotope labelling
Some elements are present in trace amounts and require sen-
Radiochemical analysis and radiolabelling
Radiochemical analysis is one of the most popular applications

analytics sitive analytical methods for their determination. The identi-

fication and quantification of molecular or ionic forms of dif-
ferent elements (speciation), which greatly determine their
toxicity, bioavailability and behaviour, is analytically challeng-
ing. One such method is isotope dilution whereby a known
amount of the same, but isotopically labelled element (or spe-
cies) is added, and then the total concentration of this element
(species) and its isotopic composition are measured. The initial
amount of the element in the sample can then be calculated.
In this way, we can assess the risk posed by potentially harmful
species, such as chromium in tea. This approach can be applied
to wide spectrum of essential and toxic elements, compounds
and element-based nanoparticles.
of nuclear techniques. Irradiation - bombardment with neu-
trons – of samples in a nuclear reactor converts chemical el-
ements into their artificial radioactive isotopes, which cannot
be produced in nature. Their concentration is then analysed by
α, β or γ counters; any further extraction procedure of target
elements from irradiated samples, if necessary, cannot induce
any contamination, therefore elements in very low concentra-
tions can be determined. Such radiolabelling can be used for
the production of radioactive tracers for tracer experiments,
for instance, to determine the uptake and fate of elements or
nanoparticles.

+ =
Cr 53Cr
50
Cr 53Cr
50
Cr 53Cr
50

Add known amount Natural unnown Measure total amount

of labelled Cr. amount of Cr isotopes. of Cr isotopes.
. .
Cal

s
pe
cu

at
to

eo is o
l

rigin Cr
al amount of
 Analytical Food analysis encompasses many metrological challenges:
complex matrices and low concentrations of analysed species
require new sample processing, validated methods, and state-
Every day, thousands of compositional, nutritional, allergen-
ic, isotopic, and contaminant profiles of all kinds of foods are
generated worldwide. These data contain a wealth of infor-
Data
challenges of-the-art facilities.

In the case of compound-specific isotope analyses and analy-

mation, however, to retrieve a custom-made, science-based
analysis from millions of data, internationally coordinated da-
tabases and software for data management and processing are
management
and sis of non-traditional stable isotopes, extraction procedures
require alterations, which do not cause isotope fractionation of
the target isotope in the analyte. Analysis often requires hy-
necessary. The task of food informatics is therefore to extract
information and manage knowledge from food research. For

data quality
example, the spatial databases of isotopic parameters of food
phenated high resolution mass spectrometric techniques with
are used to produce isotopic maps (“Isoscapes“) which will
chromatographic separation. In the case of nanoparticles in
show the isotopic composition of authentic food products.
food, methods have yet to be developed for their extraction
and characterisation.

The greatest overall challenge remains establishing stan-

dardised procedures and producing suitable certified refer-
ence materials for quality control and calibration.

Sampling + Processing + Measurement = Result

Representative Dissolution Comparison to SI ± uncertainty

Appropriate Extraction units
Contamination Dilution or conventional
Stability Labelling scale
Handling
Capacities for The Jožef Stefan Institute invests a great deal of effort and re-
sources into strengthening its capacities in terms of knowledge
and infrastructure. In 2015, the Department of Environmental
The ERA Chair for isotope techniques in food quality, safety and
traceability develops new methods and provides education
and training for food analysis and characterisation. The main
Education
tomorrow Sciences, which hosts the ERA Chair ISO-FOOD, upgraded
the Centre of Mass Spectrometry and other laboratories with
state-of-the-art research equipment. This enables the de-
research topics include:

• isotope and elemental fingerprinting and chemical profiling

through
velopment of new analytical methods required for pushing
the boundaries of interdisciplinary research in environmental
•
of food for determining the authenticity and geographical
origin
trace element speciation and fractionation
research
and food science. Moreover, recent projects funded from na-
• determination of organic contaminants and their transfor-
tional sources, through EU Regional Development Funds, and
mation products
the Horizon 2020 Twinning programme will enhance the use
• developing methods for detecting radionuclides at low
of isotopically based methodologies. Collaboration in interna-
activity concentrations
tional projects and in study programmes at the postgraduate
• nanoparticle determination and characterisation
level with the Jožef Stefan International Postgraduate School
• food composition databases and data management
keeps contact with the European and world‘s leading institu-
tions and facilitates research at the cutting edge of analytical
While focused on food safety, traceability and authenticity re-
chemistry.
search using advanced analytical methods, attention is paid to
metrology and accreditation of laboratories and methods and
preparation of a repository of data and knowledge on food
analysis and composition. The ERA Chair builds its sustainabil-
ity in tight collaboration with national and international stake-
holders from academia, science and industry.

Beside performing research and training at the PhD and post-

doctoral levels, the exchange of knowledge and best practices
is facilitated at ISO-FOOD Exploratory Workshops, ISO-FOOD
Training Courses, Summer Schools and through interlaboratory
comparison exercises.
 For more information please visit our web-pages:
www.isofood.eu and www.environment.si.
Follow us on Facebook and Twitter!
General information: info@isofood.eu
Contact:
ERA Chair ISO-FOOD
Jožef Stefan Institute
Jamova cesta 39
SI-1000 Ljubljana, Slovenia
ISO-FOOD received funding from the European Union‘s
Seventh Framework Programme for research, technological
development and demonstration under grant agreement No.
621329 (2014-2019).

Naklada: 1000 izvodov

Tekst: IJS, Odsek za znanosti o okolju
ISO
Oblikovanje in ilustracija: Irena Gubanc, Mateja Škofič (Padalci)
Fd
Food Fotografija: Miran Kambič u.d.i.a.; arhiv IJS
Tisk: Graphtech