Advanced analytical methods in the
determination of xenobiotics in
water samples
Joonas Nurmi 12.3.2009
Department of Ecological and Environmental
Sciences
Faculty of Biosciences
Outline of presentation
Background
Chemicalisation
Xenobiotics in water
Objective and methods
Chromatography
Time-of-flight mass spectrometry
Accurate mass measurements
Previous research
Non-target water analysis
Joonas Nurmi
12.3.2009
2
1. Background
- Chemicalisation
- Xenobiotics
Faculty of Biosciences
Chemicalisation
Definition: Chemicalisation signifies the increasing
use of chemicals and environmental pollution
happening in consequence of that.
Pollution caused by the use of chemicals can also be
unintentional or unconscious!
Joonas Nurmi
12.3.2009
4
How many chemicals?
Exact number is impossible to give
Chemical Abstracts Service (CAS)
43 650 433 organic and inorganic substances with CAS
Registry Number
30 206 099 commercially available chemicals
247 717 inventoried / regulated substances
www.cas.org 27.2.2009 17:06
Finnish Environment Institute (SYKE)
Over 30 000 products which contain hazardous compounds
are imported or produced in Finland
Number of compounds in these products is over 5 000
Joonas Nurmi
12.3.2009
5
What’s the problem?
Chemicals may have impacts on nature, urban
environment, working environment or households
depending on the properties of chemicals and how they
are used.
Generally - concentrations of organic compounds in
environment are poorly known
Information is limited to few compounds
New chemicals are produced all the time
Emerging contaminants?
Lack of research causes delay in legislation
Joonas Nurmi
12.3.2009
6
Small history of chemicalisation 1/2
Ancient chemistry
7000 B.C. Middle East: alkali, limestone
900 A.D. China: black powder
glass production
gun powder
Industrial revolution
Inorganic chemistry 1820
- 1823 GB: mass production of soda ash
- 1850s synthetic dyes
soap, glass
textile industry
Organic chemistry, Europe, late 19th century
- 1909 synthetic fertilizers
- 1920s chemicals for oil
II World War
Joonas Nurmi
crop yield improvement
petrochemical products
short industrial break
12.3.2009
7
Small history of chemicalisation 2/2
1950-
increased use of pesticides, organohalides,
pharmaceuticals, industrial chemicals
1962 Rachel Carson: Silent spring
”Start of environmental movement”
Dichloro diphenyl trichloroethane, DDT
organochlorine insecticide
probable human carcinogen, damages the liver nervous
system, reduces reproductive success, damages
reproductive system
Cl
Banned in Finland since1976
Cl
Cl
2001 Stockholm Convention
Cl
Joonas Nurmi
Cl
12.3.2009
8
EPA’s ”dirty dozen”
Aldrin ¹
Chlordane ¹
Dichlorodiphenyl trichloroethane (DDT) ¹
Dieldrin ¹
Endrin ¹
Heptachlor ¹
Hexachlorobenzene ¹,²
Mirex ¹
Toxaphene ¹
Polychlorinated biphenyls (PCBs) ¹,²
Polychlorinated dibenzo-p-dioxins ²(dioxins)
Polychlorinated dibenzofurans ² (furans)
Endrin
www.epa.gov
1-Intentionally produced.
2-Unintentionally produced - Result from some industrial processes and
combustion.
Joonas Nurmi
12.3.2009
9
Classification of chemicals:
1. Basic chemicals
Polymers
Mainly packing materials
- e.g. polyethylene (PE), polystyrene (PS), polyvinyl
chloride (PVC), polypropylene (PP)
Petrochemicals
Made from liquefied petroleum gas, natural gas, and crude
oil
- e.g. benzene, toluene, xylenes, methanol
Industrial chemicals
- e.g. synthetic rubber, surfactants, dyes and pigments,
turpentine, resins, carbon black, explosives and rubber
products
Joonas Nurmi
12.3.2009
10
Classification of chemicals:
1. Basic chemicals
Inorganic chemicals
Oldest chemical category
- e.g. salt, chlorine, caustic soda, soda ash, acids (such as
nitric, phosphoric, and sulfuric), titanium dioxide, and
hydrogen peroxide
Fertilizers
e.g. phosphates, ammonia, and potash chemicals
Joonas Nurmi
12.3.2009
11
Classification of chemicals:
2. Life sciences
While much smaller in volume than other chemical
sectors, their products tend to have very high prices
Product groups:
Pharmaceuticals
Animal health products
Vitamins
Crop protection chemicals
- herbicides, insecticides, and fungicides
Joonas Nurmi
12.3.2009
12
Classification of chemicals:
3. Specialty chemicals
Category of relatively high valued, rapidly growing
chemicals with diverse end product markets
Generally characterized by their innovative aspects
Sold for what they can do rather than for what chemicals
they contain
Product groups:
electronic chemicals, industrial gases, adhesives and
sealants as well as coatings, industrial and institutional
cleaning chemicals and catalysts
Joonas Nurmi
12.3.2009
13
Classification of chemicals:
4. Consumer products
Most of chemicals are used in manufacture of other items,
although a smaller number are used directly by
consumers
Major industrial customers include rubber and plastic
products, textiles, apparel, petroleum refining, pulp and
paper, and primary metals
Consumer products:
soaps
cosmetics
detergents
Joonas Nurmi
12.3.2009
14
Breakdown of the chemicals in commerce in
USA
Estimated number and categories of chemicals in
commerce registered in the USA over the past 30 years
1800
3400 1000
Industrial
8600
Food additives
82000
Cosmetics
ingredients
Pharmaceuticals
Pesticides
Derek et al. Environ. Sci. Technol. (2006) 40:7157.
Joonas Nurmi
12.3.2009
15
Xenobiotics - definitions
A xenobiotic is a chemical which is found in an organism
but which is not normally produced or expected to be
present in it. It can also cover substances which are
present in much higher concentrations than are usual
Xenobiotics is very often used in the context of pollutants
such as dioxins and polychlorinated biphenyls and their
effect on the biota, because xenobiotics are understood
as substances foreign to an entire biological system
Joonas Nurmi
12.3.2009
16
Potential emerging pollutants
Phosphorus flame retardants (PFR)
O
CF3
HO
Brominated flame retardants (BFR)
(CF2) 8
Polyfluorinated organic compounds (PFC)
O
Short chain chlorinated paraffins (CP)
O
P
O
O
Cl
Cl
Br
Cl
Cl
Cl
Cl
Br
HO
Br
Br
Joonas Nurmi
OH
12.3.2009
17
2. Objectives and methods
- Chromatography
- Mass spectometry
Faculty of Biosciences
Objectives of the study
Identification of emerging organic pollutants by non-target
screening of waste waters
Development of novel qualitative and quantitative
instrumental techniques to analyze organic xenobiotics in
different water samples
E.g. surface water, storm water, waste water
Assessment of environmental effects of organics in water
system by ecotoxicological research methods
Joonas Nurmi
12.3.2009
19
Chromatography
Collective name of laboratory separation techniques
Separation is based on the interaction of a compound
between two phases: mobile phase and stationary phase
Gas chromatography (GC):
- Mobile phase: gas, Stationary phase: liquid
- Carried out in a column
Liquid chromatography (LC):
- Mobile phase: liquid, Stationary phase: solid
- Carried out either in a column or a plane
Joonas Nurmi
12.3.2009
20
Mass spectrometry
Mass spectrometry (MS) is an analytical technique for
the determination of the elemental composition of a
sample or molecule
Sample is loaded onto ion source which splits the sample
molecules into ions
Mass analyzer sorts the ions by their masses by applying
electromagnetic fields
Detector measures the value of an indicator quantity and
thus provides data for calculating the abundances of each
ion present
Joonas Nurmi
12.3.2009
21
Time-of-flight mass spectrometry (TOF-MS)
The time-of-flight (TOF) analyzer uses an electric field to
accelerate the ions through the same potential, and then
measures the time they take to reach the detector. If the
particles all have the same charge, the kinetic energies
will be identical, and their velocities will depend only on
their masses. Lighter ions will reach the detector first.
Joonas Nurmi
12.3.2009
22
Resolution – key to accurate mass
Resolution R can be
defined as the ability
to separate and
measure the masses
of ions of similar, but
not identical,
molecular mass.
High resolution > 5000
Joonas Nurmi
12.3.2009
23
3. Previous research
- Non-target analysis
Faculty of Biosciences
Screening methods
Pre-target screening: analytes are pre-selected before
MS-data acquisition, other positives cannot be revealed
Post-target screening: all compounds eluted from column
are measured by MS and the m/z of target analytes are
extracted afterwards
Non-target screening: compounds eluted from column can
be measured and identified without any kind of selection –
search of unknown compounds, no previous information
about the analytes is taken into account
Joonas Nurmi
12.3.2009
25
Outline of non-target analysis
1. Sampling
2. Sample pretreatment
3. Separation of compounds, e.g. GC / LC
good separation efficiency
4. Detection of compounds, e.g. TOF-MS
accurate mass measurement
5. Data analysis
component deconvolution with proper software
elemental composition of components
library search of components
6. Validation of results
use of standard compounds
Joonas Nurmi
12.3.2009
26
Ionization in mass spectrometry
Electrospray ionization
positive polarity
Cl
O
[M+H]+
Cl
Protonated molecule
N
N
Imazalil
C14H14N2OCl2
Protonated Imazalil
C14H15N2OCl2
Exact mass:
C14H14N2O35Cl2: 296,0482
C14H14N2O35Cl37Cl: 298,0453
Exact mass:
C14H15N2O35Cl2: 297,0561
C14H15N2O35Cl37Cl: 299,0532
Joonas Nurmi
12.3.2009
27
Conclusions
Increased numbers of chemicals are produced and used
all the time. More information about the concentrations of
organic compounds in water system is needed to assess
impacts of chemicals on the environment.
Chemical diversity of xenobiotics requires the use various
analytical techniques.
Chromatography combined with high resolution mass
spectrometry enables the analysis of complex mixtures
and identification of unknown compounds.
Joonas Nurmi
12.3.2009
28