Republic of Serbia
Ministry of Agriculture and Environmental Protection
Serbian Environmental Protection Agency
Modern laboratory equipment for water quality
monitoring – WFD request
Country visit Serbia, Belgrade, March 22nd, 2017
History of implementation of water monitoring in Serbia
Result on reports of water quality
testing dated from 1965
source: Serbian Environmental Protection Agency
History of implementation of water monitoring in Serbia
2000 – Water Framework Directive
2008 – EQS Directive
2010 – Serbian Water Law
2011-2012 – Several bylaws adopted
2013 – new EQS Directive (August)
2014 – Regulation on limit values of priority and priority hazardous substances polluting surface
water and deadlines for their achievement (February)
Harmonization of surface water status monitoring in the Republic of Serbia with
Water Framework Directive (2000/60/ЕC) requirements
• The first Programe of surface water monitoring status in Serbia harmonized
with the WFD requirements was carried out in 2012
• In Republic of Serbia 498 surface water bodies were determined and grouped
into the following categories: rivers, heavily modified water bodies (HMWB),
artificial water bodies (AWB) and lakes
• The stations for operational and surveillance monitoring were selected (WFD,
Annex V, 1.3.1; 1.3.2)
• 50 surveillance monitoring stations represent “the basis” of water monitoring
network with main goal to provide the whole water status survey within the
catchment areas (Morava, Sava and Danube River)
Surface Water Monitoring Network in 2012-2016
During the period 2012-2016,
monitoring of surface water
status in 181 water bodies were
covered.
IPA 2012
Establisment of an integrated environmental monitoring system for air and water quality
2011 – Project idea and application
2012 - 2014 – Tender procedure and evaluation
March 2015 – Contract signed with supplier
September 2015 – Arrival of the equipment
October/November 2015 – Equipment instalation
December 2015/January2016 – Preparation of Training Program
February – November 2016 Trainings in SEPA and EU laboratories
December 2016 –Audit by Jaroslav Slobodnik, project supervisor
IPA 2012 – List of donation
2.1 Gas chromatograph - time-of-flight - mass spectrometer (GCxGC-TOF-MS) for comprehensive gas
chromatography with automated sample preparation/sample introduction facility and auto sampler system
fully compatible with SBSE and LVI techniques, LECO Instruments
2.2 Gas chromatograph – triple quadrupole - mass spectrometer GC7890/7000MSD (GC-MS/MS) with
automated sample preparation/sample introduction, Agilent Technologies
2.3 Ultra High Performance Liquid Chromatograph (UHPLC) 1290 with Diode Array UV Detector (DAD UV)
and high resolution MS/MS spectrometer 6550 with on-line SPE sample preparation system, Agilent
Technologies
2.4 Fully automated analyzer for the determination of mercury in liquid samples Mercury plus BU, Analytik
Jena AG
2.5 Gel Permeation Chromatograph 1260 for preparation of sediment and biota samples, Agilent
Technologies
IPA 2012 – List of donation
2.6 Lyophylizer for preparation of sediment and biota samples Lyoquest, Telstar Technologies, S.L.U.
2.7 Vibratory Sieve Shaker with set of sieves (0,063-2,000 mm) for preparation of sediment samples AS200,
Retsch GmbH
2.8 Mortar grinder with PTFE scraper for preparation of sediment samples RM200, Retsch GmbH
2.9 Inverted microscope with dark and light field and phase contrast with camera and image analysis for
phytoplankton analysis Axio Observer D1, Carl Zeiss GmbH
2.10 Stereomicroscope with gooseneck type cold light with camera and image analysis for analysis of
benthic invertebrates SteREO Discovery V8, Carl Zeiss GmbH
2.11 Chemicals – Standards and reagents
2.12 Trainings for 2.1, 2.2 & 2.3 in duration of 175 days
2013/39/EU – Environmental Quality Standards Directive - priority and priority
hazardous substances
No.
CAS Number
Priority substance
Analytical method
1
15972-60-8
Alachlor
GC-MS
2
120-12-7
Anthracene
GC-MS
3
1912-24-9
Atrazine
GC-MS/LC-MS
4
71-43-2
Benzene
GC-MS
5
not applicable
Brominated diphenylethers
GC-MS
6
7440-43-9
Cadmium and its compounds
AAS/ICP-MS
7
85535-84-8
Chloroalkanes, C10-13
GC-MS
8
470-90-6
Chlorfenvinphos
GC-MS
9
2921-88-2
Chlorpyrifos (Chlorpyrifos-ethyl)
GC-MS
10
107-06-2
1,2-dichloroethane
GC-MS
11
75-09-2
Dichloromethane
GC-MS
Biota
*
2013/39/EU – Environmental Quality Standards Directive - priority and priority
hazardous substances
No. CAS Number Priority substance
Analytical method
Biota
12
117-81-7
Di(2-ethylhexyl)phthalate (DEHP)
GC-MS/LC-MS
13
330-54-1
Diuron
LC-MS
14
115-29-7
Endosulfan
GC-MS
15
206-44-0
Fluoranthene
GC-MS
*
16
118-74-1
Hexachlorobenzene
GC-MS
*
17
87-68-3
GC-MS
*
18
608-73-1
Hexachlorobutadiene
Hexachlorocyclohexane
19
34123-59-6
Isoproturon
LC-MS
20
7439-92-1
Lead and its compounds
AAS/ICP-MS
21
7439-97-6
Mercury and its compounds
AAS
22
91-20-3
Naphthalene
GC-MS
GC-MS
2013/39/EU – Environmental Quality Standards Directive - priority and priority
hazardous substances
No. CAS Number
Priority substance
Analytical method
23
7440-02-0
Nickel and its compounds
AAS/ICP-MS
24
not applicable
Nonylphenols
GC-MS
25
not applicable
Octylphenols
GC-MS
26
608-93-5
Pentachlorobenzene
GC-MS
27
87-86-5
Pentachlorophenol
GC-MS
28
not applicable
Polyaromatic hydrocarbons (PAH)
GC-MS
29
122-34-9
Simazine
LC-MS
30
not applicable
Tributyltin compounds
31
12002-48-1
Trichlorobenzenes
GC-MS
32
67-66-3
Trichloromethane (chloroform)
GC-MS
33
1582-09-8
Trifluralin
GC-MS
Biota
*
2013/39/EU – Environmental Quality Standards Directive - priority and priority
hazardous substances
No. CAS Number
Priority substance
Analytical method
Biota
34
115-32-2
Dicofol
GC-MS
*
35
1763-23-1
Perfluorooctane sulfonic acid and its derivatives (PFOS)
LC-MS
*
36
124495-18-7
Quinoxyfen
GC-MS
37
not applicable
Dioxine & dioxine like compounds
GC-MS
38
74070-46-5
Aclonifen
GC-MS
39
42576-02-3
Bifenox
GC-MS
40
28159-98-0
Cybutryne
GC-MS
41
52315-07-8
Cypermethrin
GC-MS
42
62-73-7
Dichlorvos
GC-MS
43
not applicable
Hexabromocyclododecane (HBCDD)
GC-MS
*
44
76-44-8/1024-57-3 Heptachlor & heptachlor epoxide
GC-MS
*
45
886-50-0
GC-MS
Terbutryn
*
2.1 Gas chromatograph - time-of- flight - mass spectrometer (GCxGC-TOF-MS)
Gas chromatograph - time-of- flight - mass
spectrometer (GCxGC-TOF-MS) for comprehensive
gas chromatography with automated sample
preparation/sample introduction facility and
autosampler system fully compatible with SBSE and
LVI techniques
Analytical methods:
1. Pesticides
2. Industrial pollutants
3. PAHs
2.1 Gas chromatograph - time-of- flight - mass spectrometer (GCxGC-TOF-MS)- PAH
2.1 Gas chromatograph - time-of- flight - mass spectrometer (GCxGC-TOF-MS)-PAHs
PAH analytes as in the EQS Directive µg/l
Compound
Naphtalene
MAC
130
Max conc
Postive results
0.003
48
0.0002
6
0.0002
8
Phenanthrene
0.0002
16
Pyrene
0.0002
2
Benzo(a) anthracene
0.0008
30
Chrysene
0.0008
30
Fluorene
Anthracene
0.1
Benzo(b) fluoranthene
0.017
0.0007
14
Benzo(k) fluoranthene
0.017
0.0002
24
Benzo(a) pyrene
0.27
0.0008
14
0.0006
15
Indeno(1,2,3,-cd) pyrene
2.1 Gas chromatograph - time-of- flight - mass spectrometer (GCxGC-TOF-MS), Pesticides
Pesticide/biocide analytes as in the EQS Directive µg/l
Compund
Alpha HCH
MAC
Min conc
Max conc
Postive results
0.002
0.0002
0.002
30
Simazine
4
0.0002
0.001
15
Atrazine
2
0.0002
0.001
24
0.002
0.0008
3
0.002
0.0002
0.0006
0.0060
32
0.7
0.0002
0.0006
50
Heptachlor
3*10-5
0.0002
0.002
36
Terbutryn
0.034
0.0002
0.004
16
0.1
0.0002
0.0006
40
Lindane
Delta /-HCH
Alachlor
Chlorpyrifos
2.1 Gas chromatograph - time-of- flight - mass spectrometer (GCxGC-TOF-MS), Pesticide
Pesticide analytes as in the EQS Directive µg/l
Compound
MAC
Dicofol
0.0012
AA-EQS
Min conc
Max conc
Postive results
0.0002
0.003
12
0.0004
0.007
18
0.004-total
0.01-total
16
Dieldrin
0.0002
0.0016
12
Endrin
0.0002
0.0006
5
Endosulfan beta
0.0002
0.001
15
Endosulfan alfa
p,p'-DDE, p,p'-DDD, o,p'-DDT, p,p'-DDT
total 0.025
2.1 Gas chromatograph - time-of- flight - mass spectrometer (GCxGC-TOF-MS) IPS
Industrial pollutant analytes as in the EQS Directive µg/l
Compound
MAC
Min conc
Phenol, 4-(1,1,3,3-tetramethylbutyl)-
0.01 AA-EQS
Phenol, 4-(1,1-dimethylpropyl)4-Nonylphenol
2
Max conc
Postive results
0.0002
0.005
8
0.001
0.2
34
0.0006
0.002
8
2.2 Gas chromatograph – triple quadrupole - mass spectrometer (GC-MS/MS) with
automated sample preparation/sample introduction
Gas chromatograph – triple quadrupole - mass
spectrometer (GC-MS/MS) with automated sample
preparation/sample introduction facility for analysis of
WFD priority substances and identification of river basin
specific pollutants.
Analytical methods:
1. Pesticides and biocides
2. Polyaromatic hydrocarbons
3. Industrial pollutants
4. Volatile organic compounds
5. Pentachlorophenol
6. Brominated diphenylethers
7. Chloroalkanes C10-13
8. Dioxini, furani and PCB-DLs
2.2 Gas chromatograph – triple quadrupole - mass spectrometer (GC-MS/MS) with
automated sample preparation/sample introduction
Pesticides and biocides
Lindan
Analyzed compounds:
 Atrazine
 Beta-HCH
 Alachlor
 Gamma-HCH(Lindan)
 Chlorfenvinphos
 Delta-HCH
 Chlorpyrifos
 Simazine
 Aldrin
 Trifluralin
 Isodrin
 Bifenox
 Dieldrin
 Cybutryne
 Endrin
 Cypermethrin
 Endosulfan-alpha
 Dicofol
 Endosulfan-beta
 Dichlorvos
 4,4’-DDE
 Heptachlor
 4,4’-DDD
 Heptachlor epoxide
 2,4’-DDT
 Quinoxyfen
 4,4’-DDT
 Terbutryn
 Alpha-HCH
 Aclonifen
Lindan
Chlorfenvinpho
s
Chlorfenvinphos
2.2 Gas chromatograph – triple quadrupole - mass spectrometer (GC-MS/MS) with
automated sample preparation/sample introduction
Pesticide/biocide analytes as in the EQS Directive µg/l
MAC
Min conc
Max conc
Postive results
Atrazine
2.0
0.0041
0.0162
37
Alachlor
0.7
0.0412
1
Terbutryn
0.34
0.0193
2
0.0040
1
Compound
0.0024
Alpha-HCH
Beta-HCH
Gamma-HCH(Lindan)
sum 0.04
Delta-HCH
Simazine
Cypermethrin
Dicofol
4
0.0051
0.0113
6
6x10-4
0.0001
0.0056
93
NA
0.0014
0.0016
1
2.2 Gas chromatograph – triple quadrupole - mass spectrometer (GC-MS/MS) with
automated sample preparation/sample introduction
PAH analytes as in the EQS Directive µg/l
Compound
Min conc
MAC
Max conc
Postive results
Anthracene
0.1
0.001
0.006
6
Fluoranthene
0.12
0.002
0.004
3
Benzo(a)pyrene
0.27
0.001
2
Benzo(a)anthracene
/
0.001
6
Benzo(b)fluoranthene
0.017
0.001
100
2.2 Gas chromatograph – triple quadrupole - mass spectrometer (GC-MS/MS) with
automated sample preparation/sample introduction
Industrial pollutants
Analyzed compounds:
Hexachlorobenzene
 Hexachlorobenzene  4-tert-Octylphenol
 Pentachlorbenzene  DEHP (Phthalic acid,
bis-2 4-n-Nonylphenol
ethylhexylester)
 4-Nonylphenols
(technical)
Hexachlorobenzene
Industrial pollutant analytes as in the EQS Directive µg/l
Compound
DEHP (Phthalic acid, bis2-ethylhexylester)
AAC
Min conc
1,3
0.025
Max conc
8.303
Postive results
100
2.2 Gas chromatograph – triple quadrupole - mass spectrometer (GC-MS/MS) with
automated sample preparation/sample introduction
VOC analytes as in the EQS Directive µg/l
Compound
MAC
Min conc
Max conc
Postive results
Benzen
50
0.149
11.954
75
Carbon tetrachloride
NA
0.122
5.065
78
Hexachloro-1,3-butadien
0.6
0.125
0.423
78
Naphtalene
130
0.267
12.995
78
Tetrachloroethene
NA
0.154
3.754
7
Trichloromethane
NA
3.406
7.155
5
0.196
0.526
68
0.120
0.641
62
0.184
0.590
74
1,3,5-Trichlorobenzene
1,2,4-Trichlorobenzene
1,2,3-Trichlorobenzene
NA
2.2 Gas chromatograph – triple quadrupole - mass spectrometer (GC-MS/MS) with
automated sample preparation/sample introduction
Pentachlorphenol
Pentachlorphenol
Analyzed compounds:
 Pentachlorphenol
PCHP analyte as in the EQS Directive µg/l
Compound
PCHP
MAC
Max conc
Min conc
1
/
Postive results
/
0
2.2 Gas chromatograph – triple quadrupole - mass spectrometer (GC-MS/MS) with
automated sample preparation/sample introduction
Brominated diphenylethers
Analyzed compounds:
 Pentabromodiphenylether
Pentabromodiphenylether
BDE analyte as in the EQS Directive µg/l
Compound
BDE
MAC
Min conc
0.14
Max conc
0.002
Postive results
1
2.3 Ultra High Performance Liquid Chromatograph (UHPLC) with Diode Array UV Detector
(DAD UV) and high resolution MS/MS spectrometer with on-line SPE sample preparation
Ultra High Performance Liquid Chromatograph (UHPLC)
with Diode Array UV Detector (DAD UV) and high resolution
MS/MS spectrometer with on-line SPE sample preparation
system, chromatographic control and data management
station with software for analysis of WFD priority
substances and identification of river basin specific
pollutants.
Analytical methods:
1. Triazine and phenylureate herbicides
2. Neoicotinoide pesticides
3. Pharmaceuticals – macrolide antibiotics
4. Estrogens
5. Phtalates
6. PFOS
2.3 Ultra High Performance Liquid Chromatograph (UHPLC) with Diode Array UV Detector
(DAD UV) and high resolution MS/MS spectrometer with on-line SPE sample preparation
Triazine and phenylureate herbicides
Analyzed compounds:
 Desethylatrazine
 Methabenthiazuron
 Monolinuron
 Isoproturon
 Sebuthylazine
 Metobromuron
 Diuron
 Chlorotoluron
 Metazachlor
 Terbutylazine
 Linuron
 Metolachlor
 Simazine
 Hexazinon
 Metoxuron
 Cyanazine
 Atrazine
Atrazine
Isoproturon
2.3 Ultra High Performance Liquid Chromatograph (UHPLC) with Diode Array UV Detector
(DAD UV) and high resolution MS/MS spectrometer with on-line SPE sample preparation
Pesticide analytes as in the EQS Directive µg/l
Compound
Desethylatrazine
MAC
Max conc
Postive results
N/A
<0.0001
0
Simazine
4.000
<0.0001
0
Hexazinon
N/A
<0.0001
0
Metoxuron
N/A
<0.0001
0
Cyanazine
N/A
<0.0001
0
2.000
0.0041
37
Methabenzthiazuron
N/A
0.0104
0
Monolinuron
N/A
0.0065
62
Atrazin
2.3 Ultra High Performance Liquid Chromatograph (UHPLC) with Diode Array UV Detector
(DAD UV) and high resolution MS/MS spectrometer with on-line SPE sample preparation
Pesticide analytes as in the EQS Directive µg/l
Compound
Isoproturon
MAC
Max conc
Postive results
1.000
0.0076
58
Sebuthylazine
N/A
0.0015
0
Metobromuron
N/A
0.0021
0
1.800
0.0038
8
Metazachlor
N/A
0.0033
16
Terbutylazine
N/A
0.013
67
Linuron
N/A
0.014
2
Metolachlor
N/A
0.0021
25
Chlorotoluron
N/A
0.0007
12
Diuron
2.3 Ultra High Performance Liquid Chromatograph (UHPLC) with Diode Array UV Detector
(DAD UV) and high resolution MS/MS spectrometer with on-line SPE sample preparation
Neonicotinoide analytes as in the EQS Directive µg/l
Compound
PNEC
Max conc
Postive results
Thiamethoxam
0.140
0.015
4
Clothianidin
0.130
0.012
5
Imidacloprid
0.009
0.016
39 (8 over limit)
Acetamiprid
0.500
0.019
9
Thiacloprid
0.050
0.047
6
The Predicted No Effect Concentration (PNEC) is the concentration of a chemical which marks the limit at which below no
adverse effects of exposure in an ecosystem are measured
2.3 Ultra High Performance Liquid Chromatograph (UHPLC) with Diode Array UV Detector
(DAD UV) and high resolution MS/MS spectrometer with on-line SPE sample preparation
Pharmaceuticals – macrolide antibiotics
Analyzed compounds:
 Azithromycin
 Clarithromycin
 Erythromycin
 Diclofenac
Erythromycin
2.3 Ultra High Performance Liquid Chromatograph (UHPLC) with Diode Array UV Detector
(DAD UV) and high resolution MS/MS spectrometer with on-line SPE sample preparation
Pharmaceutical analytes as in the EQS Directive µg/l
Compound
PNEC
Max conc
Postive results
Azithromycin
0.09
0.0710
3
Clarithromycin
0.13
<0.0003
0
Erythromycin
0.20
<0.0003
0
Diclofenac
0.050
0.0065
35
2.3 Ultra High Performance Liquid Chromatograph (UHPLC) with Diode Array UV Detector
(DAD UV) and high resolution MS/MS spectrometer with on-line SPE sample preparation
Estrogens
Alpha and beta
estradiol
Analyzed compounds:
 Alpha estradiol
 Beta estradiol
 Estrone
 Ethynyl estradiol
Alpha estradiol
Beta estradiol
2.3 Ultra High Performance Liquid Chromatograph (UHPLC) with Diode Array UV Detector
(DAD UV) and high resolution MS/MS spectrometer with on-line SPE sample preparation
Estrogen analytes as in the EQS Directive µg/l
Compound
PNEC
Max conc
Postive results
Alpha estradiol
N/A
0.00034
55
Beta estradiol
N/A
0.00033
53
0.0036
0.00016
65
N/A
0.0007
44
Estrone
Ethynyl estradiol
2.3 Ultra High Performance Liquid Chromatograph (UHPLC) with Diode Array UV Detector
(DAD UV) and high resolution MS/MS spectrometer with on-line SPE sample preparation
PFOS
PFOS and PFOA
Analyzed compounds:
 PFOS
 PFOA
PFOS
PFOA
PFOS analytes as in the EQS Directive µg/l
Compound
MAC
Max conc
Postive results
PFOS
36
0.0034
20
PFOA
36
0.0022
26
2.4 Fully automated analyser for the determination of mercury in liquid samples, Analytik
Jena AG Mercur plus BU
Hg as in the EQS Directive µg/l
Compound
Hg
Min conc
MAC
0.07
0.005
Max conc
0.08
Postive results
1
2.9 Inverted microscope with dark and light field and phase contrast with camera and image
analysis for phytoplankton analysis, Carl Zeiss GmbH, Axio Observer D1
Biological Quality Elements investigation
-Phytoplankton (including Cyanobacteria) and phytobenthos
-Identification of pressures: eutrophication and organic pollution
-Ecological status/potential assessment according to the Water Framework Directive 2000/60/EC
Diatom indices calculation (OMNIDIA v. 5.5)
Diatom indices used in ecological status/potential assessment according to
the National Monitoring Programme
Phytoplankton (including Cyanobacteria) - eutrophication indicator
Parameters: Phytoplankton Abundance and Phytoplankton Biomass (chlorophyll a)
Planktothrix agardhii species abundance with
regard to depth of the water column in the
Garaši Reservoir in 2016
-Algal blooms in the Garaši Reservoir
caused by the species Planktothrix agardhii
in 2016 (this is potentially toxic species can
produce microcystin-LR (hepatotoxin)
0
50000
100000
150000
200000
250000
300000
350000
400000
450000
500000
550000
600000
650000
700000
аbundance (cell ml-1)
depth (m)
0,0
1,0
2,0
3,0
4,0
5,0
6,0
7,0
8,0
9,0
10,0
11,0
12,0
13,0
14,0
15,0
16,0
17,0
18,0
19,0
20,0
21,0
bad ecological potential
May
August
-The presence of Euglena species indicate high
organic pollution
2.10 Stereomicroscope with gooseneck type cold light with camera and image analysis for
analysis of benthic invertebrates, Carl Zeiss GmbH, SteREO Discovery V8
-Benthic invertebrate community composition and structure
analysis
-Identification of pressures: organic pollution and
hydromorphological degradation
-Ecological status/potential assessment according to the
Water Framework Directive 2000/60/EC
-ZEN 2 Microscope and Imaging
Software
ASTERICS v. 4.0.4 Software for benthic invertebrate metrics
-Zelinka & Marvan Saprobic Index, BMWP Score, ASPT,
Shannon-Wiener Diversity Index, Total No. of Taxa, EPT Taxa,
No. of Sensitive Taxa (Austrian list), Percentage Participation
of Oligochaeta/Tubificidae
-Sampling according to the AQEM protocol
Future – Guidelines on further harmonization of surface water monitoring to the WFD
 Defining a list of the specific polluting substances and their Environmental Quality Standards
(EQS)
 Monitoring of priority and priority hazardous substances in Biota/Sediment
 Defining a clear methodology to determine appropriate values in ecological and chemical status
assessment
 Revising the criteria considering the level of confidence and precision in ecological status
classification
 Amendment the Regulation on reference conditions of surface water types
 Amendment the Regulation on the parameters of ecological and chemical status of surface
waters and parameters of chemical status and quantitative status of groundwater
 Revision the list of parameters and ecological status class boundaries
Future – Establishing functional Regional Monitoring Centers within the SEPA
Quality Control - Quality management system
Quality management system is
established 2006 by acceptance of
general requirements specified in
SRPS ISO/IEC 17025:2006.
It covers testing and calibration
performed using standard methods,
non-standard methods and
laboratory-developed methods.
Acknowledgement
Ministry of Agriculture and Environment Protection, Republic of Serbia
Rainer Freund, ing. PhD, Project Manager, EU Delegation in Serbia
Ivan Špánik, ing. PhD, Faculty of Chemical and Food Technology, Bratislava, Slovakia
Olga Vyviurska, ing. PhD, Faculty of Chemical and Food Technology, Bratislava, Slovakia
Jaroslav Slobodnik, ing. PhD, Masaryk University, Brno, Czech Republic
Republic of Serbia
Ministry of Agriculture and Environmental Protection
Serbian Environmental Protection Agency
Thanks for your attention
Mirjana Balać, chemist
Aleksandra Djurkovic, biologist
Daliborka Popadić, physical chemist
Snežana Čadjo, biologist
Ivana Dersek Timotić, chemist
Ana Vujović, physical shemist
Katarina Nedeljković, chemist
Boris Novaković, biologist
Zoran Stojanović, chemist