Targeted and Non-Targeted Screening of Biocides in Urban Stormwater Runoff Julia Quilitzki,1 Patricia van Baar,1 Uwe Dünnbier,1 Daniel Wicke,2 Maciej Bromirski,3 Olaf Scheibner3 1Berliner Wasserbetriebe, Berlin, Germany; 2Kompetenzzentrum Wasser Berlin, Berlin, Germany; 3Thermo Fisher Scientifi c, Bremen, Germany;

Po

ster No

te 64

633

Sampling strategy

The stormwater runoff was collected with portable automatic samplers equipped with 8glass bottles (V=1.9 L). The sampler was triggered by the water level in the storm sewer(threshold: 10 cm), sample volume was 450 mL. At the beginning of a storm event,sampling intervals were shorter (5 min in the first 2 hours) to register a potential first flusheffect. After the first two hours of sampling the sampling interval was increased (every 15min). The maximum sampling duration was 4 hours.

A volume-proportional composite sample for analysis was prepared by manual mixing ofthe flow-corresponding volumes of each bottle (see Figure 3) calculated from the flow ofthe installed flow meter. In that way, the sample represents the average concentration ofthe sampled storm event.

Targeted and Non-Targeted Screening of Biocides in Urban Stormwater RunoffJulia Quilitzki1, Patricia van Baar1, Uwe Dünnbier1, Daniel Wicke², Maciej Bromirski3, Olaf Scheibner3

1Berliner Wasserbetriebe, Berlin, Germany; 2Kompetenzzentrum Wasser Berlin, Berlin, Germany; 3Thermo Fisher Scientific, Bremen, Germany; Email: olaf.scheibner@thermofisher.com

ConclusionThis study shows that suspect screening analysis is an efficient method for detection ofunknown compounds (for example simazine, nicotine, benzalkoniumchloride).The high measured concentrations of substances shows that stormwater runoff can bean important entry pathway for micropollutants to water bodies. Results also indicatethat it is necessary to consider the different urban catchment types with its specificcompounds.

References 1. Gasperi, J. et al. (2014) Micropollutants in urban stormwater: Occurrence, concen-

trations, and atmospheric contributions for a wide range of contaminants in three French catchments. Environmental Science & Pollution Research 21(8), 5282-5283

2. Zgheib, S., Moilleron, R. & Chebbo, G. (2012) Priority pollutants in urban storm-water: Part 1 - Case of separate storm sewers. Water Research 46(20), 6683-6692

OverviewPurpose: Stormwater runoff can be an important source of organic micro-pollutantsentering urban surface waters through separated sewer systems.

Methods: Screening and quantification of micropollutants was carried out for storm-water runoff samples of five different urban catchment areas in Berlin to considercatchment-specific differences.

Results: Stormwater may be a relevant source of biocides, particularly in citiesdominated by separate sewer systems.

IntroductionStormwater runoff can be an important source of organic micropollutants entering urbansurface waters through separate sewer systems (Zgheib et al. 2012). To investigate theextent of this type of pollution, a one year monitoring program was conducted in the cityof Berlin. Monitoring points were selected in five catchments of different urbanstructures to consider catchment-specific differences. Samples were analyzed for acomprehensive set of 100 micropollutants (e.g. PAHs, phthalates, phenols, heavymetals, biocides, flame retardants) that were chosen for monitoring based on literatureresearch (e.g. Gasperi et al. 2014).

MethodsSample PreparationStormwater runoff samples were filtered through 0.45 µm syringe filter before injection.

Liquid Chromatography1 mL of sample was injected directly into the online-SPE system. For pre-concentration,a C18 column, 2.1 × 20 mm with 12 µm particle size was used. For compoundseparation, a C18 column, 2.1 × 50 mm with 1.8 µm particle size was used. A water-methanol gradient both spiked with 0.1% formic acid was run from 2% to 95% in 6.7minutes. Total chromatographic cycle time including online pre-concentration was 15minutes.

Mass Spectrometry

Mass spectrometric analysis was run on a Thermo Scientific™ Q Exactive™ Focushybrid quadrupole-Orbitrap™ mass spectrometer (Figure 1).

MS-parameter for screening purposesFull Scan ESI+ and ESI– separately R = 70,000 m/z 100–1000

variable Data Independent Acquisition (vDIA) R = 70,000 CE = 30 m/z 100–205, 195–305, 295–405, 395–505, 495–1000

FIGURE 1. Q Exactive Focus with Thermo Scientific™ EQuan MAX Plus™ online SPE-system.

SamplingInvestigation areaFive sampling points were installed in separate storm sewers of five different catchmenttypes representing 85% of the connected impervious area of Berlin: one- family houseswith gardens (OFH), roads (>7500 vehicles/ day), old building (OLD, < 1930), newerbuilding (NEW, >1950) and commercial (COM) areas (Figure 2). Additionally a samplingpoint at the river “Panke” (PNK) was installed to determine contaminant concentrationsduring storm events in a stream affected by stormwater discharge.

© 2015 Thermo Fisher Scientific Inc. All rights reserved. All trademarks are the property of Thermo Fisher Scientific and its subsidiaries. This information is not intended to encourage use of these products in any manner that might infringe the intellectual property rights of others.

FIGURE 4. Suspect Screening results: Comparison between the sampling points one- family houses (OFH) and the river “Panke” (PNK).

FIGURE 2. Location of monitoring catchments in Berlin in separate sewer system (outside red line).

MS-parameter for quantitative analysisFull Scan polarity-switching (ESI+ / ESI–) R = 35,000 m/z 103–900 internal standards

FIGURE 6. Result of the pesticide/biocide concentrations for selected compounds for the different sampling locations.

Commercial areas

One-family houses with

gardens

Corresponding flow-proportional volume

5L composite sample

Chemical analysis

FIGURE 3. Sampling strategy: volume-proportional composite samples.

One- family houses(OFH) River �Panke“ (PNK)

Pharmaceuticals

Pesticides

Industrial and household chemicals

FIGURE 1. Process of data acquisition and processing for target and non-target analysis; all processing done with TraceFinder 3.3 software.

Data AnalysisQualitative and quantitative analysis was done with Thermo Scientific™ TraceFinder™software. Figure 1 shows a schematic diagram of the full workflow carried out in thisstudy. Starting with a suspect screening, micropollutants out of different classes likebiocides/ pesticides, pharmaceuticals, household and industrial chemicals containedin a homebuild database, were confirmed by using confirmation criteria such asisotopic pattern match and matching of known fragments. Selected suspects were aswell identifed as quantified by reference standards in a dilution series.

It can be seen that concentrations show distinct differences between catchment types.Whereas mecoprop (contained in bituminous sealing membranes for roofs) can befound in all catchment with roof runoff, carbendazim (fungicide, in urban contextapplied in paints and sealing compounds) and diuron (contained in exterior paints forsurface protection) are mostly found at monitoring station Old, an area of typicalhouses build <1930 with extensive renovations in recent years during which theapplication of new paints and sealing compounds is likely. On the other hand, concentrations of isoproturon (herbicide applied in gardens) are highest in catchmenttype OFH characterized by one family houses with gardens. Glyphosate (broadspectrum herbicide, �RoundUp“) was found at all monitoring stations, probably due toapplications for weed control on footpaths.

Quantitative results

A total of 19 biocides and pesticides were quantified regularly, of which 6 were addedas a result of the suspected Screening (simazine, terbuthylazine,desethylterbuthylazine, benzisothiazolinone, octylisothiazolinone and thiacloprid).Results for selected biocides and pesticides in stormwater runoff of the investigatedcatchment types can be seen in Figure 6.

std200ng_benzalkonium #364 RT: 7.33 AV:F: FTMS + p ESI Full ms2 304.30@hcd4 ...

100 200 300m/z

0

20

40

60

80

100

Relat

ive Ab

unda

nce

91.05483

304.30005

212.23746

110.11267

RT: 7.00 - 7.50

7.0 7.2 7.4Time (min)

0

20

40

60

80

100

Rela

tive

Abun

danc

e

7.33

7.41

7.16 7.277.06

RT: 7.00 - 7.50

7.0 7.2 7.4Time (min)

0

20

40

60

80

100

Rela

tive

Abun

danc

e

7.35

7.397.42

7.297.247.15

279 #432 RT: 7.39 AV: 1 NL: 8.17E6F: FTMS + p ESI Full ms2 304.30@hcd4 ...

100 200 300m/z

0

20

40

60

80

100

Relat

ive Ab

unda

nce

91.05484

304.30005

212.23746

121.48763

reference standard

sample

N+

CH3

CH3

C12H15

XIC 304.29996 (± 5 ppm) MS/MS

Time vs. flow Q[m3/ s]

ResultsSuspected Screening

The suspect screening results showed a large fraction of pesticides/biocides in thesamples indicating the relevance of stormwater as an entry pathway of biocides tosurface waters. The results of two samples are presented in figure 4: one-familiy houses(OFH) and the river “Panke” (PNK). It can be seen that the pesticide/biocide fraction ofidentified suspects (29 for OFH and 56 for PNK) is similar for both samples, whereasthe fraction of pharmaceuticals is much higher in the stream sample (PNK) due todischarges of WWTP effluent upstream the sampling location. For a suspect thefollowing confirming criteria were selected: isotopic pattern score > 85%, an accuratemass deviation < 5 ppm and at least one detected fragments.

OFH: e.g. Nicotine, 2-Hydroxybenzothiazole (OHBT), Benzothiazole (BT) and 2-Methylbenzothiazole (MTBT)

Panke: e.g. OHBT, MTBT, BT, Metoprolol, Carbamazepine, 4-formylaminoantipyrine (FAA), 4-acetylaminoantipyrine (AAA), Simazine, Terbutylazine, Ketamine, Tramadol

Suspect compound Formula [M+H]+ ∆ m/z(ppm)

Time (min)

IPS (%)

Isoproturon C12H18N2O 207.1492 -0.4 6.0 100Metolachlor C15H22ClNO2 284.1411 0.6 7.3 100Diethyltoluamide (DEET) C12H17NO 192.1383 0.0 6.4 100Benzisothiazolinone (BIT) C7H5NOS 152.0165 0.7 5.4 100

TABLE 1. Detected pesticides/biocides of sample point “new buildings” (NEW) with confirming criteria mass accuracy (Δm/z) and isotopic pattern score (IPS).

Whereas isoproturon and metolachlor are two plant protection products, diethyltoluamide(DEET) is an insect repellent and benzisothiazolinone (BIT) a biocide which is used inexterior and roof paints, caulks and varnish.

Non-Target Screening

In addition to the suspect screening an unknown screening with unbiased peak detectionwas carried out to screen for additional compounds of interest that had not come toattention yet. The first approach was to look into the 20 most intense compounds ofevery sample. Few of these masses were selected for parallel reaction monitoring (PRM)to get additional MS/MS information. The detected masses were m/z 304.29999,276.19492, 262.25311 and 248.16394. This is a sign for a homologous sequence (-CH2)of quaternary ammonium compounds.

A prominent quaternary ammonium compound is benzalkoniumchloride. Benzalkonium-chloride is used as biocide or cationic surfactant (disinfection). It has different alkyl chainlengths between C4-C18. The chosen selected masses have a chain length of C8, C9,C10 and C12. Figure 5 shows exemplary the results of benzalkoniumchloride C12, however, other chain lengths were also detected in the sample. The findings wereverified with reference standards.

In table 1 biocides detected by suspected screening of a sample of catchment NEW areshown. Beside these, further substances from other classes were found(pharmaceuticals, industrial-, and household chemicals).

FIGURE 5. Benzalkoniumchloride peak in the sample and the reference standard peak with MS/MS spectra.

C7H7+

C21H38N+

C14H30N+

Commercial Area [COM]

Newer buildings [NEW]

Street runoff [STR] – 1.3 km

Old buildings [OLD]

One-family houses with gardens [OFH]

Combined sewer area

ESI+

Sampling strategy

The stormwater runoff was collected with portable automatic samplers equipped with 8glass bottles (V=1.9 L). The sampler was triggered by the water level in the storm sewer(threshold: 10 cm), sample volume was 450 mL. At the beginning of a storm event,sampling intervals were shorter (5 min in the first 2 hours) to register a potential first flusheffect. After the first two hours of sampling the sampling interval was increased (every 15min). The maximum sampling duration was 4 hours.

A volume-proportional composite sample for analysis was prepared by manual mixing ofthe flow-corresponding volumes of each bottle (see Figure 3) calculated from the flow ofthe installed flow meter. In that way, the sample represents the average concentration ofthe sampled storm event.

Targeted and Non-Targeted Screening of Biocides in Urban Stormwater RunoffJulia Quilitzki1, Patricia van Baar1, Uwe Dünnbier1, Daniel Wicke², Maciej Bromirski3, Olaf Scheibner3

1Berliner Wasserbetriebe, Berlin, Germany; 2Kompetenzzentrum Wasser Berlin, Berlin, Germany; 3Thermo Fisher Scientific, Bremen, Germany; Email: olaf.scheibner@thermofisher.com

ConclusionThis study shows that suspect screening analysis is an efficient method for detection ofunknown compounds (for example simazine, nicotine, benzalkoniumchloride).The high measured concentrations of substances shows that stormwater runoff can bean important entry pathway for micropollutants to water bodies. Results also indicatethat it is necessary to consider the different urban catchment types with its specificcompounds.

References 1. Gasperi, J. et al. (2014) Micropollutants in urban stormwater: Occurrence, concen-

trations, and atmospheric contributions for a wide range of contaminants in three French catchments. Environmental Science & Pollution Research 21(8), 5282-5283

2. Zgheib, S., Moilleron, R. & Chebbo, G. (2012) Priority pollutants in urban storm-water: Part 1 - Case of separate storm sewers. Water Research 46(20), 6683-6692

OverviewPurpose: Stormwater runoff can be an important source of organic micro-pollutantsentering urban surface waters through separated sewer systems.

Methods: Screening and quantification of micropollutants was carried out for storm-water runoff samples of five different urban catchment areas in Berlin to considercatchment-specific differences.

Results: Stormwater may be a relevant source of biocides, particularly in citiesdominated by separate sewer systems.

IntroductionStormwater runoff can be an important source of organic micropollutants entering urbansurface waters through separate sewer systems (Zgheib et al. 2012). To investigate theextent of this type of pollution, a one year monitoring program was conducted in the cityof Berlin. Monitoring points were selected in five catchments of different urbanstructures to consider catchment-specific differences. Samples were analyzed for acomprehensive set of 100 micropollutants (e.g. PAHs, phthalates, phenols, heavymetals, biocides, flame retardants) that were chosen for monitoring based on literatureresearch (e.g. Gasperi et al. 2014).

MethodsSample PreparationStormwater runoff samples were filtered through 0.45 µm syringe filter before injection.

Liquid Chromatography1 mL of sample was injected directly into the online-SPE system. For pre-concentration,a C18 column, 2.1 × 20 mm with 12 µm particle size was used. For compoundseparation, a C18 column, 2.1 × 50 mm with 1.8 µm particle size was used. A water-methanol gradient both spiked with 0.1% formic acid was run from 2% to 95% in 6.7minutes. Total chromatographic cycle time including online pre-concentration was 15minutes.

Mass Spectrometry

Mass spectrometric analysis was run on a Thermo Scientific™ Q Exactive™ Focushybrid quadrupole-Orbitrap™ mass spectrometer (Figure 1).

MS-parameter for screening purposesFull Scan ESI+ and ESI– separately R = 70,000 m/z 100–1000

variable Data Independent Acquisition (vDIA) R = 70,000 CE = 30 m/z 100–205, 195–305, 295–405, 395–505, 495–1000

FIGURE 1. Q Exactive Focus with Thermo Scientific™ EQuan MAX Plus™ online SPE-system.

SamplingInvestigation areaFive sampling points were installed in separate storm sewers of five different catchmenttypes representing 85% of the connected impervious area of Berlin: one- family houseswith gardens (OFH), roads (>7500 vehicles/ day), old building (OLD, < 1930), newerbuilding (NEW, >1950) and commercial (COM) areas (Figure 2). Additionally a samplingpoint at the river “Panke” (PNK) was installed to determine contaminant concentrationsduring storm events in a stream affected by stormwater discharge.

© 2015 Thermo Fisher Scientific Inc. All rights reserved. All trademarks are the property of Thermo Fisher Scientific and its subsidiaries. This information is not intended to encourage use of these products in any manner that might infringe the intellectual property rights of others.

FIGURE 4. Suspect Screening results: Comparison between the sampling points one- family houses (OFH) and the river “Panke” (PNK).

FIGURE 2. Location of monitoring catchments in Berlin in separate sewer system (outside red line).

MS-parameter for quantitative analysisFull Scan polarity-switching (ESI+ / ESI–) R = 35,000 m/z 103–900 internal standards

FIGURE 6. Result of the pesticide/biocide concentrations for selected compounds for the different sampling locations.

Commercial areas

One-family houses with

gardens

Corresponding flow-proportional volume

5L composite sample

Chemical analysis

FIGURE 3. Sampling strategy: volume-proportional composite samples.

One- family houses(OFH) River �Panke“ (PNK)

Pharmaceuticals

Pesticides

Industrial and household chemicals

FIGURE 1. Process of data acquisition and processing for target and non-target analysis; all processing done with TraceFinder 3.3 software.

Commercial areas

One-family houses with

gardens

Data AnalysisQualitative and quantitative analysis was done with Thermo Scientific™ TraceFinder™software. Figure 1 shows a schematic diagram of the full workflow carried out in thisstudy. Starting with a suspect screening, micropollutants out of different classes likebiocides/ pesticides, pharmaceuticals, household and industrial chemicals containedin a homebuild database, were confirmed by using confirmation criteria such asisotopic pattern match and matching of known fragments. Selected suspects were aswell identifed as quantified by reference standards in a dilution series.

It can be seen that concentrations show distinct differences between catchment types.Whereas mecoprop (contained in bituminous sealing membranes for roofs) can befound in all catchment with roof runoff, carbendazim (fungicide, in urban contextapplied in paints and sealing compounds) and diuron (contained in exterior paints forsurface protection) are mostly found at monitoring station Old, an area of typicalhouses build <1930 with extensive renovations in recent years during which theapplication of new paints and sealing compounds is likely. On the other hand, concentrations of isoproturon (herbicide applied in gardens) are highest in catchmenttype OFH characterized by one family houses with gardens. Glyphosate (broadspectrum herbicide, �RoundUp“) was found at all monitoring stations, probably due toapplications for weed control on footpaths.

Quantitative results

A total of 19 biocides and pesticides were quantified regularly, of which 6 were addedas a result of the suspected Screening (simazine, terbuthylazine,desethylterbuthylazine, benzisothiazolinone, octylisothiazolinone and thiacloprid).Results for selected biocides and pesticides in stormwater runoff of the investigatedcatchment types can be seen in Figure 6.

std200ng_benzalkonium #364 RT: 7.33 AV:F: FTMS + p ESI Full ms2 304.30@hcd4 ...

100 200 300m/z

0

20

40

60

80

100

Relat

ive Ab

unda

nce

91.05483

304.30005

212.23746

110.11267

RT: 7.00 - 7.50

7.0 7.2 7.4Time (min)

0

20

40

60

80

100

Rela

tive

Abun

danc

e

7.33

7.41

7.16 7.277.06

RT: 7.00 - 7.50

7.0 7.2 7.4Time (min)

0

20

40

60

80

100

Rela

tive

Abun

danc

e

7.35

7.397.42

7.297.247.15

279 #432 RT: 7.39 AV: 1 NL: 8.17E6F: FTMS + p ESI Full ms2 304.30@hcd4 ...

100 200 300m/z

0

20

40

60

80

100

Relat

ive Ab

unda

nce

91.05484

304.30005

212.23746

121.48763

reference standard

sample

N+

CH3

CH3

C12H15

XIC 304.29996 (± 5 ppm) MS/MS

Time vs. flow Q[m3/ s]

ResultsSuspected Screening

The suspect screening results showed a large fraction of pesticides/biocides in thesamples indicating the relevance of stormwater as an entry pathway of biocides tosurface waters. The results of two samples are presented in figure 4: one-familiy houses(OFH) and the river “Panke” (PNK). It can be seen that the pesticide/biocide fraction ofidentified suspects (29 for OFH and 56 for PNK) is similar for both samples, whereasthe fraction of pharmaceuticals is much higher in the stream sample (PNK) due todischarges of WWTP effluent upstream the sampling location. For a suspect thefollowing confirming criteria were selected: isotopic pattern score > 85%, an accuratemass deviation < 5 ppm and at least one detected fragments.

OFH: e.g. Nicotine, 2-Hydroxybenzothiazole (OHBT), Benzothiazole (BT) and 2-Methylbenzothiazole (MTBT)

Panke: e.g. OHBT, MTBT, BT, Metoprolol, Carbamazepine, 4-formylaminoantipyrine (FAA), 4-acetylaminoantipyrine (AAA), Simazine, Terbutylazine, Ketamine, Tramadol

Suspect compound Formula [M+H]+ ∆ m/z(ppm)

Time (min)

IPS (%)

Isoproturon C12H18N2O 207.1492 -0.4 6.0 100Metolachlor C15H22ClNO2 284.1411 0.6 7.3 100Diethyltoluamide (DEET) C12H17NO 192.1383 0.0 6.4 100Benzisothiazolinone (BIT) C7H5NOS 152.0165 0.7 5.4 100

TABLE 1. Detected pesticides/biocides of sample point “new buildings” (NEW) with confirming criteria mass accuracy (Δm/z) and isotopic pattern score (IPS).

Whereas isoproturon and metolachlor are two plant protection products, diethyltoluamide(DEET) is an insect repellent and benzisothiazolinone (BIT) a biocide which is used inexterior and roof paints, caulks and varnish.

Non-Target Screening

In addition to the suspect screening an unknown screening with unbiased peak detectionwas carried out to screen for additional compounds of interest that had not come toattention yet. The first approach was to look into the 20 most intense compounds ofevery sample. Few of these masses were selected for parallel reaction monitoring (PRM)to get additional MS/MS information. The detected masses were m/z 304.29999,276.19492, 262.25311 and 248.16394. This is a sign for a homologous sequence (-CH2)of quaternary ammonium compounds.

A prominent quaternary ammonium compound is benzalkoniumchloride. Benzalkonium-chloride is used as biocide or cationic surfactant (disinfection). It has different alkyl chainlengths between C4-C18. The chosen selected masses have a chain length of C8, C9,C10 and C12. Figure 5 shows exemplary the results of benzalkoniumchloride C12, however, other chain lengths were also detected in the sample. The findings wereverified with reference standards.

In table 1 biocides detected by suspected screening of a sample of catchment NEW areshown. Beside these, further substances from other classes were found(pharmaceuticals, industrial-, and household chemicals).

FIGURE 5. Benzalkoniumchloride peak in the sample and the reference standard peak with MS/MS spectra.

C7H7+

C21H38N+

C14H30N+

Commercial Area [COM]

Newer buildings [NEW]

Street runoff [STR] – 1.3 km

Old buildings [OLD]

One-family houses with gardens [OFH]

Combined sewer area

ESI+

Sampling strategy

The stormwater runoff was collected with portable automatic samplers equipped with 8glass bottles (V=1.9 L). The sampler was triggered by the water level in the storm sewer(threshold: 10 cm), sample volume was 450 mL. At the beginning of a storm event,sampling intervals were shorter (5 min in the first 2 hours) to register a potential first flusheffect. After the first two hours of sampling the sampling interval was increased (every 15min). The maximum sampling duration was 4 hours.

A volume-proportional composite sample for analysis was prepared by manual mixing ofthe flow-corresponding volumes of each bottle (see Figure 3) calculated from the flow ofthe installed flow meter. In that way, the sample represents the average concentration ofthe sampled storm event.

Targeted and Non-Targeted Screening of Biocides in Urban Stormwater RunoffJulia Quilitzki1, Patricia van Baar1, Uwe Dünnbier1, Daniel Wicke², Maciej Bromirski3, Olaf Scheibner3

1Berliner Wasserbetriebe, Berlin, Germany; 2Kompetenzzentrum Wasser Berlin, Berlin, Germany; 3Thermo Fisher Scientific, Bremen, Germany; Email: olaf.scheibner@thermofisher.com

ConclusionThis study shows that suspect screening analysis is an efficient method for detection ofunknown compounds (for example simazine, nicotine, benzalkoniumchloride).The high measured concentrations of substances shows that stormwater runoff can bean important entry pathway for micropollutants to water bodies. Results also indicatethat it is necessary to consider the different urban catchment types with its specificcompounds.

References 1. Gasperi, J. et al. (2014) Micropollutants in urban stormwater: Occurrence, concen-

trations, and atmospheric contributions for a wide range of contaminants in three French catchments. Environmental Science & Pollution Research 21(8), 5282-5283

2. Zgheib, S., Moilleron, R. & Chebbo, G. (2012) Priority pollutants in urban storm-water: Part 1 - Case of separate storm sewers. Water Research 46(20), 6683-6692

OverviewPurpose: Stormwater runoff can be an important source of organic micro-pollutantsentering urban surface waters through separated sewer systems.

Methods: Screening and quantification of micropollutants was carried out for storm-water runoff samples of five different urban catchment areas in Berlin to considercatchment-specific differences.

Results: Stormwater may be a relevant source of biocides, particularly in citiesdominated by separate sewer systems.

IntroductionStormwater runoff can be an important source of organic micropollutants entering urbansurface waters through separate sewer systems (Zgheib et al. 2012). To investigate theextent of this type of pollution, a one year monitoring program was conducted in the cityof Berlin. Monitoring points were selected in five catchments of different urbanstructures to consider catchment-specific differences. Samples were analyzed for acomprehensive set of 100 micropollutants (e.g. PAHs, phthalates, phenols, heavymetals, biocides, flame retardants) that were chosen for monitoring based on literatureresearch (e.g. Gasperi et al. 2014).

MethodsSample PreparationStormwater runoff samples were filtered through 0.45 µm syringe filter before injection.

Liquid Chromatography1 mL of sample was injected directly into the online-SPE system. For pre-concentration,a C18 column, 2.1 × 20 mm with 12 µm particle size was used. For compoundseparation, a C18 column, 2.1 × 50 mm with 1.8 µm particle size was used. A water-methanol gradient both spiked with 0.1% formic acid was run from 2% to 95% in 6.7minutes. Total chromatographic cycle time including online pre-concentration was 15minutes.

Mass Spectrometry

Mass spectrometric analysis was run on a Thermo Scientific™ Q Exactive™ Focushybrid quadrupole-Orbitrap™ mass spectrometer (Figure 1).

MS-parameter for screening purposesFull Scan ESI+ and ESI– separately R = 70,000 m/z 100–1000

variable Data Independent Acquisition (vDIA) R = 70,000 CE = 30 m/z 100–205, 195–305, 295–405, 395–505, 495–1000

FIGURE 1. Q Exactive Focus with Thermo Scientific™ EQuan MAX Plus™ online SPE-system.

SamplingInvestigation areaFive sampling points were installed in separate storm sewers of five different catchmenttypes representing 85% of the connected impervious area of Berlin: one- family houseswith gardens (OFH), roads (>7500 vehicles/ day), old building (OLD, < 1930), newerbuilding (NEW, >1950) and commercial (COM) areas (Figure 2). Additionally a samplingpoint at the river “Panke” (PNK) was installed to determine contaminant concentrationsduring storm events in a stream affected by stormwater discharge.

© 2015 Thermo Fisher Scientific Inc. All rights reserved. All trademarks are the property of Thermo Fisher Scientific and its subsidiaries. This information is not intended to encourage use of these products in any manner that might infringe the intellectual property rights of others.

FIGURE 4. Suspect Screening results: Comparison between the sampling points one- family houses (OFH) and the river “Panke” (PNK).

FIGURE 2. Location of monitoring catchments in Berlin in separate sewer system (outside red line).

MS-parameter for quantitative analysisFull Scan polarity-switching (ESI+ / ESI–) R = 35,000 m/z 103–900 internal standards

FIGURE 6. Result of the pesticide/biocide concentrations for selected compounds for the different sampling locations.

Commercial areas

One-family houses with

gardens

Corresponding flow-proportional volume

5L composite sample

Chemical analysis

FIGURE 3. Sampling strategy: volume-proportional composite samples.

One- family houses(OFH) River �Panke“ (PNK)

Pharmaceuticals

Pesticides

Industrial and household chemicals

FIGURE 1. Process of data acquisition and processing for target and non-target analysis; all processing done with TraceFinder 3.3 software.

Data AnalysisQualitative and quantitative analysis was done with Thermo Scientific™ TraceFinder™software. Figure 1 shows a schematic diagram of the full workflow carried out in thisstudy. Starting with a suspect screening, micropollutants out of different classes likebiocides/ pesticides, pharmaceuticals, household and industrial chemicals containedin a homebuild database, were confirmed by using confirmation criteria such asisotopic pattern match and matching of known fragments. Selected suspects were aswell identifed as quantified by reference standards in a dilution series.

It can be seen that concentrations show distinct differences between catchment types.Whereas mecoprop (contained in bituminous sealing membranes for roofs) can befound in all catchment with roof runoff, carbendazim (fungicide, in urban contextapplied in paints and sealing compounds) and diuron (contained in exterior paints forsurface protection) are mostly found at monitoring station Old, an area of typicalhouses build <1930 with extensive renovations in recent years during which theapplication of new paints and sealing compounds is likely. On the other hand, concentrations of isoproturon (herbicide applied in gardens) are highest in catchmenttype OFH characterized by one family houses with gardens. Glyphosate (broadspectrum herbicide, �RoundUp“) was found at all monitoring stations, probably due toapplications for weed control on footpaths.

Quantitative results

A total of 19 biocides and pesticides were quantified regularly, of which 6 were addedas a result of the suspected Screening (simazine, terbuthylazine,desethylterbuthylazine, benzisothiazolinone, octylisothiazolinone and thiacloprid).Results for selected biocides and pesticides in stormwater runoff of the investigatedcatchment types can be seen in Figure 6.

std200ng_benzalkonium #364 RT: 7.33 AV:F: FTMS + p ESI Full ms2 304.30@hcd4 ...

100 200 300m/z

0

20

40

60

80

100

Relat

ive Ab

unda

nce

91.05483

304.30005

212.23746

110.11267

RT: 7.00 - 7.50

7.0 7.2 7.4Time (min)

0

20

40

60

80

100

Rela

tive

Abun

danc

e

7.33

7.41

7.16 7.277.06

RT: 7.00 - 7.50

7.0 7.2 7.4Time (min)

0

20

40

60

80

100

Rela

tive

Abun

danc

e

7.35

7.397.42

7.297.247.15

279 #432 RT: 7.39 AV: 1 NL: 8.17E6F: FTMS + p ESI Full ms2 304.30@hcd4 ...

100 200 300m/z

0

20

40

60

80

100

Relat

ive Ab

unda

nce

91.05484

304.30005

212.23746

121.48763

reference standard

sample

N+

CH3

CH3

C12H15

XIC 304.29996 (± 5 ppm) MS/MS

Time vs. flow Q[m3/ s]

ResultsSuspected Screening

The suspect screening results showed a large fraction of pesticides/biocides in thesamples indicating the relevance of stormwater as an entry pathway of biocides tosurface waters. The results of two samples are presented in figure 4: one-familiy houses(OFH) and the river “Panke” (PNK). It can be seen that the pesticide/biocide fraction ofidentified suspects (29 for OFH and 56 for PNK) is similar for both samples, whereasthe fraction of pharmaceuticals is much higher in the stream sample (PNK) due todischarges of WWTP effluent upstream the sampling location. For a suspect thefollowing confirming criteria were selected: isotopic pattern score > 85%, an accuratemass deviation < 5 ppm and at least one detected fragments.

OFH: e.g. Nicotine, 2-Hydroxybenzothiazole (OHBT), Benzothiazole (BT) and 2-Methylbenzothiazole (MTBT)

Panke: e.g. OHBT, MTBT, BT, Metoprolol, Carbamazepine, 4-formylaminoantipyrine (FAA), 4-acetylaminoantipyrine (AAA), Simazine, Terbutylazine, Ketamine, Tramadol

Suspect compound Formula [M+H]+ ∆ m/z(ppm)

Time (min)

IPS (%)

Isoproturon C12H18N2O 207.1492 -0.4 6.0 100Metolachlor C15H22ClNO2 284.1411 0.6 7.3 100Diethyltoluamide (DEET) C12H17NO 192.1383 0.0 6.4 100Benzisothiazolinone (BIT) C7H5NOS 152.0165 0.7 5.4 100

TABLE 1. Detected pesticides/biocides of sample point “new buildings” (NEW) with confirming criteria mass accuracy (Δm/z) and isotopic pattern score (IPS).

Whereas isoproturon and metolachlor are two plant protection products, diethyltoluamide(DEET) is an insect repellent and benzisothiazolinone (BIT) a biocide which is used inexterior and roof paints, caulks and varnish.

Non-Target Screening

In addition to the suspect screening an unknown screening with unbiased peak detectionwas carried out to screen for additional compounds of interest that had not come toattention yet. The first approach was to look into the 20 most intense compounds ofevery sample. Few of these masses were selected for parallel reaction monitoring (PRM)to get additional MS/MS information. The detected masses were m/z 304.29999,276.19492, 262.25311 and 248.16394. This is a sign for a homologous sequence (-CH2)of quaternary ammonium compounds.

A prominent quaternary ammonium compound is benzalkoniumchloride. Benzalkonium-chloride is used as biocide or cationic surfactant (disinfection). It has different alkyl chainlengths between C4-C18. The chosen selected masses have a chain length of C8, C9,C10 and C12. Figure 5 shows exemplary the results of benzalkoniumchloride C12, however, other chain lengths were also detected in the sample. The findings wereverified with reference standards.

In table 1 biocides detected by suspected screening of a sample of catchment NEW areshown. Beside these, further substances from other classes were found(pharmaceuticals, industrial-, and household chemicals).

FIGURE 5. Benzalkoniumchloride peak in the sample and the reference standard peak with MS/MS spectra.

C7H7+

C21H38N+

C14H30N+

Commercial Area [COM]

Newer buildings [NEW]

Street runoff [STR] – 1.3 km

Old buildings [OLD]

One-family houses with gardens [OFH]

Combined sewer area

ESI+

2 Targeted and Non-Targeted Screening of Biocides in Urban Stormwater Runoff

Sampling strategy

The stormwater runoff was collected with portable automatic samplers equipped with 8glass bottles (V=1.9 L). The sampler was triggered by the water level in the storm sewer(threshold: 10 cm), sample volume was 450 mL. At the beginning of a storm event,sampling intervals were shorter (5 min in the first 2 hours) to register a potential first flusheffect. After the first two hours of sampling the sampling interval was increased (every 15min). The maximum sampling duration was 4 hours.

A volume-proportional composite sample for analysis was prepared by manual mixing ofthe flow-corresponding volumes of each bottle (see Figure 3) calculated from the flow ofthe installed flow meter. In that way, the sample represents the average concentration ofthe sampled storm event.

Targeted and Non-Targeted Screening of Biocides in Urban Stormwater RunoffJulia Quilitzki1, Patricia van Baar1, Uwe Dünnbier1, Daniel Wicke², Maciej Bromirski3, Olaf Scheibner3

1Berliner Wasserbetriebe, Berlin, Germany; 2Kompetenzzentrum Wasser Berlin, Berlin, Germany; 3Thermo Fisher Scientific, Bremen, Germany; Email: olaf.scheibner@thermofisher.com

ConclusionThis study shows that suspect screening analysis is an efficient method for detection ofunknown compounds (for example simazine, nicotine, benzalkoniumchloride).The high measured concentrations of substances shows that stormwater runoff can bean important entry pathway for micropollutants to water bodies. Results also indicatethat it is necessary to consider the different urban catchment types with its specificcompounds.

References 1. Gasperi, J. et al. (2014) Micropollutants in urban stormwater: Occurrence, concen-

trations, and atmospheric contributions for a wide range of contaminants in three French catchments. Environmental Science & Pollution Research 21(8), 5282-5283

2. Zgheib, S., Moilleron, R. & Chebbo, G. (2012) Priority pollutants in urban storm-water: Part 1 - Case of separate storm sewers. Water Research 46(20), 6683-6692

OverviewPurpose: Stormwater runoff can be an important source of organic micro-pollutantsentering urban surface waters through separated sewer systems.

Methods: Screening and quantification of micropollutants was carried out for storm-water runoff samples of five different urban catchment areas in Berlin to considercatchment-specific differences.

Results: Stormwater may be a relevant source of biocides, particularly in citiesdominated by separate sewer systems.

IntroductionStormwater runoff can be an important source of organic micropollutants entering urbansurface waters through separate sewer systems (Zgheib et al. 2012). To investigate theextent of this type of pollution, a one year monitoring program was conducted in the cityof Berlin. Monitoring points were selected in five catchments of different urbanstructures to consider catchment-specific differences. Samples were analyzed for acomprehensive set of 100 micropollutants (e.g. PAHs, phthalates, phenols, heavymetals, biocides, flame retardants) that were chosen for monitoring based on literatureresearch (e.g. Gasperi et al. 2014).

MethodsSample PreparationStormwater runoff samples were filtered through 0.45 µm syringe filter before injection.

Liquid Chromatography1 mL of sample was injected directly into the online-SPE system. For pre-concentration,a C18 column, 2.1 × 20 mm with 12 µm particle size was used. For compoundseparation, a C18 column, 2.1 × 50 mm with 1.8 µm particle size was used. A water-methanol gradient both spiked with 0.1% formic acid was run from 2% to 95% in 6.7minutes. Total chromatographic cycle time including online pre-concentration was 15minutes.

Mass Spectrometry

Mass spectrometric analysis was run on a Thermo Scientific™ Q Exactive™ Focushybrid quadrupole-Orbitrap™ mass spectrometer (Figure 1).

MS-parameter for screening purposesFull Scan ESI+ and ESI– separately R = 70,000 m/z 100–1000

variable Data Independent Acquisition (vDIA) R = 70,000 CE = 30 m/z 100–205, 195–305, 295–405, 395–505, 495–1000

FIGURE 1. Q Exactive Focus with Thermo Scientific™ EQuan MAX Plus™ online SPE-system.

SamplingInvestigation areaFive sampling points were installed in separate storm sewers of five different catchmenttypes representing 85% of the connected impervious area of Berlin: one- family houseswith gardens (OFH), roads (>7500 vehicles/ day), old building (OLD, < 1930), newerbuilding (NEW, >1950) and commercial (COM) areas (Figure 2). Additionally a samplingpoint at the river “Panke” (PNK) was installed to determine contaminant concentrationsduring storm events in a stream affected by stormwater discharge.

© 2015 Thermo Fisher Scientific Inc. All rights reserved. All trademarks are the property of Thermo Fisher Scientific and its subsidiaries. This information is not intended to encourage use of these products in any manner that might infringe the intellectual property rights of others.

FIGURE 4. Suspect Screening results: Comparison between the sampling points one- family houses (OFH) and the river “Panke” (PNK).

FIGURE 2. Location of monitoring catchments in Berlin in separate sewer system (outside red line).

MS-parameter for quantitative analysisFull Scan polarity-switching (ESI+ / ESI–) R = 35,000 m/z 103–900 internal standards

FIGURE 6. Result of the pesticide/biocide concentrations for selected compounds for the different sampling locations.

Commercial areas

One-family houses with

gardens

Corresponding flow-proportional volume

5L composite sample

Chemical analysis

FIGURE 3. Sampling strategy: volume-proportional composite samples.

One- family houses(OFH) River �Panke“ (PNK)

Pharmaceuticals

Pesticides

Industrial and household chemicals

FIGURE 1. Process of data acquisition and processing for target and non-target analysis; all processing done with TraceFinder 3.3 software.

Data AnalysisQualitative and quantitative analysis was done with Thermo Scientific™ TraceFinder™software. Figure 1 shows a schematic diagram of the full workflow carried out in thisstudy. Starting with a suspect screening, micropollutants out of different classes likebiocides/ pesticides, pharmaceuticals, household and industrial chemicals containedin a homebuild database, were confirmed by using confirmation criteria such asisotopic pattern match and matching of known fragments. Selected suspects were aswell identifed as quantified by reference standards in a dilution series.

It can be seen that concentrations show distinct differences between catchment types.Whereas mecoprop (contained in bituminous sealing membranes for roofs) can befound in all catchment with roof runoff, carbendazim (fungicide, in urban contextapplied in paints and sealing compounds) and diuron (contained in exterior paints forsurface protection) are mostly found at monitoring station Old, an area of typicalhouses build <1930 with extensive renovations in recent years during which theapplication of new paints and sealing compounds is likely. On the other hand, concentrations of isoproturon (herbicide applied in gardens) are highest in catchmenttype OFH characterized by one family houses with gardens. Glyphosate (broadspectrum herbicide, �RoundUp“) was found at all monitoring stations, probably due toapplications for weed control on footpaths.

Quantitative results

A total of 19 biocides and pesticides were quantified regularly, of which 6 were addedas a result of the suspected Screening (simazine, terbuthylazine,desethylterbuthylazine, benzisothiazolinone, octylisothiazolinone and thiacloprid).Results for selected biocides and pesticides in stormwater runoff of the investigatedcatchment types can be seen in Figure 6.

std200ng_benzalkonium #364 RT: 7.33 AV:F: FTMS + p ESI Full ms2 304.30@hcd4 ...

100 200 300m/z

0

20

40

60

80

100

Relat

ive Ab

unda

nce

91.05483

304.30005

212.23746

110.11267

RT: 7.00 - 7.50

7.0 7.2 7.4Time (min)

0

20

40

60

80

100

Rela

tive

Abun

danc

e

7.33

7.41

7.16 7.277.06

RT: 7.00 - 7.50

7.0 7.2 7.4Time (min)

0

20

40

60

80

100

Rela

tive

Abun

danc

e

7.35

7.397.42

7.297.247.15

279 #432 RT: 7.39 AV: 1 NL: 8.17E6F: FTMS + p ESI Full ms2 304.30@hcd4 ...

100 200 300m/z

0

20

40

60

80

100

Relat

ive Ab

unda

nce

91.05484

304.30005

212.23746

121.48763

reference standard

sample

N+

CH3

CH3

C12H15

XIC 304.29996 (± 5 ppm) MS/MS

Time vs. flow Q[m3/ s]

ResultsSuspected Screening

The suspect screening results showed a large fraction of pesticides/biocides in thesamples indicating the relevance of stormwater as an entry pathway of biocides tosurface waters. The results of two samples are presented in figure 4: one-familiy houses(OFH) and the river “Panke” (PNK). It can be seen that the pesticide/biocide fraction ofidentified suspects (29 for OFH and 56 for PNK) is similar for both samples, whereasthe fraction of pharmaceuticals is much higher in the stream sample (PNK) due todischarges of WWTP effluent upstream the sampling location. For a suspect thefollowing confirming criteria were selected: isotopic pattern score > 85%, an accuratemass deviation < 5 ppm and at least one detected fragments.

OFH: e.g. Nicotine, 2-Hydroxybenzothiazole (OHBT), Benzothiazole (BT) and 2-Methylbenzothiazole (MTBT)

Panke: e.g. OHBT, MTBT, BT, Metoprolol, Carbamazepine, 4-formylaminoantipyrine (FAA), 4-acetylaminoantipyrine (AAA), Simazine, Terbutylazine, Ketamine, Tramadol

Suspect compound Formula [M+H]+ ∆ m/z(ppm)

Time (min)

IPS (%)

Isoproturon C12H18N2O 207.1492 -0.4 6.0 100Metolachlor C15H22ClNO2 284.1411 0.6 7.3 100Diethyltoluamide (DEET) C12H17NO 192.1383 0.0 6.4 100Benzisothiazolinone (BIT) C7H5NOS 152.0165 0.7 5.4 100

TABLE 1. Detected pesticides/biocides of sample point “new buildings” (NEW) with confirming criteria mass accuracy (Δm/z) and isotopic pattern score (IPS).

Whereas isoproturon and metolachlor are two plant protection products, diethyltoluamide(DEET) is an insect repellent and benzisothiazolinone (BIT) a biocide which is used inexterior and roof paints, caulks and varnish.

Non-Target Screening

In addition to the suspect screening an unknown screening with unbiased peak detectionwas carried out to screen for additional compounds of interest that had not come toattention yet. The first approach was to look into the 20 most intense compounds ofevery sample. Few of these masses were selected for parallel reaction monitoring (PRM)to get additional MS/MS information. The detected masses were m/z 304.29999,276.19492, 262.25311 and 248.16394. This is a sign for a homologous sequence (-CH2)of quaternary ammonium compounds.

A prominent quaternary ammonium compound is benzalkoniumchloride. Benzalkonium-chloride is used as biocide or cationic surfactant (disinfection). It has different alkyl chainlengths between C4-C18. The chosen selected masses have a chain length of C8, C9,C10 and C12. Figure 5 shows exemplary the results of benzalkoniumchloride C12, however, other chain lengths were also detected in the sample. The findings wereverified with reference standards.

In table 1 biocides detected by suspected screening of a sample of catchment NEW areshown. Beside these, further substances from other classes were found(pharmaceuticals, industrial-, and household chemicals).

FIGURE 5. Benzalkoniumchloride peak in the sample and the reference standard peak with MS/MS spectra.

C7H7+

C21H38N+

C14H30N+

Commercial Area [COM]

Newer buildings [NEW]

Street runoff [STR] – 1.3 km

Old buildings [OLD]

One-family houses with gardens [OFH]

Combined sewer area

ESI+

Sampling strategy

The stormwater runoff was collected with portable automatic samplers equipped with 8glass bottles (V=1.9 L). The sampler was triggered by the water level in the storm sewer(threshold: 10 cm), sample volume was 450 mL. At the beginning of a storm event,sampling intervals were shorter (5 min in the first 2 hours) to register a potential first flusheffect. After the first two hours of sampling the sampling interval was increased (every 15min). The maximum sampling duration was 4 hours.

A volume-proportional composite sample for analysis was prepared by manual mixing ofthe flow-corresponding volumes of each bottle (see Figure 3) calculated from the flow ofthe installed flow meter. In that way, the sample represents the average concentration ofthe sampled storm event.

Targeted and Non-Targeted Screening of Biocides in Urban Stormwater RunoffJulia Quilitzki1, Patricia van Baar1, Uwe Dünnbier1, Daniel Wicke², Maciej Bromirski3, Olaf Scheibner3

1Berliner Wasserbetriebe, Berlin, Germany; 2Kompetenzzentrum Wasser Berlin, Berlin, Germany; 3Thermo Fisher Scientific, Bremen, Germany; Email: olaf.scheibner@thermofisher.com

ConclusionThis study shows that suspect screening analysis is an efficient method for detection ofunknown compounds (for example simazine, nicotine, benzalkoniumchloride).The high measured concentrations of substances shows that stormwater runoff can bean important entry pathway for micropollutants to water bodies. Results also indicatethat it is necessary to consider the different urban catchment types with its specificcompounds.

References 1. Gasperi, J. et al. (2014) Micropollutants in urban stormwater: Occurrence, concen-

trations, and atmospheric contributions for a wide range of contaminants in three French catchments. Environmental Science & Pollution Research 21(8), 5282-5283

2. Zgheib, S., Moilleron, R. & Chebbo, G. (2012) Priority pollutants in urban storm-water: Part 1 - Case of separate storm sewers. Water Research 46(20), 6683-6692

OverviewPurpose: Stormwater runoff can be an important source of organic micro-pollutantsentering urban surface waters through separated sewer systems.

Methods: Screening and quantification of micropollutants was carried out for storm-water runoff samples of five different urban catchment areas in Berlin to considercatchment-specific differences.

Results: Stormwater may be a relevant source of biocides, particularly in citiesdominated by separate sewer systems.

IntroductionStormwater runoff can be an important source of organic micropollutants entering urbansurface waters through separate sewer systems (Zgheib et al. 2012). To investigate theextent of this type of pollution, a one year monitoring program was conducted in the cityof Berlin. Monitoring points were selected in five catchments of different urbanstructures to consider catchment-specific differences. Samples were analyzed for acomprehensive set of 100 micropollutants (e.g. PAHs, phthalates, phenols, heavymetals, biocides, flame retardants) that were chosen for monitoring based on literatureresearch (e.g. Gasperi et al. 2014).

MethodsSample PreparationStormwater runoff samples were filtered through 0.45 µm syringe filter before injection.

Liquid Chromatography1 mL of sample was injected directly into the online-SPE system. For pre-concentration,a C18 column, 2.1 × 20 mm with 12 µm particle size was used. For compoundseparation, a C18 column, 2.1 × 50 mm with 1.8 µm particle size was used. A water-methanol gradient both spiked with 0.1% formic acid was run from 2% to 95% in 6.7minutes. Total chromatographic cycle time including online pre-concentration was 15minutes.

Mass Spectrometry

Mass spectrometric analysis was run on a Thermo Scientific™ Q Exactive™ Focushybrid quadrupole-Orbitrap™ mass spectrometer (Figure 1).

MS-parameter for screening purposesFull Scan ESI+ and ESI– separately R = 70,000 m/z 100–1000

variable Data Independent Acquisition (vDIA) R = 70,000 CE = 30 m/z 100–205, 195–305, 295–405, 395–505, 495–1000

FIGURE 1. Q Exactive Focus with Thermo Scientific™ EQuan MAX Plus™ online SPE-system.

SamplingInvestigation areaFive sampling points were installed in separate storm sewers of five different catchmenttypes representing 85% of the connected impervious area of Berlin: one- family houseswith gardens (OFH), roads (>7500 vehicles/ day), old building (OLD, < 1930), newerbuilding (NEW, >1950) and commercial (COM) areas (Figure 2). Additionally a samplingpoint at the river “Panke” (PNK) was installed to determine contaminant concentrationsduring storm events in a stream affected by stormwater discharge.

© 2015 Thermo Fisher Scientific Inc. All rights reserved. All trademarks are the property of Thermo Fisher Scientific and its subsidiaries. This information is not intended to encourage use of these products in any manner that might infringe the intellectual property rights of others.

FIGURE 4. Suspect Screening results: Comparison between the sampling points one- family houses (OFH) and the river “Panke” (PNK).

FIGURE 2. Location of monitoring catchments in Berlin in separate sewer system (outside red line).

MS-parameter for quantitative analysisFull Scan polarity-switching (ESI+ / ESI–) R = 35,000 m/z 103–900 internal standards

FIGURE 6. Result of the pesticide/biocide concentrations for selected compounds for the different sampling locations.

Commercial areas

One-family houses with

gardens

Corresponding flow-proportional volume

5L composite sample

Chemical analysis

FIGURE 3. Sampling strategy: volume-proportional composite samples.

One- family houses(OFH) River �Panke“ (PNK)

Pharmaceuticals

Pesticides

Industrial and household chemicalsIndustrial and household chemicals

FIGURE 1. Process of data acquisition and processing for target and non-target analysis; all processing done with TraceFinder 3.3 software.

Data AnalysisQualitative and quantitative analysis was done with Thermo Scientific™ TraceFinder™software. Figure 1 shows a schematic diagram of the full workflow carried out in thisstudy. Starting with a suspect screening, micropollutants out of different classes likebiocides/ pesticides, pharmaceuticals, household and industrial chemicals containedin a homebuild database, were confirmed by using confirmation criteria such asisotopic pattern match and matching of known fragments. Selected suspects were aswell identifed as quantified by reference standards in a dilution series.

It can be seen that concentrations show distinct differences between catchment types.Whereas mecoprop (contained in bituminous sealing membranes for roofs) can befound in all catchment with roof runoff, carbendazim (fungicide, in urban contextapplied in paints and sealing compounds) and diuron (contained in exterior paints forsurface protection) are mostly found at monitoring station Old, an area of typicalhouses build <1930 with extensive renovations in recent years during which theapplication of new paints and sealing compounds is likely. On the other hand, concentrations of isoproturon (herbicide applied in gardens) are highest in catchmenttype OFH characterized by one family houses with gardens. Glyphosate (broadspectrum herbicide, �RoundUp“) was found at all monitoring stations, probably due toapplications for weed control on footpaths.

Quantitative results

A total of 19 biocides and pesticides were quantified regularly, of which 6 were addedas a result of the suspected Screening (simazine, terbuthylazine,desethylterbuthylazine, benzisothiazolinone, octylisothiazolinone and thiacloprid).Results for selected biocides and pesticides in stormwater runoff of the investigatedcatchment types can be seen in Figure 6.

std200ng_benzalkonium #364 RT: 7.33 AV:F: FTMS + p ESI Full ms2 304.30@hcd4 ...

100 200 300m/z

0

20

40

60

80

100

Relat

ive Ab

unda

nce

91.05483

304.30005

212.23746

110.11267

RT: 7.00 - 7.50

7.0 7.2 7.4Time (min)

0

20

40

60

80

100

Rela

tive

Abun

danc

e

7.33

7.41

7.16 7.277.06

RT: 7.00 - 7.50

7.0 7.2 7.4Time (min)

0

20

40

60

80

100

Rela

tive

Abun

danc

e

7.35

7.397.42

7.297.247.15

279 #432 RT: 7.39 AV: 1 NL: 8.17E6F: FTMS + p ESI Full ms2 304.30@hcd4 ...

100 200 300m/z

0

20

40

60

80

100

Relat

ive Ab

unda

nce

91.05484

304.30005

212.23746

121.48763

reference standard

sample

N+

CH3

CH3

C12H15

XIC 304.29996 (± 5 ppm) MS/MS

Time vs. flow Q[m3/ s]

ResultsSuspected Screening

The suspect screening results showed a large fraction of pesticides/biocides in thesamples indicating the relevance of stormwater as an entry pathway of biocides tosurface waters. The results of two samples are presented in figure 4: one-familiy houses(OFH) and the river “Panke” (PNK). It can be seen that the pesticide/biocide fraction ofidentified suspects (29 for OFH and 56 for PNK) is similar for both samples, whereasthe fraction of pharmaceuticals is much higher in the stream sample (PNK) due todischarges of WWTP effluent upstream the sampling location. For a suspect thefollowing confirming criteria were selected: isotopic pattern score > 85%, an accuratemass deviation < 5 ppm and at least one detected fragments.

OFH: e.g. Nicotine, 2-Hydroxybenzothiazole (OHBT), Benzothiazole (BT) and 2-Methylbenzothiazole (MTBT)

Panke: e.g. OHBT, MTBT, BT, Metoprolol, Carbamazepine, 4-formylaminoantipyrine (FAA), 4-acetylaminoantipyrine (AAA), Simazine, Terbutylazine, Ketamine, Tramadol

Suspect compound Formula [M+H]+ ∆ m/z(ppm)

Time (min)

IPS (%)

Isoproturon C12H18N2O 207.1492 -0.4 6.0 100Metolachlor C15H22ClNO2 284.1411 0.6 7.3 100Diethyltoluamide (DEET) C12H17NO 192.1383 0.0 6.4 100Benzisothiazolinone (BIT) C7H5NOS 152.0165 0.7 5.4 100

TABLE 1. Detected pesticides/biocides of sample point “new buildings” (NEW) with confirming criteria mass accuracy (Δm/z) and isotopic pattern score (IPS).

Whereas isoproturon and metolachlor are two plant protection products, diethyltoluamide(DEET) is an insect repellent and benzisothiazolinone (BIT) a biocide which is used inexterior and roof paints, caulks and varnish.

Non-Target Screening

In addition to the suspect screening an unknown screening with unbiased peak detectionwas carried out to screen for additional compounds of interest that had not come toattention yet. The first approach was to look into the 20 most intense compounds ofevery sample. Few of these masses were selected for parallel reaction monitoring (PRM)to get additional MS/MS information. The detected masses were m/z 304.29999,276.19492, 262.25311 and 248.16394. This is a sign for a homologous sequence (-CH2)of quaternary ammonium compounds.

A prominent quaternary ammonium compound is benzalkoniumchloride. Benzalkonium-chloride is used as biocide or cationic surfactant (disinfection). It has different alkyl chainlengths between C4-C18. The chosen selected masses have a chain length of C8, C9,C10 and C12. Figure 5 shows exemplary the results of benzalkoniumchloride C12, however, other chain lengths were also detected in the sample. The findings wereverified with reference standards.

In table 1 biocides detected by suspected screening of a sample of catchment NEW areshown. Beside these, further substances from other classes were found(pharmaceuticals, industrial-, and household chemicals).

FIGURE 5. Benzalkoniumchloride peak in the sample and the reference standard peak with MS/MS spectra.

C7H7+

C21H38N+

C14H30N+

Commercial Area [COM]

Newer buildings [NEW]

Street runoff [STR] – 1.3 km

Old buildings [OLD]

One-family houses with gardens [OFH]

Combined sewer area

ESI+

Sampling strategy

The stormwater runoff was collected with portable automatic samplers equipped with 8glass bottles (V=1.9 L). The sampler was triggered by the water level in the storm sewer(threshold: 10 cm), sample volume was 450 mL. At the beginning of a storm event,sampling intervals were shorter (5 min in the first 2 hours) to register a potential first flusheffect. After the first two hours of sampling the sampling interval was increased (every 15min). The maximum sampling duration was 4 hours.

A volume-proportional composite sample for analysis was prepared by manual mixing ofthe flow-corresponding volumes of each bottle (see Figure 3) calculated from the flow ofthe installed flow meter. In that way, the sample represents the average concentration ofthe sampled storm event.

Targeted and Non-Targeted Screening of Biocides in Urban Stormwater RunoffJulia Quilitzki1, Patricia van Baar1, Uwe Dünnbier1, Daniel Wicke², Maciej Bromirski3, Olaf Scheibner3

1Berliner Wasserbetriebe, Berlin, Germany; 2Kompetenzzentrum Wasser Berlin, Berlin, Germany; 3Thermo Fisher Scientific, Bremen, Germany; Email: olaf.scheibner@thermofisher.com

ConclusionThis study shows that suspect screening analysis is an efficient method for detection ofunknown compounds (for example simazine, nicotine, benzalkoniumchloride).The high measured concentrations of substances shows that stormwater runoff can bean important entry pathway for micropollutants to water bodies. Results also indicatethat it is necessary to consider the different urban catchment types with its specificcompounds.

References 1. Gasperi, J. et al. (2014) Micropollutants in urban stormwater: Occurrence, concen-

trations, and atmospheric contributions for a wide range of contaminants in three French catchments. Environmental Science & Pollution Research 21(8), 5282-5283

2. Zgheib, S., Moilleron, R. & Chebbo, G. (2012) Priority pollutants in urban storm-water: Part 1 - Case of separate storm sewers. Water Research 46(20), 6683-6692

OverviewPurpose: Stormwater runoff can be an important source of organic micro-pollutantsentering urban surface waters through separated sewer systems.

Methods: Screening and quantification of micropollutants was carried out for storm-water runoff samples of five different urban catchment areas in Berlin to considercatchment-specific differences.

Results: Stormwater may be a relevant source of biocides, particularly in citiesdominated by separate sewer systems.

IntroductionStormwater runoff can be an important source of organic micropollutants entering urbansurface waters through separate sewer systems (Zgheib et al. 2012). To investigate theextent of this type of pollution, a one year monitoring program was conducted in the cityof Berlin. Monitoring points were selected in five catchments of different urbanstructures to consider catchment-specific differences. Samples were analyzed for acomprehensive set of 100 micropollutants (e.g. PAHs, phthalates, phenols, heavymetals, biocides, flame retardants) that were chosen for monitoring based on literatureresearch (e.g. Gasperi et al. 2014).

MethodsSample PreparationStormwater runoff samples were filtered through 0.45 µm syringe filter before injection.

Liquid Chromatography1 mL of sample was injected directly into the online-SPE system. For pre-concentration,a C18 column, 2.1 × 20 mm with 12 µm particle size was used. For compoundseparation, a C18 column, 2.1 × 50 mm with 1.8 µm particle size was used. A water-methanol gradient both spiked with 0.1% formic acid was run from 2% to 95% in 6.7minutes. Total chromatographic cycle time including online pre-concentration was 15minutes.

Mass Spectrometry

Mass spectrometric analysis was run on a Thermo Scientific™ Q Exactive™ Focushybrid quadrupole-Orbitrap™ mass spectrometer (Figure 1).

MS-parameter for screening purposesFull Scan ESI+ and ESI– separately R = 70,000 m/z 100–1000

variable Data Independent Acquisition (vDIA) R = 70,000 CE = 30 m/z 100–205, 195–305, 295–405, 395–505, 495–1000

FIGURE 1. Q Exactive Focus with Thermo Scientific™ EQuan MAX Plus™ online SPE-system.

SamplingInvestigation areaFive sampling points were installed in separate storm sewers of five different catchmenttypes representing 85% of the connected impervious area of Berlin: one- family houseswith gardens (OFH), roads (>7500 vehicles/ day), old building (OLD, < 1930), newerbuilding (NEW, >1950) and commercial (COM) areas (Figure 2). Additionally a samplingpoint at the river “Panke” (PNK) was installed to determine contaminant concentrationsduring storm events in a stream affected by stormwater discharge.

© 2015 Thermo Fisher Scientific Inc. All rights reserved. All trademarks are the property of Thermo Fisher Scientific and its subsidiaries. This information is not intended to encourage use of these products in any manner that might infringe the intellectual property rights of others.

FIGURE 4. Suspect Screening results: Comparison between the sampling points one- family houses (OFH) and the river “Panke” (PNK).

FIGURE 2. Location of monitoring catchments in Berlin in separate sewer system (outside red line).

MS-parameter for quantitative analysisFull Scan polarity-switching (ESI+ / ESI–) R = 35,000 m/z 103–900 internal standards

FIGURE 6. Result of the pesticide/biocide concentrations for selected compounds for the different sampling locations.

Commercial areas

One-family houses with

gardens

Corresponding flow-proportional volume

5L composite sample

Chemical analysis

FIGURE 3. Sampling strategy: volume-proportional composite samples.

One- family houses(OFH) River �Panke“ (PNK)

Pharmaceuticals

Pesticides

Industrial and household chemicals

FIGURE 1. Process of data acquisition and processing for target and non-target analysis; all processing done with TraceFinder 3.3 software.

Data AnalysisQualitative and quantitative analysis was done with Thermo Scientific™ TraceFinder™software. Figure 1 shows a schematic diagram of the full workflow carried out in thisstudy. Starting with a suspect screening, micropollutants out of different classes likebiocides/ pesticides, pharmaceuticals, household and industrial chemicals containedin a homebuild database, were confirmed by using confirmation criteria such asisotopic pattern match and matching of known fragments. Selected suspects were aswell identifed as quantified by reference standards in a dilution series.

It can be seen that concentrations show distinct differences between catchment types.Whereas mecoprop (contained in bituminous sealing membranes for roofs) can befound in all catchment with roof runoff, carbendazim (fungicide, in urban contextapplied in paints and sealing compounds) and diuron (contained in exterior paints forsurface protection) are mostly found at monitoring station Old, an area of typicalhouses build <1930 with extensive renovations in recent years during which theapplication of new paints and sealing compounds is likely. On the other hand, concentrations of isoproturon (herbicide applied in gardens) are highest in catchmenttype OFH characterized by one family houses with gardens. Glyphosate (broadspectrum herbicide, �RoundUp“) was found at all monitoring stations, probably due toapplications for weed control on footpaths.

Quantitative results

A total of 19 biocides and pesticides were quantified regularly, of which 6 were addedas a result of the suspected Screening (simazine, terbuthylazine,desethylterbuthylazine, benzisothiazolinone, octylisothiazolinone and thiacloprid).Results for selected biocides and pesticides in stormwater runoff of the investigatedcatchment types can be seen in Figure 6.

std200ng_benzalkonium #364 RT: 7.33 AV:F: FTMS + p ESI Full ms2 304.30@hcd4 ...

100 200 300m/z

0

20

40

60

80

100

Relat

ive Ab

unda

nce

91.05483

304.30005

212.23746

110.11267

RT: 7.00 - 7.50

7.0 7.2 7.4Time (min)

0

20

40

60

80

100

Rela

tive

Abun

danc

e

7.33

7.41

7.16 7.277.06

RT: 7.00 - 7.50

7.0 7.2 7.4Time (min)

0

20

40

60

80

100

Rela

tive

Abun

danc

e

7.35

7.397.42

7.297.247.15

279 #432 RT: 7.39 AV: 1 NL: 8.17E6F: FTMS + p ESI Full ms2 304.30@hcd4 ...

100 200 300m/z

0

20

40

60

80

100

Relat

ive Ab

unda

nce

91.05484

304.30005

212.23746

121.48763

reference standard

sample

N+

CH3

CH3

C12H15

XIC 304.29996 (± 5 ppm) MS/MS

Time vs. flow Q[m3/ s]

ResultsSuspected Screening

The suspect screening results showed a large fraction of pesticides/biocides in thesamples indicating the relevance of stormwater as an entry pathway of biocides tosurface waters. The results of two samples are presented in figure 4: one-familiy houses(OFH) and the river “Panke” (PNK). It can be seen that the pesticide/biocide fraction ofidentified suspects (29 for OFH and 56 for PNK) is similar for both samples, whereasthe fraction of pharmaceuticals is much higher in the stream sample (PNK) due todischarges of WWTP effluent upstream the sampling location. For a suspect thefollowing confirming criteria were selected: isotopic pattern score > 85%, an accuratemass deviation < 5 ppm and at least one detected fragments.

OFH: e.g. Nicotine, 2-Hydroxybenzothiazole (OHBT), Benzothiazole (BT) and 2-Methylbenzothiazole (MTBT)

Panke: e.g. OHBT, MTBT, BT, Metoprolol, Carbamazepine, 4-formylaminoantipyrine (FAA), 4-acetylaminoantipyrine (AAA), Simazine, Terbutylazine, Ketamine, Tramadol

Suspect compound Formula [M+H]+ ∆ m/z(ppm)

Time (min)

IPS (%)

Isoproturon C12H18N2O 207.1492 -0.4 6.0 100Metolachlor C15H22ClNO2 284.1411 0.6 7.3 100Diethyltoluamide (DEET) C12H17NO 192.1383 0.0 6.4 100Benzisothiazolinone (BIT) C7H5NOS 152.0165 0.7 5.4 100

TABLE 1. Detected pesticides/biocides of sample point “new buildings” (NEW) with confirming criteria mass accuracy (Δm/z) and isotopic pattern score (IPS).

Whereas isoproturon and metolachlor are two plant protection products, diethyltoluamide(DEET) is an insect repellent and benzisothiazolinone (BIT) a biocide which is used inexterior and roof paints, caulks and varnish.

Non-Target Screening

In addition to the suspect screening an unknown screening with unbiased peak detectionwas carried out to screen for additional compounds of interest that had not come toattention yet. The first approach was to look into the 20 most intense compounds ofevery sample. Few of these masses were selected for parallel reaction monitoring (PRM)to get additional MS/MS information. The detected masses were m/z 304.29999,276.19492, 262.25311 and 248.16394. This is a sign for a homologous sequence (-CH2)of quaternary ammonium compounds.

A prominent quaternary ammonium compound is benzalkoniumchloride. Benzalkonium-chloride is used as biocide or cationic surfactant (disinfection). It has different alkyl chainlengths between C4-C18. The chosen selected masses have a chain length of C8, C9,C10 and C12. Figure 5 shows exemplary the results of benzalkoniumchloride C12, however, other chain lengths were also detected in the sample. The findings wereverified with reference standards.

In table 1 biocides detected by suspected screening of a sample of catchment NEW areshown. Beside these, further substances from other classes were found(pharmaceuticals, industrial-, and household chemicals).

FIGURE 5. Benzalkoniumchloride peak in the sample and the reference standard peak with MS/MS spectra.

C7H7+

C21H38N+

C14H30N+

Commercial Area [COM]

Newer buildings [NEW]

Street runoff [STR] – 1.3 km

Old buildings [OLD]

One-family houses with gardens [OFH]

Combined sewer area

ESI+

Sampling strategy

The stormwater runoff was collected with portable automatic samplers equipped with 8glass bottles (V=1.9 L). The sampler was triggered by the water level in the storm sewer(threshold: 10 cm), sample volume was 450 mL. At the beginning of a storm event,sampling intervals were shorter (5 min in the first 2 hours) to register a potential first flusheffect. After the first two hours of sampling the sampling interval was increased (every 15min). The maximum sampling duration was 4 hours.

A volume-proportional composite sample for analysis was prepared by manual mixing ofthe flow-corresponding volumes of each bottle (see Figure 3) calculated from the flow ofthe installed flow meter. In that way, the sample represents the average concentration ofthe sampled storm event.

Targeted and Non-Targeted Screening of Biocides in Urban Stormwater RunoffJulia Quilitzki1, Patricia van Baar1, Uwe Dünnbier1, Daniel Wicke², Maciej Bromirski3, Olaf Scheibner3

1Berliner Wasserbetriebe, Berlin, Germany; 2Kompetenzzentrum Wasser Berlin, Berlin, Germany; 3Thermo Fisher Scientific, Bremen, Germany; Email: olaf.scheibner@thermofisher.com

ConclusionThis study shows that suspect screening analysis is an efficient method for detection ofunknown compounds (for example simazine, nicotine, benzalkoniumchloride).The high measured concentrations of substances shows that stormwater runoff can bean important entry pathway for micropollutants to water bodies. Results also indicatethat it is necessary to consider the different urban catchment types with its specificcompounds.

References 1. Gasperi, J. et al. (2014) Micropollutants in urban stormwater: Occurrence, concen-

trations, and atmospheric contributions for a wide range of contaminants in three French catchments. Environmental Science & Pollution Research 21(8), 5282-5283

2. Zgheib, S., Moilleron, R. & Chebbo, G. (2012) Priority pollutants in urban storm-water: Part 1 - Case of separate storm sewers. Water Research 46(20), 6683-6692

OverviewPurpose: Stormwater runoff can be an important source of organic micro-pollutantsentering urban surface waters through separated sewer systems.

Methods: Screening and quantification of micropollutants was carried out for storm-water runoff samples of five different urban catchment areas in Berlin to considercatchment-specific differences.

Results: Stormwater may be a relevant source of biocides, particularly in citiesdominated by separate sewer systems.

IntroductionStormwater runoff can be an important source of organic micropollutants entering urbansurface waters through separate sewer systems (Zgheib et al. 2012). To investigate theextent of this type of pollution, a one year monitoring program was conducted in the cityof Berlin. Monitoring points were selected in five catchments of different urbanstructures to consider catchment-specific differences. Samples were analyzed for acomprehensive set of 100 micropollutants (e.g. PAHs, phthalates, phenols, heavymetals, biocides, flame retardants) that were chosen for monitoring based on literatureresearch (e.g. Gasperi et al. 2014).

MethodsSample PreparationStormwater runoff samples were filtered through 0.45 µm syringe filter before injection.

Liquid Chromatography1 mL of sample was injected directly into the online-SPE system. For pre-concentration,a C18 column, 2.1 × 20 mm with 12 µm particle size was used. For compoundseparation, a C18 column, 2.1 × 50 mm with 1.8 µm particle size was used. A water-methanol gradient both spiked with 0.1% formic acid was run from 2% to 95% in 6.7minutes. Total chromatographic cycle time including online pre-concentration was 15minutes.

Mass Spectrometry

Mass spectrometric analysis was run on a Thermo Scientific™ Q Exactive™ Focushybrid quadrupole-Orbitrap™ mass spectrometer (Figure 1).

MS-parameter for screening purposesFull Scan ESI+ and ESI– separately R = 70,000 m/z 100–1000

variable Data Independent Acquisition (vDIA) R = 70,000 CE = 30 m/z 100–205, 195–305, 295–405, 395–505, 495–1000

FIGURE 1. Q Exactive Focus with Thermo Scientific™ EQuan MAX Plus™ online SPE-system.

SamplingInvestigation areaFive sampling points were installed in separate storm sewers of five different catchmenttypes representing 85% of the connected impervious area of Berlin: one- family houseswith gardens (OFH), roads (>7500 vehicles/ day), old building (OLD, < 1930), newerbuilding (NEW, >1950) and commercial (COM) areas (Figure 2). Additionally a samplingpoint at the river “Panke” (PNK) was installed to determine contaminant concentrationsduring storm events in a stream affected by stormwater discharge.

© 2015 Thermo Fisher Scientific Inc. All rights reserved. All trademarks are the property of Thermo Fisher Scientific and its subsidiaries. This information is not intended to encourage use of these products in any manner that might infringe the intellectual property rights of others.

FIGURE 4. Suspect Screening results: Comparison between the sampling points one- family houses (OFH) and the river “Panke” (PNK).

FIGURE 2. Location of monitoring catchments in Berlin in separate sewer system (outside red line).

MS-parameter for quantitative analysisFull Scan polarity-switching (ESI+ / ESI–) R = 35,000 m/z 103–900 internal standards

FIGURE 6. Result of the pesticide/biocide concentrations for selected compounds for the different sampling locations.

Commercial areas

One-family houses with

gardens

Corresponding flow-proportional volume

5L composite sample

Chemical analysis

FIGURE 3. Sampling strategy: volume-proportional composite samples.

One- family houses(OFH) River �Panke“ (PNK)

Pharmaceuticals

Pesticides

Industrial and household chemicals

FIGURE 1. Process of data acquisition and processing for target and non-target analysis; all processing done with TraceFinder 3.3 software.

Data AnalysisQualitative and quantitative analysis was done with Thermo Scientific™ TraceFinder™software. Figure 1 shows a schematic diagram of the full workflow carried out in thisstudy. Starting with a suspect screening, micropollutants out of different classes likebiocides/ pesticides, pharmaceuticals, household and industrial chemicals containedin a homebuild database, were confirmed by using confirmation criteria such asisotopic pattern match and matching of known fragments. Selected suspects were aswell identifed as quantified by reference standards in a dilution series.

It can be seen that concentrations show distinct differences between catchment types.Whereas mecoprop (contained in bituminous sealing membranes for roofs) can befound in all catchment with roof runoff, carbendazim (fungicide, in urban contextapplied in paints and sealing compounds) and diuron (contained in exterior paints forsurface protection) are mostly found at monitoring station Old, an area of typicalhouses build <1930 with extensive renovations in recent years during which theapplication of new paints and sealing compounds is likely. On the other hand, concentrations of isoproturon (herbicide applied in gardens) are highest in catchmenttype OFH characterized by one family houses with gardens. Glyphosate (broadspectrum herbicide, �RoundUp“) was found at all monitoring stations, probably due toapplications for weed control on footpaths.

Quantitative results

A total of 19 biocides and pesticides were quantified regularly, of which 6 were addedas a result of the suspected Screening (simazine, terbuthylazine,desethylterbuthylazine, benzisothiazolinone, octylisothiazolinone and thiacloprid).Results for selected biocides and pesticides in stormwater runoff of the investigatedcatchment types can be seen in Figure 6.

std200ng_benzalkonium #364 RT: 7.33 AV:F: FTMS + p ESI Full ms2 304.30@hcd4 ...

100 200 300m/z

0

20

40

60

80

100

Relat

ive Ab

unda

nce

91.05483

304.30005

212.23746

110.11267

RT: 7.00 - 7.50

7.0 7.2 7.4Time (min)

0

20

40

60

80

100

Rela

tive

Abun

danc

e

7.33

7.41

7.16 7.277.06

RT: 7.00 - 7.50

7.0 7.2 7.4Time (min)

0

20

40

60

80

100

Rela

tive

Abun

danc

e

7.35

7.397.42

7.297.247.15

279 #432 RT: 7.39 AV: 1 NL: 8.17E6F: FTMS + p ESI Full ms2 304.30@hcd4 ...

100 200 300m/z

0

20

40

60

80

100

Relat

ive Ab

unda

nce

91.05484

304.30005

212.23746

121.48763

reference standard

sample

N+

CH3

CH3

C12H15

XIC 304.29996 (± 5 ppm) MS/MS

Time vs. flow Q[m3/ s]

ResultsSuspected Screening

The suspect screening results showed a large fraction of pesticides/biocides in thesamples indicating the relevance of stormwater as an entry pathway of biocides tosurface waters. The results of two samples are presented in figure 4: one-familiy houses(OFH) and the river “Panke” (PNK). It can be seen that the pesticide/biocide fraction ofidentified suspects (29 for OFH and 56 for PNK) is similar for both samples, whereasthe fraction of pharmaceuticals is much higher in the stream sample (PNK) due todischarges of WWTP effluent upstream the sampling location. For a suspect thefollowing confirming criteria were selected: isotopic pattern score > 85%, an accuratemass deviation < 5 ppm and at least one detected fragments.

OFH: e.g. Nicotine, 2-Hydroxybenzothiazole (OHBT), Benzothiazole (BT) and 2-Methylbenzothiazole (MTBT)

Panke: e.g. OHBT, MTBT, BT, Metoprolol, Carbamazepine, 4-formylaminoantipyrine (FAA), 4-acetylaminoantipyrine (AAA), Simazine, Terbutylazine, Ketamine, Tramadol

Suspect compound Formula [M+H]+ ∆ m/z(ppm)

Time (min)

IPS (%)

Isoproturon C12H18N2O 207.1492 -0.4 6.0 100Metolachlor C15H22ClNO2 284.1411 0.6 7.3 100Diethyltoluamide (DEET) C12H17NO 192.1383 0.0 6.4 100Benzisothiazolinone (BIT) C7H5NOS 152.0165 0.7 5.4 100

TABLE 1. Detected pesticides/biocides of sample point “new buildings” (NEW) with confirming criteria mass accuracy (Δm/z) and isotopic pattern score (IPS).

Whereas isoproturon and metolachlor are two plant protection products, diethyltoluamide(DEET) is an insect repellent and benzisothiazolinone (BIT) a biocide which is used inexterior and roof paints, caulks and varnish.

Non-Target Screening

In addition to the suspect screening an unknown screening with unbiased peak detectionwas carried out to screen for additional compounds of interest that had not come toattention yet. The first approach was to look into the 20 most intense compounds ofevery sample. Few of these masses were selected for parallel reaction monitoring (PRM)to get additional MS/MS information. The detected masses were m/z 304.29999,276.19492, 262.25311 and 248.16394. This is a sign for a homologous sequence (-CH2)of quaternary ammonium compounds.

A prominent quaternary ammonium compound is benzalkoniumchloride. Benzalkonium-chloride is used as biocide or cationic surfactant (disinfection). It has different alkyl chainlengths between C4-C18. The chosen selected masses have a chain length of C8, C9,C10 and C12. Figure 5 shows exemplary the results of benzalkoniumchloride C12, however, other chain lengths were also detected in the sample. The findings wereverified with reference standards.

In table 1 biocides detected by suspected screening of a sample of catchment NEW areshown. Beside these, further substances from other classes were found(pharmaceuticals, industrial-, and household chemicals).

FIGURE 5. Benzalkoniumchloride peak in the sample and the reference standard peak with MS/MS spectra.

C7H7+

C21H38N+

C14H30N+

Commercial Area [COM]

Newer buildings [NEW]

Street runoff [STR] – 1.3 km

Old buildings [OLD]

One-family houses with gardens [OFH]

Combined sewer area

ESI+

PN64633-EN 0915S

Sampling strategy

The stormwater runoff was collected with portable automatic samplers equipped with 8glass bottles (V=1.9 L). The sampler was triggered by the water level in the storm sewer(threshold: 10 cm), sample volume was 450 mL. At the beginning of a storm event,sampling intervals were shorter (5 min in the first 2 hours) to register a potential first flusheffect. After the first two hours of sampling the sampling interval was increased (every 15min). The maximum sampling duration was 4 hours.

A volume-proportional composite sample for analysis was prepared by manual mixing ofthe flow-corresponding volumes of each bottle (see Figure 3) calculated from the flow ofthe installed flow meter. In that way, the sample represents the average concentration ofthe sampled storm event.

Targeted and Non-Targeted Screening of Biocides in Urban Stormwater RunoffJulia Quilitzki1, Patricia van Baar1, Uwe Dünnbier1, Daniel Wicke², Maciej Bromirski3, Olaf Scheibner3

1Berliner Wasserbetriebe, Berlin, Germany; 2Kompetenzzentrum Wasser Berlin, Berlin, Germany; 3Thermo Fisher Scientific, Bremen, Germany; Email: olaf.scheibner@thermofisher.com

ConclusionThis study shows that suspect screening analysis is an efficient method for detection ofunknown compounds (for example simazine, nicotine, benzalkoniumchloride).The high measured concentrations of substances shows that stormwater runoff can bean important entry pathway for micropollutants to water bodies. Results also indicatethat it is necessary to consider the different urban catchment types with its specificcompounds.

References 1. Gasperi, J. et al. (2014) Micropollutants in urban stormwater: Occurrence, concen-

trations, and atmospheric contributions for a wide range of contaminants in three French catchments. Environmental Science & Pollution Research 21(8), 5282-5283

2. Zgheib, S., Moilleron, R. & Chebbo, G. (2012) Priority pollutants in urban storm-water: Part 1 - Case of separate storm sewers. Water Research 46(20), 6683-6692

OverviewPurpose: Stormwater runoff can be an important source of organic micro-pollutantsentering urban surface waters through separated sewer systems.

Methods: Screening and quantification of micropollutants was carried out for storm-water runoff samples of five different urban catchment areas in Berlin to considercatchment-specific differences.

Results: Stormwater may be a relevant source of biocides, particularly in citiesdominated by separate sewer systems.

IntroductionStormwater runoff can be an important source of organic micropollutants entering urbansurface waters through separate sewer systems (Zgheib et al. 2012). To investigate theextent of this type of pollution, a one year monitoring program was conducted in the cityof Berlin. Monitoring points were selected in five catchments of different urbanstructures to consider catchment-specific differences. Samples were analyzed for acomprehensive set of 100 micropollutants (e.g. PAHs, phthalates, phenols, heavymetals, biocides, flame retardants) that were chosen for monitoring based on literatureresearch (e.g. Gasperi et al. 2014).

MethodsSample PreparationStormwater runoff samples were filtered through 0.45 µm syringe filter before injection.

Liquid Chromatography1 mL of sample was injected directly into the online-SPE system. For pre-concentration,a C18 column, 2.1 × 20 mm with 12 µm particle size was used. For compoundseparation, a C18 column, 2.1 × 50 mm with 1.8 µm particle size was used. A water-methanol gradient both spiked with 0.1% formic acid was run from 2% to 95% in 6.7minutes. Total chromatographic cycle time including online pre-concentration was 15minutes.

Mass Spectrometry

Mass spectrometric analysis was run on a Thermo Scientific™ Q Exactive™ Focushybrid quadrupole-Orbitrap™ mass spectrometer (Figure 1).

MS-parameter for screening purposesFull Scan ESI+ and ESI– separately R = 70,000 m/z 100–1000

variable Data Independent Acquisition (vDIA) R = 70,000 CE = 30 m/z 100–205, 195–305, 295–405, 395–505, 495–1000

FIGURE 1. Q Exactive Focus with Thermo Scientific™ EQuan MAX Plus™ online SPE-system.

SamplingInvestigation areaFive sampling points were installed in separate storm sewers of five different catchmenttypes representing 85% of the connected impervious area of Berlin: one- family houseswith gardens (OFH), roads (>7500 vehicles/ day), old building (OLD, < 1930), newerbuilding (NEW, >1950) and commercial (COM) areas (Figure 2). Additionally a samplingpoint at the river “Panke” (PNK) was installed to determine contaminant concentrationsduring storm events in a stream affected by stormwater discharge.

© 2015 Thermo Fisher Scientific Inc. All rights reserved. All trademarks are the property of Thermo Fisher Scientific and its subsidiaries. This information is not intended to encourage use of these products in any manner that might infringe the intellectual property rights of others.

FIGURE 4. Suspect Screening results: Comparison between the sampling points one- family houses (OFH) and the river “Panke” (PNK).

FIGURE 2. Location of monitoring catchments in Berlin in separate sewer system (outside red line).

MS-parameter for quantitative analysisFull Scan polarity-switching (ESI+ / ESI–) R = 35,000 m/z 103–900 internal standards

FIGURE 6. Result of the pesticide/biocide concentrations for selected compounds for the different sampling locations.

Commercial areas

One-family houses with

gardens

Corresponding flow-proportional volume

5L composite sample

Chemical analysis

FIGURE 3. Sampling strategy: volume-proportional composite samples.

One- family houses(OFH) River �Panke“ (PNK)

Pharmaceuticals

Pesticides

Industrial and household chemicals

FIGURE 1. Process of data acquisition and processing for target and non-target analysis; all processing done with TraceFinder 3.3 software.

Data AnalysisQualitative and quantitative analysis was done with Thermo Scientific™ TraceFinder™software. Figure 1 shows a schematic diagram of the full workflow carried out in thisstudy. Starting with a suspect screening, micropollutants out of different classes likebiocides/ pesticides, pharmaceuticals, household and industrial chemicals containedin a homebuild database, were confirmed by using confirmation criteria such asisotopic pattern match and matching of known fragments. Selected suspects were aswell identifed as quantified by reference standards in a dilution series.

It can be seen that concentrations show distinct differences between catchment types.Whereas mecoprop (contained in bituminous sealing membranes for roofs) can befound in all catchment with roof runoff, carbendazim (fungicide, in urban contextapplied in paints and sealing compounds) and diuron (contained in exterior paints forsurface protection) are mostly found at monitoring station Old, an area of typicalhouses build <1930 with extensive renovations in recent years during which theapplication of new paints and sealing compounds is likely. On the other hand, concentrations of isoproturon (herbicide applied in gardens) are highest in catchmenttype OFH characterized by one family houses with gardens. Glyphosate (broadspectrum herbicide, �RoundUp“) was found at all monitoring stations, probably due toapplications for weed control on footpaths.

Quantitative results

A total of 19 biocides and pesticides were quantified regularly, of which 6 were addedas a result of the suspected Screening (simazine, terbuthylazine,desethylterbuthylazine, benzisothiazolinone, octylisothiazolinone and thiacloprid).Results for selected biocides and pesticides in stormwater runoff of the investigatedcatchment types can be seen in Figure 6.

std200ng_benzalkonium #364 RT: 7.33 AV:F: FTMS + p ESI Full ms2 304.30@hcd4 ...

100 200 300m/z

0

20

40

60

80

100

Relat

ive Ab

unda

nce

91.05483

304.30005

212.23746

110.11267

RT: 7.00 - 7.50

7.0 7.2 7.4Time (min)

0

20

40

60

80

100

Rela

tive

Abun

danc

e

7.33

7.41

7.16 7.277.06

RT: 7.00 - 7.50

7.0 7.2 7.4Time (min)

0

20

40

60

80

100

Rela

tive

Abun

danc

e

7.35

7.397.42

7.297.247.15

279 #432 RT: 7.39 AV: 1 NL: 8.17E6F: FTMS + p ESI Full ms2 304.30@hcd4 ...

100 200 300m/z

0

20

40

60

80

100

Relat

ive Ab

unda

nce

91.05484

304.30005

212.23746

121.48763

reference standard

sample

N+

CH3

CH3

C12H15

XIC 304.29996 (± 5 ppm) MS/MS

Time vs. flow Q[m3/ s]

ResultsSuspected Screening

The suspect screening results showed a large fraction of pesticides/biocides in thesamples indicating the relevance of stormwater as an entry pathway of biocides tosurface waters. The results of two samples are presented in figure 4: one-familiy houses(OFH) and the river “Panke” (PNK). It can be seen that the pesticide/biocide fraction ofidentified suspects (29 for OFH and 56 for PNK) is similar for both samples, whereasthe fraction of pharmaceuticals is much higher in the stream sample (PNK) due todischarges of WWTP effluent upstream the sampling location. For a suspect thefollowing confirming criteria were selected: isotopic pattern score > 85%, an accuratemass deviation < 5 ppm and at least one detected fragments.

OFH: e.g. Nicotine, 2-Hydroxybenzothiazole (OHBT), Benzothiazole (BT) and 2-Methylbenzothiazole (MTBT)

Panke: e.g. OHBT, MTBT, BT, Metoprolol, Carbamazepine, 4-formylaminoantipyrine (FAA), 4-acetylaminoantipyrine (AAA), Simazine, Terbutylazine, Ketamine, Tramadol

Suspect compound Formula [M+H]+ ∆ m/z(ppm)

Time (min)

IPS (%)

Isoproturon C12H18N2O 207.1492 -0.4 6.0 100Metolachlor C15H22ClNO2 284.1411 0.6 7.3 100Diethyltoluamide (DEET) C12H17NO 192.1383 0.0 6.4 100Benzisothiazolinone (BIT) C7H5NOS 152.0165 0.7 5.4 100

TABLE 1. Detected pesticides/biocides of sample point “new buildings” (NEW) with confirming criteria mass accuracy (Δm/z) and isotopic pattern score (IPS).

Whereas isoproturon and metolachlor are two plant protection products, diethyltoluamide(DEET) is an insect repellent and benzisothiazolinone (BIT) a biocide which is used inexterior and roof paints, caulks and varnish.

Non-Target Screening

In addition to the suspect screening an unknown screening with unbiased peak detectionwas carried out to screen for additional compounds of interest that had not come toattention yet. The first approach was to look into the 20 most intense compounds ofevery sample. Few of these masses were selected for parallel reaction monitoring (PRM)to get additional MS/MS information. The detected masses were m/z 304.29999,276.19492, 262.25311 and 248.16394. This is a sign for a homologous sequence (-CH2)of quaternary ammonium compounds.

A prominent quaternary ammonium compound is benzalkoniumchloride. Benzalkonium-chloride is used as biocide or cationic surfactant (disinfection). It has different alkyl chainlengths between C4-C18. The chosen selected masses have a chain length of C8, C9,C10 and C12. Figure 5 shows exemplary the results of benzalkoniumchloride C12, however, other chain lengths were also detected in the sample. The findings wereverified with reference standards.

In table 1 biocides detected by suspected screening of a sample of catchment NEW areshown. Beside these, further substances from other classes were found(pharmaceuticals, industrial-, and household chemicals).

FIGURE 5. Benzalkoniumchloride peak in the sample and the reference standard peak with MS/MS spectra.

C7H7+

C21H38N+

C14H30N+

Commercial Area [COM]

Newer buildings [NEW]

Street runoff [STR] – 1.3 km

Old buildings [OLD]

One-family houses with gardens [OFH]

Combined sewer area

ESI+

Sampling strategy

The stormwater runoff was collected with portable automatic samplers equipped with 8glass bottles (V=1.9 L). The sampler was triggered by the water level in the storm sewer(threshold: 10 cm), sample volume was 450 mL. At the beginning of a storm event,sampling intervals were shorter (5 min in the first 2 hours) to register a potential first flusheffect. After the first two hours of sampling the sampling interval was increased (every 15min). The maximum sampling duration was 4 hours.

A volume-proportional composite sample for analysis was prepared by manual mixing ofthe flow-corresponding volumes of each bottle (see Figure 3) calculated from the flow ofthe installed flow meter. In that way, the sample represents the average concentration ofthe sampled storm event.

Targeted and Non-Targeted Screening of Biocides in Urban Stormwater RunoffJulia Quilitzki1, Patricia van Baar1, Uwe Dünnbier1, Daniel Wicke², Maciej Bromirski3, Olaf Scheibner3

1Berliner Wasserbetriebe, Berlin, Germany; 2Kompetenzzentrum Wasser Berlin, Berlin, Germany; 3Thermo Fisher Scientific, Bremen, Germany; Email: olaf.scheibner@thermofisher.com

ConclusionThis study shows that suspect screening analysis is an efficient method for detection ofunknown compounds (for example simazine, nicotine, benzalkoniumchloride).The high measured concentrations of substances shows that stormwater runoff can bean important entry pathway for micropollutants to water bodies. Results also indicatethat it is necessary to consider the different urban catchment types with its specificcompounds.

References 1. Gasperi, J. et al. (2014) Micropollutants in urban stormwater: Occurrence, concen-

trations, and atmospheric contributions for a wide range of contaminants in three French catchments. Environmental Science & Pollution Research 21(8), 5282-5283

2. Zgheib, S., Moilleron, R. & Chebbo, G. (2012) Priority pollutants in urban storm-water: Part 1 - Case of separate storm sewers. Water Research 46(20), 6683-6692

OverviewPurpose: Stormwater runoff can be an important source of organic micro-pollutantsentering urban surface waters through separated sewer systems.

Methods: Screening and quantification of micropollutants was carried out for storm-water runoff samples of five different urban catchment areas in Berlin to considercatchment-specific differences.

Results: Stormwater may be a relevant source of biocides, particularly in citiesdominated by separate sewer systems.

IntroductionStormwater runoff can be an important source of organic micropollutants entering urbansurface waters through separate sewer systems (Zgheib et al. 2012). To investigate theextent of this type of pollution, a one year monitoring program was conducted in the cityof Berlin. Monitoring points were selected in five catchments of different urbanstructures to consider catchment-specific differences. Samples were analyzed for acomprehensive set of 100 micropollutants (e.g. PAHs, phthalates, phenols, heavymetals, biocides, flame retardants) that were chosen for monitoring based on literatureresearch (e.g. Gasperi et al. 2014).

MethodsSample PreparationStormwater runoff samples were filtered through 0.45 µm syringe filter before injection.

Liquid Chromatography1 mL of sample was injected directly into the online-SPE system. For pre-concentration,a C18 column, 2.1 × 20 mm with 12 µm particle size was used. For compoundseparation, a C18 column, 2.1 × 50 mm with 1.8 µm particle size was used. A water-methanol gradient both spiked with 0.1% formic acid was run from 2% to 95% in 6.7minutes. Total chromatographic cycle time including online pre-concentration was 15minutes.

Mass Spectrometry

Mass spectrometric analysis was run on a Thermo Scientific™ Q Exactive™ Focushybrid quadrupole-Orbitrap™ mass spectrometer (Figure 1).

MS-parameter for screening purposesFull Scan ESI+ and ESI– separately R = 70,000 m/z 100–1000

variable Data Independent Acquisition (vDIA) R = 70,000 CE = 30 m/z 100–205, 195–305, 295–405, 395–505, 495–1000

FIGURE 1. Q Exactive Focus with Thermo Scientific™ EQuan MAX Plus™ online SPE-system.

SamplingInvestigation areaFive sampling points were installed in separate storm sewers of five different catchmenttypes representing 85% of the connected impervious area of Berlin: one- family houseswith gardens (OFH), roads (>7500 vehicles/ day), old building (OLD, < 1930), newerbuilding (NEW, >1950) and commercial (COM) areas (Figure 2). Additionally a samplingpoint at the river “Panke” (PNK) was installed to determine contaminant concentrationsduring storm events in a stream affected by stormwater discharge.

© 2015 Thermo Fisher Scientific Inc. All rights reserved. All trademarks are the property of Thermo Fisher Scientific and its subsidiaries. This information is not intended to encourage use of these products in any manner that might infringe the intellectual property rights of others.

FIGURE 4. Suspect Screening results: Comparison between the sampling points one- family houses (OFH) and the river “Panke” (PNK).

FIGURE 2. Location of monitoring catchments in Berlin in separate sewer system (outside red line).

MS-parameter for quantitative analysisFull Scan polarity-switching (ESI+ / ESI–) R = 35,000 m/z 103–900 internal standards

FIGURE 6. Result of the pesticide/biocide concentrations for selected compounds for the different sampling locations.

Commercial areas

One-family houses with

gardens

Corresponding flow-proportional volume

5L composite sample

Chemical analysis

FIGURE 3. Sampling strategy: volume-proportional composite samples.

One- family houses(OFH) River �Panke“ (PNK)

Pharmaceuticals

Pesticides

Industrial and household chemicals

FIGURE 1. Process of data acquisition and processing for target and non-target analysis; all processing done with TraceFinder 3.3 software.

Data AnalysisQualitative and quantitative analysis was done with Thermo Scientific™ TraceFinder™software. Figure 1 shows a schematic diagram of the full workflow carried out in thisstudy. Starting with a suspect screening, micropollutants out of different classes likebiocides/ pesticides, pharmaceuticals, household and industrial chemicals containedin a homebuild database, were confirmed by using confirmation criteria such asisotopic pattern match and matching of known fragments. Selected suspects were aswell identifed as quantified by reference standards in a dilution series.

It can be seen that concentrations show distinct differences between catchment types.Whereas mecoprop (contained in bituminous sealing membranes for roofs) can befound in all catchment with roof runoff, carbendazim (fungicide, in urban contextapplied in paints and sealing compounds) and diuron (contained in exterior paints forsurface protection) are mostly found at monitoring station Old, an area of typicalhouses build <1930 with extensive renovations in recent years during which theapplication of new paints and sealing compounds is likely. On the other hand, concentrations of isoproturon (herbicide applied in gardens) are highest in catchmenttype OFH characterized by one family houses with gardens. Glyphosate (broadspectrum herbicide, �RoundUp“) was found at all monitoring stations, probably due toapplications for weed control on footpaths.

Quantitative results

A total of 19 biocides and pesticides were quantified regularly, of which 6 were addedas a result of the suspected Screening (simazine, terbuthylazine,desethylterbuthylazine, benzisothiazolinone, octylisothiazolinone and thiacloprid).Results for selected biocides and pesticides in stormwater runoff of the investigatedcatchment types can be seen in Figure 6.

std200ng_benzalkonium #364 RT: 7.33 AV:F: FTMS + p ESI Full ms2 304.30@hcd4 ...

100 200 300m/z

0

20

40

60

80

100

Relat

ive Ab

unda

nce

91.05483

304.30005

212.23746

110.11267

RT: 7.00 - 7.50

7.0 7.2 7.4Time (min)

0

20

40

60

80

100

Rela

tive

Abun

danc

e

7.33

7.41

7.16 7.277.06

RT: 7.00 - 7.50

7.0 7.2 7.4Time (min)

0

20

40

60

80

100

Rela

tive

Abun

danc

e

7.35

7.397.42

7.297.247.15

279 #432 RT: 7.39 AV: 1 NL: 8.17E6F: FTMS + p ESI Full ms2 304.30@hcd4 ...

100 200 300m/z

0

20

40

60

80

100

Relat

ive Ab

unda

nce

91.05484

304.30005

212.23746

121.48763

reference standard

sample

N+

CH3

CH3

C12H15

XIC 304.29996 (± 5 ppm) MS/MS

Time vs. flow Q[m3/ s]

ResultsSuspected Screening

The suspect screening results showed a large fraction of pesticides/biocides in thesamples indicating the relevance of stormwater as an entry pathway of biocides tosurface waters. The results of two samples are presented in figure 4: one-familiy houses(OFH) and the river “Panke” (PNK). It can be seen that the pesticide/biocide fraction ofidentified suspects (29 for OFH and 56 for PNK) is similar for both samples, whereasthe fraction of pharmaceuticals is much higher in the stream sample (PNK) due todischarges of WWTP effluent upstream the sampling location. For a suspect thefollowing confirming criteria were selected: isotopic pattern score > 85%, an accuratemass deviation < 5 ppm and at least one detected fragments.

OFH: e.g. Nicotine, 2-Hydroxybenzothiazole (OHBT), Benzothiazole (BT) and 2-Methylbenzothiazole (MTBT)

Panke: e.g. OHBT, MTBT, BT, Metoprolol, Carbamazepine, 4-formylaminoantipyrine (FAA), 4-acetylaminoantipyrine (AAA), Simazine, Terbutylazine, Ketamine, Tramadol

Suspect compound Formula [M+H]+ ∆ m/z(ppm)

Time (min)

IPS (%)

Isoproturon C12H18N2O 207.1492 -0.4 6.0 100Metolachlor C15H22ClNO2 284.1411 0.6 7.3 100Diethyltoluamide (DEET) C12H17NO 192.1383 0.0 6.4 100Benzisothiazolinone (BIT) C7H5NOS 152.0165 0.7 5.4 100

TABLE 1. Detected pesticides/biocides of sample point “new buildings” (NEW) with confirming criteria mass accuracy (Δm/z) and isotopic pattern score (IPS).

Whereas isoproturon and metolachlor are two plant protection products, diethyltoluamide(DEET) is an insect repellent and benzisothiazolinone (BIT) a biocide which is used inexterior and roof paints, caulks and varnish.

Non-Target Screening

In addition to the suspect screening an unknown screening with unbiased peak detectionwas carried out to screen for additional compounds of interest that had not come toattention yet. The first approach was to look into the 20 most intense compounds ofevery sample. Few of these masses were selected for parallel reaction monitoring (PRM)to get additional MS/MS information. The detected masses were m/z 304.29999,276.19492, 262.25311 and 248.16394. This is a sign for a homologous sequence (-CH2)of quaternary ammonium compounds.

A prominent quaternary ammonium compound is benzalkoniumchloride. Benzalkonium-chloride is used as biocide or cationic surfactant (disinfection). It has different alkyl chainlengths between C4-C18. The chosen selected masses have a chain length of C8, C9,C10 and C12. Figure 5 shows exemplary the results of benzalkoniumchloride C12, however, other chain lengths were also detected in the sample. The findings wereverified with reference standards.

In table 1 biocides detected by suspected screening of a sample of catchment NEW areshown. Beside these, further substances from other classes were found(pharmaceuticals, industrial-, and household chemicals).

FIGURE 5. Benzalkoniumchloride peak in the sample and the reference standard peak with MS/MS spectra.

C7H7+

C21H38N+

C14H30N+

Commercial Area [COM]

Newer buildings [NEW]

Street runoff [STR] – 1.3 km

Old buildings [OLD]

One-family houses with gardens [OFH]

Combined sewer area

ESI+

Sampling strategy

The stormwater runoff was collected with portable automatic samplers equipped with 8glass bottles (V=1.9 L). The sampler was triggered by the water level in the storm sewer(threshold: 10 cm), sample volume was 450 mL. At the beginning of a storm event,sampling intervals were shorter (5 min in the first 2 hours) to register a potential first flusheffect. After the first two hours of sampling the sampling interval was increased (every 15min). The maximum sampling duration was 4 hours.

A volume-proportional composite sample for analysis was prepared by manual mixing ofthe flow-corresponding volumes of each bottle (see Figure 3) calculated from the flow ofthe installed flow meter. In that way, the sample represents the average concentration ofthe sampled storm event.

Targeted and Non-Targeted Screening of Biocides in Urban Stormwater RunoffJulia Quilitzki1, Patricia van Baar1, Uwe Dünnbier1, Daniel Wicke², Maciej Bromirski3, Olaf Scheibner3

1Berliner Wasserbetriebe, Berlin, Germany; 2Kompetenzzentrum Wasser Berlin, Berlin, Germany; 3Thermo Fisher Scientific, Bremen, Germany; Email: olaf.scheibner@thermofisher.com

ConclusionThis study shows that suspect screening analysis is an efficient method for detection ofunknown compounds (for example simazine, nicotine, benzalkoniumchloride).The high measured concentrations of substances shows that stormwater runoff can bean important entry pathway for micropollutants to water bodies. Results also indicatethat it is necessary to consider the different urban catchment types with its specificcompounds.

References 1. Gasperi, J. et al. (2014) Micropollutants in urban stormwater: Occurrence, concen-

trations, and atmospheric contributions for a wide range of contaminants in three French catchments. Environmental Science & Pollution Research 21(8), 5282-5283

2. Zgheib, S., Moilleron, R. & Chebbo, G. (2012) Priority pollutants in urban storm-water: Part 1 - Case of separate storm sewers. Water Research 46(20), 6683-6692

OverviewPurpose: Stormwater runoff can be an important source of organic micro-pollutantsentering urban surface waters through separated sewer systems.

Methods: Screening and quantification of micropollutants was carried out for storm-water runoff samples of five different urban catchment areas in Berlin to considercatchment-specific differences.

Results: Stormwater may be a relevant source of biocides, particularly in citiesdominated by separate sewer systems.

IntroductionStormwater runoff can be an important source of organic micropollutants entering urbansurface waters through separate sewer systems (Zgheib et al. 2012). To investigate theextent of this type of pollution, a one year monitoring program was conducted in the cityof Berlin. Monitoring points were selected in five catchments of different urbanstructures to consider catchment-specific differences. Samples were analyzed for acomprehensive set of 100 micropollutants (e.g. PAHs, phthalates, phenols, heavymetals, biocides, flame retardants) that were chosen for monitoring based on literatureresearch (e.g. Gasperi et al. 2014).

MethodsSample PreparationStormwater runoff samples were filtered through 0.45 µm syringe filter before injection.

Liquid Chromatography1 mL of sample was injected directly into the online-SPE system. For pre-concentration,a C18 column, 2.1 × 20 mm with 12 µm particle size was used. For compoundseparation, a C18 column, 2.1 × 50 mm with 1.8 µm particle size was used. A water-methanol gradient both spiked with 0.1% formic acid was run from 2% to 95% in 6.7minutes. Total chromatographic cycle time including online pre-concentration was 15minutes.

Mass Spectrometry

Mass spectrometric analysis was run on a Thermo Scientific™ Q Exactive™ Focushybrid quadrupole-Orbitrap™ mass spectrometer (Figure 1).

MS-parameter for screening purposesFull Scan ESI+ and ESI– separately R = 70,000 m/z 100–1000

variable Data Independent Acquisition (vDIA) R = 70,000 CE = 30 m/z 100–205, 195–305, 295–405, 395–505, 495–1000

FIGURE 1. Q Exactive Focus with Thermo Scientific™ EQuan MAX Plus™ online SPE-system.

SamplingInvestigation areaFive sampling points were installed in separate storm sewers of five different catchmenttypes representing 85% of the connected impervious area of Berlin: one- family houseswith gardens (OFH), roads (>7500 vehicles/ day), old building (OLD, < 1930), newerbuilding (NEW, >1950) and commercial (COM) areas (Figure 2). Additionally a samplingpoint at the river “Panke” (PNK) was installed to determine contaminant concentrationsduring storm events in a stream affected by stormwater discharge.

© 2015 Thermo Fisher Scientific Inc. All rights reserved. All trademarks are the property of Thermo Fisher Scientific and its subsidiaries. This information is not intended to encourage use of these products in any manner that might infringe the intellectual property rights of others.

FIGURE 4. Suspect Screening results: Comparison between the sampling points one- family houses (OFH) and the river “Panke” (PNK).

FIGURE 2. Location of monitoring catchments in Berlin in separate sewer system (outside red line).

MS-parameter for quantitative analysisFull Scan polarity-switching (ESI+ / ESI–) R = 35,000 m/z 103–900 internal standards

FIGURE 6. Result of the pesticide/biocide concentrations for selected compounds for the different sampling locations.

Commercial areas

One-family houses with

gardens

Corresponding flow-proportional volume

5L composite sample

Chemical analysis

FIGURE 3. Sampling strategy: volume-proportional composite samples.

One- family houses(OFH) River �Panke“ (PNK)

Pharmaceuticals

Pesticides

Industrial and household chemicals

FIGURE 1. Process of data acquisition and processing for target and non-target analysis; all processing done with TraceFinder 3.3 software.

Data AnalysisQualitative and quantitative analysis was done with Thermo Scientific™ TraceFinder™software. Figure 1 shows a schematic diagram of the full workflow carried out in thisstudy. Starting with a suspect screening, micropollutants out of different classes likebiocides/ pesticides, pharmaceuticals, household and industrial chemicals containedin a homebuild database, were confirmed by using confirmation criteria such asisotopic pattern match and matching of known fragments. Selected suspects were aswell identifed as quantified by reference standards in a dilution series.

It can be seen that concentrations show distinct differences between catchment types.Whereas mecoprop (contained in bituminous sealing membranes for roofs) can befound in all catchment with roof runoff, carbendazim (fungicide, in urban contextapplied in paints and sealing compounds) and diuron (contained in exterior paints forsurface protection) are mostly found at monitoring station Old, an area of typicalhouses build <1930 with extensive renovations in recent years during which theapplication of new paints and sealing compounds is likely. On the other hand, concentrations of isoproturon (herbicide applied in gardens) are highest in catchmenttype OFH characterized by one family houses with gardens. Glyphosate (broadspectrum herbicide, �RoundUp“) was found at all monitoring stations, probably due toapplications for weed control on footpaths.

Quantitative results

A total of 19 biocides and pesticides were quantified regularly, of which 6 were addedas a result of the suspected Screening (simazine, terbuthylazine,desethylterbuthylazine, benzisothiazolinone, octylisothiazolinone and thiacloprid).Results for selected biocides and pesticides in stormwater runoff of the investigatedcatchment types can be seen in Figure 6.

std200ng_benzalkonium #364 RT: 7.33 AV:F: FTMS + p ESI Full ms2 304.30@hcd4 ...

100 200 300m/z

0

20

40

60

80

100

Relat

ive Ab

unda

nce

91.05483

304.30005

212.23746

110.11267

RT: 7.00 - 7.50

7.0 7.2 7.4Time (min)

0

20

40

60

80

100

Rela

tive

Abun

danc

e

7.33

7.41

7.16 7.277.06

RT: 7.00 - 7.50

7.0 7.2 7.4Time (min)

0

20

40

60

80

100

Rela

tive

Abun

danc

e

7.35

7.397.42

7.297.247.15

279 #432 RT: 7.39 AV: 1 NL: 8.17E6F: FTMS + p ESI Full ms2 304.30@hcd4 ...

100 200 300m/z

0

20

40

60

80

100

Relat

ive Ab

unda

nce

91.05484

304.30005

212.23746

121.48763

reference standard

sample

N+

CH3

CH3

C12H15

XIC 304.29996 (± 5 ppm) MS/MS

Time vs. flow Q[m3/ s]

ResultsSuspected Screening

The suspect screening results showed a large fraction of pesticides/biocides in thesamples indicating the relevance of stormwater as an entry pathway of biocides tosurface waters. The results of two samples are presented in figure 4: one-familiy houses(OFH) and the river “Panke” (PNK). It can be seen that the pesticide/biocide fraction ofidentified suspects (29 for OFH and 56 for PNK) is similar for both samples, whereasthe fraction of pharmaceuticals is much higher in the stream sample (PNK) due todischarges of WWTP effluent upstream the sampling location. For a suspect thefollowing confirming criteria were selected: isotopic pattern score > 85%, an accuratemass deviation < 5 ppm and at least one detected fragments.

OFH: e.g. Nicotine, 2-Hydroxybenzothiazole (OHBT), Benzothiazole (BT) and 2-Methylbenzothiazole (MTBT)

Panke: e.g. OHBT, MTBT, BT, Metoprolol, Carbamazepine, 4-formylaminoantipyrine (FAA), 4-acetylaminoantipyrine (AAA), Simazine, Terbutylazine, Ketamine, Tramadol

Suspect compound Formula [M+H]+ ∆ m/z(ppm)

Time (min)

IPS (%)

Isoproturon C12H18N2O 207.1492 -0.4 6.0 100Metolachlor C15H22ClNO2 284.1411 0.6 7.3 100Diethyltoluamide (DEET) C12H17NO 192.1383 0.0 6.4 100Benzisothiazolinone (BIT) C7H5NOS 152.0165 0.7 5.4 100

TABLE 1. Detected pesticides/biocides of sample point “new buildings” (NEW) with confirming criteria mass accuracy (Δm/z) and isotopic pattern score (IPS).

Whereas isoproturon and metolachlor are two plant protection products, diethyltoluamide(DEET) is an insect repellent and benzisothiazolinone (BIT) a biocide which is used inexterior and roof paints, caulks and varnish.

Non-Target Screening

In addition to the suspect screening an unknown screening with unbiased peak detectionwas carried out to screen for additional compounds of interest that had not come toattention yet. The first approach was to look into the 20 most intense compounds ofevery sample. Few of these masses were selected for parallel reaction monitoring (PRM)to get additional MS/MS information. The detected masses were m/z 304.29999,276.19492, 262.25311 and 248.16394. This is a sign for a homologous sequence (-CH2)of quaternary ammonium compounds.

A prominent quaternary ammonium compound is benzalkoniumchloride. Benzalkonium-chloride is used as biocide or cationic surfactant (disinfection). It has different alkyl chainlengths between C4-C18. The chosen selected masses have a chain length of C8, C9,C10 and C12. Figure 5 shows exemplary the results of benzalkoniumchloride C12, however, other chain lengths were also detected in the sample. The findings wereverified with reference standards.

In table 1 biocides detected by suspected screening of a sample of catchment NEW areshown. Beside these, further substances from other classes were found(pharmaceuticals, industrial-, and household chemicals).

FIGURE 5. Benzalkoniumchloride peak in the sample and the reference standard peak with MS/MS spectra.

C7H7+

C21H38N+

C14H30N+

Commercial Area [COM]

Newer buildings [NEW]

Street runoff [STR] – 1.3 km

Old buildings [OLD]

One-family houses with gardens [OFH]

Combined sewer area

ESI+

Africa +43 1 333 50 34 0Australia +61 3 9757 4300Austria +43 810 282 206Belgium +32 53 73 42 41Canada +1 800 530 8447China 800 810 5118 (free call domestic)

400 650 5118

Denmark +45 70 23 62 60Europe-Other +43 1 333 50 34 0Finland +358 10 3292 200France +33 1 60 92 48 00Germany +49 6103 408 1014India +91 22 6742 9494Italy +39 02 950 591

Japan +81 45 453 9100Korea +82 2 3420 8600Latin America +1 561 688 8700Middle East +43 1 333 50 34 0Netherlands +31 76 579 55 55New Zealand +64 9 980 6700Norway +46 8 556 468 00

Russia/CIS +43 1 333 50 34 0Singapore +65 6289 1190Spain +34 914 845 965Sweden +46 8 556 468 00Switzerland +41 61 716 77 00UK +44 1442 233555USA +1 800 532 4752

www.thermoscientific.com©2015 Thermo Fisher Scientifi c Inc. All rights reserved. ISO is a trademark of the International Standards Organization. All other trademarks are the property of Thermo Fisher Scientifi c and its subsidiaries. This information is presented as an example of the capabilities of Thermo Fisher Scientifi c products. It is not intended to encourage use of these products in any manners that might infringe the intellectual property rights of others. Specifi cations, terms and pricing are subject to change. Not all products are available in all countries. Please consult your local sales representative for details.

Thermo Fisher Scientifi c, San Jose, CA USA is ISO 13485 Certifi ed.

ISO 13485