Table S1: Sample preparation, instrumental analysis, validation parameters and application of sent standard mixture for laboratory coded 1.
COMPOUND
IF
SAMPLE PREP
INTRUM ANALYSIS
VALIDATION PARAM
Sample preparation
(sample pre-treatment: pH adjustment, filter pore
size, material and trade name, extraction method
and conditions, additional clean-up, derivatisation,
internal standard …)
Instrumental analysis
(operational parameters: separation and
detection condition,…)
Validation parameters
(sensitivity, accuracy, recovery,
reproducibility,
repeatability, …)
1º) pH adjustment to 2 with HCl
2º) Addition of isotopically labelled surrogate
standards (SS)
3º) Dilution of samples B, C and D 1:1 with HPLCwater
4º) Filtration through 0.45 µm cellulose acetate
membranes
5º) Concentration by on-line SPE-LC-MS/MS:
sample volume, 5 mL; cartridge, PLRP-s (10×2
mm, Spark Holland, Emmem, The Netherlands);
washing, 0.5 mL HPLC-water
- Analysis by LC-MS/MS
- Equipment: SymbiosisTM Pico extraction
system (Spark Holland, Emmem, The
Netherlands) coupled on-line with a
4000QTRAP hybrid quadrupole-linear
ion trap mass spectrometer equipped
with a Turbo Ion Spray source from
Applied Biosystems-Sciex (Foster City,
California, USA).
- LC separation on a reversed-phase
column Purospher STAR RP-18e (125 x
2 mm, 5 µm particle size) from Merck
(Darmstadt, Germany), maintained at 25
ºC. Ultrapure water (A) and methanol
(B), both containing 0.1% of formic
acid, were employed as mobile phase
(flow-rate 0.2 mL min-1). LC gradient:
0–1 min, 5% B; 2 min, 20% B; 12 min,
80% B; 25–30 min, 100% B; 35–40 min,
5% B.
- MS/MS conditions: positive ESI; SRM
- Quantification by the
isotope dilution method
with calibration curves
prepared in HPLC-water
- Linearity: determination
coefficient r2 = 0.9995
- Sensitivity: method limit of
quantification (s/n 10) = 2.0
ng L-1 in surface water and
in wastewater.
- Accuracy: relative recovery
(calculated with respect to
the SS, at 20, 500 and 5,000
ng L-1 spiking levels, in
surface and wastewater,
n=5) = 89-107%.
- Repeatability: relative
standard deviation (RSD, in
surface and wastewater,
spiking levels 20, 500 and
5,000 ng L-1, n=5) = 1 -
Application of
sent mixture
a. Standard
mixture was
used for
calibration
b. Standard
mixture was
used for
check-up
c.Standard
mixture was
not used
b
quantification transition, 261.1 > 92.0;
SRM confirmation transition, 261.1 >
154.0; collision energy, 35 eV and 31
eV, respectively; declustering potential,
81 V.
CP
1º) pH adjustment to 2 with HCl
2º) Addition of isotopically labelled surrogate
standards (SS)
3º) Dilution of samples B, C and D 1:1 with HPLCwater
4º) Filtration through 0.45 µm cellulose acetate
membranes
5º) Concentration by on-line SPE-LC-MS/MS:
sample volume, 5 mL; cartridge, PLRP-s (10×2
mm, Spark Holland, Emmem, The Netherlands);
washing, 0.5 mL HPLC-water.
ETO
1º) pH adjustment to 2 with HCl
2º) Addition of isotopically labelled surrogate
standards (SS)
13 %.
- Analysis by LC-MS/MS
- Equipment: SymbiosisTM Pico extraction
system (Spark Holland, Emmem, The
Netherlands) coupled on-line with a
4000QTRAP hybrid quadrupole-linear
ion trap mass spectrometer equipped
with a Turbo Ion Spray source from
Applied Biosystems-Sciex (Foster City,
California, USA).
- LC separation on a reversed-phase
column Purospher STAR RP-18e (125 x
2 mm, 5 µm particle size) from Merck
(Darmstadt, Germany), maintained at 25
ºC. Ultrapure water (A) and methanol
(B), both containing 0.1% of formic
acid, were employed as mobile phase
(flow-rate 0.2 mL min-1). LC gradient:
0–1 min, 5% B; 2 min, 20% B; 12 min,
80% B; 25–30 min, 100% B; 35–40 min,
5% B.
- MS/MS conditions: positive ESI; SRM
quantification transition, 261.1 > 140.0;
SRM confirmation transition, 261.1 >
106.1; collision energy, 33 eV and 25
eV, respectively; declustering potential,
86 V.
- Quantification by the
isotope dilution method
with calibration curves
prepared in HPLC-water
- Linearity: determination
coefficient r2 = 0.9999
- Sensitivity: method limit of
quantification (s/n 10) = 2.0
ng L-1 in surface water and
3.0 ng L-1 in wastewater.
- Accuracy: relative recovery
(calculated with respect to
the SS, at 20, 500 and 5,000
ng L-1 spiking levels, in
surface and wastewater,
n=5) = 83-115%.
- Repeatability: relative
standard deviation (RSD, in
surface and wastewater,
spiking levels 20, 500 and
5,000 ng L-1, n=5) = 1 13 %.
b
- Analysis by LC-MS/MS
- Equipment: SymbiosisTM Pico extraction
system (Spark Holland, Emmem, The
Netherlands) coupled on-line with a
4000QTRAP hybrid quadrupole-linear
ion trap mass spectrometer equipped
- Quantification by the
isotope dilution method
with calibration curves
prepared in HPLC-water
- Linearity: determination
b
3º) Dilution of samples B, C and D 1:1 with HPLCwater
4º) Filtration through 0.45 µm cellulose acetate
membranes
5º) Concentration by on-line SPE-LC-MS/MS:
sample volume, 5 mL; cartridge, PLRP-s (10×2
mm, Spark Holland, Emmem, The Netherlands);
washing, 0.5 mL HPLC-water.
GEM
1º) pH adjustment to 2 with HCl
2º) Addition of isotopically labelled surrogate
standards (SS)
3º) Dilution of samples B, C and D 1:1 with HPLCwater
4º) Filtration through 0.45 µm cellulose acetate
membranes
5º) Concentration by on-line SPE-LC-MS/MS:
sample volume, 5 mL; cartridge, PLRP-s (10×2
mm, Spark Holland, Emmem, The Netherlands);
washing, 0.5 mL HPLC-water
with a Turbo Ion Spray source from
Applied Biosystems-Sciex (Foster City,
California, USA).
- LC separation on a reversed-phase
column Purospher STAR RP-18e (125 x
2 mm, 5 µm particle size) from Merck
(Darmstadt, Germany), maintained at 25
ºC. Ultrapure water (A) and methanol
(B), both containing 0.1% of formic
acid, were employed as mobile phase
(flow-rate 0.2 mL min-1). LC gradient:
0–1 min, 5% B; 2 min, 20% B; 12 min,
80% B; 25–30 min, 100% B; 35–40 min,
5% B.
- MS/MS conditions: positive ESI; SRM
quantification transition, 589.0 > 229.0;
SRM confirmation transition, 589.0 >
185.0; collision energy, 15 eV and 10
eV, respectively; declustering potential,
71 V.
coefficient r2 = 0.9997
- Sensitivity: method limit of
quantification (s/n 10) = 43
ng L-1 in surface water and
65 ng L-1 in wastewater.
- Accuracy: relative recovery
(calculated with respect to
the SS, at 500 and 5,000 ng
L-1 spiking levels, in surface
and wastewater, n=5) = 76106%.
- Repeatability: relative
standard deviation (RSD, in
surface and wastewater,
spiking levels 500 and 5,000
ng L-1, n=5) = 2-12 %.
- Analysis by LC-MS/MS
- Equipment: SymbiosisTM Pico extraction
system (Spark Holland, Emmem, The
Netherlands) coupled on-line with a
4000QTRAP hybrid quadrupole-linear
ion trap mass spectrometer equipped
with a Turbo Ion Spray source from
Applied Biosystems-Sciex (Foster City,
California, USA).
- LC separation on a reversed-phase
column Purospher STAR RP-18e (125 x
2 mm, 5 µm particle size) from Merck
(Darmstadt, Germany), maintained at 25
ºC. Ultrapure water (A) and methanol
(B), both containing 0.1% of formic
acid, were employed as mobile phase
(flow-rate 0.2 mL min-1). LC gradient:
0–1 min, 5% B; 2 min, 20% B; 12 min,
- Quantification by the
isotope dilution method
with calibration curves
prepared in HPLC-water
- Linearity: determination
coefficient r2 = 0.9993
- Sensitivity: method limit of
quantification (s/n 10) = 9.3
ng L-1 in surface water and
in wastewater.
- Accuracy: relative recovery
(calculated with respect to
the SS, at 20, 500 and 5000
ng L-1 spiking levels, in
surface and wastewater,
n=5) = 93-115%.
- Repeatability: relative
standard deviation (RSD, in
b
MTX
1º) pH adjustment to 2 with HCl
2º) Addition of isotopically labelled internal
standards (IS)
3º) Dilution of samples B, C and D 1:1 with HPLCwater
4º) Filtration through 0.45 µm cellulose acetate
membranes
5º) Concentration by on-line SPE-LC-MS/MS:
sample volume, 5 mL; cartridge, PLRP-s (10×2
mm, Spark Holland, Emmem, The Netherlands);
washing, 0.5 mL HPLC-water.
80% B; 25–30 min, 100% B; 35–40 min,
5% B.
- MS/MS conditions: positive ESI; SRM
quantification transition, 264.2 > 112.0;
SRM confirmation transition, 264.2 >
95.0; collision energy, 25 eV and 63 eV,
respectively; declustering potential, 71
V.
surface and wastewater,
spiking levels 20, 500 and
5,000 ng L-1, n=5) = 1 15 %.
- Analysis by LC-MS/MS
- Equipment: SymbiosisTM Pico extraction
system (Spark Holland, Emmem, The
Netherlands) coupled on-line with a
4000QTRAP hybrid quadrupole-linear
ion trap mass spectrometer equipped
with a Turbo Ion Spray source from
Applied Biosystems-Sciex (Foster City,
California, USA).
- LC separation on a reversed-phase
column Purospher STAR RP-18e (125 x
2 mm, 5 µm particle size) from Merck
(Darmstadt, Germany), maintained at 25
ºC. Ultrapure water (A) and methanol
(B), both containing 0.1% of formic
acid, were employed as mobile phase
(flow-rate 0.2 mL min-1). LC gradient:
0–1 min, 5% B; 2 min, 20% B; 12 min,
80% B; 25–30 min, 100% B; 35–40 min,
5% B.
- MS/MS conditions: positive ESI; SRM
quantification transition, 455.2 > 308.2;
SRM confirmation transition, 455.2 >
175.1; collision energy, 33 eV and 59
eV, respectively; declustering potential,
91 V.
- Quantification by the
isotope dilution method
with calibration curves
prepared in HPLC-water
- Linearity: determination
coefficient r2 = 0.9995
- Sensitivity: method limit of
quantification (s/n 10) = 2.0
ng L-1 in surface water and
in wastewater.
- Accuracy: relative recovery
(calculated with respect to
the SS, at 20, 500 and 5000
ng L-1 spiking levels, in
surface and wastewater,
n=5) = 84-104%.
- Repeatability: relative
standard deviation (RSD, in
surface and wastewater,
spiking levels 20, 500 and
5,000 ng L-1, n=5) = 115 %.
b