Table S9: Sample preparation, instrumental analysis, validation parameters and application of sent standard mixture for laboratory coded 9
COMPOUND / SAMPLE PREP / INTRUM ANALYSIS / VALIDATION PARAM / Application of sent mixtureSample 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,…) / a. Standard mixture was used for calibration
b. Standard mixture was used for check-up
c.Standard mixture was not used
5-FU / Filtration of samples: initially filtered
through glass microfibre filters (Machery Nagel, Dueren, Germany),
and then through 1.2 μm cellulose nitrate filters (Whatman, Dassel, Germany).
WW sample preparation protocol: employed for determination
of 5-FU in trace level concentrations in actual wastewater samples. 1 g/6 mL Isolute ENV+ cartridges (Biotage AB, Uppsala, Sweden) were conditioned with 6 mL methanol, equilibrated with 6 mL deionised water, then enriched with 100 mL wastewater samples (pH 6), vacuum-dried and eluted with 3 mL × 2 mL of methanol. The extracts were dried under nitrogen stream and dissolved in 150 μL ethyl acetate.
SW sample preparation protocol: was employed for determination of 5-FU in trace level concentrations in surface water samples. This protocol is identical to “WW sample preparation protocol”, apart from the sorbent mass which was in the latter case 500 mg of Isolute ENV+ sorbent per 6 mL cartridge.
Derivatization step: 30 μL MTBSTFA at 80 ◦C for 1 h and analysed with GC–MS/MS.
Internal standard: 5-Chlorouracil and deuterated 5-FU / An Agilent 450-GC hyphenated with an ion trap 240-MS mass
spectrometer was employed to determine 5-FU. The GC oven was programmed as follows: an initial temperature of 65 ◦C was held for 2 min, then ramped at 30 ◦C/min to 180 ◦C, ramped at 7 ◦C/min to 220 ◦C, at 40 ◦C/min to 300 ◦C, and finally held for 3 min at this temperature. The total GC run time was 16.55 min. A DB-5 MS 30 m × 0.25 mm × 0.25 μm (Agilent J&W, CA, USA) capillary column was used, with He as the carrier gas (37 cm s−1). One-microlitre samples were injected at 250 ◦C in splitless mode, kept until 3 min, and then split at 1/30. The ion trap MS was operated in electron impact (EI) ionisation mode with an external ionisation source at 15μAmps emission current and a target ion count of 20,000. All MS/MS transitions were performed in a resonant waveform type at following excitation amplitudes: 1.19 V for 5-FU, 1.20 V for 5-FU-d1 and 1.24 V for 5-CU.
MS Workstation v6.9.3 software was used for control, automation
and processing. / WW:
SPE recovery: 53 ± 28%
R2: 0.966-0.993
Accurracy: 10%
LOD: ng L-1
SW:
SPE recovery: 93 ± 2%
R2: 0.947-0.997
Accurracy: 7.8%
LOD: ng L-1 / c
IF / 100 mL of hospital wastewater samples and 200 mL of wastewater treatment plant (WWTP) effluent were filtered through glass microfiber filters (minimal pore size 0.5 μm, Machery Nagel, Dueren, Germany) to remove any solids and through cellulose-nitrate filters (0.45 μm; Sartorius Stedim Biotech, Göttingen, Germany) to remove bacteria. The samples were then pre-concentrated using Oasis HLB (60 mg, 3 cc) cartridges (Waters, Massachusetts, USA). Each cartridge was conditioned with 3 mL of ethyl acetate, 3 mL of methanol and equilibrated with 3 mL of water. The samples were then extracted at a flow rate of 3 mL min−1. The sorbents were further dried under vacuum before being eluted with 3 mL of ethyl acetate (3 x 1 mL). The remaining organic solvent was removed under a gentle stream of nitrogen.
100 µL of TFAA and 100 µL of ethyl acetate were added to the dried extract and left to derivatize for 1 h at 60 °C. Afterwards, the samples were dried under nitrogen to remove any trifluoroacetic acid formed during derivatization before being re-dissolved in 250 µL of ethyl acetate prior to analysis by GC-MS.
The same procedure was employed for SW samples, with the exception of sample V for SPE, which was 400 mL. / The samples were analysed using a HP 6890 series (Hewlett-Packard, Waldbron, Germany) gas chromatograph with a single quadrupole mass selective detector. The capillary column was a DB-5 MS 30 m × 0.25 mm × 0.25 µm (Agilent J&W, CA, US). The carrier gas was He set at a flow rate of 1 mL min-1. One microlitre of the sample extract was injected in splitless mode at 270 °C. The temperature programme was as follows: an initial temperature of 65 °C was ramped at 30 °C min-1 to 300 °C, where it was held for 2 min. Total runtime was 9.83 min. The MS was operated in electron impact (EI) mode with an ionization voltage of 70 eV. For qualitative and quantitative determination selective ion monitoring (SIM) was used and following ions were monitored: m/z 309, 307 and 150 for CP, m/z 307, 181 and 150 for IF and m/z 313 and 315 for CP-d6. Instrumental control and data processing were performed using ChemStation software. / WW hospital
SPE recovery: 93±1.0 %
R2: 0.999
Accurracy: 6.8%
LOD: 2.8 ng L-1
WW effluent
SPE recovery: 95±3.8 %
R2: 0.997
Accurracy:2.3%
LOD: 0.36 ng L-1
SW
SPE recovery: 96±0.9 %
R2: 0.995
Accurracy: 3.0%
LOD: 2.5 ng L-1 / c
CP / 100 mL of hospital wastewater samples and 200 mL of wastewater treatment plant (WWTP) effluent were filtered through glass microfiber filters (minimal pore size 0.5 μm, Machery Nagel, Dueren, Germany) to remove any solids and through cellulose-nitrate filters (0.45 μm; Sartorius Stedim Biotech, Göttingen, Germany) to remove bacteria. The samples were then pre-concentrated using Oasis HLB (60 mg, 3 cc) cartridges (Waters, Massachusetts, USA). Each cartridge was conditioned with 3 mL of ethyl acetate, 3 mL of methanol and equilibrated with 3 mL of water. The samples were then extracted at a flow rate of 3 mL min−1. The sorbents were further dried under vacuum before being eluted with 3 mL of ethyl acetate (3 x 1 mL). The remaining organic solvent was removed under a gentle stream of nitrogen.
100 µL of TFAA and 100 µL of ethyl acetate were added to the dried extract and left to derivatize for 1 h at 60 °C. Afterwards, the samples were dried under nitrogen to remove any trifluoroacetic acid formed during derivatization before being re-dissolved in 250 µL of ethyl acetate prior to analysis by GC-MS.
The same procedure was employed for SW samples, with the exception of sample V for SPE, which was 400 mL. / The samples were analysed using a HP 6890 series (Hewlett-Packard, Waldbron, Germany) gas chromatograph with a single quadrupole mass selective detector. The capillary column was a DB-5 MS 30 m × 0.25 mm × 0.25 µm (Agilent J&W, CA, US). The carrier gas was He set at a flow rate of 1 mL min-1. One microlitre of the sample extract was injected in splitless mode at 270 °C. The temperature programme was as follows: an initial temperature of 65 °C was ramped at 30 °C min-1 to 300 °C, where it was held for 2 min. Total runtime was 9.83 min. The MS was operated in electron impact (EI) mode with an ionization voltage of 70 eV. For qualitative and quantitative determination selective ion monitoring (SIM) was used and following ions were monitored: m/z 309, 307 and 150 for CP, m/z 307, 181 and 150 for IF and m/z 313 and 315 for CP-d6. Instrumental control and data processing were performed using ChemStation software. / WW hospital
SPE recovery: 99±0.42 %
R2: 0.996
Accurracy: 3.2%
LOD: 0.78 ng L-1
WW effluent
SPE recovery: 97± 1.6%
R2: 0.998
Accurracy: 1.5%
LOD: 6.55 ng L-1
SW
SPE recovery: 97±3.1 %
R2: 0.995
Accurracy: 3.0%
LOD: 4.5ng L-1 / c
Cis-Pt/Pt / Samples were filtered through 0.45 μm filters (Sartorius Stedim Biotech, Göttingen, Germany)
Than samples were acidified with 0.1 mL of HNO3 per 100 mL of sample for total Pt determination. / Total Pt concentrations were determined by ICP-MS, model 7700x, from Agilent Technologies (Tokyo, Japan).
ICP-MS parameters:
Forward power: 1500 W
Plasma gas flow: 15.0 L min-1 Carrier gas flow: 0.25 L min-1 Dilution gas flow: 0.92 L min-1 Isotope monitored: 195 Pt / *LOD: 0.6 ng Pt L-1 or 1 ng cis-Pt L-1
*LOQ:2.0 ng Pt L-1 or 3.33 ng cis-Pt L-1 / c
*conc. of cis-Pt was estimated from the determination of the total Pt conc. in samples analysed