Appendix 1. Conditions for GC/MS analyses of phthalate diesters

Chromatography / Apparatus / Injector / Column / Ovenparameters / Vectorgas
GC/MS-EI / Agilent GC (7890)-MS (5975) / splitless 290°C; 1 µL / ZB-5MS (30 m, 250 µm ID, 0.25µm film thickness from Phenomenex) with a deactivated silica pre-column
(1 m; 0.25 mm ID) / 50°C for 1 min, then 30°C/min to 280°C and 15°C/min to 310°C for 4 min / Helium 1mL/min

Appendix 2. Parameters for GC/MS analyses of phthalate diesters

Compound / m/z quantification ion / m/z qualification / Retention time / Apparatus / IDLs
pginjected / LOQs Fish tissue ng g-1dw / LOQs River water ng L-1
DMP / 163 / 194 / 6.6 / Source: 230°C
Interface: 200°C
Quadrupole: 150°C / 2 / 0.6 / 15
DEP / 149 / 177 / 7.3 / 4 / 0.4 / 29
DiBP / 149 / 104 / 8.2 / 4 / 1 / 46
DnBP / 149 / 223 / 8.6 / 4 / 2 / 54
BBP / 149 / 104 / 9.9 / 2 / 9 / 28
DEHP / 149 / 279 / 10.4 / 5 / 10 / 50
DnOP / 149 / 279 / 11.2 / 17 / 8 / 53
DPP (IS) / 149 / 104 / 9.2
Benzyl benzoate (SrS) / 105 / 212 / 7.9

Appendix 3.LC gradient (flow: 0.3 ml/min)

Time (min) / 0 / 1 / 14 / 21 / 22 / 23 / 30
% acetonitrile + 0.1% formic acid / 5 / 15 / 25 / 35 / 100 / 5 / 5
% H2O + 0.1% formic acid / 95 / 85 / 75 / 35 / 0 / 95 / 95

Appendix4.Parameters for LC-MS/MS detection and limits of quantification (LOQ) of phthalate monoesters inroach tissues

Compound / Internal
standard / Precursor
ion / Product
ion / Fragmentor
energy (V) / Collision energy (V) / Retention time (min) / IDLs / LOQs
pginjected / Plasma pg/mL / Bile pg/mL / Liverng/g dw
MMP / MnBP-13C12 / 179 / 77 / 80 / 16 / 7.9 / 2 / 9 / 12 / 2
MEP / MnBP-13C12 / 193 / 77 / 110 / 20 / 11.4 / 2 / 8 / 24 / 3
MiBP / MnBP-13C12 / 221 / 77 / 110 / 16 / 19.7 / 0.6 / 16 / 22 / 6
MEHHP / MnBP-13C12 / 293 / 145 / 110 / 16 / 19.8 / 2 / 19 / 6 / 14
MEHHP / MnBP-13C12 / 121 / 110 / 16
MnBP / MnBP-13C12 / 221 / 77 / 80 / 8 / 19.9 / 0.5 / 16 / 22 / 6
MEOHP / MEHP-13C12 / 291 / 143 / 110 / 8 / 19.9 / 0.3 / 15 / 6 / 4
MEOHP / MEHP-13C12 / 121 / 110 / 12
MBzP / MEHP-13C12 / 255 / 77 / 80 / 16 / 20.4 / 0.1 / 4 / 5 / 2
MEHP / MEHP-13C12 / 277 / 134 / 50 / 25 / 23.9 / 6 / 16 / 7 / 13
MnOP / MiNP-13C12 / 277 / 125 / 110 / 12 / 24.1 / 2 / 28 / 5 / 24
MnBP-13C12 / 225 / 79 / 80 / 12 / 19.9
MEHP-13C12 / 281 / 137 / 80 / 8 / 23.8
MiNP-13C12 / 295 / 250 / 80 / 8 / 24
4-MU / 4-MU-13C12 / 175 / 147 / 110 / 20 / 9.1 / 2 / 12 / 11 / 5
133 / 110 / 20
119 / 110 / 24
4-MU-13C12 / 179 / 152 / 80 / 24 / 9.1
136 / 80 / 20
125 / 80 / 20

Appendix 5.Instrumental performances for phthalate monoesters and diesters

Phthalate monoesters
Compound / Calibration range (ng mL-1) / Range (ng injected) / R²
MMP / 31.25 - 1000 / 0.3 - 10 / 0.997
MEP / 31.25 - 1000 / 0.3 - 10 / 0.998
MiBP / 31.25 - 1000 / 0.3 - 10 / 0.997
MnBP / 31.25 - 1000 / 0.3 - 10 / 0.998
MBzP / 31.25 - 1000 / 0.3 - 10 / 0.999
MEHP / 31.25 - 1000 / 0.3 - 10 / 0.998
MEHHP / 31.25 - 1000 / 0.3 - 10 / 0.998
MEOHP / 31.25 - 1000 / 0.3 - 10 / 0.998
4MU / 0.2-200 / 0.002 - 2 / 0.999
Phthalate diesters
Compound / Calibration range (ng mL-1) / Range (ng injected) / R²
DMP / 31.25 - 8000 / 0.03 - 8 / 0.993
DEP / 31.25 - 8000 / 0.03 - 8 / 0.991
DiBP / 31.25 - 8000 / 0.03 - 8 / 0.989
DnBP / 31.25 - 8000 / 0.03 - 8 / 0.991
BBP / 31.25 - 8000 / 0.03 - 8 / 0.998
DEHP / 31.25-8000 / 0.03 - 8 / 0.991
DnOP / 31.25 - 8000 / 0.03 - 8 / 0.997

Appendix 6. Laboratory blank values and recoveries

Phthalate diesters / DMP / DEP / DiBP / DnBP / BBP / DEHP / DnOP
Blank values (ng) / 11 / 24 / 76 / 11 / 52 / 36 / 4
Recovery (%) / 69 / 64 / 72 / 76 / 57 / 87 / 74
Phthalate monoesters / MMP / MEP / MiBP / MnBP / MBzP / MEHP / MnOP / MEHHP / MEOHP
Blank values (ng) / 0.14 / 0.46 / 1.5 / 0.76 / 0.04 / 0.37 / 0.59 / 0.06 / 0.07
Recovery (%) / 58 / 94 / 66 / 64 / 90 / 65 / 82 / 56 / 65

1