Xin Zhang1, Hongxia Zhao1*, Juan Du1, Yixuan Qu2, Chen Shen1, Feng Tan1, Jingwen Chen1

Supplementary material

Occurrence, removal and risk assessment of antibiotics in twelve wastewater treatment plants from Dalian, China

Xin Zhang1, Hongxia Zhao1*, Juan Du1, Yixuan Qu2, Chen Shen1, Feng Tan1, Jingwen Chen1, Xie Quan1

1 Key Laboratory of Industrial Ecology and Environmental Engineering (Ministry of Education), School of Environmental Science and Technology, Dalian University of Technology, Linggong Road 2, Dalian 116024, China

2 Dalian Hai Xin Detection Technology Co., Ltd, Dalian 116020, China

*Corresponding author

Phone: +86-411-84707965; Fax: +86-411-84707965;

Email:

This Supplementary Material Contains the Following Sections:

Figure S1: Correlation analysis between (a) the concentrations in WWTP waste water and the obtained parameter, inhabitant served; (b) the removal efficiency in in WWTP waste water and the hydraulic retention time.

Table S1: Parameters of WWTPs.

Table S2: MS/MS parameters for the 31 target antibiotics.

Table S2: Recovery and LOQ of 31 antibiotics in this study.

Table S4: Detection frequency in all sample.

Table S5: Concentrations of antibiotics in the influents and effluents of WWTPs from different countries/regions

Table S6: Toxicity data and the calculated PNECs values.

Table S7: The calculated RQs of antibiotics for three aquatic organisms (algae, invertebrate and fish).

Figure S1 Correlation analysis between (a) the concentrations in wastewater and the obtained parameter, inhabitant served; (b) the removal efficiency in wastewater and the hydraulic retention time

Table S1 Parameters of WWTPs

WWTPs / Inhabitants served (×104) / Type of water treated / Second treatment process / Disinfection method / Hydraulic retention time (h) / Daily flow (×104,m3d-1) / Effluent discharge
S1 / 20 / D / CAST / UV / 6 / 6 / sea
S2 / - / D,I / AAO / UV / - / - / sea
S3 / 24 / D, I / BFOR / UV / 5 / 12 / sea, reused
S4 / 20 / D, I / AS / NaClO / 7 / 8 / sea, reused
S5 / 22 / D / BFOR / UV, NaClO / 3.5 / 8 / sea, reused
S6 / 1 / D, I / A/O / ClO2 / 4 / 1 / sea
S7 / 35 / D, I / BFOR / UV / 4 / 12 / sea
S8 / 27 / D / CAST / UV / 4 / 8 / sea
S9 / - / D / CAST / ClO2 / - / 3.5 / sea
S10 / 5 / D, I / AS / UV / 6 / 3 / sea
S11 / 25 / D,I / BFOR / UV / 4 / 10 / sea
S12 / - / D / AS / - / - / 1 / sea

D: domestic; I: industrial; CAST: cyclic activated sludge technology; AAO: anaerobic-aerobic-oxic; BFOR: biological filtration oxygenated reactor; AS: activated sludge; A/O: anoxic/oxic

Table S2 MS/MS parameters for the 31 target antibiotics

No / Analyte / RT
(min) / Precursor ion(m/z) / Product ion(m/z) / Cone voltage (V) / Collision energy
(eV) / Internal standard / No / Analyte / RT
(min) / Precursor ion(m/z) / Product ion(m/z) / Cone voltage (V) / Collision energy
(eV) / Internal standard
1 / Sulfaguanidine / SGD / 0.8 / 215 / 156 / 30 / 15 / Sulfamethoxazole-d4 (SMX-d4) / 2 / Sulfadimethoxine / SDM / 7.0 / 311 / 155.9* / 25 / 20 / Sulfamethoxazole-d4 (SMX-d4)
108 / 25 / 108 / 35
92.0* / 25 / 92.1 / 35
3 / Sulfapyridine / SP / 1.6 / 250.3 / 155.9* / 25 / 15 / 4 / Sulfadiazine / SD / 5.0 / 251.3 / 156.1* / 30 / 15
108 / 25 / 92.1 / 25
92 / 25 / 108 / 20
5 / Sulfamethoxazole / SMX / 2 / 254 / 155.8* / 25 / 15 / 6 / Sulfathiazole / SFT / 6.3 / 256 / 155.9* / 25 / 20
108 / 25 / 108 / 30
92 / 25 / 92 / 30
7 / Sulfamethyldiazine / SMD / 2.5 / 265 / 156 / 30 / 15 / 8 / Sulfamethazine / SMT / 3.4 / 279 / 156.0* / 25 / 20
108.0* / 30 / 92 / 30
92 / 30 / 108 / 20
9 / Sulfachloropyridazine / SCP / 4.1 / 285 / 156.0* / 30 / 15 / 10 / Sulfamonomethoxine / SMM / 4.3 / 281.3 / 156 / 25 / 15
92.1 / 25 / 108.1 / 25
108 / 25 / 92.1* / 25
11 / Trimethoprim / TMP / 3.2 / 291 / 230.1* / 30 / 25 / Trimethoprim-d3 (TMP-d3) / 12 / Doxycycline / DOX / 4 / 445.2 / 428.1 / 30 / 20 / Atrazine-d5 (ATR-d5)
261 / 25 / 410* / 20
123.1* / 30 / 154 / 30
13 / Tetracycline / TCY / 4.0 / 445.2 / 428.1 / 30 / 20 / Atrazine-d5 (ATR-d5) / 14 / Oxytetracycline / OXY / 3.8 / 461.1 / 201 / 30 / 35
410* / 20 / 426* / 20
154 / 30 / 444 / 30

Table S2 continued

No / Analyte / RT
(min) / Precursor ion(m/z) / Product ion(m/z) / Cone voltage (V) / Collision energy
(eV) / Internal standard / No / Analyte / RT
(min) / Precursor ion(m/z) / Product ion(m/z) / Cone voltage (V) / Collision energy
(eV) / Internal standard
15 / Chlortetracycline / CTE / 5.9 / 479 / 154 / 30 / 30 / Atrazine-d5 (ATR-d5) / 16 / Erythromycin-H2O / ERY / 11.9 / 716.5 / 158.2* / 25 / 25 / Atrazine-d5 (ATR-d5)
462.0* / 15 / 116 / 35
444.1 / 20 / 558 / 20
17 / Clarithromycin / CLR / 12.7 / 748.5 / 590.3 / 25 / 15 / Atrazine-d5 (ATR-d5) / 18 / Azithromycin / AZM / 12.7 / 749.9 / 591.4 / 25 / 25
158.0* / 25 / 115.9 / 35
116 / 35 / 158.0* / 40
19 / Roxithromycin / ROX / 5.09 / 837 / 679.2 / 30 / 25 / 20 / Norfloxacin / NFX / 4.0 / 320 / 302.2* / 25 / 20 / Ciprofloxacin-d8 (CIP-d8)
158.0* / 35 / 276 / 20
116.2 / 35 / 231.2* / 35
21 / Ciprofloxacin / CIP / 4.2 / 332.1 / 314.1* / 30 / 20 / Ciprofloxacin-d8 (CIP-d8) / 22 / Enrofloxacin / ENR / 4.75 / 360.4 / 342.1* / 25 / 20
288 / 15 / 316.1 / 15
23 / Ofloxacin / OFX / 3.9 / 362 / 318.2* / 30 / 15 / 245.1 / 25
344.1 / 20 / 24 / Enoxacin / ENO / 3.6 / 321.3 / 231.1 / 25 / 35
261.1 / 25 / 303.2* / 20
25 / Chloramphenicol / CAP / 6.0 / 320.9 / 257 / 30 / 10 / Chloramphenicol-d5 (CAP-d5) / 232.1 / 30
151.9* / 15 / 26 / Thiamphenicol / TAP / 2.9 / 354 / 289.8 / 30 / 30 / Chloramphenicol-d5 (CAP-d5)
121 / 25 / 184.9* / 20
27 / Florfenicol / FFC / 4.7 / 356 / 337* / 30 / 15 / 119.1 / 30
184.9 / 20 / 28 / Lincomycin / LIN / 2.7 / 407.5 / 126.2* / 30 / 25 / Atrazine-d5 (ATR-d5)
119.1 / 35 / 359 / 15

Table S2 continued

Table S3 Recovery and LOQ of 31 antibiotics in this study

Compound / LOQ
(ng/L) / Spiked water / Compound / LOQ
(ng/L) / Spiked water
Spiked
(ng/L) / Recovery
(%) / RSD
(%) / Spiked
(ng/L) / Recovery
(%) / RSD
(%) / Spiked
(ng/L) / Recovery
(%) / RSD
(%) / Spiked
(ng/L) / Recovery
(%) / RSD
(%) / Spiked
(ng/L) / Recovery
(%) / RSD
(%) / Spiked
(ng/L) / Recovery
(%) / RSD
(%)
SGD / 0.5 / 10 / 90.2 / 10.3 / 40 / 86 / 8.2 / 200 / 74.9 / 5.6 / SDM / 0.3 / 2.5 / 73 / 7.1 / 10 / 71 / 5 / 50 / 87.3 / 1.8
SP / 0.6 / 10 / 98 / 12.2 / 40 / 76 / 6 / 200 / 64.3 / 4.5 / SD / 1.8 / 10 / 101.8 / 12.4 / 40 / 68.8 / 6.5 / 200 / 76.5 / 2.1
SMX / 0.5 / 10 / 93.2 / 4.8 / 40 / 80.3 / 1.4 / 200 / 91.3 / 2.2 / SFT / 0.7 / 25 / 80.7 / 9.2 / 100 / 81.4 / 7.4 / 500 / 90.5 / 1.2
SMD / 0.2 / 10 / 84.3 / 8 / 40 / 93.5 / 4.6 / 200 / 102.9 / 3.5 / SMT / 0.3 / 10 / 81.1 / 6.1 / 40 / 80.2 / 2.2 / 200 / 88.9 / 3
SCP / 0.2 / 5 / 99 / 10.7 / 20 / 103.8 / 3.2 / 100 / 123.7 / 0.4 / SMM / 0.2 / 10 / 59.2 / 6.8 / 40 / 65.2 / 8 / 200 / 77.3 / 2.8
TMP / 0.2 / 1.3 / 94.3 / 5.1 / 5 / 73.7 / 2 / 25 / 82.8 / 5.6 / DOX / 0.3 / 2.5 / 96.2 / 5.6 / 10 / 102.4 / 4.8 / 50 / 109.6 / 6.4
TCY / 0.5 / 2.5 / 104.2 / 6.6 / 10 / 108.3 / 8.8 / 50 / 101.3 / 4.3 / OXY / 0.4 / 2.5 / 101.3 / 4.2 / 10 / 90.6 / 5.6 / 50 / 96.4 / 8.2
CTE / 0.3 / 2.5 / 99.6 / 5.6 / 10 / 105.2 / 7.6 / 50 / 110.6 / 8.1 / ERY / 0.3 / 1.3 / 116.8 / 2.7 / 5 / 113.8 / 12.3 / 25 / 109.1 / 8.7
CLR / 0.3 / 1.3 / 128.5 / 3.7 / 5 / 103.5 / 7.3 / 25 / 109.1 / 12.9 / AZM / 0.3 / 2.5 / 104.2 / 0.9 / 10 / 133.4 / 11.8 / 50 / 100.5 / 12.7
ROX / 0.3 / 0.5 / 117.7 / 3.5 / 2 / 104.6 / 17.8 / 10 / 117.7 / 0.5 / NFX / 1.7 / 5 / 128.9 / 17.2 / 20 / 103.5 / 4.6 / 100 / 109.9 / 11.2
CIP / 0.3 / 2.5 / 119.8 / 11.7 / 10 / 98.4 / 4.9 / 50 / 105 / 14 / ENR / 0.5 / 2.5 / 131.2 / 9 / 10 / 65 / 3 / 50 / 92.7 / 15.7
OFX / 1.2 / 1.3 / 103.5 / 10 / 5 / 117 / 9.8 / 25 / 100.8 / 12.1 / ENO / 0.2 / 2.5 / 115.5 / 10.6 / 10 / 108.1 / 2 / 50 / 116.8 / 17.2
CAP / 1.7 / 5 / 121.3 / 19.1 / 20 / 103.7 / 8.1 / 100 / 115.2 / 3.2 / TAP / 3.8 / 5 / 73.8 / 3.7 / 20 / 111.8 / 17.5 / 100 / 108.7 / 9.2
FFC / 2.2 / 5 / 81.1 / 3.8 / 20 / 107.9 / 5.8 / 100 / 88.6 / 14.1 / LIN / 0.5 / 2.5 / 98.6 / 6.6 / 10 / 90.2 / 9.2 / 50 / 83.5 / 8.1
VCM / 9.6 / 10 / 103.4 / 5.9 / 40 / 71 / 7.9 / 200 / 93.4 / 16.9 / OLA / 0.6 / 2.5 / 80.3 / 8.2 / 10 / 88.6 / 4.9 / 50 / 96.2 / 6.7
RIF / 1.1 / 5 / 115.4 / 12.9 / 20 / 83 / 8.1 / 100 / 76.8 / 3.2

Table S4 Detection frequency in all samples

Sample / Detection frequency / Sample / Detection frequency / Sample / Detection frequency
effluent / influent / effluent / influent / effluent / influent
SGD / 25.0 / 50.0 / SDM / 8.3 / 66.7 / SP* / 83.3 / 91.7
SD* / 91.7 / 91.7 / SMX* / 100.0 / 100.0 / SFT / 25.0 / 33.3
SMD / 25.0 / 75.0 / SMT* / 58.3 / 83.3 / SCP / 16.7 / 58.3
SMM / 66.7 / 66.7 / TMP* / 91.7 / 100.0 / DOX / 0.0 / 66.7
TCY* / 75.0 / 91.7 / OXY* / 91.7 / 91.7 / ERY* / 100.0 / 100.0
CLR* / 100.0 / 100.0 / AZM* / 100.0 / 100.0 / NFX* / 100.0 / 100.0
CIP* / 91.7 / 100.0 / ENR* / 25.0 / 83.3 / OFX* / 100.0 / 100.0
ENO* / 91.7 / 100.0 / CAP / 58.3 / 91.7 / TAP / 33.3 / 50.0
FFC / 58.3 / 75.0 / LIN* / 91.7 / 100.0 / VCM / 0.0 / 8.3
OLA / 0.0 / 33.3 / RIF / 66.7 / 41.7

*: Represent the compound with high detection rate (>80%).

Table S5 Concentration of individual antibiotics in the influents and effluents of WWTPs from different countries/regions

Reference / Sample site / Analyte / Range(ng/L) / Reference / Sample site / Analyte / Range(ng/L)
Miao et al (2004) / Canada / Effluents / SAs / SP / --/81/228 / Xu et al (2007) / China / Influents / MLs / ROX / 75/129/164
SD / --/19/19 / FQs / NFX / 54/204/263
SMX / --/243/871 / OFX / 80/248/368
SMT / --/363/363 / CAPs / CAP / 0/0/31
TCs / DOX / --/38/46 / Effluents / SAs / SD / 0/0/36
TCY / --/151/977 / SMX / 9/14/78
MLs / ERY / --/80/838 / SMT / 0/6/346
CLA / --/87/536 / MLs / ERY / 216/344.5/2054
ROX / --/8/18 / ROX / 35/36.5/278
FQs / NFX / --/50/112 / FQs / NFX / 27/53/85
CIP / --/118/400 / OFX / 41/92.5/165
OFX / --/94/506 / CAPs / CAP / 0/0/17
Batt et al (2006) / USA / Effluents / SAs / SMX / 370/900/6000 / Watkinson et al (2007) / Australia / Effluent / SAs / SMX / --/270/320
TMP / 0/155/530 / SFT / --/0/5
TCs / TCY / 0/0/560 / TMP / --/50/70
FQs / CIP / 91/360/5600 / TCs / DOX / --/0/40
Xu et al (2007) / China / Influents / SAs / SD / 0/0/72 / TCY / --/0/30
SMX / 10/20.5/118 / OXY / --/0/20
SMT / 0/12.5/696 / CTE / --/0/5
MLs / ERY / 253/610/1978 / MLs / ERY / --/0/0

Table S5 continued

Reference / Sample site / Analyte / Range(ng/L) / Reference / Sample site / Analyte / Range(ng/L)
Watkinson et al (2007) / Australia / Effluent / MLs / ROX / --/0/100 / Gulkowska et al (2008) / China / Influents / SAs / TMP / 120/200/320
FQs / NFX / --/25/40 / TCs / TCY / 96/550/1300
CIP / --/640/720 / MLs / ERY / 470/590/810
ENR / --/10/10 / FQs / NFX / 110/280/460
Others / LIN / --/50/60 / Effluents / SAs / TMP / 120/155/230
Influents / SAs / SMX / --/360/500 / TCs / TCY / 180/440/620
SFT / --/2/40 / MLs / ERY / 510/560/850
TMP / --/340/930 / FQs / NFX / 85/105/320
TCs / DOX / --/0/65 / Karthikeyan and Meyer (2006) / USA / Influents / SAs / SMX / 0/150/1250
TCY / --/0/35 / SMT / 0/0/210
OXY / --/0/0 / TMP / 50/325/1300
CTE / --/0/0 / TCs / TCY / 0/495/1200
MLs / ERY / --/0/0 / MLs / ERY / 0/125/1200
ROX / --/0/18 / FQs / CIP / 0/0/210
FQs / NFX / --/170/210 / Effluents / SAs / SMX / 0/85/370
CIP / --/3800/4600 / SMT / 0/0/0
ENR / --/10/100 / TMP / 0/35/550
Others / LIN / --/60/80 / TCs / TCY / 0/165/850
MLs / ERY / 0/0/300
FQs / CIP / 0/0/140

Table S5 continued

Reference / Sample site / Analyte / Range(ng/L) / Reference / Sample site / Analyte / Range(ng/L)
Kasprzyk-Hordern et al (2009) / UK / Influents / SAs / SP / 26/--/12397 / Watkinson et al (2009) / Australia / Influents / SAs / SFT / --/0/300
SMX / 0/--/274 / TMP / --/430/4300
TMP / 464/--/6796 / Other / LIN / --/20/500
MLs / ERY / 114/--/10025 / Effluents / FQs / NFX / --/25/250
CAPs / CAP / 0/--/452 / ENR / --/2/50
Effluents / SAs / SP / 94/--/1112 / CIP / --/0/0
SMX / 0/--/44 / MLs / ERY / --/0/0
TMP / 385/--/3052 / ROX / --/20/500
MLs / ERY / 23/--/2841 / TCs / DOX / --/10/150
CAPs / CAP / 0/--/69 / OXY / --/0/70
Watkinson et al (2009) / Australia / Influents / FQs / NFX / --/60/220 / TCY / --/0/20
ENR / --/0/40 / CTE / --/5/250
CIP / --/600/1100 / SAs / SMX / --/50/200
MLs / ERY / --/0/0 / SFT / --/0/600
ROX / --/8/500 / TMP / --/10/250
TCs / DOX / --/20/650 / Other / LIN / --/3/300
OXY / --/0/350 / Minh et al (2009) / China / Effluents / SAs / SMX / 0/--/278
TCY / --/0/100 / SFT / 0/--/0
CTE / --/6/200 / SMT / 0/--/0
SAs / SMX / --/250/3000 / TMP / 59/--/465
TCs / TCY / 0/--/1420

Table S5 continued