Kiwao Kadokami, Masashi Yanagita,
and Keita Mochinaga
The University of Kitakyushu, JAPAN
June 10, 2015
UOK, Lab for Envi Pre Anal
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
Background










Introduction of Kitakyushu City
Automated Identification and Quantification System (AIQS)
for GC-MS and LC-TOF-MS
Comprehensive analytical methods using two AIQSs
Objectives
Outline of results
Major chemical groups detected
Emission amounts
Change in concentrations during treatment
processes
Risk evaluation
Conclusions
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Population: 957,597 (April
2015)
Area: 487.89km2 (Oct 2010)
GDP: 29.6 billion US$ (2014)
Nippon Steel & Sumitomo Metal
Kyushu Electric Power Co.,Inc.
Toyota Motor Corporation
Nissan Motor Co., Ltd.
Mitsubishi Chemical Corporation
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TOTO LTD
4
Yahata Steel Works (Japan’s first state-owned steel works) began operation in 1901.
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1960s
Today
Theorder
Nation-wide
survey by Ministry
of the
In
to examine
pollution
Environment, Japan shows that a large
by
chemicals,
have
to
number
of chemicals we
pollute
the environment.
 measure
More than thousands
chemicals
including
as many
substances
unintentionally created chemicals seem to
as
possible.
exist
in the aquatic environment.

Chemicals
Surveyed
Water
Sediment
Fish
Air
Diet
Others
Total
1095
1026
473
455
27
26
1250
391
470
288
322
21
13
705
Highly
efficient
analytical
Detection
36
46
61 necessary.
71
78
50
56
Ratio, % methods
are
Chemicals
Detected
Summary of results from 1974 to 2013.
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UOK, Lab for Envi Pre Anal
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Automated Identification and Quantification
System with a DataBase
Mass spectra
%
%
100.0
100.0
198
198
75.0
75.0
50.0
50.0
25.0
25.0
0.0
0.0
44
44
77
51 77
51
255
255
127
127
110
110
206
206
100
100
275
275
296
296
167
167
200
200
Calibration curves
No use standard substances
Retention times
(IS)
442
442
300
300
Reduce(PTRI)
time, cost and waste
Simultaneous measurement
–ʐ
Ï”ä (x100)
443
443
365
365
0.175
400
400
0.150
0.125
0.100
0.075
0.050
0.025
Results
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UOK, Lab for Envi Pre Anal
0.000
0.0
5.0
”Z“x”ä
12
GC-MS
[GC]
Column
Column oven
temp.
Injection temp.
Injection mode
Carrier gas
Linear velocity
[MS]
Interface temp.
Ion source temp.
Ionization method
Tuning method
Measurement
method
June 10, 2015
： GC-Quadrupole MS
： J&W DB-5 MS (30 m×0.25 mm i.d, 0.25 µm film)
： 40 ℃(2 min) - 8 ℃/min - 310 ℃(5 min)
：
：
：
：
250 ℃
Splitless, 1 min for purge-off time
He
40 cm/s, constant flow mode
： 300 ℃
： 200 ℃
： EI
： Target tuning for US EPA method 625
： SIM/Scan (m/z 45 to 600)
UOK, Lab for Envi Pre Anal
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Class 1
Number
Chemicals consisting of
C and H
195
Chemicals consisting of
C, H and O
150
Class 2
Number
PAHs
79
PCBs
62
Others
54
Phenols
50
Others
100
Aromatic amines
43
Nitro compounds
42
Others
28
Chemicals
containing N
113
Chemicals containing S
12
12
Chemicals containing P
8
8
PPCPs
14
14
Pesticides
Total
June 10, 2015
451
Insecticide
184
Herbicide
118
Fungicide
116
Others
33
943
UOK, Lab for Envi Pre Anal
January 1, 2014
15
Item
Prediction of
retention times (RT)
Measurement
reproducibility
Instrumental
detection limit (IDL)
June 10, 2015
Performance
Difference between
actual and Predicted RT
is less than 3 s.
Reproducibility (RSD) of
78% of the registered
compounds is below 20%.
IDL of 86% of the
registered compounds is
below 10 pg (scan mode).
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J. Chromatogr A, 1089 pp219-226, 2005.
Bunseki, 60, pp543-556. 2011.
16
Sample
1000 mL, pH 7.0, NaCl, Surrogates
Liquid-liquid Extraction
Dichloromethane, twice
Dehydration
Na2SO4
Concentration
RE, N2 stream, 1 mL, IS
GC/MS
SIM/Scan
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Bar shows 95% confidence interval of average.
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J. Environ. Chem, 19, pp351-360, 2009.
Anal. Sci., 29, pp483-486, 2013.
19
PAHs
GC-MS
TIM
10
GC-MS
SIM
1
GC-MSMS
-
Sterols
32-320
8-320
-
Compounds
PCBs
1.6-40 0.4-1.6 0.4-1.6
Pesticides
20
2
0.1-16
Others
5-500
-
-
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UOK, Lab for Envi Pre Anal
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LC
Column
Column temp.
Flow rate
Agilent 1200
GL Science ODS-4 HP (150mm, 2.1mm, 3µm)
40℃
0.3mL/min
A: 5mM CH3COONH4 in H2O
Mobile phase
B: 5mM CH3COONH4 in CH3OH
Time[min]
0
30
50
Gradient profile
B[%]
5
95
95
MS
Agilent 6220 Time of Flight LC/MS
Ionization
ESI-positive
Measurement mode
Scan
Scan range
50-1000
Reference ion
m/z=121.050873,922.009798
Drying gas
10L/min at 325℃
Nebulizer gas
50psi
pressure
Vcap voltage
3500V
Fragmentor voltage
100V (100, 150, 200 and 250V)
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Class
Number
Example
Industrial
chemicals
6
Flame retardant, etc.
PPCPs
95
Pesticides
207
Total
308
Antibiotic, Analgesic,
etc.
Fungicide, Herbicide,
etc.
PPCPs: Pharmaceutical and Personal Care Products
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Sample
200 mL, pH 7.0, Surrogates
Filtration
Whatman GF/C
SPE
Waters PS2 and AC2
Dry
N2 stream, 40 min
Elution
MeOH 5 ml, DCM 5 mL, each
Concentration
N2 stream, 1 mL, IS
LC-TOF-MS
TIM
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Chemosphere, 122, pp125-130, 2015.
25
Conc: 0.2 mg/L, n=7
Bar shows 95% confidence interval of average.
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Method Detection Limit of Water, ng/l
MDL, ng/L
No of comp
2.5
55 (44％)
5.0
32 (25％)
10
23 (18％)
20
7 (6％)
50
6 (5％)
100
3 (2％)
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Objectives
• To clarify the types and quantities of
organic micro-pollutants (OMPs)
discharged from domestic activities
• To clarify emission amounts per capita
• To clarify treatment efficiency of STPs
by the activated sludge method
• To clarify the types and quantities of
OMPs discharged to the environment
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Study Sewage Treatment Plants
Kitaminato STP
Treatment population：91,796
Treatment capacity：44,000m3/d
Hiagari STP
Shinmachi STP
Treatment population：34,7367
Treatment capacity：263,000m3/d
Treatment population：102,151
Treatment capacity: 64,000m3/d
Sampling
date: Oct 29,
2014
A sewerage
coverage
ratio in
Kitakyushu is
99.8%.
Kogasaki STP
Treatment population：298,298
Treatment capacity：177,000m3/d
Sone STP
Activated
sludge
treatment
method
Treatment population：140,108
Treatment capacity： 73,000m3/d
https://www.city.kitakyushu.lg.jp/files/000096297.pdf
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Drain
Grit
camber
Influent
Sea
Sludge treatment
facility
Primary
sedimentation
tank
Activated sludge
Final
tank
sedimentation
tank
Primary
treatment
water
Chlorine
disinfection
Effluent
https://www.city.kitakyushu.lg.jp/suidou/04600128.html
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substances
Number of detected
140
120
Pesticides
100
PPPCPs
CHP
80
CHS
60
CHN
40
CHO
20
0
CH
A-GC A-PST A-FST A-GC A-PST A-FST A-GC A-PST A-FST A-GC A-PST A-FST A-GC A-PST A-FST A-GC:
Sone
Shinmachi
Hiagari
Kitaminato
Kogasaki
900
Concentration, ug/L
800
700
600
After grid chamber
A-PST:
After primary
sedimentation tank
A-FST:
After final
sedimentation tank
Pesticides
500
PPCPs
400
CHP
300
CHS
200
CHN
100
CHO
0
A-GC A-PST A-FST A-GC A-PST A-FST A-GC A-PST A-FST A-GC A-PST A-FST A-GC A-PST A-FST
Sone
June 10, 2015
Shinmachi
Hiagari
Kitaminato
UOK, Lab for Envi Pre Anal
CH
Kogasaki
35
Effluent
(after grid chamber)
(after final
sedimentation tank)
Removal
ratio, %
Chemicals consisting of C and H
(190)
36 (44)
0.39 (28)
98.9
Chemicals consisting of C, H, and
O (159)
532 (37)
4.3 (22)
99.2
Chemicals containing N (118)
1.8 (3)
0.54 (4)
70
Chemicals containing S (12)
0.29 (2)
0.48 (3)
-65.5
Chemicals containing P (9)
0.47 (3)
0.27 (3)
42.6
Pharmaceuticals and personal
care products (108)
95 (30)
11 (24)
88.4
Pesticides (574)
0.11 (5)
0.11 (6)
0
Total (1170)
665 (124)
16.9 (90)
97.5
Influent
Target substance
June 10, 2015
UOK, Lab for Envi Pre Anal
(Number of substances)
36
Influent
No
Compound
Effluent
Conc,
ug/L
Compound
Conc,
ug/L
1
Cholesterol
136.6
Metformin
2.87
2
Coprostanol
64.1
Bis(2-ethylhexyl)phthalate
1.96
3
2-Phenoxy-ethanol
35.9
Cholesterol
1.95
4
beta-Sitosterol
35.4
Crotamiton
1.84
5
3-&4-Methylphenol
33.7
Azithromycin
1.05
6
Epicoprostanol
29.7
Sulpiride
0.98
7
Campesterol
27.2
Etodolac
0.67
8
2-Butoxyethanol
23.2
Coprostanol
0.65
9
L-Menthol
21.0
Diethyl phthalate
0.63
20.1
Clarithromycin
0.59
10 Caffeine
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Effluent
Influent
Compound
Conc. ug/L
Amount
kg/d
Compound
Conc. ug/L
Amount
kg/d
Acetaminophen
12.60
5.86
Analgesics/antiinflammatorics
Metformin
2.87
1.10
Antidiabetics
Metformin
12.16
5.98
Antidiabetics
Crotamiton
1.84
0.79
Anti-itch
medication
Etodolac
6.01
2.81
Analgesics/antiinflammatorics
Azithromycin
1.05
0.45
Antibiotics
Azithromycin
3.36
1.46
Antibiotics
Sulpiride
0.98
0.42
Antidepressant
Theophylline
2.85
1.37
Bronchodilator
Etodolac
0.67
0.27
Analgesics/antiinflammatorics
Bezafibrate
2.66
1.36
Lipid regulators
Clarithromycin
0.59
0.26
Antibiotics
Crotamiton
2.24
0.94
Lidocaine
0.49
0.21
Local anesthetic
Ibuprofen
1.79
0.55
Sulfamethoxazole
0.31
0.17
Antibiotics
Ketoprofen
1.61
0.67
Atenolol
0.37
0.15
Beta-blockers
Clarithromycin
0.86
0.39
Bezafibrate
0.30
0.14
Lipid regulators
Class
Anti-itch
medication
Analgesics/antiinflammatorics
Analgesics/antiinflammatorics
Antibiotics
Class
Concentration is the average of the 5 STPs.
Amount is the sum of the 5 STPs.
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Name
Sone
Shinmachi
Hiagari
Kitaminato
Kogasaki
Whole city
June 10, 2015
Point
All substances,
kg/day
PPCPs,
kg/day
A-GC
A-PST
A-FST
A-GC
A-PST
A-FST
A-GC
A-PST
A-FST
A-GC
A-PST
A-FST
A-GC
A-PST
A-FST
37.7
31.6
0.76
34.3
27.9
0.90
106.2
88.6
3.25
25.8
25.2
0.79
75.5
61.9
1.52
5.32
4.38
0.52
4.75
4.53
0.48
18.2
16.6
1.83
3.29
3.61
0.57
10.5
10.4
1.10
A-GC
A-PST
A-FST
279.5
235.2
7.22
42.1
39.5
4.50
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Water volume,
ton/day
47,964
45,413
181,388
39,706
137,217
451,688
A-GC:
After grid
chamber
A-PST:
After primary
sedimentation
tank
A-FST:
After final
sedimentation
tank
40
Amounts, mg/capita/day
All
substances
PPCPs
Influent
292
44
Effluent
7.5
4.7
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Effluent conc > Influent conc
Substance
Removal rate, %
45
STP
40
1,2,3-Trimethoxybenzene
5/5
35
2,6-Di-tert-butyl-4benzoquinone
5/5
30
2-Methylaniline
5/5
Sulfamethoxazole
5/5
Sulpiride
5/5
Candesartan
5/5
Carbamazepin
5/5
1,3-Dicyclohexylurea
4/5
5
Roxithromycin
4/5
0
Carbendazim
4/5
Diuron
4/5
Anthraquinone
3/5
n-C25H52
2/5
n-C26H54
2/5
2(3H)-Benzothiazolone
2/5
2-(Methylthio)-benzothiazol
2/5
Tris(2-chloroethyl) phosphate
2/5
Dicyclohexylamine
2/5
Sulfapyridine
2/5
42
25
20
15
10
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UOK, Lab for Envi Pre Anal
Compound Sulfamethoxazole Clarithromycin
Azithromycin
Max
19.9
11.8
9.3
Min
6.1
6.4
4.8
Average
11.6
8.4
7.0
PNEC*, mg/L
0.027
0.07
0.15
* Sci Total Environ, 429, pp123-155, 2012.
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Conclusions
• 133 out of 1170 analytes with 665 mg/L were detected
in influents. Detected substances were comprised of
sterols, PPCPs, phenols, phthalates and PAHs.
• Emission quantities of all analytes and PPCPs are 292
and 44 mg/capita/day, respectively.
• Activated sludge method can remove over 97% of the
detected substances except for PPCPs and
organophosphorus flame retardants.
• 7.22 kg of analytes and 4.50 kg of PPCPs were
discharged from the STPs to the aquatic environment.
Several PPCPs have a potential to give adverse
effects on aquatic organisms.
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Thanks for your
attention!!