Bearer CF. 1998. Biomarkers in pediatric environmental health: A crosscutting issue.
Environ Health Perspect.106 Suppl 3:813-816.
Bouchard MF, Chevrier J, Harley KG, Kogut K, Vedar M, Calderon N, Trujillo C,
Johnson C, Bradman A, Barr DB, Eskenazi B. 2011. Prenatal exposure to
organophosphate pesticides and IQ in 7-year-old children. Environ Health Perspect.
119:1189-1195.
Bouchard
MF,
Bellinger
DC,
Attention-deficit/hyperactivity
Wright
disorder
RO,
and
Weisskopf
urinary
MG.
metabolites
2010.
of
organophosphate pesticides. Pediatrics. 125:1270-1277.
Chen M, Tao L, McLean J, Lu C. 2014. Quantitative analysis of neonicotinoid
insecticide residues in foods: implication for dietary exposures. J Agric Food Chem.
62:6082-6090.
Cimino AM, Boyles AL, Thayer KA, Perry MJ. 2017. Effects of Neonicotinoid
Pesticide Exposure on Human Health: A Systematic Review. Environ Health
Perspect. 125:155-162.
Claudio L, Kwa WC, Russell AL, Wallinga D. 2000. Testing methods for
developmental neurotoxicity of environmental chemicals. Toxicol Appl Pharmacol.
164:1-14.
Costa LG, Giordano G, Guizzetti M, Vitalone A. 2008. Neurotoxicity of pesticides: a
brief review. Front Biosci. 13:1240-1249.
Davies D. 2004. Bag urine specimens still not appropriate in diagnosing urinary tract
infections in infants. Can J Infect Dis Med Microbiol. 15:210-211.
Ford KA, Casida JE. 2006a. Chloropyridinyl neonicotinoid insecticides: diverse
- 20 -
molecular substituents contribute to facile metabolism in mice. Chem Res Toxicol.
19: 944-951.
Ford KA, Casida JE. 2006b. Unique and common metabolites of thiamethoxam,
clothianidin, and dinotefuran in mice. Chem Res Toxicol. 19: 1549-1556.
González-Alzaga B, Lacasaña M, Aguilar-Garduño C, Rodríguez-Barranco M,
Ballester F, Rebagliato M, Hernández AF. 2014. A systematic review of
neurodevelopmental effects of prenatal and postnatal organophosphate pesticide
exposure. Toxicol Lett. 230:104-121.
Harada KH, Tanaka K, Sakamoto H, Imanaka M, Niisoe T, Hitomi T, Kobayashi H,
Okuda H, Inoue S, Kusakawa K, Oshima M, Watanabe K, Yasojima M, Takasuga T,
Koizumi A. 2016. Biological Monitoring of Human Exposure to Neonicotinoids
Using Urine Samples, and Neonicotinoid Excretion Kinetics. PLoS One.
11:e0146335.
Koureas M, Tsakalof A, Tsatsakis A, Hadjichristodoulou C. 2012. Systematic review of
biomonitoring studies to determine the association between exposure to
organophosphorus and pyrethroid insecticides and human health outcomes. Toxicol
Lett. 210:155–168.
Li P, Ann J, Akk G. 2011. Activation and modulation of human α4β2 nicotinic
acetylcholine receptors by the neonicotinoids clothianidin and imidacloprid. J
Neurosci Res. 89:1295-1301.
Matsuda K, Buckingham SD, Kleier D, Rauh JJ, Grauso M, Sattelle DB. 2001.
Neonicotinoids: insecticides acting on insect nicotinic acetylcholine receptors.
Trends Pharmacol Sci. 22:573-580.
National Institute for Environmental Studies, WebKis-Plus database [accessed 2019
- 21 -
Jun 26] http://w-chemdb.nies.go.jp
Osaka A, Ueyama J, Kondo T, Nomura H, Sugiura Y, Saito I, Nakane K, Takaishi A,
Ogi H, Wakusawa S, Ito Y, Kamijima M. 2016. Exposure characterization of three
major insecticide lines in urine of young children in Japan-neonicotinoids,
organophosphates, and pyrethroids. Environ Res. 147:89-96.
Oya N, Ito Y, Hioki K, Asai Y, Aoi A, Sugiura Y, Ueyama J, Oguri T, Kato S, Ebara T,
Kamijima M. 2017. Quantitative analysis of organophosphate insecticide
metabolites in urine extracted from disposable diapers of toddlers in Japan. Int J
Hyg Environ Health. 220:209-216.
Richardson JR, Fitsanakis V, Westerink RHS, Kanthasamy AG. 2019. Neurotoxicity of
pesticides. Acta Neuropathol. 138:343-362.
Saito S, Ueyama J, Kondo T, Saito I, Shibata E, Gotoh M, Nomura H, Wakusawa S,
Nakai K, Kamijima M. 2014. A non-invasive biomonitoring method for assessing
levels of urinary pyrethroid metabolites in diapered children by gas
chromatography-mass spectrometry. J Expo Sci Environ Epidemiol. 24:200-207.
Stewart SD, Lorenz GM, Catchot AL, Gore J, Cook D, Skinner J, Mueller TC, Johnson
DR, Zawislak J, Barber J. 2014. Potential exposure of pollinators to neonicotinoid
insecticides from the use of insecticide seed treatments in the Mid-Southern United
States. Environ Sci Technol. 48:9762-9769.
Stone WW, Gilliom RJ, Ryberg KR. 2014. Pesticides in U.S. streams and rivers:
Occurrence and trends during 1992−2011. Environ Sci Technol. 48:11025-11030.
Tilson HA. 1998. Developmental neurotoxicology of endocrine disrupters and
pesticides: Identification of information gaps and research needs. Environ Health
Perspect.106 Suppl 3:807-811.
- 22 -
Tomizawa M, Casida JE. 2000. Imidacloprid, thiacloprid, and their imine derivatives
up-regulate the alpha 4 beta 2 nicotinic acetylcholine receptor in M10 cells. Toxicol
Appl Pharmacol. 169:114-120.
Tomizawa M, Casida JE. 2003. Selective toxicity of neonicotinoids attributable to
specificity of insect and mammalian nicotinic receptors. Ann Rev Entomol.
48:339-364.
Ueyama J, Nomura H, Kondo T, Saito I, Ito Y, Osaka A, Kamijima M. 2014. Biological
monitoring method for urinary neonicotinoid insecticides using LC-MS/MS and its
application to Japanese adults. J Occup Health. 56:461-468.
U.S. Food and Drug Administration. 2018. Guidance for Industry, Bioanalytical
Method Validation. [accessed, 2019 Jul 21] http://w-chemdb.nies.go.jp
Wagner-Schuman M, Richardson JR, Auinger P, Braun JM, Lanphear BP, Epstein JN,
Yolton K, Froehlich TE. 2015. Association of pyrethroid pesticide exposure with
attention-deficit/hyperactivity disorder in a nationally representative sample of U.S.
children. Environ Health. 14:44.
Watanabe M, Ueyama J, Ueno E, Ueda Y, Oda M, Umemura Y, Tanahashi T, Ikai Y,
Saito I. 2018. Effects of processing and cooking on the reduction of dinotefuran
concentration in Japanese rice samples. Food Addit Contam Part A Chem Anal
Control Expo Risk Assess. 23:1-8.
Weiss B. 2000. Vulnerability of children and the developing brain to neurotoxic
hazards. Environ Health Perspect. 108 Suppl 3:375-381.
Yusa V, Millet M, Coscolla C, Roca M. 2015. Analytical methods for human
biomonitoring of pesticides. A review. Anal Chim Acta. 891:15-31.
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Figure legends
Figure 1. Chemical structures of the six NEOs and the acetamiprid metabolite
measured in this study.
Figure 2. Method for extraction of urine from a disposable diaper.
Figure 3. Analytical procedure for analysis of NEOs from diaper urine.
Figure 4. MRM chromatograms of NEOs extracted from urine absorbent, which
contained pooled human urine samples spiked with spiked with NEO standards: blank,
0.8 and 1.5 µg L-1 for acetamiprid and N-desmethylacetamiprid; blank, 1.5 and 3 µg
L-1 for thiacloprid; and blank, 0.6 and 1.3 µg L-1 for dinotefuran, clothianidin,
imidacloprid, and thiamethoxam. Y- and X-axis represent counts per second (cps) of
ions and retention time (min), respectively.
- 24 -
Table 1. Supplementary explanation of figure 1 (SPE column procedure at-a-glance).
SPE column procedure 1
(for dinoteturan)
SPE column procedure 2
(for the other neonicotinoids)
1. Condition Bond Elut PCX
-0.5 mL methanol and acetonitrile (1:1, v/v)
containing 5% NH 3
-0.5 mL H 2 O
2. Apply sample
3. Collect pass through liquid into a new glass
tube
3. Wash Bond Elute PCX cartridge with 0.5 ml
formic acid solution (2%)
4. Transfer liquid from 3. into ISOLUTE SLE+
4. Elute into a new glass tube with 0.5 ml
methanol
5. Wait 30 min
5. Dry with N 2 gas at 40oC
6. Elute with 20 ml ethyl acetate
6. Add 30 µl acetonitrile and 60 µl H 2 O
7. Dry with N 2 gas at 40oC
7. Mix well and sonicate 10 min
8. Add 30 µl acetonitrile and 60 µl H2O
9. Mix well and sonicate 10 min
Combine extracts from both procedures
Analyze by LC-MS/MS
Table 2. Compound-specific mass spectrometer settings.
compounds
fragmentor
(V)
collision energy
(eV)
precursor ion
(m/z)
product ion
(m/z)
retention time
(min)
acetamiprid
120
18
223
8.3
clothianidin
80
16
250
dinotefuran
90
203
imidacloprid
140
10
256
thiacloprid
160
22
253
thiamethoxam
70
292
N-desmethylacetamiprid
110
20
209
acetamiprid-d 6
100
24
229
126 (Q)
56 (C)
169 (Q)
131 (C)
129 (Q)
157 (C)
209 (Q)
175 (C)
126 (Q)
99 (C)
211 (Q)
132 (C)
126 (Q)
90 (C)
126 (Q)
62 (C)
Abbreviations: C, confirmation ion. Q, quantification ion. IS, internal standard.
8.0
6.3
8.1
8.8
7.4
7.9
8.3
40
35
33
43
3.0
42
48
25
41
36
35
37
0.6
60
65
36
51
51
51
71
1.3
51
58
45
49
47
49
73
2.5
64
62
47
53
51
53
60
1.3
88
96
72
76
83
86
113
5.0
94
66
66
83
80
92
101
0.8
5.5
11.2
8.2
10.1
7.6
8.7
6.9
1.5
6.4
10.9
12.4
7.2
6.7
12.7
3.3
3.0
3.9
12.1
10.7
5.3
5.8
8.4
5.3
0.8
5.1
11.4
17.8
11.6
11.3
13.8
7.1
1.5
5.3
15.4
14.8
14.1
11
14.8
6.8
3.0
5.5
13.5
17.1
13.9
11.9
19.5
4.7
5.7
5.6
9.8
18.1
16.7
4.3
14.9
6.2
0.6
0.1
13.6
-a
-a
7.9
0.9
15.8
1.3
13.4
0.7
4.4
0.7
0.9
10.0
6.9
3.0
4.0
13.8
9.2
7.6
2.9
12.1
4.5
10.0
12.1
15.0
16.4
3.3
7.8
14.5
2.4
slope
51.1
717.3
203.4
1015.8
76.7
168.0
83.6
intercept
0.01
0.05
0.43
0.39
0.24
0.07
0.13
r2
0.994
0.988
0.983
0.985
0.996
0.990
0.993
0.07
0.75
0.34
0.40
0.08
0.21
0.08
0.62
0.23
N-desmethyl
acetamiprid
19
thiacloprid
50
imidacloprid
40
dinotefuran
clothianidin
0.8
acetamiprid
concentration
(µg L-1 urine)
thiamethoxam
Table 3. Precision, recovery rate, linearity, LOD, and LOQ data of the analytical procedure.
absolute recovery rate (%)
all of the protocol
extraction rate of
neonicotinoids from diaper
by acetone
IS-corrected recovery rate (%)
all of the protocol
within-run
precision (%RSD)
between-run
precision (%RSD)
relative error (%)
calibration curve
(for environmental PYR exposure)
LOD (µg L-1) (S/N = 3)
0.22
2.25
1.02
1.21
0.23
LOQ (µg L-1) (S/N = 10)
Abbreviations: n, number of observations. RSD, relative standard deviation. LOD, limit of detection.
LOQ, limit of quantitation. Note: amore than 20%.
storage
time
acetamiprid
clothianidin
dinotefuran
imidacloprid
thiacloprid
thiamethoxam
0.8
6h
101
103
103
89
100
101
102
0.8
12 h
99
96
101
105
97
99
107
0.8
24 h
102
96
99
81
98
95
97
6h
103
106
100
101
101
93
103
12 h
106
103
109
120
102
97
104
24 h
105
99
103
101
98
94
99
0.8
109
102
92
109
106
88
103
100
104
94
105
102
93
107
0.8
2 weeks
101
106
97
102
94
96
97
0.8
1 month
101
94
102
93
101
103
92
2 weeks
103
98
95
94
98
103
99
1 month
101
97
104
97
93
100
91
N-desmethyl
acetamiprid
concentration
(µg L-1 urine)
Table 4. NEO stabilities in diaper and diaper urine.
stability in diapers at 37oC (%)
stability in extracted urine from diaper
three freeze-thaw cycles (%)
stability at -80 °C (%)
Abbreviation: n, number of observations.
Table 5. Detection rates, geometric means, and percentiles of urinary NEOs concentrations (µg L-1).
>LOD (%)a
Selected percentiles
GM
Max.
5th
25th
50th
75th
95th
acetamiprid
10
NCb
<LOD
<LOD
<LOD
<LOD
0.13
0.70
clothianidin
18
NCb
<LOD
<LOD
<LOD
<LOD
1.21
1.99
dinotefuran
84
2.01
<LOD
0.82
1.99
4.44
24.30
77.79
<LOD
<LOD
<LOD
<LOD
<LOD
<LOD
imidacloprid
NC
thiacloprid
NC
<LOD
<LOD
<LOD
<LOD
<LOD
0.08
thiamethoxam
24
NCb
<LOD
<LOD
<LOD
<LOD
0.62
1.10
N-desmethylacetamiprid
78
0.34
<LOD
0.08
0.26
1.53
3.09
14.56
Abbreviations: LOD, limit of detection. GM, geometric mean. NC, not calculated. <LOD, Lower than level of
limit of detection.
Notes: aPercent of detection frequency. bGM was not calculated due to low detection rate (less than 60% of
the samples).
Figure 1
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Figure 2
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Figure 3
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Figure 4
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