CH3NCl2 Formation Resulting from

Chlorination of Carbamate Insecticides in

Water
Yue E1, Qian Yang1, Yang Guo1, Lushi Lian1, Jing Li 1,*, Ernest R. Blatchley III2,3

1
Department of Applied Chemistry, China Agricultural University, Beijing 100193,
China

2
Lyles School of Civil Engineering, Purdue University, West Lafayette, IN 47907, USA
3
Division of Environmental & Ecological Engineering, Purdue University, West
Lafayette, IN 47907, USA

*Corresponding author telephone: +86-10-62736957; fax: +86-10-62733474; email:

jli@cau.edu.cn

Supporting Information

(13 pages, 6 Figures, 3Tables)

S1
TABLE S1.Carbamate Insecticides Used in This Study
Solubility Toxicity pKa
Compounds Structure
in water LD50 (SciFinder)
H 3CNH SCH3 21
Methomyl O N 58 g/L mg/kg 13.270.46
O CH3
(oral, rat)
H3C O
N 8–14
H O
Carbofuran O
CH 3 320 mg/L mg/kg 12.280.46
CH 3 (oral, rat)
O
H3C
N O
230
Carbaryl H 110 mg/L mg/kg 12.020.46
(oral, rat)

H3C O
CH 3 56.2
H 3C S N
Thiodicarb NO S ON
S CH 3 35 mg/L mg/kg -1.790.07
H 3C N
O CH 3 (oral, rat)
(1) Hassall, K. A. The Biochemistry and Uses of Pesticides: structure,
metabolism, mode of action, and uses in crop protection; 2nd ed.; VCH: New York,
1990.

S2
 400000
400000 a. 98 b.
83

Abundance
98
Abudance
300000
300000
85
200000
200000
62
62
100000 100000
84
86
0 0
50 60 70 80 90 100 110 120 50 60 70 80 90 100 110 120
m/z m/z
8000 98 d. 98
c. 12000

Abundance
10000
Abudance

6000
8000
4000 62 6000
4000 62
2000
83 2000
84
0 0 86
50 60 70 80 90 100 110 120 50 60 70 80 90 100 110 120
m/z m/z

FIGURE S1. Mass spectra of chlorinated samples of aqueous solutions of carbamate

insecticides: (a) methomyl, (b) carbofuran, (c) carbaryl, and (d) thiodicarb after 24 h.

Methomyl, carbofuran, and carbaryl were present at an initial concentration of 2.0×10-

4
M, while thiodicarb was present at an initial concentration of 2.0×10-5 M. For all

reactions, initial Cl/P = 8.0, pH =7.0, and T = 25℃.

S3
 0.5 1.2
Concentration(mg/l as Cl2)

Concentration(mg/l as Cl2)
ratio 1 CH3NCl2 1.0 ratio 2
0.4
NCl3
free chlroine 0.8
0.3
0.6
0.2
0.4
0.1 0.2
0.0 0.0
0 20 40 60 80 100 0 20 40 60 80 100
5 reaction time / h 10 reaction time / h
Concentration(mg/l as Cl2)

Concentration(mg/l as Cl2)
4 ratio 4 8 ratio 8

3 6

2 4

1 2

0 0
0 20 40 60 80 100 0 20 40 60 80 100
reaction time / h reaction time / h

FIGURE S2. CH3NCl2 concentration as a function of time at different Cl/P molar ratios

resulting from chlorination of aqueous solutions containing methomyl (2×10-5M) at pH

7.0 and 25℃. (a) Cl/P molar ratio = 1.0; (b) Cl/P molar ratio = 2.0; (c) Cl/P molar ratio

= 4.0; (d) Cl/P molar ratio = 8.0.

S4
FIGURE S3. Evaluation of pseudo first-order kinetics of CH3NCl2 formation from four

carbamates at pH 7.0 and 25℃.

S5
FIGURE S4. Effects of pH on the formation of DCMA resulting from the chlorination

of CH3NH2 at 25℃. The initial concentration of CH3NH2 is 4×10-5M, Cl/P = 8.0.

S6
TABLE S2. Activation energy and pre-exponential factor (A) for chlorination of the
four carbamate insecticides.
Carbamate Pesticide Ea (kJ.mol-1) A (M-1s-1)
Methomyl 57.15  0.57 (1.1 0.01) ×109
Thiodicarb 66.40  2.66 (1.2 0.06) ×1011
Carbofuran 21.87  0.43 (7.6 0.15) ×101
Carbaryl 32.59  1.63 (5.2 0.26) ×103

S7
FIGURE S5. CH3NCl2 decay as a function of time for various of Cl/P molar ratios after

48 h chlorination of an initial concentration 4 mg/L as Cl2 at pH 7.0 and 25℃.

S8
FIGURE S6. Vibrio fisheri bioluminescence inhibition assay for CH3NCl2 (a), CHCl3

(b) and NaOCl (c). Fraction of A549 retaining viability after 48 h exposure to CH3NCl2

(d) and CHCl3 (e).

S9
The procedure of SRB assay:

Human lung cancer cell A549 cells were cultured in RPMI 1640 medium (Gibco)

containing 10% fetal bovine serum (Gibco) and 2 mM L-glutamine (Gibco).

Logarithmic growth phase cells were diluted with RPMI 1640 medium to achieve a cell

concentration of 7.5 × 105 / mL. 100μL of the cell suspension was added to each well

in a 96-well microtiter plate, then the plates were incubated at 37° C, 5% CO2, and 100%

relative humidity for 24 h. An aliquot of 100 μL of each test compound concentration

was then added to each well.

There were three groups for the test, the test group, menstruum control group, and

parallel control group. The cells in the test group were treated with different

concentrations of the test substance for 48 h, then 50 μL of 50% TCA (10% final

concentration) was added and fixed at 4 ° C for 60 min. The parallel control group was

fixed by 25 μL TCA immediately before addition of the test substance.

The plate supernatant was decanted from each well, then the well was washed with

deionized water 5 times and air-dried. SRB was formulated as a 0.4% solution in 1%

acetic acid, and 100 μL was added to each well and left at room temperature for 10

minutes. After washing off the unbound SRB with 1% acetic acid 5 times, the wells

were air dried, followed by addition of 150 μL of 10 mM unbuffered Tris base（Gibco）.

Absorbance was read on a BMG POLAR star Omega microplate reader at a wavelength

of 515 nm.

The parallel control group OD value was recorded as Tz, the menstruum control

group OD value was recorded as C, the test group OD value recorded as Ti:

S10
1) Ti> / = Tz, indicated that the menstruum did not kill the cells.

Percentage growth = [(Ti-Tz) / (C-Tz)] × 100

2) Ti <Tz, indicated that that the menstruum killed the cells.

Percentage growth = [(Ti-Tz) / Tz] × 100

The curve was fitted with GraphPad software and LogGI50 was estimated.

S11
TABLE S3. Volatile DBP Measurement in Tap Water Samples and Surface Water Samples.
t (h) CH3NCl2 (μg/L Cl2) NCl3 (mg/L Cl2) Free Chlorine (mg/L Cl2)
TOC
Chlorine dose pH
1.0 5.0 10.0 1.0 5.0 10.0 1.0 5.0 10.0 (mg/L)
(mg/L Cl2)
1 ND ND 0.7 ND 0.02 0.03 0.5 4.4 9.3
Surface water 5 0.7 1.4 1.4 0.05 0.06 0.07 0.1 4.2 9.3
7.83 0.89
63# 10 0.7 2.1 3.1 0.03 0.07 0.14 ND 4.0 7.9
24 1.0 3.6 3.8 0.04 0.09 0.08 ND 1.1 5.9
1 ND 1.6 2.3 0.01 0.12 0.24 ND 2.2 9.7
Surface water 5 ND 4.0 4.0 0.06 0.26 0.32 ND 0.3 8.6
7.51 4.74
70# 10 0.7 4.3 5.1 0.06 0.22 0.37 ND ND 7.6
24 1.0 3.3 4.5 0.04 0.18 0.32 ND ND 4.7
1 ND 0.9 0.7 0.04 0.06 0.06 0.6 4.9 9.7
Surface water 5 0.7 1.6 1.8 0.06 0.09 0.10 0.1 3.6 10.4
8.24 1.95
77# 10 1.0 2.4 2.4 0.05 0.08 0.10 ND 3.9 7.6
24 1.4 3.8 4.5 0.06 0.13 0.17 ND 1.0 6.2
1 ND 0.7 0.7 0.01 0.02 0.02 0.7 4.9 9.2
Surface water 5 0.6 1.1 1.4 0.02 0.05 0.07 0.5 4.7 9.3
7.24 1.77
210# 10 0.7 1.7 2.0 0.02 0.08 0.11 0.4 4.8 9.4
24 1.3 2.7 3.1 0.03 0.12 0.11 0.2 2.7 6.6
1 ND 0.9 1.0 0.05 0.03 0.02 0.8 4.9 9.7
Tap water 5 0.6 1.0 1.1 0.03 0.03 0.06 0.6 4.8 8.9
6.68 1.99
2# 10 0.7 1.1 1.4 0.02 0.02 0.06 0.4 4.9 9.0
24 1.0 1.6 2.3 ND 0.07 0.09 0.1 1.0 6.1

S12
Paddy field water samples in Table 3 were collected from Beijing (Haidian district),

Hubei province (Huanggang), and Henan province (Huangchuan). Water depth in the

paddy fields was kept at about 15 cm. Paddy field water samples were collected by

grabbing samples in 500 mL bottles with screw caps. Other surface water samples were

randomly chosen from 200 water samples collected from different locations in Beijing.

Water samples were transported back to the lab and stored at -20C until use in

experiments. Water samples were brought back to room temperature and filtered before

chlorination. The chlorination was conducted without phosphate buffer, and the pH was

fairly stable during the chlorination.

S13