Thenylchlor

Materials to be Rice grain, soil and water

analyzed
Instrumentation Gas-chromatographic determination for rice grain, water
and soil

1 Introduction

Chemical name 2-Chloro-N-(3-methoxy-2-thenyl)-2’,6’-dimethyl-acet-

(IUPAC) anilide

Structural formula Cl
N

O O
S

Empirical formula C16 H18 ClNO2 S

Molar mass 323.8
Melting point 72–74 ◦ C
Boiling point 173–175 ◦ C/0.5 mmHg
Vapor pressure 2.8 × 10−2 mPa at 25 ◦ C
Solubility Water 11 mg L−1 at 20 ◦ C
Stability Decomposition at 260 ◦ C and by ultraviolet (UV) irradi-
ation (400 nm, 8 h).
Stable to acid and base in the pH range 3–8.
Use pattern Thenylchlor is a chloroacetamide herbicide for the pre-
emergence and post-planting control of annual grass
and broad-leaved weeds in paddy rice
Regulatory position The residue definition is for the parent, thenylchlor,
only

2 Outline of method
Milled rice grain is soaked with water and allowed to stand at room temperature for
2 h. The soaked sample is mixed with acetone. The extract is filtered, the acetone in
the extract is evaporated and the aqueous residue is transferred into a C18 cartridge.

Handbook of Residue Analytical Methods for Agrochemicals.


C 2003 John Wiley & Sons Ltd.
586 Individual compounds

The eluate from the C18 cartridge is concentrated by rotary evaporation and the
residue is dissolved in n-hexane and then subjected to a cleanup procedure using
a Florisil cartridge. The eluate is dried and analyzed by gas chromatography (GC)
with nitrogen–phosphorus detection (NPD).

3 Apparatus
High-speed blender fitted with a leak-proof glass jar and explosion-proof motor
Laboratory mechanical shaker
Filter paper, 6-cm diameter (Kiriyama 5A filter paper)
Glass filter paper, 6-cm diameter
Erlenmeyer flask, 300-mL
Filtration flask, 300-mL
Round-bottom flasks, 100-, 300- and 500-mL
Rotary vacuum evaporator, 40 ◦ C bath temperature
Separatory funnel, 500-mL
Gas chromatograph equipped with a nitrogen–phosphorus detector
Water-bath: electrically heated, temperature 45 ◦ C

4 Reagents
Acetonitrile, n-hexane, acetone, methanol and dichloromethane, reagent grade
Sodium chloride
Sodium sulfate, anhydrous
C18 cartridge: Bond Elut, 1-g/6-mL (Varian)
Florisil cartridge: Sep-Pak Plus Florisil cartridge (Waters)
Silica gel: Wako gel C-200, 100–200 mesh
Alumina cartridge: Sep-Pak Plus Alumina N cartridge (Waters)
Nitrogen, repurified
Hydrogen, repurified

5 Sampling and preparation

Mill the unpolished rice grain with an ultracentrifuge mill in the presence of dry-ice
and sieve through a 42-mesh screen.

6 Procedure

6.1 Extraction
6.1.1 Rice grain
Weigh 10 g of the milled sample in a 300-mL Erlenmeyer flask and soak in 20 mL
of distilled water for 2 h. After adding 100 mL of acetone and shaking vigorously
with a mechanical shaker for 30 min, filter the extract by suction through overlapping
 Thenylchlor 587

filter paper and glass filter paper. Add 50 mL of acetone to the residue, shake and
filter the extract as before. Combine the filtrates in a 300-mL round-bottom flask and
concentrate by rotary evaporation.

6.1.2 Soil
Mix 10 g of the air-dried soil with 100 mL of acetone and shake the mixture with a
mechanical shaker for 30 min. Filter the mixture through a fluted filter paper into a
300-mL round-bottom flask. Wash the residue on the filter with 50 mL of acetone.
Combine the filtrates and concentrate by rotary evaporation.

6.1.3 Water
Transfer 200 mL of water into a 500-mL separatory funnel. Extract the sample twice
with 50 mL of dichloromethane for 30 min with a mechanical shaker and collect the
extracts in a 300-mL Erlenmeyer flask. Filter the combined extracts, together with the
washings from the collecting flask, through anhydrous sodium sulfate into a 300-mL
round-bottom flask. Remove the dichloromethane by rotary evaporation. Dissolve the
residue in 10 mL of n-hexane.

6.2 Cleanup
6.2.1 Rice grain
First cleanup: Transfer the concentrate into a C18 cartridge preconditioned with 5 mL
of methanol, followed by 5 mL of water. Rinse the cartridge with 5 mL of acetonitrile–
water (3 : 7, v/v). Elute thenylchlor with 10 mL of acetonitrile. Concentrate the eluate
to dryness by rotary evaporation at 40 ◦ C. Dissolve the residue in 5 mL of n-hexane.
Second cleanup: Transfer the n-hexane solution into a Florisil cartridge precondi-
tioned with 5 mL of n-hexane and elute thenylchlor with 10 mL of acetone–n-hexane
(1 : 9, v/v). Concentrate the eluate to dryness by rotary evaporation at 40 ◦ C and
dissolve the residue in 5 ml of acetone for GC analysis.

6.2.2 Soil
First cleanup: Transfer the concentrate into a C18 cartridge preconditioned with 5 mL
of methanol, followed by 5 mL of water. Rinse the cartridge with 5 mL of acetonitrile–
water (3 : 7, v/v). Elute thenylchlor with 10 mL of acetonitrile. Concentrate the eluate
to dryness by rotary evaporation at 40 ◦ C. Dissolve the residue in 5 mL of n-hexane.
Second cleanup: Transfer the n-hexane solution into an alumina cartridge precon-
ditioned with 5 mL of n-hexane. Rinse the cartridge with 10 mL of acetone–n-hexane
(1 : 9, v/v) and elute thenylchlor with 7 mL of acetone–n-hexane (3 : 17, v/v). Con-
centrate the eluate to dryness by rotary evaporation at 40 ◦ C and dissolve the residue
in 5 mL of acetone for GC analysis.

6.2.3 Water
Transfer the n-hexane solution into a glass column packed with 5 g of Florisil saturated
with n-hexane. Rinse the column, first with 2 mL of n-hexane and then with 50 mL
588 Individual compounds

of acetone–n-hexane (1 : 19, v/v). Elute thenylchlor with 50 mL of acetone–n-hexane

(1 : 9, v/v). Concentrate the eluate to dryness by rotary evaporation at 40 ◦ C and
dissolve the residue in 5 mL of acetone for GC analysis.

6.3 Determination (rice grain, soil and water)

Inject an aliquot of the solution into the gas chromatograph.
Operating conditions
Gas chromatograph HP 5890A, Hewlett-Packard
Sample injector Solventless sample injector
Column DB-1 (J&W Scientific), 0.53-mm i.d. × 15 m, film
thickness 1.5-µm
Column temperature 235 ◦ C
Injection port temperature 250 ◦ C
Detector Nitrogen–phosphorus detector
Detector temperature 280 ◦ C
Gas flow rates Helium carrier gas, 10 mL min−1
Hydrogen, 3.5 mL min−1
Air, 115 mL min−1
Injection volume 2 µL
Retention time of thenylchlor 4.5 min

7 Evaluation
7.1 Method
7.1.1 Rice grain and soil
Quantitation is performed by the calibration technique. Construct a new calibration
curve with thenylchlor standard solutions for each set of analyses. The thenylchlor
peak usually appears at a retention time around 4.5 min. Plot the peak area against the
injected amount of thenylchlor. The injection volume (2 µL) should be kept constant
as the peak area varies with the injection volume with NPD. Before injecting the
sample solutions, check the stability of sensitivity of the GC system by injecting more
than one standard solution containing ca 0.05–2 ng of thenylchlor. Recommendation:
inject standard solutions and sample solutions alternately rather than constructing the
calibration curve in advance.

7.2 Recoveries and limit of detection

7.2.1 Rice grain
The recoveries from untreated control samples fortified with thenylchlor at levels of
0.2 mg kg−1 ranged from 75 to 100%. The limit of detection was 0.01 mg kg−1 .
 Thenylchlor 589

7.2.2 Soil
The recoveries from untreated control soil fortified with thenylchlor at levels between
0.3 and 3.0 mg kg−1 ranged from 91 to 95%. The limit of detection was 0.01 mg kg−1 .

7.3 Calculation of residues

The amount of thenylchlor residue (R, mg kg−1 ) in the sample is calculated using the
following equation:

R = (Wi × Vf )/(Vi × G)

where
G = sample weight (g)
Vi = injection volume into the gas chromatograph (µL)
Vf = final sample volume (mL)
Wi = amount of thenylchlor for Vi read from the calibration curve (ng)

8 Important points
To avoid degradation of the analytes, rice grain samples must be homogenized and
milled in the presence of dry-ice. During evaporation of organic solvents, the tem-
perature of the water-bath should be kept at 40 ◦ C or lower.
Thenylchlor residues in rice grain fortified with thenylchlor at levels of 0.5 mg kg−1
are stable, with more than 95% remaining after approximately 45 days of storage at
−20 ◦ C (personal data).

References
1. ‘Persistency in Agricultural Products and Water Pollution of Pesticides,’ No. 3, The Chemical
Daily Co., Tokyo, pp. 492–494 (1998) (in Japanese).
2. ‘Analytical Method for Thenylchlor,’ Ministry of the Environment, Tokyo (1993).
3. K. Kobayashi, M. Onoe and H. Sugiyama, Weed Res. (Jpn), 39, 160 (1994).

Hiroko Kobayashi
Research Institute of Japan Plant Protection Association, Ushiku, Japan