7.7.06
Prepare
AOAC Official Method 979.04
Ethion in Pesticide Formulations
Liquid Chromatographic Method
First Action 1979
Final Action 1980
(Applicable to dry and liquid formulations containing ethion as
only active ingredient.)
A. Apparatus
(a) Liquid chromatograph.—With 254 nm UV detector. Typical
operating conditions: eluant flow rate 1 mL/min (ca 1100 psi),
detector sensitivity 0.2 A unit full scale, ambient temperature,
injection volume 10 mL. Adjust operating conditions to elute ethion
peak in 6 ± 2 min. Column condition and H2O content of methanol
eluant can change retention times. Ethion peak must be separated
completely from internal standard peak which normally elutes in ca
7 min.
(b) Liquid chromatographic column.—A 250 ´ 4.6 mm C18
column. [Study used either (1) Waters mBondapak C18, 300 ´
3.9 mm id; or (2) Du Pont ODS Permaphase, 0.5 m ´ 2.1 mm id].
B. Reagents
(a) Eluant.—Either (1) degassed methanol–H2O (90 + 10), UV
cutoff <230 nm, or (2) degassed acetonitrile–H2O (40 + 60), UV
cutoff <230 nm.
(b) Light mineral oil.—USP, viscosity 38.1 centi stokes at
37.8°C.
( c ) I n t e r n a l s t a n d a rd s o l u t i o n . — ( 1 ) F o r Wa t e r s
column.—Accurately weigh ca 0.24 g pentachloronitrobenzene
(PCNB), reference grade, with no interfering peaks on LC, into
200 mL volumetric flask. Dilute to volume with methanol and mix.
(2) For Du Pont column.—Using CH3CN as solvent, vary amount
PCNB in internal standard to give peak height approximately same
as ethion peak.
(d) Ethion standard solutions.—(1) For Waters column.—Stock
solution.—Accurately weigh amount of standard equivalent to
250 mg ethion, 95+% pure (available from Chemical and Biological
Investigations, U.S. Environmental Protection Agency, Beltsville,
MD 20705, USA) into 25 mL volumetric flask, dilute to volume with
methanol, and mix. Working solution.—Pipet 10 mL stock solution
into 50 mL volumetric flask, pipet 10 mL internal standard solution,
(c)(1), into flask, dilute to volume with methanol, and mix.
standard and samples daily. (2) For Du Pont column.—Prepare as
above, us ing CH 3 CN in stead of meth a nol. (3) For oil
formulations.—Pipet 10 mL 1% stock solution (1) or (2) into 50 mL
volumetric flask containing ca same weight of light mineral oil as
test portion. Add 20 mL methanol (or CH3CN) and proceed as in
C(c) beginning with “Stopper and agitate¼”.
C. Preparation of Test Solution
(a) Dry powder.—Accurately weigh test portion containing ca
100 mg ethion into 250 mL glass-stoppered flask. Pipet in 40 mL
methanol (or CH3CN) and 10 mL internal standard solution. Shake
30 min on mechanical shaker and centrifuge to separate phases.
(b) Liquid concentrates.—Prepare test solution as in B(d).
(c) Oil formulations.—Accurately weigh test portion containing
ca 100 mg ethion into 50 mL volumetric flask. Add 30 mL methanol
(or CH3CN). Stopper and agitate vigorously 1 min, with side to side
action, keeping mixture in main body of flask. Pipet in 4 mL H2O
and repeat vigorous mixing 1 min. Dilute to approximate volume
with methanol (or CH3CN). Cool to ambient temperature and dilute
to volume. Mix thoroughly by inverting 10 times and swirling
vigorously each time. Centrifuge to separate phases.
D. Determination
Use high-pressure liquid syringe or injection loop to inject 10 mL
portions of standard until 2 peak height ratios agree within ±1%.
Alternately inject two 10 mL portions each of test and standard
solutions. Measure peak heights and calculate average peak height
ratios for both standard and test solutions. Adjust attenuation or
amount injected for convenient size peaks (60–80% full scale).
Measure peak heights from baseline between ethion and internal
standard peaks.
Ethion, % = (R/R¢) ´ (W¢/W) ´ P
where R and R¢ = average peak height ratios for test and standard
solutions, respectively; W¢ = mg ethion in working standard solution
(ca 100 mg); W = mg test portion in final dilution; and P = % purity
of standard.
References: JAOAC 62, 11(1979); 63, 302(1980).
CAS-563-12-2 (ethion)
ã 2005 AOAC INTERNATIONAL