AZOXYSTROBIN 571

AZOXYSTROBIN
571

N N

O O
CN O O
CH3 CH3
O

ISO common name Azoxystrobin

Chemical name Methyl (E)-2-{2-[6-(2-cyanophenoxy)pyrimidin-4-yl-
oxy]phenyl}-3-methoxyacrylate (IUPAC) methyl (E)-
2-[[6-(2-cyanophenoxy)-4-pyrimidinyl]oxy]-α-(methoxy-
methylene)benzeneacetate (CA, 131860-33-8)
Empirical formula C22H17N3O5
RMM. 403.4
b.p. Decomposes at 345 ºC
m.p. 116 ºC
v.p. 1.1 × 10-4 Pa at 20 ºC
Solubility In water: 6 mg/l at 20 ºC; low solubility in hexane,
n-octanol; moderate solubility in methanol, toluene,
acetone; high solubility in ethyl acetate, acetonitrile,
dichloromethane
Description Off-white to light brown or yellowish solid
Stability Stable to hydrolysis. DT50 for aqueous photolysis 14 d
Formulations Water dispersible granules and suspension concentrates

10
 AZOXYSTROBIN 571

AZOXYSTROBIN TECHNICAL
*
571/TC/M/-

1 Sampling. Take at least 100g.

2 Identity tests
2.1 GLC. Use the capillary GC method below. The relative retention time of
azoxystrobin with respect to the internal standard for the sample solution should
not deviate by more than 1% from that of the calibration solution (Fig 5).
2.2 Infrared. Take 1 mg of sample and 500 mg of dry KBr powder to form a
KBr disc and scan the disc from 4000 – 500 cm-1. The spectrum produced from
the sample should not differ significantly from that of the standard (Fig 4).

3 Azoxystrobin
OUTLINE OF METHOD The sample is dissolved in acetone containing an
internal standard and the azoxystrobin content determined by capillary gas
chromatography.

REAGENTS
Azoxystrobin standard, of known purity.
Acetone glass distilled
3-(2-Pyridyl)-5,6-diphenyl-1,2,4-triazine internal standard. Must not contain
impurities with the same retention time as azoxystrobin.
Internal standard solution. Dissolve 0.25 g of 3-(2-pyridyl)-5,6-diphenyl-1,2,4-
triazine in acetone (100 ml). Ensure that a sufficient volume is prepared for all
samples to be analysed.
Calibration solution. Prepare calibration solutions in duplicate. Weigh (to the
nearest 0.1 mg) 45 - 55 mg (s mg) of standard azoxystrobin into a suitable
container (about 15 ml). Add by pipette or calibrated dispenser internal
standard solution (10.0) ml. Cap the flask and place it in an ultrasonic bath for
2 min to dissolve the azoxystrobin (solutions CA and CB).

APPARATUS
Gas chromatograph equipped with a split/splitless injection system and a flame
ionisation detector
Capillary column fused silica, 25 m × 0.32 mm (i.d.), coated with crosslinked
14% phenyl 86% dimethyl polysiloxane (CP-Sil 13CB), film thickness: 0.2 µm
Electronic integrator or data system

*
CIPAC method 2008. Prepared by PAC UK. Based on a method submitted by Syngenta Crop Protection AG.

11
 AZOXYSTROBIN 571

PROCEDURE

(a) Gas chromatographic conditions (typical):

Column Fused silica, 25 m × 0.32 mm (i.d.) coated with
CP-Sil 13CB phase, film thickness 0.2 µm
(crosslinked 14% phenyl 86% dimethyl poly-
siloxane)
Injection system
Injector split injection
Injection volume 1 µl
Split ratio 50:1
Detector flame ionisation
Temperatures
Injection port 275 ºC
Detector 325 ºC
Oven programme temp 1: 240 ºC, hold 0 min, ramp rate 3 ºC/min
temp 2: 270 ºC, hold 0 min, ramp rate 30 ºC/min*
temp 3: 320 ºC, hold 5 min
*Note: Azoxystrobin should elute below 270°C to
prevent conversion to the Z-isomer (see Fig 5).
Gas flow rates
Hydrogen (carrier) 75 cm/s (typically 7.7 kPa at 240 ºC), run at
constant pressure
or helium 75 cm/s (typically 18 kPa at 240 ºC), run at
constant pressure
Air 400 ml/min
Hydrogen 30 ml/min
Nitrogen (make up) to 30 ml/min
Retention times internal standard: 6.8 – 7.2 min
azoxystrobin: 8.1 – 8.5 min

(b) System equilibration. Prepare two calibration solutions. Inject 1 µl portions

of the first one until the response factors obtained for two consecutive injections
differ by less than 1.0%. Then inject a 1 µl portion of the second solution. The
response factor for this solution should not deviate by more than 1.0% from that
of the first calibration solution, otherwise prepare new calibration solutions. If
the peak retention times are not within the indicated time windows adjust the
carrier pressure.

12
 AZOXYSTROBIN 571

(c) Sample preparation. Prepare solutions in duplicate for each sample. Weigh
(to the nearest 0.1 mg) sufficient sample to contain 45-55 mg (w mg) of
azoxystrobin into a suitable container (flask or bottle, about 15 ml). Add by
pipette internal standard solution (10.0 ml). Cap the flask and place it in
ultrasonic bath for 2 min to dissolve the sample (solutions SA and SB).

(e) Determination. Inject in duplicate 1µl portions of each sample solution

bracketing them with injections of the calibration solutions as follows:
calibration solution CA, sample solution SA, sample solution SA, calibration
solution CB, sample solution SB, sample solution SB, calibration solution CA, and
so on. Measure the relevant peak areas.

(f) Calculation. Calculate the mean value of each pair of response factors
bracketing the two injections of a sample and use this value for calculating the
azoxystrobin contents of the bracketed sample injections.

Ir ´ s ´ P
fi =
Hs

Content of azoxystrobin Procymidone content =

f ´ Hw g/kg
g/kg
Iq ´ w

where:
fi = individual response factor
f = mean response factor
Hs = peak area of azoxystrobin in the calibration solution
Hw = peak area of azoxystrobin in the sample solution
Ir = peak area of the internal standard in the calibration solution
Iq = peak area of the internal standard in the sample solution
s = mass of the azoxystrobin in the calibration solution (mg)
w = mass of sample taken (mg)
P = purity of azoxystrobin standard (g/kg)

Repeatability r = 8.5 g/kg at g/kg 993active ingredient content

Repeatability r = 15 g/kg at 971 g/kg active ingredient content
Reproducibility R = 17 g/kg at 993 g/kg active ingredient content
Reproducibility R = 22 g/kg at 973 g/kg active ingredient content

13
 AZOXYSTROBIN 571

AZOXYSTROBIN WATER DISPERSABLE GRANULES

*
571/WG/M/-

1 Sampling. Take at least 1 kg.

2 Identity tests
2.1 GLC. As for azoxystrobin technical 571/TC/M/2.2 (Fig 5).
2.2 Infrared. Take sufficient sample to contain about 0.25 g of azoxystrobin,
and shake vigorously for 2 minutes with acetone (2 ml). Filter to remove any
residue. To 1 ml of the clear filtrate add water (9 ml) and mix. Centrifuge the
precipitate of azoxystrobin. Remove the supernatant and add water (10 ml) to
the residue. Shake the mixture and again centrifuge. Remove the supernatant
and dry the residue at 80 ºC under vacuum for several hours. Proceed as for
azoxystrobin technical 571/TC/M/2.2.

3 Azoxystrobin. As for azoxystrobin technical 571/TC/M/3 except:

add at:
APPARATUS
Sample filtering device with a membrane filtration unit compatible with organic
solvents and a 0.45 µm pore diameter.
and replace '(c) Sample preparation' by:
(c) Sample preparation Grind the sample prior to analysis. Prepare solutions in
duplicate for each sample. Weigh (to the nearest 0.1 mg) sufficient sample
(w mg) to contain about 50 mg of azoxystrobin into a suitable container (flask or
bottle, about 15 ml). Add by pipette internal standard solution (10.0 ml) to the
weighed aliquots. Cap the flask and place it in ultrasonic bath for 2 min to
dissolve the active ingredient (solutions SA and SB). Filter each solution prior to
analysis.

Repeatability r = 4.8 g/kg at 506 g/kg active ingredient content

Reproducibility R = 14 g/kg at 506 g /kg active ingredient content

4 Suspensibility (Draft method)

REAGENTS AND APPARATUS As for 571/TC/M/3 and MT 184.

PROCEDURE

(a) Preparation of suspension and determination of sedimentation. MT 184.

*
CIPAC method 2008. Prepared by PAC UK. Based on a method submitted by Syngenta Crop Protection AG.

14
 AZOXYSTROBIN 571

(b) Determination of azoxystrobin in the bottom 25 ml of suspension. After

removal of the top 225 ml of suspension transfer the 25 ml remaining in the
cylinder to a separatory funnel and extract with ethyl acetate (1.5 ml in case of a
0.3% test concentration or 1.0 ml in case of a 0.2% test concentration at
500 g/kg active ingredient content). Evaporate the ethyl acetate layer almost to
dryness and dissolve the residue in internal standard solution (3.5 ml in case of a
0.3% test concentration or 2.5 ml in case of a 0.2% test concentration).
Determine the mass of azoxystrobin (Q g) by 571/TC/M/3.

(c) Calculation

111 ( c - Q )
Suspensibility = %
c
where:
c = mass of azoxystrobin in the sample taken for the preparation of the
suspension (g)
Q = mass of azoxystrobin in the bottom 25 ml of suspension (g)

AZOXYSTROBIN SUSPENSION CONCENTRATES

*
571/SC/M/-

1 Sampling. Take at least 1l.

2 Identity tests
2.1 GLC. As for technical 571/TC/M/2.1.
2.1 Infrared. Take 1ml of sample and shake vigorously for 2 minutes with of
acetone (2 ml). Filter to remove any residue. To 1 ml of the clear filtrate add
water (9 ml) and mix. Centrifuge the precipitate of azoxystrobin. Remove the
supernatant and add 10 ml of water to the residue. Shake the mixture and again
centrifuge. Remove the supernatant and dry the residue at 80 ºC under vacuum
for several hours. Proceed as for azoxystrobin technical 571/TC/M/2.2.

3Azoxystrobin. As for azoxystrobin technical 571 TC/M/3 except:

*
CIPAC method 2008. Prepared by PAC UK. Based on a method submitted by Syngenta Crop Protection AG.

15
 AZOXYSTROBIN 571

(c) Sample preparation. Prepare solutions in duplicate for each sample. Weigh
(to the nearest 0.1 mg) sufficient sample to contain about 50 mg (w mg) of
azoxystrobin into a suitable container (flask or bottle, about 15 ml). Add by
pipette internal standard solution (10.0 ml). Cap the flask and place it in
ultrasonic bath for 2 min (solutions SA and SB)..

Repeatability r = 5 to 7 g/kg at 229 g/kg active ingredient content

Reproducibility R = 7 to 8 g/kg at 229 g/kg active ingredient content

4 Suspensibility (Draft method)

REAGENTS AND APPARATUS As for 571/TC/M/3 and MT 184.

PROCEDURE

(a) Preparation of suspension and determination of sedimentation. MT 184.

(b) Determination of azoxystrobin in the bottom 25 ml of suspension. After
removal of the top 225 ml of suspension transfer the 25 ml remaining in the
cylinder to a separatory funnel and extract with ethyl acetate (3.0 ml in case of a
6% test concentration or 1.0 ml in case of a 0.2% test concentration at 250 g/l
active ingredient content). Evaporate the ethyl acetate layer almost to dryness
and dissolve the residue in internal standard solution (7.5 ml in case of a 0.3%
test concentration or 0.25 ml in case of a 0.2% test concentration). Determine
the mass of azoxystrobin (Q g) by 571/TC/M/3.

(c) Calculation

111 ( c - Q )
Suspensibility = %
c

where:
c = mass of azoxystrobin in the sample taken for the preparation of the
suspension (g)
Q = mass of azoxystrobin in the bottom 25 ml of suspension (g)

16
 AZOXYSTROBIN 571

Peak Wavelength cm-1 Peak Wavelength cm-1

1 2949 7 1487
2 2233 8 1436
3 1710 9 1382
4 1635 10 1252
5 1591 11 1144
6 1564 12 991
13 556

Fig 4 Infrared spectrum of azoxystrobin technical

Z Isomer

Fig 5 Chromatogram of azoxystrobin technical

17