CYPHENOTHRIN 804

CYPHENOTHRIN
804

CH3 CH3
O

CH3
O
C
CH3 O H CN

ISO common name Cyphenothrin

Chemical name (RS)-α-Cyano-3-phenoxybenzyl (1RS, 3RS;
1RS, 3SR)- 2,2-
dimethyl-3-(2-methylprop-1-enyl)-
cyclopropanecarboxylate (IUPAC);
cyano(3-phenoxyphenyl)methyl 2,2-dimethyl-3-
(2-methyl-1-propenyl)cyclopropanecarboxylate
(CA; 39515-40-7)
Empirical formula C24H25NO3
RMM 375.47
v.p. <7.52 × 10-10 Pa (20, 35, 45 °C)
Solubility In water: <10 × 10-9 g/l (25 °C at pH7); soluble in
organic solvents
Description Yellow to yellowish brown oil or yellow waxy
solid

Note: Cyphenothrin is the ISO common name for the mixture of eight
isomers. The isomers are designated as follows:
S,1R-trans, S,1R-cis, R,1R-trans, R,1R-cis, S,1S-trans, S,1S-cis,
R,1S-trans, and R,1S-cis.

44
 CYPHENOTHRIN 804

CYPHENOTHRIN TECHNICAL
*
804/TC/(M)/-

SCOPE The method is suitable for the determination of cyphenothrin as

well as for isomer mixtures enriched with one – e.g. the S,1R-trans –
isomer (Note 1).

1 Sampling. Take at least 100 g.

2 Identity tests
2.1 GLC. Use the GLC method below. The relative retention time of peak
A with respect to the internal standard for the sample solution should not
deviate by more than 1% from that for the calibration solution.
2.2 HPLC. The retention time of for the S,1R-trans isomer in the sample
solution should not deviate by more than 5% from that for the S,1R-trans
isomer of the standard solution and intensities of the isomers should give
the same pattern as in the standard solution.

REAGENTS
Hexane HPLC grade
1-Butanol HPLC grade
Cyphenothrin enriched working standard product of certified purity and
composition. Store refrigerated.
Mobile phase hexane –1-butanol, 300 +1 (v/v); add 1-butanol (1 ml) by
pipette to hexane (300 ml); degas before use
Standard solution. Weigh 20 mg of cyphenothrin enriched working
standard into a stoppered flask (50 ml). Add by measuring cylinder
mobile phase (20 ml) and dissolve completely.

APPARATUS
High performance liquid chromatograph with a detector suitable for
operation at 278 nm and an injector capable of delivering 5 µl
Column stainless steel, 250 × 4.6 mm (i.d.), packed with Sumichiral
OA-2000, 5 µm (obtainable from Sumika Chemical Analysis Service)
Electric integrator or data system

*
Provisional CIPAC method 2006. Prepared by the Japanese committee (JAPAC). Based on a method
supplied by Sumitomo Chemical Co, Japan.

45
 CYPHENOTHRIN 804

PROCEDURE
(a) Liquid chromatographic conditions (typical):
Mobile phase hexane –1-butanol, 300 + 1 (v/v)
Column two columns connected in series, each
stainless steel, 250 × 4.6 mm (i.d.), packed
with Sumichiral OA-2000, 5 µm
Flow rate 1.0 ml/min
Column temperature ambient
Injection volume 5 µl
Detector wavelength 278 nm
Retention times S, 1R-cis isomer: about 39 min
R, 1R-trans isomer: about 41 min
S, 1S-trans isomer: about 43 min
S, 1R-trans isomer: about 45 min

(b) System equilibration. Inject 5 µl portions of a working standard

solution until the retention times of S,1R-trans isomer obtained for two
consecutive injections differ by less than 5%.

(c) Preparation of sample solution. Weigh 20 mg of sample into a

stoppered flask (50 ml). Add by measuring cylinder mobile phase (20 ml)
and dissolve.

(d) Determination. Inject 5 µl portion of each sample solution. Measure the

relevant peak areas.

(e) Calculation.

Trans isomer percentage in the enriched cyphenothrin =

H SST + H RRT + H SRT
× 100 %
H SST + H RRT + H SRT + H SRC

1R isomer percentage in the acid moiety of the enriched cyphenothrin =

H SRC + H RRT + H SRT
× 100 %
H SRC + H RRT + H SRT + H SST

S isomer percentage in the alcohol moiety of the enriched cyphenothrin =

H SRC + H SST + H SRT
× 100 %
H SRC + H SST + H SRT + H RRT

46
 CYPHENOTHRIN 804

where:
HSRC = peak area of S, 1R-cis isomer
HRRT = peak area of R, 1R-trans isomer
HSST = peak area of S, 1S-trans isomer
HSRT = peak area of S, 1R-trans isomer

3 Cyphenothrin

OUTLINE OF METHOD Cyphenothrin is determined by capillary gas

chromatography using flame ionisation detection and triphenyl phosphate
as internal standard.

REAGENTS

Acetone
Cyphenothrin enriched working standard product of certified purity and
composition. Store refrigerated.
Triphenyl phosphate internal standard. Must not show a peak with the
same retention time as peak A, peak B or peak C.
Internal standard solution. Dissolve triphenyl phosphate (2.0 g) in acetone
(100 ml). Ensure that a sufficient quantity of this solution is prepared for
all samples and calibration standards to be analysed.
Calibration solution. Homogenise the cyphenothrin enriched working
standard by stirring or by warming it to melting and by stirring when it is
waxy solid or partly waxy solid. Prepare calibration solutions in
duplicate. Weigh (to the nearest 0.1 mg) 90 to 110 mg (s mg) of
cyphenothrin enriched working standard into a volumetric flask (50 ml).
Add by pipette internal standard solution (5.0 ml) and dissolve. Make up
to volume with acetone and mix well. (Solutions CA and CB)

APPARATUS
Gas chromatograph equipped with a split/splitless injection and a flame
ionisation detector.
Capillary column fused silica, 30 m × 0.25 × mm (i.d.), film thickness:
0.25 µm, coated with crosslinked 50% phenyl 50% dimethyl polysiloxane
(DB-17 or equivalent)
Electric integrator or data system

47
 CYPHENOTHRIN 804

PROCEDURE
(a) Gas chromatographic conditions (typical):
Column fused silica, 30 m × 0.25 mm, film thickness:
0.25 µm, coated with crosslinked 50% phenyl
50% dimethyl polysiloxane (DB-17 or
equivalent)
Injection system
Injector split injection
Split flow approximately 100 ml/min
Injection volume 1 µl
Detector flame ionisation
Temperatures
Column oven 260°C
Injection port 280°C
Detector 280°C
Carrier gas helium, 30 cm/sec
Retention times triphenyl phosphate: about 8.9 min
enriched cyphenothrin:
peak A: about 11.1 min
peak B: about 11.3 min
peak C: about 11.7 min
Note: peak A consists of the S,1R-trans, R,1S-trans, R,1R-trans and
S,1S-trans isomers
peak B consists of the R,1R-cis and S,1S-cis isomers: this peak is
usually not detected since the amounts of these isomers are very
low.
peak C consists of the S,1R-cis and R,1S-cis isomers;

(b) Linearity check. Check the linearity of the detector response by

injecting 1 µl of solutions with cyphenothrin enriched working standard
concentrations 0.5, 1 and 2 times that of the calibration solution before
conducting analysis.

(c) 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 for the first calibration solution,
otherwise prepare new calibration solutions.

48
 CYPHENOTHRIN 804

(d) Preparation of sample solution. Homogenise the sample by stirring or

by warming it to melting and by stirring when it is a waxy solid or a partly
waxy solid. Prepare sample solutions in duplicate for each sample. Weigh
(to the nearest 0.1 mg) 90 to 110 mg (w mg) of cyphenothrin enriched
mixture into a volumetric flask (50 ml). Add by pipette internal standard
solution (5 ml) and dissolve completely. Make up to volume with acetone
and mix well. (Solutions SA and SB).

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

bracketing them by 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 cyphenothrin working standard contents of the bracketed sample
injections.

Ir × s × P
fi =
Hs

f × Hw
Content of enriched-cyphenothrin = g/kg
Iq × w

where:
fi = individual response factor
f = mean response factor
Hs = peak area of enriched cyphenothrin (peak A+B+C) in the
calibration solution
Hw = total peak area of enriched cyphenothrin (peak A+B+C) in the
sample solution
HA = peak area of the trans isomers peak (A) in the sample solution
HB = peak area of peak B in the sample solution
HC = peak area of peak C 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 enriched-cyphenothrin cyphenothrin working standard in
the calibration solution (mg)
w = mass of sample taken (mg)
P = purity of enriched cyphenothrin working standard (g/kg)

49
 CYPHENOTHRIN 804

Repeatability r = 14 g/kg at 946 g/kg active ingredient content

= 16 g/kg at 944 g/kg active ingredient content
Reproducibility R = 22 g/kg at 946 g/kg active ingredient content
= 26 g/kg at 944 g/kg active ingredient content
Note 1 The method was collaboratively tested with a technical mixture of
isomers enriched with the S,1R-trans isomer as specified by the
WHO specification for d,d-trans cyphenothrin.

CYPHNOTHRIN EMULSIFIABLE CONCENTRATES

*
804/EC/(M)/-

1 Sampling. Take at least 100 g.

2 Identity tests
2.1 GLC. As for 804/TC/(M)/2.
2.2 HPLC. As for 804/TC/(M)/2. except:
(c) Preparation of sample solution. Weigh sufficient sample to contain
20 mg of the cyphenothrin enriched mixture into a stoppered flask (50 ml).
Add by measuring cylinder mobile phase (20 ml) and mix well. Filter the
supernatant through a 0.45 µm filter.

3 Cyphenothrin. As for 804/TC/(M)/3 except:

(a) Gas chromatographic conditions:
Temperatures
Column oven 260°C (use a short temperature program to
remove formulation components, if necessary)

(b) Preparation of sample solution. Weigh (to the nearest 0.1mg) sufficient
sample to contain 90 to 110 mg (w mg) of the cyphenothrin enriched
mixture into a volumetric flask (50 ml). Add by pipette internal standard
solution (5 ml) and dissolve completely. Make up to volume with acetone
and mix well (Solutions SA and SB).

Repeatability r = 1.0 to 1.2 g/kg at 59.2 g/kg active ingredient content

Reproducibility R = 1.9 to 2.2 g/kg at 59.2 g/kg active ingredient content

*
Provisional CIPAC method 2006. Prepared by the Japanese committee (JAPAC). Based on a method
supplied by Sumitomo Chemical Co, Japan.

50
 CYPHENOTHRIN 804

Fig 15 HPLC chromatogram of technical cyphenothrin

composition:
R, 1R-trans 39.25%
S, 1R-trans 39.25%
R, 1R-cis 8.95%
S, 1R-cis 8.95%
R, 1S-trans 1.45%
S, 1S-trans 1.45%
R, 1S-cis 0.35%
S, 1S-cis 0.35%

51
 CYPHENOTHRIN 804

Fig 16 HPLC chromatogram of technical S,1R-trans enriched

cyphenothrin EC

triphenyl
Acetone
phosphate

Peak A

Peak B

Peak C

Fig 17 Gas chromatogram of technical S,1R-trans enriched

cyphenothrin

52
 CYPHENOTHRIN 804

Acetone &
formulants

triphenyl phosphate
(internal standard)

Peak A

Peak B

Peak C

Fig 18 Gas chromatogram of S,1R-trans enriched cyphenothrin EC

53