ALPHACYPERMETHRIN WHO/SIT/32

TECHNICAL

TECHNICAL ALPHACYPERMETHRIN
Full specification: WHO/SIT/32
Adopted 10 December 1999

1. Specification
1.1 Description

The material shall consist of a homogeneous mixture of technical alphacypermethrin

together with related manufacturing impurities and shall be in the form of a white to
cream crystalline powder, free from visible extraneous matter and added modifying
agents.

1.2 Chemical and physical requirements

The material sampled from any part of the consignment (see method WHO/M/1.R1),
shall comply with the requirements of section 1.1 and with the following
requirements.

1.2.1 Identity test

Where the identity of the active ingredient is in doubt, then it shall comply with at least
one additional test. (CIPAC method 454/TC/(M)/2, CIPAC Handbook H, p.15).

1.2.2 Alphacypermethrin cis 2 ([IR cis] S and [IS cis] R isomers) content (g/kg
basis)

The alphacypermethrin cis 2 ([IR cis] S and [IS cis] R isomers) content shall be
declared (not less than 910 g/kg), and when determined by the method described in
section 2.1 the mean measured content obtained shall not be lower than the declared
content.

1.2.3 Total pyrethroid content

The combined content of the cis and trans isomers of alpha-cyano-3-phenoxybenzyl-

2,2-dimethyl-3-(2,2-dichlorovinyl-)cyclopropanecarboxylate, when determined by the
method described in section 2.2, shall not be less than 975 g/kg.
.
1.2.4 Volatile hydrocarbon solvent content

The maximum volatile hydrocarbon solvent content of the material, when determined
by the method described in section 2.3 shall not be higher than 1g/kg.

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1.2.5 Triethylamine content

The maximum triethylamine content of the material, when determined by the method
described in section 2.4 shall not be higher than 1 g/kg.

1.3 Packing and marking of packages

The technical alphacypermethrin shall be packed in suitable clean containers, as

specified in the order:

All packages shall bear, durably and legibly marked on the container, the following:

Manufacturers name
Technical alphacypermethrin
Batch or reference number, and date of test
Net weight of contents
Date of manufacture

and the following cautionary notice:

Alphacypermethrin is a pyrethroid that acts predominantly on the central nervous system; high dosages
have been found to cause tonic seizures in experimental animals. A high concentration in air may be an
irritant and contact with the concentrated product may induce a temporary tingling sensation, particularly
on the face. It may be hazardous if swallowed. Do not inhale spray mist. Avoid skin contact; wear
protective gloves, clean protective clothing, and a face mask (surgical type) when handling the product.
Wash hands and exposed skin thoroughly after using.

Keep containers out of reach of children and well away from foodstuffs and animal feed and their
containers.

Alphacypermethrin is toxic to aquatic wildlife. Avoid accidental contamination of water.

If poisoning occurs, call a physician. Treatment is symptomatic.

2. Methods of determining chemical and physical properties

2.1 Alphacypermethrin content

2.1.1 Outline of method

The sample is dissolved in tetrahydrofuran with di-octyl phthalate used as the internal
standard. Separation is carried out in the split injection mode using a dimethyl
polysiloxane WCOT fused silica capillary column with automatic injection and flame
ionisation detector. The alphacypermethrin is determined by comparison with
calibration solutions.

2.1.2 Apparatus

Gas chromatograph. Capable of operating over the range 100 to 300C with a flame
ionisation detector, split/splitless injector and autosampler.

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ALPHACYPERMETHRIN WHO/SIT/32
TECHNICAL

Column. 30 m WCOT fused silica 0.25 mm id and 0.25 m film thickness coated
with dimethyl polysiloxane (chemically bonded), or equivalent.

Injector system. Injector split/splitless with fused silica liner containing a 1 cm plug
of glass wool. An automatic injector is strongly recommended to ensure the best
analytical results.

It is important to ensure that the split liner is acid treated, thoroughly deactivated and
conditioned before use, to ensure that alphacypermethrin does not epimerise during
analysis.
Split ratio 75-100:1 (split vent at 75 mL/min)
Injection Volume 1.0 L using a autosampler

Detector system. Type FID, with no special requirements.

Automatic digital integrator or chromatography data system. Compatible with the

gas chromatograph.

100 mL volumetric flasks.

2.1.3 Reagents

Tetrahydrofuran, analytical grade.

Citric acid. 5% citric acid solution.

Dissolve citric acid (25 g) in water (500 mL).

Di-octyl phthalate, di-(2-ethylhexyl)phthalate, Internal standard. Select for use a batch

which, when chromatographed under the conditions below for the
determination of alphacypermethrin gives no peak with a similar retention
time to the alphacypermethrin isomers.

Cypermethrin analytical standard of known purity (containing a mixture of trans 3,

trans 4, cis 1 and cis 2 cypermethrin).

Alphacypermethrin working standard, of known alphacypermethrin content .

2.1.4 Preparation of standard solutions

Internal standard solution

Dissolve di-octyl phthalate (5.0 g) in tetrahydrofuran (500 mL) (solution I). Ensure
sufficient quantity of this solution is prepared for all samples and calibration standards
being analysed. The same internal standard solution must be used for standards and
samples in the same analytical run.

Citric acid. 5% citric acid solution.

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Dissolve citric acid (25 g) in water (500 mL).

Alphacypermethrin calibration solution

Weigh in duplicate (to the nearest 0.1 mg) about 0.1 g of alphacypermethrin (CA and
CB, g) into 100 mL volumetric flasks. Add to each, 70 mL of tetrahydrofuran and
agitate the flask gently to dissolve. Add by pipette 10.0 mL of internal standard
solution (solution I) and 10 mL of 5% citric acid solution from a measuring cylinder.
Make to volume with tetrahydrofuran and mix well. (Solutions CA and CB).

Note: Citric acid is added to stop epimerisation of alphacypermethrin in solution.

Prepare a solution without internal standard, by dissolving about 0.1 g of

alphacypermethrin standard into a 100 mL volumetric flask. Add approximately 70
mL of tetrahydrofuran and swirl to dissolve. Add by pipette 10 mL of 5% citric acid
solution from a measuring cylinder. Make to volume with tetrahydrofuran and mix
well. (Solution CO).

Cypermethrin calibration check solution

Prepare a solution of cypermethrin, by dissolving about 0.1 g of cypermethrin
standard into a 100 mL volumetric flask. Add approximately 70 mL of
tetrahydrofuran and swirl to dissolve. Add by pipette 10mL of the internal standard
solution to volume with tetrahydrofuran and mix well (Solution CC).

2.1.5 Operating conditions

The conditions given below are typical values and may have to be adjusted to obtain
optimum results from a given apparatus.

Temperatures

Column oven 230C

Injector 260C
Detector 300C

Adjust the column oven temperature if required to obtain retention time windows for
di-octyl (approximately 13.5 minutes), alphacypermethrin cis 2 isomer
(approximately 27.0 minutes) and alphacypermethrin cis 1 isomer (approximately
25.2 minutes). The trans 3 and trans 4 isomers elute at approximately 26.1 and 27.5
minutes respectively.

Gas flow rates

Helium carrier approximately 0.8 mL min-1

Helium make up gas 60 mL min-1 or optimum for instrument

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Septum purge 2 mL min-1

Hydrogen as recommended for the instrument
Air as recommended for the instrument

All gases should be purified through molecular sieves. The carrier gas should be
further purified through an oxygen trap.

2.1.6 Sample preparation

Sampling

Homogenize the bulk material before taking the sample. Take at least 25 g. Re-
homogenize the bulk before taking a sub-sample for analysis.

Preparation of the sample solutions.

Weigh in duplicate (to the nearest 0.1 mg) into 100 mL volumetric flasks sufficient
sample (w, g) to contain 0.1 g of alphacypermethrin. Add approximately 70 mL of
tetrahydrofuran and agitate the flask gently to dissolve.

Add by pipette to each flask 10 mL of di-octyl phthalate solution I and by measuring

cylinder, 10 mL of 5% citric acid solution. Make to volume with tetrahydrofuran and
mix well (Solutions SA and SB).

Prepare a solution without internal standard, by dissolving about 0.1 g of

alphacypermethrin standard into a 100 mL volumetric flask. Add approximately 70
mL of tetrahydrofuran and swirl to dissolve. Add by pipette 10 mL of 5% citric acid
solution from a measuring cylinder. Make to volume with tetrahydrofuran and mix
well. (Solution SO).

2.1.7 Equilibration of the system.

Carry out alternate 1.0 L injections of solutions I, Co and So and check whether there
are any interfering peaks from impurities. If there are make any necessary corrections
but do not use external calibration.

Inject calibration solutions CA and CB to equilibrate the system and use the data from
these chromatograms to set the integration parameters. Calculate the response factors
for these injections to check the stability of the instrument. Response factors should
not differ by more than 1% of the mean.

Inject the solution of cypermethrin in tetrahydrofuran to ensure complete separation

of the alphacypermethrin isomers.

2.1.8 Analysis of sample

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Carry out 1.0 L injections of calibration solutions CA and CB and sample solutions
SA and SB in the following sequence and record the integrated areas of the peaks.

Injection sequence CA1, SA1, SA2, CB1, CA2, SB1, SB2, CB2.

2.1.9. Calculation

Calculate the relative response factors (f1, f2 etc) for the pair of calibration injections
which bracket the sample injections eg use CA1 and CB1 for sample injections SA1, SA2
etc and obtain the mean response factor, f. Sample analysis should be repeated if
calibration response factors f1 and f2 differ by more than 2% of the mean f.

Relative response factor = Hs

Ir x s x P

Where:
Hs = area of alphacypermethrin cis 2 peak in the calibration solution.
Ir = area of di-octyl phthalate peak in the calibration solution.
s = mass of alphacypermethrin analytical standard in calibration
solution (g).
P = purity of the alphacypermethrin standard (g/kg).

For each sample injection eg SA1, calculate the alphacypermethrin cis 2 content.

Alphacypermethrin content = Hw
f x Iq x w
Where:
f = mean relative response factor
Hw = mean area of alphacypermethrin cis 2 peak in the sample solution
Iq = area of di-octyl phthalate peak in the sample solution
w = mass of sample (g)

Calculate the alphacypermethrin cis 2 content of the sample as the mean of the four
determinations as follows:

Sample injection Use relative response factor Alphacypermethrin cis 2

from (g/kg)

SA1 CA1 and CB1 Q )

) X
SA2 CA1 and CB1 R )

SB1 CA2 and CB2 S )

) Y
SB2 CA2 and CB2 T )

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Q and R, S and T should agree to within 0.5% of their mean values (X and Y). X
and Y should agree within 1% of their mean values.

Take the mean of the two values x and y as the alphacypermethrin content.

2.2 Total pyrethroid content

2.2.1 Outline of method

The sample is dissolved in a chloroform/heptane mixture containing 0.2%

tetrahydrofuran. Separation is carried out by normal phase high performance liquid
chromatography with ultra violet detection. The total pyrethroid content is
determined by comparison with calibration solutions.

2.2.2 Apparatus

Liquid chromatograph equipped with a 10 L loop injector. UV detector capable of

operating at 278 nm.

Column stainless steel, 250 x 4.6 mm (i.d.) Zorbax SIL normal phase column, or
equivalent.

Guard column 15 x 3.2 mm (i.d.) 7 m silica cartridge or equivalent.

Injector system capable of reproducible 10 L injections.

Detector system UV, with no special requirements.

Automatic digital integrator or chromatography data system compatible with the

liquid chromatograph

50 mL, 100 mL volumetric flasks.

2.2.3 Reagents

Cis and trans cypermethrin Analytical standard of known purity

Heptane HPLC grade
Chloroform HPLC grade (containing 1% ethanol as a preservative)
Tetrahydrofuran (THF) ACS reagent (contains butylated hydroxytoluene, BHT, as a
preservative)
Mobile phase Heptane and chloroform (9 to 1 volume ratio) with 0.2% (v/v)
tetrahydrofuran. Measure heptane (2250 mL) and chloroform (250 mL) using a
graduated cylinder and add by pipette THF (5 mL).

2.2.4 Preparation of standard solutions

Cis and trans cypermethrin calibration solution

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Weigh in duplicate (to the nearest 0.1 mg) about 0.05 g each of cis and trans
cypermethrin (CC and CD, g) into 100 mL volumetric flasks. Add to each, 5 mL of
chloroform to dissolve and dilute to volume with heptane and mix well. (Solutions
CC and CD).

2.2.5 Operating Conditions

The conditions given below are typical values and may have to be adjusted to obtain
optimum results from a given apparatus.

Eluting solvent flow rate 2 mL/min

Temperature 35C
Injection volume 10 L
Wavelength 278 nm
Retention times Cis 1 cypermethrin 15-18 minutes
Cis 2 cypermethrin 16-20 minutes
Trans 3 cypermethrin 17-25 minutes
Trans 4 cypermethrin 22-27 minutes

2.2.6 Sample preparation

Sampling

Homogenise the bulk material before taking the sample. Take at least 25 g. Re-
homogenise the bulk before taking a sub-sample for analysis.

Preparation of the sample solutions.

Weigh in duplicate (to the nearest 0.1 mg) into 50 mL volumetric flasks sufficient
sample (w, g) to contain 0.1 g of technical grade cypermethrin. Add approximately 5
mL of chloroform to dissolve the sample and dilute with heptane. (Solutions SC and
SD).

2.2.7 Equilibration of the system

Inject calibration solutions CC and CD to equilibrate the system and use the data from
these chromatograms to set the integration parameters. Calculate the response factors
for these injections to check the stability of the instrument. Response factors should
not differ by more than 1% of the mean.

2.2.8 Analysis of sample

Carry out 10 L injections of calibration solutions CC and CD and sample solutions SC

and SD in the following sequence and record the integrated areas of the peaks.

Injection sequence CC1, SC1, SC2, CD1, CC2, SD1, SD2, CD2

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TECHNICAL

2.2.9 Calculation

Calculate the relative response factors (f1, f2 etc.) for the pair of calibration injections
which bracket the sample injections e.g. use CC1 and CD1 for sample injections SC1,
SC2 etc. and obtain the mean response factor, f. Sample analysis should be repeated if
calibration response factors f3 and f4 differ by more than 2% of the mean f.

Hs
Relative response factor f =
s x P

Where:
Hs = area of the four cypermethrin peaks in the calibration solution
s = mass of the cypermethrin analytical standard in calibration
solution (g)
P = purity of the cypermethrin standard (g/kg)

For each sample injection eg SC1, calculate the cypermethrin isomer content

Hw
Cypermethrin isomer content=
f x w

Where:
f = mean relative response factor
Hw = mean area of the four cypermethrin isomer peaks in the sample
solution
w = mass of sample (g)

2.3 Volatile hydrocarbon solvent content

2.3.1 Outline of method

The volatile hydrocarbon solvent content is determined by measuring the difference in

the weight of the sample before and after drying in a vacuum oven.

2.3.2 Apparatus

Vacuum oven capable of maintaining a temperature of 35-45C with a vacuum of

about 2430" Hg.

Desiccator: Note it is recommended to allow the sample to reach room temperature

in a desiccator, after removal from the drying oven, to minimise adsorption of
atmospheric moisture.

Petri Dishes Pyrex, 100 x 15 mm

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TECHNICAL

2.3.3 Reagents

Desiccant e.g. Indicating Drierite Absorbent, or equivalent.

2.3.4 Procedure

In duplicate, accurately weigh Petri dishes (x g). Accurately weigh 5-10 g of the
technical material into the Petri dish. (y g). Place the sample in the oven set at a
temperature of about 40C and adjust the vacuum to about 2430" Hg. Dry the sample
for about 2.5 hours. Remove the sample from the oven and allow it cool in the
desiccator and reweigh (z g). Dry the sample for an additional 30 minutes and re-
determine the weight. If the weight is constant, i.e., the relative change between two
weighings is less than 0.5%, calculate the percentage volatile hydrocarbon content.
Otherwise, continue drying the sample to constant weight.

100 (y-z)
Percentage ( w/w ) volatile hydrocarbon content =
y-x

2.4 Triethylamine content

2.4.1 Outline of method

This method is used to determine the content of triethylamine ( TEA ) in

alphacypermethrin technical material. The sample is dissolved in a mixture of glacial
acetic acid and chlorobenzene and potentiometrically titrated with standard acetous
perchloric acid. The amount of TEA is calculated from the equivalence point of the
titration curve.

2.4.2 Apparatus

Potentiometric Titrator e.g. Mettler DL25 titrator equipped with a combination glass
electrode ( Mettler DG 112), or equivalent. An Epson FX-85 or other printer to record
the results.

Titrator cups 100 mL to fit the titrator.

5 mL, 100 mL Volumetric pipettes (Class A)

1L and 100 mL Volumetric flasks (Class A)

2.4.3 Reagents

Acetous perchloric acid. 1 M solution of standard perchloric acid in glacial acetic

acid
Glacial acetic acid. Reagent grade
Potassium hydrogen phthalate A.C.S. Primary standard (99.95-100.05%)
Chlorobenzene HPLC Grade

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Acetous perchloric acid 0.01 M. Pipette 100 mL of the 0.1M acetous perchloric acid
into a 1L volumetric flask. Dilute to mark with glacial acetic acid.
Solution Accurately weigh 204 mg of potassium hydrogen phthalate into a 100 mL
volumetric flask. Record the weight to the nearest 0.01 mg. Dissolve in glacial acetic
acid.

2.4.4 Procedure

Blank determination (Repeat in triplicate)

Add 10mL of chlorobenzene to a titrator and fill to 100 mL with glacial acetic acid.
Determine the equivalence point and record the volume at this point. Average the
recorded volumes together and use this as the blank value.

Standardization of 0.01M Perchloric (Repeat in triplicate)

Pipette 5 mL of the potassium hydrogen phthalate solution into a titrator cup. Add 10
mL of chlorobenzene and fill to 100 mL with glacial acetic acid. Attach vessel to
titrator, and determine the equivalence point and record the volume at this point.
Determine the concentration of the perchloric solution using the calculation found in
section 2.4.7. Average the results.

Analysis of samples

All weighings should be accurate to within 0.1 mg.

Accurately weigh 3 to 4 g of technical material into the titrator vessel. Record the
weight to the nearest 0.1 mg. Add 10 mL of chlorobenzene and fill to 100 mL with
glacial acetic acid. Attach the vessel to titrator, and turn on stirrer to dissolve sample.
Determine the equivalence point and record the volume at this point.

2.4.5 Calculation

The concentration of the potassium hydrogen phthalate standard solution is calculated

by the following equation:

m1
C1 =
204.22 x V1
where:

C1 = concentration of potassium hydrogen phthalate (mmoles/mL)

m1 = mass of potassium hydrogen phthalate (mg)
V1 = volume of potassium hydrogen phthalate solution (mL)

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The concentration of the perchloric solution is determined with the following

equation:

C1V1
C2 =
V3 - V4

where:

C2 = concentration of standard perchloric solution (mmoles/mL)

C1 = concentration of potassium hydrogen phthalate solution
(mmoles/mL)
V2 = volume of standard potassium hydrogen phthalate solution (mL)
V3 = volume of perchloric to reach equivalence point (mL)
V4 = volume average blank value (mL)

The weight percent of triethylamine is determined with the following equation:

(V5-V4) x C2 x 101.2 x 100

weight % triethylamine =
m2
where:

V5 = volume of perchloric used to reach equivalence point (mL)

V4 = volume of average blank value (mL)
C2 = concentration of perchloric solution (mmol/mL)
m2 = mass of alphacypermethrin in sample (mg)

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ALPHACYPERMETHRIN WHO/SIF/61
AQUEOUS SUSPENSION CONCENTRATE

ALPHACYPERMETHRIN
AQUEOUS SUSPENSION CONCENTRATE
Full specification: WHO/SIF/61
Adopted 10 December 1999

1. Specification
1.1 Description

The material consists of a suspension of fine particles of technical alphacypermethrin,

complying with the requirements of the WHO specification WHO/SIT/32, in an
aqueous phase together with suitable formulants; After gentle agitation the material
shall be homogeneous and suitable for further dilution in water.

1.2 Chemical and physical requirements

The material sampled from any part of the consignment (see method WHO/M/1.R1),
shall comply with the requirements of section 1.1 and with the following
requirements.

1.2.1 Alphacypermethrin cis 2 ([IR cis] S and [IS cis] R isomers) content (g/kg
basis)

The content of alphacypermethrin cis 2 ([IR cis] S and [IS cis] R isomers) content
(g/kg basis), determined by the method described in section 2.1 shall not differ from
the declared content by more than the following amount:

Declared content Tolerance permitted

Up to 25 g/kg 15% of the declared content

Above 25 up to 100 10% of the declared content
g/kg

Higher declared contents are not currently available.

The average content of all samples taken shall not be lower than the declared content.

1.2.2 Alphacypermethrin cis 1 ([IR cis] R and [IS cis] S isomer) content

The alphacypermethrin cis 1:cis 2 isomer ratio determined by the method described in
section 2.1.10, shall not be higher than 5:95.

1.2.3 pH range

The pH of the concentrate, determined by the CIPAC method MT 75 (CIPAC

Handbook F, p.205), shall be between 7.0 and 8.7.

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1.2.4 Rinsability

The maximum non volatile rinsed residue as determined by CIPAC MT 148.1 (in
press)1 shall not exceed 0.05%.

1.2.5 Spontaneity of dispersion at 30C

A minimum of 60% of the alphacypermethrin content found under 1.2.1 shall be in

suspension after 5 min in WHO standard hard water C at 30 2C as determined by
CIPAC method MT 160 (CIPAC Handbook F, p.391).

1.2.6 Suspensibility after heat stability treatment

In WHO standard hard water. When tested by the method described in section 2.2, a
minimum of 70% of the alphacypermethrin content (0.63 g/L) shall be in suspension
30 minutes after agitating a suspension containing (0.9 g/L) of alphacypermethrin
prepared in standard hard water at 30 2C from the suspension concentrate subjected
to the heat stability treatment described in section 2.3.

1.2.7 Wet sieve test

Not less than 98% of the concentrate shall pass through a 75 m sieve when tested by
the CIPAC method MT 59 (CIPAC Handbook F, p.177).

1.2.8 Persistent foam

The persistent foam at the top of a 100 mL of suspension prepared in WHO standard
soft water, shall not exceed 20 mL after 12 minutes when tested by the CIPAC method
described MT 47.2 (CIPAC Handbook F, p.152)

1.2.9 Heat stability

The suspension after treatment as described in section 2.3 must comply with the
requirements of sections 1.2.1, 1.2.2, 1.2.3, 1.2.4, 1.2.5, and 1.2.7 of this specification.

1.2.10 Cold stability

After storage testing at 0 1C for 7 days, according to the CIPAC method MT 39.1
(CIPAC Handbook F, p.128), the product shall continue to comply with 1.2.5, 1.2.6
and 1.2.7.

1.3 Packing and marking of packages

The alphacypermethrin suspension concentrate shall be packed in suitable clean drums

as specified in the order. The shall be made of high density polyethylene or any
suitable materials compatible with the product..

1
Method can be obtained on request from CIPAC secretariat, t Gotink 7, 7261 VE Ruurlo,
Netherlands.

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AQUEOUS SUSPENSION CONCENTRATE

All packages shall bear, durably and legibly marked on the container, the following:
Manufacturers name
Alphacypermethrin suspension concentrate
Alphacypermethrin.......g/l
Batch or reference number, and date of test
Net weight of contents
Date of manufacture
Instruction for use

and the following minimum cautionary notice:

Alphacypermethrin is a pyrethroid that acts predominantly on the central nervous system; high
dosages have been found to cause tonic seizures in experimental animals. A high
concentration in air may be an irritant and contact with the concentrated product may induce a
temporary tingling sensation, particularly on the face. It may be hazardous if swallowed. Do
not inhale spray mist. Avoid skin contact; wear protective gloves, clean protective clothing,
and a face mask (surgical type) when handling the product. Wash hands and exposed skin
thoroughly after using.

Keep containers out of reach of children and well away from foodstuffs and animal feed and
their containers.

Alphacypermethrin is toxic to aquatic wildlife. Avoid accidental contamination of water.

If poisoning occurs, call a physician. Treatment is symptomatic.

2 Methods of determining chemical and physical properties

2.1 Alphacypermethrin content

2.1.1 Outline of method

The sample is extracted in tetrahydrofuran with di-octyl phthalate used as the internal
standard. Separation is carried out in the split injection mode using a dimethyl
polysiloxane WCOT fused silica capillary column with automatic injection and flame
ionisation detector. The alphacypermethrin is determined by comparison with
calibration solutions.

2.1.2 Apparatus

Gas chromatograph. Capable of operating over the range 100 to 300C with a flame
ionisation detector, split/splitless injector and autosampler.

Column. 30 m WCOT fused silica 0.25 mm id and 0.25 m film thickness coated
with dimethyl polysiloxane (chemically bonded), or equivalent.

Injector system. Injector Split/splitless with fused silica liner containing a 1 cm plug
of glass wool. An automatic injector is strongly recommended to ensure the best
analytical results.

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ALPHACYPERMETHRIN WHO/SIF/61
AQUEOUS SUSPENSION CONCENTRATE

It is important to ensure that the split liner is acid treated, thoroughly deactivated and
conditioned before use, to ensure that alphacypermethrin does not epimerise during
analysis.

Split ratio 75-100:1 (split vent at 75 mL/min)

Injection Volume 1.0 L using a autosampler.

Detector system. Type FID, with no special requirements.

Automatic digital integrator or chromatography data system. Compatible with the gas
chromatograph

100 mL volumetric flasks.

2.1.3 Reagents

Tetrahydrofuran.

Citric acid. 5% citric acid solution.

Di-octyl phthalate, di-(2-ethylhexyl)phthalate, Internal standard. Select for use a batch
which, when chromatographed under the conditions below for the
determination of alphacypermethrin gives no peak with a similar retention time
to the alphacypermethrin isomers.
Cypermethrin analytical standard (containing a mixture of trans 3, trans 4, cis 1 and
cis 2 cypermethrin) of known purity
Alphacypermethrin working standard, of known alphacypermethrin content

2.1.4 Preparation of standard solutions

Internal standard solution.

Dissolve di-octyl phthalate (5.0 g) in tetrahydrofuran (500 mL) (solution I). Ensure
sufficient quantity of this solution is prepared for all samples and calibration standards
being analysed.

Citric acid. 5% citric acid solution.

Dissolve citric acid (25 g) in water (500 mL).

Alphacypermethrin calibration solution.

Weigh in duplicate (to the nearest 0.1 mg) about 0.1 g of alphacypermethrin (CA and
CB, g) into 100 mL volumetric flasks. Add to each, 70 mL of tetrahydrofuran and
agitate the flask gently to dissolve. Add by pipette 10.0 mL of internal standard
solution (solution I) and 10mL of 5% citric acid solution from a measuring cylinder.
Make to volume with tetrahydrofuran and mix well. (Solutions CA and CB).

Note: Citric acid is added to stop epimerisation of alphacypermethrin in solution.

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ALPHACYPERMETHRIN WHO/SIF/61
AQUEOUS SUSPENSION CONCENTRATE

Prepare a solution without internal standard, by dissolving about 0.1 g of

alphacypermethrin standard into a 100 mL volumetric flask. Add approximately 70
mL of tetrahydrofuran and swirl to dissolve. Add by pipette 10 mL of 5% citric acid
solution from a measuring cylinder. Make to volume with tetrahydrofuran and mix
well. (Solution CO).

Cypermethrin calibration check solution

Prepare a solution of cypermethrin, by dissolving about 0.1 g of cypermethrin standard

into a 100 mL volumetric flask. Add approximately 70 mL of tetrahydrofuran and
swirl to dissolve. Add by pipette 10 mL of the internal standard solution to volume
with tetrahydrofuran and mix well (Solution CC).

2.1.5 Operating conditions

The conditions given below are typical values and may have to be adjusted to obtain
optimum results from a given apparatus.

Temperatures

Column oven 230C

Injector 260C
Detector 300C

Adjust the column oven temperature if required to obtain retention time windows for
di-octyl (approximately 13.5 minutes), alphacypermethrin cis 2 isomer (approximately
27.0 minutes) and alphacypermethrin cis 1 isomer (approximately 25.2 minutes). The
trans 3 and trans 4 isomers elute at approximately 26.1 and 27.5 minutes respectively.

Gas flow rates

Helium carrier approximately 0.8mL min-1

Helium make up gas 60mL min-1 or optimum for instrument
Septum purge 2mL min-1
Hydrogen as recommended for the instrument
Air as recommended for the instrument

All gases should be purified through molecular sieves. The carrier gas should be
further purified through an oxygen trap.

2.1.6 Sample Preparation

Sampling

Homogenize the bulk material before taking the sample. Take at least 250 g. Re-
homogenize the bulk before taking a sub-sample for analysis.

Preparation of the sample solutions.

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Weigh accurately in duplicate (to the nearest 0.1mg) into 100 mL volumetric flasks
sufficient sample (w,g) to contain 0.1 g of alphacypermethrin. Add by measuring
cylinder 10 mL of the 5% citric acid solution to each of the flasks and swirl to fully
disperse the formulation. Add approximately 70 mL of tetrahydrofuran in
approximately 10 to 15 mL portions, swirling in between each addition to fully
disperse the sample. Place in an ultrasonic bath for 15 minutes with occasional
swirling. Add by pipette 10 mL of the di-octyl phthalate, internal standard solution I
and make to volume with tetrahydrofuran and mix well. Filter through a suitable paper
eg GF/A or filter unit eg PTFE 0.45 m to give clear solutions. (Solutions SA and SB).

Prepare a solution without internal standard, by dissolving sufficient sample to contain

0.1 g of alphacypermethrin into a 100 mL volumetric flask. Add by measuring
cylinder 10 mL of the 5% citric acid solution to each of the flasks and swirl to fully
disperse the formulation. Add approximately 70 mL of tetrahydrofuran in
approximately 10 to 15 mL portions, swirling in between each addition to fully
disperse the sample. Place in an ultrasonic bath for 15 minutes with occasional
swirling. Make to volume with tetrahydrofuran and mix well. Filter through a suitable
paper eg GF/A or filter unit eg PTFE 0.45 m to give a clear solution. (Solution SO).

2.1.7 Equilibration of the system.

Carry out alternate 1.0 L injections of solutions I, Co and So and check whether there
are any interfering peaks from impurities. If there are make any necessary corrections
but do not use external calibration.

Inject calibration solutions CA and CB to equilibrate the system and use the data from
these chromatograms to set the integration parameters. Calculate the response factors
for these injections to check the stability of the instrument. Response factors should
not differ by more than 1% of the mean.

Inject the solution of cypermethrin in tetrahydrofuran (Cc) to ensure complete

separation of the alphacypermethrin isomers.

2.1.8 Analysis of sample

Carry out 1.0 L injections of calibration solutions CA and CB and sample solutions SA
and SB in the following sequence and record the integrated areas of the peaks.

Injection sequence CA1, SA1, SA2, CB1, CA2, SB1, SB2, CB2.

2.1.9 Calculation.

Calculate the relative response factors (f1, f2 etc) for the pair of calibration injections
which bracket the sample injections eg use CA1 and CB1 for sample injections SA1, SA2
etc and obtain the mean response factor, f. Sample analysis should be repeated if
calibration response factors f1 and f2 differ by more than 2% of the mean f.

Hs
Relative response factor =
Ir x s x P

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where:

Hs = area of alphacypermethrin cis 2 peak in the calibration solution.

Ir = area of di-octyl phthalate peak in the calibration solution.
s = mass of alphacypermethrin analytical standard in calibration
solution (g).
P = purity of the alphacypermethrin standard (g/kg).

For each sample injection eg SA1, calculate the alphacypermethrin cis 2 content.

Hw
Alphacypermethrin content =
f x Iq x w

where:

f = mean relative response factor

Hw = mean area of alphacypermethrin cis 2 peak in the sample solution
Iq = area of di-octyl phthalate peak in the sample solution
w = mass of sample (g)

Calculate the alphacypermethrin cis 2 content of the sample as the mean of the four
determinations as follows:

Sample injection Use relative response factor Alphacypermethrin cis 2

from (g/kg)

SA1 CA1 and CB1 Q )

) X
SA2 CA1 and CB1 R )

SB1 CA2 and CB2 S )

) Y
SB2 CA2 and CB2 T )

Q and R, S and T should agree to within 0.5% of their mean values (X and Y). X
and Y should agree within 1% of their mean values.

Take the mean of the two values x and y as the alphacypermethrin content.

2.1.10 Determination of the Cis 1 to Cis 2 ratio

.
To determine the cis 1 to cis 2 percentage ratio in a sample

Peak area cis 1 x 100

cis 1 % =
Total peak area of cis 1 and cis 2

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Peak area cis 2 x 100

cis 2 % =
Total peak area of cis 1 and cis 2

2.2 Suspensibility

2.2.1 Outline of method

A suspension of known concentration of alphacypermethrin in standard hard water is

prepared, poured into a 250 mL graduated cylinder, maintained at constant
temperature and allowed to remain undisturbed for 30 minutes. The top 9/10ths are
drawn off and the content of alphacypermethrin in the bottom 1/10th is determined, so
allowing to evaluate the active ingredient mass still in suspension after 30 minutes.

2.2.2 Apparatus

A 250 m1 graduated cylinder with a ground-glass stopper and a distance of 20.0-21.5

cm between the bottom and the 250 mL calibration mark.

A glass tube, about 40 cm long and about 5 mm in internal diameter, pointed at one
end to an opening of 2-3 mm, the other end being connected to a suitable source of
suction.

2.2.3 Reagents

WHO standard hard water. See method WHO/M/29

2.2.4 Procedure

Weigh (to the nearest 1 mg) into 100 mL beaker an amount of the sample to form 250
m1 of a suspension containing 0.9 g/L of alphacypermethrin. Add a volume of water
at 30 2C equal to at least twice the mass of the sample taken. Allow to stand for 30
seconds and then stir by hand for 30 seconds with a glass rod 4-6 mm in diameter, at
not more than four revolutions per second, making no deliberate attempt to break up
any lumps. The immediately transfer the mixture quantitatively to the 250 mL
graduated cylinder, using water at 30 2C for rinsing, and again avoiding mechanical
disintegration of lumps.

Immediately add sufficient water at 30 2C to bring the volume up to the 250 mL

mark. Insert the stopper and invert the cylinder end over end 30 times at the rate of
one complete cycle every 2 seconds. This operation should be carried out as smoothly
as possible, keeping the axis of rotation fixed. Allow the graduated cylinder to stand
for 30 minutes in a water-bath at 30 2C taking care that the bath is free from
vibrations.

Should excessive flocculation occur during the test, the material is unsatisfactory.

At the end of the 30 minutes settling period, insert the glass tube into the cylinder and,
with a minimum of disturbance, withdraw nine-tenths, of the suspension (i.e. 225 mL)

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by means of the suction tube in a period of 10-15 seconds. This is achieved by

maintaining the tip of the glass tube just below the sinking surface of the suspension.
Discard the suspension withdrawn.

2.2.5 Determination of alphacypermethrin in the retained one-tenth of the

suspension

Quantitatively transfer the bottom one-tenth of suspension (25 mL) from the
suspensibility test into a 100 mL volumetric flask with approximately 50 mL of
tetrahydrofuran. Add 10 mL internal standard by pipette and 5 mL citric acid solution
by measuring cylinder. Dilute to volume with tetrahydrofuran. Place in an ultrasonic
bath for 10 minutes. Filter through a suitable disc filter.

Determine the alphacypermethrin content of the filtered solution by gas

chromatography as described in section 2.1.8 injecting duplicate 1 .0 L aliquots of
sample and calibration solutions.

2.2.6 Calculation

Mass of alphacypermethrin (g) in the bottom one-tenth of suspension =

f2 x P x w

3
f1 x 2 x 10
where:
f1 = mean response ratio for the calibration solution
f2 = mean response ratio for the sample solution
w = mass (g) of alphacypermethrin standard in the calibration solution
P = purity (g/kg) of alphacypermethrin standard

10 (b-a) x 100 = 111(b-a)

Suspensibility % =
9 b b

where:

a= mass (g) of alphacypermethrin in bottom one-tenth of suspension

b= mass (g) of alphacypermethrin in the sample used in the suspensibility test
(calculated from the mass of sample and its percentage alphacypermethrin content).

2.3 Heat stability treatment

54C 2C for 14 days (CIPAC method MT 46.1, CIPAC Handbook F, p.149), unless
other temperatures and times are requested (FAO Manual on the development and use of
FAO specifications for plant protection products, n 149, p.33).

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After completion of the heat stability treatment, the samples should not be exposed to
heat, bright sunshine , or atmospheric humidity.

If required the test should be conducted in commercial type pack.

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