CARBOSULFAN 417

CARBOSULFAN
417

O CH3
C4H9
O C13
N S N
O C4H9
CH3
CH3

ISO Common Name Carbosulfan

Chemical name 2,3-Dihydro-2,2-dimethyl-benzofuran-7-yl (dibutylamino-
thio) methylcarbamate (IUPAC); 2,3-dihydro-2,2-dimethyl
7-benzofuranyl [(di-n-butylamino)thio] methylcarbamate
(CA; 55285-14)
Empirical formula C20H32N2O3S
RMM 380.5
d20 1.056
v.p. 0.041 mPa at 25 C
Solubility In water: 0.3 mg/l at 25 C
Description Orange to brown clear viscous liquid with a mild phenolic
odour.
Formulations Wettable powders, water dispersible granules, granules and
emulsifiable concentrates.

CARBOSULFAN TECHNICAL
*
417/TC/M/-

1 Sampling. Take at least 100 g.

2 Identity test
2.1 HPLC. Use the HPLC method below. The relative retention time of carbosulfan
with respect to the internal standard for the sample solution should not deviate by
more than 1% from that for the standard solution.

*
CIPAC method 1992. Based on a method supplied by FMC Corporation USA.

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 CARBOSULFAN 417

3 Carbosulfan
OUTLINE OF METHOD. The sample is dissolved in an internal standard solution
containing nonaphenone. The constituents are separated by high performance liquid
chromatography (HPLC) on a reverse phase column (C8) with methanol/water
(88+12) mobile phase.
REAGENTS
Methanol HPLC grade
Acetonitrile HPLC grade
Water HPLC grade
Mobile phase methanol/water, 88+12 (v/v)
Carbosulfan analytical standard of known purity
Nonaphenone internal standard, free from components which co-elute with
carbosulfan under HPLC conditions described in Section (a)
Internal standard solution. Weigh (to the nearest 0.1 mg) 3.6 g of n-nonaphenone (r g)
into a 1000 ml volumetric flask, dissolve in acetonitrile, and dilute to 1000 ml with
acetonitrile.
Calibration solution. Weigh (to the nearest 0.1 mg) about 100 - 120 mg (s g) of
carbosulfan analytical standard into a 40 ml sample vial. Accurately pipette 25 ml
of the internal standard solution, cap the vial and shake to dissolve the
carbosulfan. Pipette 5 ml of the solution into a 25 ml volumetric flask and make
up to volume with acetonitrile. Prepare a second calibration solution of similar
concentration. Prepare new calibration solutions daily.

APPARATUS
HPLC system consisting of :
High precision HPLC pump
Injection valve with 10 µl loop
Stainless steel column : 250 mm × 4.6 mm packed with Zorbax C8, 5 µm, or
equivalent C8 column.
UV detector set at 280 nm with linear response.
Data system with suitable electronic integrator for peak evaluation.
PROCEDURE
(a) Operating conditions (typical) :
Column Stainless steel 250 mm × 4.6 mm
Stationary phase Zorbax® C8, 5 m, pre-packed, or equivalent C8
column. If a C8 column is not available, a C18 reverse
phase LC column may be substituted.

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 CARBOSULFAN 417

Mobile phase Methanol/water 88+12 (v/v)

Injection volume 10 µl
Flow rate 1.0 ml/min
Column temperature Ambient
Detection UV detector at 280 nm
Chart speed 0.5 cm/min
Retention times Nonaphenone: 8.5 min
Carbosulfan: 11.5 min
The ideal retention time for nonaphenone is 8-9 min.
Minor adjustments to mobile phase composition or
flow rate may be required to maintain the above
retention times especially if a C18 reverse phase
column is substituted.

(b) Equilibration of system Inject 10 µl portions of both the calibration solutions.

Inject each calibration solution at least two times and calculate the average response
factor. The individual values should not deviate from the mean by more than 0.5%,
otherwise repeat the calibration.

(c) Preparation of sample solution Weigh (to the nearest 0.1 mg) sufficient sample
to contain about 100 - 120 mg (w g) of carbosulfan into a 40 ml sample vial. Pipette
25 ml of internal standard solution. Hand shake the sample to dissolve the
carbosulfan. Pipette 5 ml of each sample into a 25 ml volumetric flask and dilute to
volume with acetonitrile. Prepare three weights for each sample.

(d) Determination Inject in duplicate 10 µl portions of each sample solution

bracketing them by injections of the calibration solutions as follows: calibration
solution I, calibration solution I, sample solution 1, sample solution 1, calibration
solution II, calibration solution II, sample solution 2, sample solution 2, calibration
solution I, calibration solution I, sample solution 3, sample solution 3, calibration
solution II, calibration solution II, and so on for other sample solutions. Measure and
record the relevant peaks areas.
Calculate the mean value of each pair of calibration factors bracketing the two
sample injections and use this value for calculating the two bracketed sample runs.

(e) Calculation
Ir s P
f
Hs r

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 CARBOSULFAN 417

Hw r f
Carbosulfan content g/kg
Ig w
where:
f = relative response factor
r = mass of internal standard in the calibration and sample solution (g)
s = mass of carbosulfan standard in the calibration solution (g)
w = mass of sample taken (g)
Ir = peak area of the internal standard in the calibration solution
Ig = peak area of the internal standard in the sample solution
Hs = peak area of carbosulfan in the calibration solution
Hw = peak area of carbosulfan in the sample solution
P = purity of the carbosulfan standard (g/kg)
Calculate the carbosulfan content of the sample as the mean value of three
determinations as follows:
Sample Mean bracketed Content of
Mean
injection response factor carbosulfan
SA1 C1 and C2 Q
SA2 C1 and C2 R } X g/kg

SB1 C1 and C2 S
SB2 C1 and C2 T } Y g/kg

SC1 C1 and C2 U
SC2 C1 and C2 V } Z g/kg

Repeatability r = 7.4 g/kg at 882.4 g/kg active ingredient content

Reproducibility R = 13.0 g/kg at 882.4 g/kg active ingredient content

CARBOSULFAN WETTABLE POWDERS

*
417/WP/M/-
1 Sampling. Take at least 500 g.
2 Identity test
HPLC. As for carbosulfan technical 417/TC/M/2.

*
CIPAC method 1992. Based on a method supplied by FMC Corporation USA.

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 CARBOSULFAN 417

3 Carbosulfan. As for carbosulfan technical 417/TC/M/3 except:

(c) Preparation of sample solution. Weigh (to the nearest 0.1 mg) sufficient sample
to contain about 100-120 mg (w g) of carbosulfan into a 40 ml sample vial. Pipette
25 ml of internal standard solution. Extract the sample for 20 minutes using a
reciprocating shaker and allow the solid material to settle. Continue according to
417/TC/M/3 (c) beginning at : "Pipette 5 ml ..."

Repeatability r = 11.9 g/kg at 257.6 g/kg active ingredient content

Reproducibility R = 13.3 g/kg at 257.6 g/kg active ingredient content

4 Suspensibility

APPARATUS AND REAGENTS As for 417/TC/M/3 and MT 15.

PROCEDURE

(a) Preparation of suspension : MT 15.1 (i)

(b) Determination of sedimentation : MT 15.1 (ii)
(c) Determination of carbosulfan in the bottom 25 ml of suspension. After removal
of the top 225 ml of suspension, determine the amount of carbosulfan in the bottom
25 ml of suspension by 417/TC/M/3.
Hw q f
Mass of carbosulfan (Q)
Iq
where:

Hw = peak area of carbosulfan in the sample solution

Iq = peak area of the internal standard in the sample solution
f = relative response factor
q = mass of internal standard in the calibration and sample solution (g)

(e) Calculation
111 ( c - Q )
Suspensibility =
c
where:

c = mass of carbosulfan in the sample taken for preparing the suspension

(mg)
Q = mass of carbosulfan in the in the bottom 25 ml of suspension (mg)

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 CARBOSULFAN 417

CARBOSULFAN WATER DISPERSABLE GRANULES

*
417/WG/M/-

1 Sampling. Take at least 1 kg.

2 Identity test

HPLC. As for carbosulfan technical 417/TC/M/2.

3 Carbosulfan. As for carbosulfan wettable powders 417/WP/M/3.

4 Suspensibility
APPARATUS AND REAGENTS As for 417/TC/M/3 and MT 161 CIPAC F p. 394

PROCEDURE

(a) Preparation of suspension and determination of sedimentation. MT 161

(b) Determination of carbosulfan in the bottom 25 ml of suspension. As for
417/WP/M/4(c)

CARBOSULFAN EMULSIFIABLE CONCENTRATE

*
417/EC/M/-

1 Sampling. Take at least 500 ml.

2 Identity test

HPLC. As for carbosulfan technical 417/TC/M/2.

3 Carbosulfan. As for carbosulfan technical 417/TC/M/3.

Repeatability r = 4.0 g/kg at 275.2 g/kg active ingredient content

4.5 g/kg at 484.0 g/kg active ingredient content

Reproducibility R = 6.8 g/kg at 275.2 g/kg active ingredient content

11.3 g/kg at 484.0 g/kg active ingredient content
*
CIPAC method 1992. Based on a method supplied by FMC Corporation USA.

40
 CARBOSULFAN 417

CARBOSULFAN GRANULES
*
417/GR/M/-

1 Sampling. Take at least 1 kg.

2 Identity test
HPLC. As for carbosulfan technical 417/TC/M/2.

3 Carbosulfan : As for carbosulfan wettable powders 417/WP/M/3.

Repeatability r = 3.4 g/kg at 104.2 g/kg active ingredient content

Reproducibility R = 3.4 g/kg at 104.2 g/kg active ingredient content

*
CIPAC method 1992. Based on a method supplied by FMC Corporation USA.

41