### Page 1 of 17 Study Title

Adsorption Coefficient of

1,1’-(1,6-hexanediyldiimino)bis[1-oxo-methanesulfonic acid], sodium salt (1:2)

Data requirements

REACH requirement EC/1907/2006

Author

M. T. Garcia-Sanchez

Study completion date

2012-02-16

LANXESS Deutschland GmbH

51369 Leverkusen

Germany

Testing facility

CURRENTA GmbH & Co. OHG

Analytik

51368 Leverkusen

Germany

Laboratory Project Identification

Study No.: 2011/0125/04

### Page 2 of 17 CURRENTA GmbH & Co. OHG

Analytik Study No.: 2011/0125/04

This page is intentionally left blank for the purpose of submitting administrative

information that is required by regulations promulgated by various countries.

### Page 3 of 17 CURRENTA GmbH & Co. OHG

Analytik Study No.: 2011/0125/04

CONTENTS

Page



### 1. GLP DECLARATION 4

2. ARCHIVING 4

3. QUALITY ASSURANCE STATEMENT _ 6

4. STUDY TIME TABLE 6

5. GLP CERTIFICATE 7

6. SUMMARY _ 8

7. EXPERIMENTAL PROCEDURE 9

7.1. Methods, guidelines and documents 9

7.2. Principle of the test 9



### 8. TEST MATERIAL 11

8.1. Sample description 11

8.2. Calibration substances 11



### 9. METHODS AND RESULTS 12

9.1. Determination of the pH-value of the aqueous part of the mobile

phase 12

9.2. Determination of the adsorption coefficient 12

9.2.1. Preparation of the test solutions 12

9.2.2. HPLC-method parameters 12

9.2.3. Example chromatograms 14

9.2.4. Calibration 15

9.2.5. Determination of the test item 16Page 4 of 1 7 CURRENTA GmbH & Co. OHG Study No.: 201110125104

Analytik



### 1. GLP DECLARATION

This study was conducted in compliance with the OECD principles of Good

Laboratory Practice (1999) and with the Principles of Good Laboratory Practice

according to Annex 1, German Chemical Law (2008).

Date 

### I Signature

Study Director

<

M. 

### T. Garcia-Sanchez

Date 

### I Signature

Head of Test Facility



### 2. ARCHIVING

The original report, the study plan, and all raw data pertaining to this study are stored

in the "GLP Archive, CURRENTA GmbH & Co. OHG, Analytik , CHEMPARK

, Building Q 18, D-51368 Leverkusen". A sample of the test item is stored in

"GLP-Sample Store, CURRENTA GmbH & Co. OHG, Analytik

, CHEMPARK

, Building DA 1, D-41538 Dormagen".

### Page 5 of 17 CURRENTA GmbH & Co. OHG Study No.: 2011/0125/04

Analytik

This page is intentionally left blank for the purpose of submitting administrative

information that is required by regulations promulgated by various countries.Pag e 6 of 17 CURRENTA GmbH & Co. OHG

Analytik Study No. : 201110125104



### 3. QUALITY A S SURANC E S TAT E M E NT

This report was audited by the Quality Assurance Unit Currenta, Analytik, Quality

Management at Currenta GmbH & Co . OHG and this statement confirms that the

final report reflects the raw data

.

The dates of Quality Assurance inspections and audits are given below.

Audits Dates of QAU

Inspections Dates of

Reports

study plan rev i ew

process based inspection

final report review (draft)

final report review 2012-01-12

2011-12-20

2012-01-23



### . 2l'JI 

### 2 0 

### 2 I 

## G 2012-01-12

2011-12-20

2012-01-23



### J..-o/2 o;: 

### -IG

Date 

### I Signature

012 

### 02 

## ·IIG 

# // dfl-·A '(

Ms Senic 

### I Brl3err-B-asl:l-baA§e-1-Br-Netlpert



## 4. STUDY TIM E TABL E

Study initiation d a te: 2012-01-12

Study completion date: 2012-02-16

St a rt of experimental t e sts: 2012-01-18

End of ex p e rim e nt a l t es t s: 2012-01-20

### CURRENTA GmbH & Co. OHG

Analytik Study No.: 2011/0125/04

Page 7 of 17 

### 5. GLP CERTIFICATE

Please note:

Effective January 1st, 2008, the company name Bayer Industry Services GmbH & Co.

OHG was changed to CURRENTA GmbH & Co. OHG.

### CURRENTA GmbH & Co. OHG

Analytik Study No.: 2011/0125/04

Page 8 of 17 

### 6. SUMMARY

For 1,1’-(1,6-hexanediyldiimino)bis[1-oxo-methanesulfonic acid], sodium salt (1:2),

batch no.: RJE 76081 the adsorption coefficient was estimated by the HPLC-method

according OECD TG 121.

The adsorption coefficient of the test item is estimated to be:

log K

oc 

### = < - 3.6

Extrapolated value.

Retention time of the test item set to 2.0 min. for the calculations (see 9.2.5)

### CURRENTA GmbH & Co. OHG

Analytik Study No.: 2011/0125/04

Page 9 of 17 

### 7. EXPERIMENTAL PROCEDURE

7.1. Methods, guidelines and documents

The determination of the partition coefficient by the HPLC-method was based on the

following guidelines:

OECD Guidelines for Testing of Chemicals No.: 121 “Estimation of the Adsorption

Coefficient (Koc) on Soil and on Sewage Sludge using High Performance Liquid

Chromatography (HPLC)” (adopted, January 2001)

SOP 00193 Version 3, this SOP corresponds to Council Regulations EC No

440/2008, Guideline Part C – Methods for the Determination of Ecotoxicity, C.19.

“Estimation of the Adsorption Coefficient (Koc) on Soil and on Sewage Sludge using

High Performance Liquid Chromatography (HPLC)” (adopted, May 2008).

SOP 00178 Version 2 for the determination of the pH-value from the aqueous part of

the mobile phase.

7.2. Principle of the test

The adsorption coefficient (K) is a partition coefficient and is defined as the ratio of

the equilibrium concentrations of a dissolved substance in a two-phase system

consisting of a sorbent (soil or sewage sludge) and an aqueous phase.

K

oc 

### = C soil (sludge)



### C

water



### HPLC is performed on analytical columns packed with a commercially available

cyano solid phase chemically bound onto silica. Chemicals injected onto such a

column move along it by partitioning between the mobile solvent phase and the

stationary cyano-phase. The chemicals are retained in proportion to their solid

phase/water partition coefficient, with water-soluble chemicals eluted first and soil

(sludge)-soluble chemicals last. This enables the relationship between the retention

time on a cyano phase column and the soil(sludge)/water adsorption coefficient to be

established. The adsorption coefficient is deduced from the capacity factor (k), given

by the expression where, t R 

### is the retention time of the test item, and t 0 

### is the deadtime, i.e. the average time a solvent molecule needs to pass the column. Quantitative

analytical methods are not required; only the determination of retention times is

necessary.

The HPLC method enables adsorption coefficients to be estimated in the log K

oc 

### range between 1.5 and 5. The pH value has a significant influence on sorption

behaviour in particular for polar substances. For agricultural soils or tanks of sewage

treatment plants pH normally varies between pH 5.5 and 7.5. For this reason the test

is performed at pH 6 (e.g. by adding citrate buffer to the mobile phase).

### CURRENTA GmbH & Co. OHG

Analytik Study No.: 2011/0125/04

Page 10 of 17 In case more than 10 % of the test item is dissociated at pH 6, a second test has to

be performed at a pH where the test item is undissociated.

The test substance is a sodium salt of an aliphatic sulfonic acid, these salts are

dissociated in water. Therefore there is no pH range where the nonionic form of the

test item exists. The test was performed at pH 6 (environmental relevant)

### CURRENTA GmbH & Co. OHG

Analytik Study No.: 2011/0125/04

Page 11 of 17 8.1. 

### 8.

Sample description 

### TEST MATERIAL

Test item: 1,1’-(1,6-hexanediyldiimino)bis[1-oxomethanesulfonic acid], sodium salt (1:2)

CAS name: 1,1’-(1,6-hexanediyldiimino)bis[1-oxomethanesulfonic acid], sodium salt (1:2)

CAS number: 38632-47-2

Empirical formula: C 8 

### H 14 

### N 2 

### O 8 

### S 2. 

### 2Na

Molecular mass: 376.3 g/mol

Batch number: RJE 76081

Sample number: 1452

Date of receipt: 2011-10-07

Expiry date: 2013-10-10

Storage conditions: Room temperature

Structural formula:

2-

O O

O H O

–

S N

– O H

O N S

O

2 Na

O



### 8.2. Calibration substances

The reference substances used for calibration are mentioned in paragraph 9.2.4.

### CURRENTA GmbH & Co. OHG

Analytik Study No.: 2011/0125/04

Page 12 of 17 

### 9. METHODS AND RESULTS

9.1. Determination of the pH-value of the aqueous part of the mobile

phase

Method: SOP 00178 version 2

Supervisor: J. Leibold

Procedure:

The pH determination of the aqueous part of the mobile phase was done with a pH

meter equipped with a calibrated single-rod glass electrode.

Result: pH 6.3

9.2. Determination of the adsorption coefficient

Method: SOP 00193 Version 3

Supervisor: J. Leibold

9.2.1. Preparation of the test solutions

The calibration substances solutions were dissolved in a mixture of 30 % water and

70 % acetonitrile. The concentrations of the calibration substances after dilution with

acetonitrile to the measuring concentration were between 0.05 mg /ml and 0.17

mg/ml.

The solution for the determination of the dead time was prepared by dissolving

sodium nitrate in a mixture of 30 % water and 70 % acetonitrile. The sodium nitrate

solution was diluted to a final concentration of 0.11 mg/ml with acetonitrile.

The test item was dissolved in water. The concentration of the test item after dilution

with acetonitrile to the measuring concentration was 0.31 mg/ml.

9.2.2. HPLC-method parameters

Equipment: HPLC Agilent 1100

Column: LiChrospher 100 CN, 250 mm length,

4 mm internal diameter, 5 μm particle size

Eluent A: 835 ml water + 165 ml buffer pH 6

### CURRENTA GmbH & Co. OHG

Analytik Study No.: 2011/0125/04

Page 13 of 17 Eluent B: acetonitrile

Solvent program: Isocratic 70 % A and 30 % B

Flow: 1 ml/min

Column temperature: 40 °C

Injection volume: 5 μL

Detector: UV, 210 nm

Acetonitrile, Fa. Merck, Lichrosolve, Art-Nr.: 1.00030

Buffer pH 6, Fa. Sigma Aldrich, Art-Nr.: 33545

Demineralized water

Remark:

In order to ensure a stable performance of the HPLC column, the column temperature was set to 40°C.

The reference Koc-values in the OECD guideline presumably refer to 25°C. Even though the column

temperature was 40°C, this temperature has no influ ence on the result of the determination. Therefore

the Koc -value of the test item determined within this study also refers to 25 °C

### CURRENTA GmbH & Co. OHG

Analytik Study No.: 2011/0125/04

Page 14 of 17 9.2.3. Example chromatograms

Figure 1: HPLC-chromatogram of the calibration substances and the test item

Figure 2: HPLC-chromatogram of the test item.

### CURRENTA GmbH & Co. OHG

Analytik Study No.: 2011/0125/04

Page 15 of 17 Results

9.2.4. Calibration

Table 1: Calibration substances (individual values)

Calibration substance Retention time

t R in min

Test 1 Retention time

t R in min

Test 2 Retention time

t R in min

Test 3 Retention time

t R in min

Mean value

Sodium nitrate (dead time (t 0 )) 1.907 1.907 1.908 1.907

Acetanilide 3.848 3.848 3.849 3.848

Methylbenzoate 5.150 5.149 5.151 5.150

Thiazoxide 6.473 6.473 6.472 6.473

Naphthalene 8.530 8.526 8.523 8.526

1,2,3-Trichlorobenzene 9.803 9.796 9.792 9.797

Phenanthrene 14.216 14.195 14.185 14.199



### Table 2: Calibration substances (mean values)

Calibration substance Retention time

t R in min

Mean value Capacity

factor k’ log k’ log K

oc *

Sodium nitrate (dead time (t 0 )) 1.907

- - -

Acetanilide 3.848 1.018 0.008 1.25

Methylbenzoate 5.150 1.700 0.230 1.80

Thiazoxide 6.473 2.393 0.379 2.44

Naphthalene 8.526 3.470 0.540 2.75

1,2,3-Trichlorobenzene 9.797 4.137 0.617 3.16

Phenanthrene 14.199 6.444 0.809 4.09

* OECD Guidelines for Testing of Chemicals No.: 121, table 3, recommended reference

substances.

### CURRENTA GmbH & Co. OHG

Analytik Study No.: 2011/0125/04

Page 16 of 17 Test item Retention

time t R in

min

Test 1 Retention time

t R in min

Test 2 Retention time

t R in min

Test 3 Retention time

t R in min

Mean value

1,1’-(1,6-hexanediyldiimino)bis[1-

oxo-methanesulfonic acid],

sodium salt (1:2) 1.671 1.671 1.672 1.671 

### a = - 0.2985

b = 0.282

r = 0.989 Capacity factor k’ = (t R 

### – t 0 

### ) / t 0



### t R 

### : retention time of the substance

t 0 

### : dead time

Effective correlation log k’ = a + b log Koc

Linear regression gives:

where a and b are the regression factors and r is the correlation factor.

9.2.5. Determination of the test item

Table 3: Test item (individual values)

Table 4: Test item evaluation

Test item Retention

time t R in

min

Mean value Capacity

factor k’ log k’ log K

oc

1,1’-(1,6-hexanediyldiimino)bis[1-

oxo-methanesulfonic acid],

sodium salt (1:2) < 2.0 < 0.049 < - 1.313 < - 3.59

### Page 17 of 17 CURRENTA GmbH & Co. OHG Study No.: 2011/0125/04

Analytik

Remark:

No reference substance with a log K

oc 

### value under the value for the test item is

available. The result was extrapolated.

The test item showed a lower retention time (t

r 

### =1.67 min.) than the dead time marker

sodium nitrate (t

r 

### =1.91 min.). For the calculation of the adsorption coefficient, the

retention time for the test item is set to t

r 

### = 2.0 min. Otherwise a calculation would

have not been possible.

Figure 3: Graphic of the linear regression log k’ / log K

oc 

### of test item

and calibration substances

The adsorption coefficient of the test item is estimated to be:

log K

oc 

### < - 3.6

Extrapolated value.

Retention time of the test item set to 2.0 min. for the calculations