Registration Dossier

Data platform availability banner - registered substances factsheets

Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.

The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.

Diss Factsheets

Some information in this page has been claimed confidential.

Administrative data

Endpoint:
mode of degradation in actual use
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2002
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
test procedure in accordance with generally accepted scientific standards and described in sufficient detail
Cross-reference
Reason / purpose for cross-reference:
reference to other study
Reference
Endpoint:
biodegradation in water: ready biodegradability
Type of information:
experimental study
Adequacy of study:
key study
Study period:
29 August 2001 - 26 September 2001
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 301 C (Ready Biodegradability: Modified MITI Test (I))
GLP compliance:
yes (incl. QA statement)
Oxygen conditions:
aerobic
Inoculum or test system:
activated sludge (adaptation not specified)
Details on inoculum:
ACTIVATED SLUDGE
1) Mixed liquor suspended solid (MLSS) : 4500 mg/L
2) Source : Chemical Evaluation and Research Institute, Japan
3) Date of receipt : July 19, 2001
Duration of test (contact time):
28 d
Initial conc.:
30 mg/L
Based on:
test mat.
Remarks:
measured with HPLC
Parameter followed for biodegradation estimation:
DOC removal
Details on study design:
The test substance was exposed to the activated sludge in a closed-system oxygen consumption measuring apparatus. The biochemical oxygen demand (BOD) was measured over a 28 day period, After this period, the concentrations of the dissolved organic carbon (DOC) and the residual test substance in the test bottles were measured. The biodegradability of the test substance was evaluated from these results.

ACTIVATED SLUDGE
1) Mixed liquor suspended solid (MLSS) : 4500 mg/L
2) Source : Chemical Evaluation and Research Institute ,Japan
3) Date of receipt : July 19, 2001

EXPOSURE CONDITIONS_
1) Temperature : 25 +/- 1˚C
2) Exposure period : 28 days
3) Test volume : 300 mL
4) Concentration : test substance (bottles 3-6) : 100 mg/L
aniline* (bottle 1) : 100 mg/L
activated sludge (bottles 1-5) : 30 mg/L
* Reference substance : KANTO CHEMICAL Co., INC. Lot No.212G1294

Test bottle contents :
Bottle 1 : Aniline + activated sludge + basal medium 29 µL (30 mg) of aniline was added to the basal medium*, then activated sludge was added.
Bottle 2 : Activated sludge + basal medium
Activated sludge was added to the basal medium**.
Bottles 3-5 : Test substance + activated sludge + basal medium
30 mg of the test substance was added to the basal medium, then activated sludge was added.
Bottle 6 : Test substance + purified water
30 mg of the test substance was added to 300 mL of purified water***.
** The total volume of the basal medium and the sludge was held fixed at 300 mL.
*** Grade A4, Japanese Industrial Standards KO557

BOD MEASUREMENT
The BOD was measured for 28 days
1)Apparatus : closed system oxygen consumption measuring apparatus Ohkura Electric Co., Model OM-2001 (M.S.I. ID NO.D).

pH MEASUREMENT
After the exposure period, 20 mL of the test solution in each test bottle was transferred into a 20-mL glass beaker for pH measurement. After pH measurement, the test solution was returned to each bottle for measurement of residual. test substance concentration.
1)Apparatus : pH meter, ORION Research, Model 720

DOC MEASUREMEN2
The concentration of the DOC was measured as follows.
1 ) Apparatus : TOC analyzer, Shimaclzu Co., model TOC-5000A
2 Conditions : Furnace temperature : 680˚C (TC)
Air flow rate : 250 mL/min.
Sensitivity: x5
Injection volume : 50 ILL
3) Calibration curve
The following standard solutions were injected into the TOC analyzer
The calibration curve was prepared by the data processor of the analyzer.
Standard solutions
TC (total carbon) 20 and 50 mg C/L aqueous solutions of potassium biphthalate.
IC(inorganic carbon) 0 mg C/1 purified water (purified by Milli-Q) and 10 mg C/1 aqueous solution of sodium hydrogen carbonate and sodium carbonate.
4) Measurement of DOC in the bottles
Ten millilitre of the test solution in each bottle was transferred into a 10-mL centrifuge tube and centrifuged at 3000 rpm for 10 minutes. Five millilitre of supernatant was used for the DOC measurement. The remaining supernatant and the precipitate were returned to each bottle for the measurement of the residual test substance concentration.
Reference substance:
aniline
Key result
Parameter:
other:
Value:
>= 11 - <= 48
Sampling time:
28 d
Remarks on result:
other: Degradability based on residual test substance
Details on results:
OBSERVATION OF TEST BOTTLES AFTER EXPOSURE PERIOD
The solution in the bottles 1 and 5 were cloudy, in the bottles 2, 3, 4 and 6 were colorless. Growth of the sludge was observed in bottle I, in contrast with the control bottle (bottle 2). No growth was observed in bottles 3, 4 and 5.

pH MEASUREMKNT
After 28 days of exposure, the pi1 was determined to be 6.9, 7.1, 6.9 and 8.0 for bottles 3, 4, 5 and 6, respectively.

ACTIVITY OF SLUDGE
The degradability of aniline, based on the BOD measurements was 62 % after 7 days. The activity of the sludge was thus shown to be satisfactory.

DEGRADABILITY BASED ON BOD
BODk1 in bottles 3, 4, and 5 (as corrected with the value in bottle 2) were 6.7, -1.0 and 3.9 mg, and BOD in bottle 6 was 0.0 mg, respectively.
(*I Maximum theoretical value = 88.9 mg)
The degree of degradability based on the BOD measurements were 8, O(ca1culated value = -1) and 4 % for bottles 3, 4, and 5, respectively.

DEGRADABILITY BASED ON DOC
DOC*> in bottles 3, 4 and 5 (as corrected with the value in bottle 2) were 18.0, 3.9, and 23.0 mg/L, respectively. DOC in bottle 6 was 2.0 mg/L.
(*2 Maximum theoretical value = 78.9 mg/L)

Degradability was not calculated because the test substance was insoluble in water.

DEGRADABILITY BASED ON THE RESIDUAL TEST SUBSTANCE CONCENTRATION
The test substance*' was detected at concentrations of 67.2, 91.8, 53.5 are 103.6 mg/L in bottles 3, 4, 5 and 6, respectively.

(*3 Initial concentration = 100 mg/l)
The degree of degradability based on the residual test substance concentration was calculated to be 35, 11 and 48 for bottles 3, 4 and 5, respectively.
A new peak was detected on the HPLC chromatograms for bottles 3-5 with retention time of about 2.2 min.
Key result
Parameter:
BOD5
Value:
>= 0 - <= 8 other: mg/L

SUMMARY OF DOC MEASUREMENT

Degradability based on BOD

 

Bottle No.

Test substance

ThOD mg

Day 7

Day 14

Day 21

Day 28

BOD mg

% degradability

BOD mg

% degradability

BOD mg

% degradability

BOD mg

% degradability

1

Aniline

90.2

59.7

62

70.5

71

71.1

71

71.1

71

2

-

-

3.6

-

6.3

-

6.8

-

6.7

-

3

ANOX BF

88.9

3.4

0

6.3

0

7.0

0

13.4

8

4

ANOX BF

88.9

2.1

0 (-2)

5.2

0 (-1)

5.8

0 (-1)

5.7

0 (-1)

5

ANOX BF

88.9

2.1

0 (-2)

4.7

0 (-2)

5.0

0 (-2)

10.6

4

6

ANOX BF

88.9

0.0

-

0.0

-

0.0

-

0.0

-

Where % degradability was calculated to be negative, this value is shown in parentheses

 

pH measurement

Bottle No.

pH

Day 0

Day 28

1

-

7.7

2

7.0

7.1

3

-

6.9

4

-

7.1

5

-

6.9

6

7.9

8.0

 

Results of DOC measurement

Bottle No.

Organic carbon

DOC mg/L

2

-

1.1

3

78.9

19.1

4

78.9

5.0

5

78.9

24.1

6

78.9

2.0

 

Degradability based on residual test substance

Bottle No.

Concentration mg/L

Degradability %

2

< 1

-

3

67.2

35

4

91.8

11

5

53.5

48

6

103.6

-

 

RESULT OF DOC MEASUREMENT

 

Bottle 2

Bottle 3

Bottle 4

Bottle 5

Bottle 6

Calibration curve

 

TC

TC

TC

TC

TC

TC standard area

 

Area

 mg/L

Area

 mg/L

Area

 mg/L

Area

 mg/L

Area

 mg/L

20 mg/L

50 mg/L

Measure 1

878

1.279

13457

19.610

3618

5.271

16743

24.390

1400

2.040

13860

34435

Measure 2

893

1.301

13159

19.170

3585

5.223

16689

24.310

1378

2.008

13880

34437

Measure 3

892

1.300

13423

19.560

3586

5.225

16905

24.630

1401

2.041

13968

34610

Mean

888

1.293

13346

19.447

3596

5.240

16779

24.443

1393

2.030

13909

34494

 

 

Bottle 2

Bottle 3

Bottle 4

Bottle 5

Bottle 6

Calibration curve

 

IC

IC

IC

IC

IC

IC standard area

 

Area

 mg/L

Area

 mg/L

Area

 mg/L

Area

 mg/L

Area

 mg/L

0 mg/L

10 mg/L

Measure 1

111

0.157

260

0.368

175

0.248

227

0.321

0

0

0

7034

Measure 2

145

0.205

262

0.371

186

0.263

242

0.343

0

0

0

7072

Measure 3

126

0.178

275

0.389

205

0.290

229

0.324

0

0

0

7082

Mean

127

0.180

266

0.376

189

0.267

233

0.329

0

0

0

7063

 

 

TC (mg/L)

IC (mg/L)

TOC (mg/L)

Bottle 2

1.293

0.180

1.113

Bottle 3

19.447

0.376

19.071

Bottle 4

5.240

0.267

4.973

Bottle 5

24.443

0.329

24.114

Bottle 6

2.030

0.000

2.030

 

TOC-5000A Conditions

TC Furnace Temp. : 680°C

TC Catalyst: 25L

Springe Size: Normal

 

Measurement Mode: TOC

 

Air flow r a t e: 150 ml/min

 

No of Injects: 3

 

Range: x 5

 

Inj vol: 50µL

 

 

CALCULATION OF RECOVERY AND DETECTION LIMIT

 

Peak area mAb.sec (A)

Concentration in solution mg/L

Recovered mass mg (D)

Added mass mg (E)

Recovery & (F)

Detection limit mg/L (G)

 

For HPLC analysis (B)

In bottle (C)

Recovery test 1

4413292

1433.4

95.6

28.7

30.0

96

-

Recovery test 2

4529207

1471.0

98.1

29.4

30.0

98

-

Blank

<2000

<0.7

<0.1

-

0.0

-

1

 

 

 

 

 

 

Average recovery = 97%

 

 

Standard solution

Concentration (H) : 1500 mg/L

Peak area (I) : 4618494 mAbs. sec

Volume of solution for HPLC analysis (J) : 0.020 L

Volume of test solution for HPLC analysis (K) : 0.300 L

Volume of test solution in bottle (L) : 0.300 L

 

Equation: B = H x A ÷ I C = H x A ÷ I x J ÷ K      D = C x L                      F = D ÷ E x 100 G: raise decimal fractions to units

 

Calculation of the detection limit

Retention time (min)

1500 mg/L std.

Blank

Peak area mAb. sec

Rate of peak area (%)

Peak area mAb. sec

2.5

275638

6.0

-

5.7

116385

2.5

-

6.3

129733

2.8

-

7.0

2325708

50.4

< 1000

7.9

1771030

38.3

-

Total

4618494

100.0

< 2000

 

The minimum detectable peak area of the test substance was calculated by the following equation based on the minimum detectable peak area of 1000 mAbs-sec for the largest peak (retention time 7.0 min. ) in HPLC chromatogram.

 

Minimum detectable peak area (mAbs. sec) 4618494 x (1000 (mAbs-sec) ÷ 2325708 (dbs. sec) = 1986 (mAbs-sec) (< 2000 (mAbs.sec))

 

RESIDUAL TEST SUBSTANCE CONCENTRATION IN TEST BOTTLE

Bottle No.

Peak area mAb. sec (A)

Concentration in solution (mg/L)

For HPLC analysis (B)

In bottle (C)

2

< 2000

< 0.7

< 1

3

3146970

977.9

67.2

4

4299421

1336.1

91.8

5

2506017

778.8

53.5

6

4849234

1506.9

103.6

 

Standard solution

Concentration (D) : 1500 mg/L

Peak area (E) : 4826956 mAbs. sec

Volume of solution for HPLC analysis (F) : 0.020 L

Volume of test solution for HPLC analysis (G) : 0.300 L

Average recovery (H) : 97%

 

Equation: B = D x A ÷ E             C = D x A ÷ E x F ÷ G ÷ H x 100

Validity criteria fulfilled:
yes
Interpretation of results:
not readily biodegradable
Conclusions:
From the degradability results based on the BOD and the residual test substance concentration, it is concluded that the test substance was not readily biodegradable under the conditions of this test.
As a new peak was detected on the HPLC chromatograms for bottles 3-5, it is suggested that the test substance has been transformed during the exposure period.
Executive summary:

The test substance was exposed to the activated sludge in a closed-system oxygen consumption measuring apparatus. The biochemical oxygen demand (BOD) was measured over a 28 day period, After this period, the concentrations of the dissolved organic carbon (DOC) and the residual test substance in the test bottles were measured. The biodegradability of the test substance was evaluated from these results. Study was conducted in accordance with "Ready biodegradability, Modified MITI test" in OECD Guidelines for Testing of Chemicals No. 301C.

Results of the study are summarised as follows:

Measured values (day 28)

 

Bottle No.

 

3*

4*

5*

6

Theoretical value

BOD (mg)

6.7

-1.0

3.9

0.0

88.9

DOC (mg/L)

18.0

3.9

23.0

2.0

78.9

Test substance (mg/L)

67.2

91.8

53.5

103.6

100.0

* value corrected with BOD or DOC value of Bottle 2.

Degradabilities (%)

 

Bottle No.

 

3

4

5

Average

BOD

8

0 (-1)**

4

4

DOC

NA***

NA***

NA***

-

Test substance

35

11

48

31

** where % degradability was calculated to be negative, this value is shown in parentheses.

*** degradability was not calculated because the test substance was insoluble in water.

From the degradability results based on the BOD and the residual test substance concentration, it is concluded that the test substance was not readily biodegradable under the conditions of this test. As a new peak was detected on the HPLC chromatograms for bottles 3-5, it is suggested that the test substance has been transformed during the exposure

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2002
Report date:
2002

Materials and methods

Test guideline
Qualifier:
no guideline followed
Guideline:
other: The transformed product was analyzed by high performance liquid chromatography - mass spectrometry (LC-MS) to estimate the structure.
Principles of method if other than guideline:
ESTIMATION OF THE TRANSFORMED PRODUCT BY LC-MS
The test solution for measurement of the residual test substance was analyzed by LC-MS under the following conditions.
6.1 CONDITIONS OF LC-MS
1)Apparatus : LC-MS: Agilent Technologies Model 1100 (ID No.1)
Binary Pump : Agilent Technologies Model G1312A
Auto Injector : Agilent Technologies Model G1313A
UV-VIS Detector: Agilent Technologies Model G1315B
Column Oven : Agilent Technologies Model G1316A
Degasser : Agilent Technologies Model G1322A
Mass spectrometer: Agilent Technologies Model G1946D
Workstation : Agilent 1100 series Chemistation(Windows NT)

2)HPLC Conditions
Column : GL Science Co. Inertsil ODs-3, 2.1 mm i.d.X 150 mm
Mobile phase : Acetonitrile : Formic acid buffer* = 97 : 3
Flow rate : 0.4 ml/min.
Wavelength : 240 nm
Injection volume : 5 p1
Oven temperature : 40 "C
* 20 mM Ammonium formate : Formic acid = 1000 : 1

3) Mass conditions
Ionization : API-ES
Fragmentor : lOOV
Nebulizer : N2 (30psi)
Drying gas : N2 (lO.OL/min, 300°C)
Polarity : Negative
Mode : scan
Mass range : m/z 100-500
GLP compliance:
no
Type of study / information:
LC-MS assessment of transformation product.

Test material

Constituent 1
Test material form:
liquid
Details on test material:
Batch* : Lot No. 010009POI
Purity* : 97.3 % ( Impurity : Benzenepropanoic acid, 3,5-bis (1,1-dimethylethyl) -4-hydroxy, methylester 2.66 % )
Appearance : Clear yellow viscous liquid
Date of receipt : June 12, 2001
Specific details on test material used for the study:
As above.

Results and discussion

Any other information on results incl. tables

A peak derived from transformed product was detected at the retention time of approximately 1.2 minutes under the conditions of ESI negative mode on the HPLC chromatograms of bottles 3, 4 and 5. The molecular ion peak of m/z 277 (M-H) was detected on the MS Spectra. From these results, the structure of the transformed product was estimated to be 3-[3,5-Bis(tert-butyl)-4-hydroxyphenyl]propionic acid, MW 278.39.  Image as below.

Applicant's summary and conclusion

Conclusions:
A peak derived from transformed product was detected at the retention time of approximately 1.2 minutes under the conditions of ESI negative mode on the HPLC chromatograms of bottles 3, 4 and 5. The molecular ion peak of m/z 277 (M-H) was detected on the MS Spectra. From these results, the structure of the transformed product was estimated to be 3-[3,5-Bis(tert-butyl)-4-hydroxyphenyl]propionic acid, MW 278.39. Image as above.
Executive summary:

The structure of the transformed product was estimated to be 3-[3,5-Bis(tert-butyl)-4-hydroxyphenyl]propionic acid, MW 278.39