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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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

Environmental fate & pathways

Biodegradation in water: screening tests

Currently viewing:

Administrative data

Link to relevant study record(s)

Referenceopen allclose all

Endpoint:
biodegradation in water: ready biodegradability
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
3 (not reliable)
Rationale for reliability incl. deficiencies:
other: Study not performed to GLP and test parameters are only similar to international test guidelines. The parameters are sufficient to accept the data. Initial concentration not reported.
Qualifier:
according to guideline
Guideline:
other: Swedish Standard SS 02 81 43 Edition 2 (17.10.1990)
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 301 D (Ready Biodegradability: Closed Bottle Test)
GLP compliance:
not specified
Oxygen conditions:
aerobic
Inoculum or test system:
not specified
Details on inoculum:
No Data
Duration of test (contact time):
28 d
Initial conc.:
1 g/L
Based on:
test mat.
Parameter followed for biodegradation estimation:
O2 consumption
Parameter:
% degradation (O2 consumption)
Value:
1.8
Sampling time:
28 d
Conclusions:
For a test period of 28 days, di-pentaerythritol was found to biodegrade by 1.8% under aerobic conditions.
Executive summary:

The biodegradability of di-pentaerythritol was assessed according to Swedish Standards SS 02 81 43 Edition 2 (17/10/1990), similar to OECD Method 301D (Closed Bottle Test). BOD28 value was determined to be 27 mg/g di-pentaerythritol. Based on the theoretical oxygen demand of 1510 mg O2/g,

di-pentaerythritol

was determined to biodegrade by 1.8% after 28 days.
Endpoint:
biodegradation in water: inherent biodegradability
Type of information:
experimental study
Adequacy of study:
key study
Study period:
02 Sept 2015 - 27 Feb 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 302 B (Inherent biodegradability: Zahn-Wellens/EMPA Test)
Version / remarks:
1992
GLP compliance:
yes
Specific details on test material used for the study:
Batch Number: 1500300133
Purity: 94.6%
Appearance: white crystalline powder
Receipt date: 26 March 2015
Expiry date: 03 March 2017
Storage conditions: ambient
Oxygen conditions:
aerobic
Inoculum or test system:
activated sludge (adaptation not specified)
Details on inoculum:
A sample of activated sewage sludge was obtained from Haddington Municipal Sewage Works (a local sewage processing plant situated in Haddington, East Lothian, UK, which handles predominantly domestic sewage) on 03 May 2016.

On arrival at the laboratory, the sludge was already settled so the overlying supernatant was siphoned off. A portion of this supernatant was used to determine the BOD (biological oxygen demand) of the sludge prior to washing using a 5 day BOD test method (Eaton et. al (1995)). The BOD was determined to be 24.98 mg/L over a 7 Day period. Test was conducted over a 7 day period in error, as the final results were below <25 mg/L there was no impact.

The activated sludge was then suspended in mineral medium, the contents were portioned out to centrifuge bottles and spun at 1000 g for 5 minutes. This was regarded as the first mineral medium wash. Supernatant from each centrifuge bottle was then discarded, and the wash cycle was repeated once more. The resultant washed, concentrated solids were then combined into one bottle, made up to ca 3L with mineral medium and stored at ambient conditions until use.

In order to determine the solid content sludge was shaken by hand to homogenise thoroughly. Sub-samples (5 mL) of the homogenised sludge were dried in an oven at approximately 105oC and the suspended solids content determined as 8.474 g/L.
Duration of test (contact time):
28 d
Details on study design:
Cylindrical, 2000 mL capacity Pyrex glass bottles, fitted with two port screw cap system were used for the test. The inlet sides were connected to a manifold through which purified, CO2- free air was supplied. Air was supplied to each vessel by a glass tube at a depth of approximately 1 cm from the base of each flask. All connections were made with PVC tubing.

Each flask was mixed using a magnetic stirrer and a stir bar (8 cm long). All flasks were labelled with the study number, bioreactor number and treatment. Six flasks were prepared for the test; two control, two test item, one reference item (as a procedural control) and a toxicity control (as a check of potential toxicity). Each test flask was prepared by combining 500 mL of mineral medium, with either test item or reference item stocks at the appropriate concentrations. Microbial inoculums at a rate of 1.0 g dry matter/L final volume were then added and the flasks were made up to a final volume of 2000 mL with mineral medium. The ratio between inoculums (as dry weight solids) and test or reference material (as DOC) was 2.5:1.

Three test item stock solutions were prepared as follows in mineral medium. The entire stock solution was then added to the appropriate bioreactor. The flask was then rinsed with 500 mL mineral medium.

A 20 mg/mL stock solution of reference item was prepared by adding 3999.92 mg of Aniline to 200 mL of mineral medium. Aliquots of 51.7 mL were added to the reference and toxicity control flasks. Flow rates were adjusted until bioreactors were gently bubbling. The pH of each flask was measured at the beginning of the test, no pH adjustments were necessary. The flasks were connected to the manifold and purified CO2-free air passed through the system for 28 days

The test was conducted in a laboratory where the temperature was monitored over the duration of the test using a Brannan digital in/out calibrated thermometer. The temperature was maintained at 20.7-22.1°C. The bioreactors were covered with aluminium foil to exclude light.

Duplicate ca 40 mL samples were removed from each flask for DOC measurement on: Days 0 (ca 3 hours), 3, 7, 10, 14, 17, 21, 24, 27 and 28. At the start of the test and after each sampling, the liquid level was marked on each flask. Evaporation losses were made up with deionised water immediately prior to each sampling. Dissolved oxygen concentrations and pH were measured at each sampling. The pH was adjusted to 6.5-8.0 with 50 g/L H2SO4 or 40 g/L NaOH where necessary. On every sampling occasion samples were filtered using vacuum filtration equipment (Millipore®, glass) and 0.45µm Whatman nylon filters (Lot: A1008977). The first 5 mL of each filtrate was discarded. Filtered samples were stored at ca -20°C until analysis of DOC was made using a total organic carbon (TOC) analyser (Shimadzu TOC-L). Samples were analysed in duplicates.
Reference substance:
aniline
Remarks:
The reference material used was Aniline (C6H5NH2, Batch No. STBF2812V, Expiry: Aug 2020, Purity: ≥99.5%), a light yellow liquid known to be biodegradable and recommended in the guidelines. This was introduced to the test system at a target concentration o
Test performance:
The results of an initial test were rejected due to there being less microbial inoculum added then required due to a calculation error. The results from the initial test are not reported, however the data will be archived with the rest of the study data.

Filtered samples were stored in freezers set to maintain ca -20°C, not -18°C as the protocol requires. This had no impact on the integrity or the validity of the test.
Key result
Parameter:
% degradation (DOC removal)
Value:
ca. 25
Sampling time:
28 d
Details on results:
The percentage biodegradation of the test item was calculated based on the mg DOC measured from samples removed from each bioreactor at 3h ± 0.5 h (Section 6.5.8). The Day 28 value taken as a mean of Day 27 and 28 to account for fluctuations observed between Day 27 and 28 for Test bioreactor 2.

Dipentaerythritol was not inherently biodegradable under the test conditions within 28 days as the mean cumulative percentage biodegradation was 25.0% at the end of the test.

The reference item Aniline was inherently biodegradable under the test conditions within 28 days as 99.5% biodegradation was achieved by the end of the test.

The test item was not considered to be inhibitory to the microbial inoculum as 55.2% biodegradation was observed in the toxicity control bioreactor (test and reference items) by Day 28. This accounts for the biodegradation of Aniline, and the biodegradation seen in the test item.

The dissolved oxygen and pH values for all bioreactors are presented in Table 3 and Table 4. The dissolved oxygen concentration was maintained at or above 1 mg/L for the duration of the test.

The test met all validity criteria as the reference compound biodegraded by at least 70% (>99%) within 14 days and the test item was not adsorbed by the activated sludge as the TOC values at Day 0 (ca 3 h) were greater than the addition rate (400 mg DOC/L).
Validity criteria fulfilled:
yes
Interpretation of results:
not inherently biodegradable
Conclusions:
The test item Dipentaerythritol was not inherently biodegradable under the conditions of the test within the 28 day period. A mean total of 25.0% biodegradation was achieved by Day 28.
Executive summary:

The inherent biodegradability of Dipentaerythritol was assessed over a 28 day period by the Modified Zahn-Wellens Test. The test was designed to comply with the requirements of OECD (1992) Guideline 302B.  Dipentaerythritol was introduced to the test system at an addition rate of 400 mg DOC/L (Dissolved Organic Carbon/L). The reference item (aniline) was introduced to the test system at 400 mg DOC/L. The reference substance (aniline) was degraded by 99.5 % within 28 days. The test item is not considered to be inhibitory to the microbial inoculum as 55.2% biodegradation was observed in the toxicity control bioreactor (containing 400 mg DOC/L of test and reference items) on Day 28. Dipentaerythritol did not show signs of inherent biodegradability under the conditions of the test as the test item biodegraded by 25.0% during the 28 day test.


 


The test met all validity criteria as the reference compound biodegraded by at least 70% (>99%) within 14 days and the test item was not adsorbed by the activated sludge as the TOC values at Day 0 (ca 3 h) were greater than the addition rate (400 mg DOC/L).


 


According to the guideline, Dipentaerythritol is not inherently biodegradable.

Endpoint:
biodegradation in water: ready biodegradability
Type of information:
experimental study
Adequacy of study:
key study
Study period:
May 15 to June 12 2008
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
test procedure in accordance with national standard methods with acceptable restrictions
Qualifier:
according to guideline
Guideline:
OECD Guideline 301 A (Ready Biodegradability: DOC Die Away Test)
Deviations:
not specified
Qualifier:
according to guideline
Guideline:
other: SS-EN ISO 7827:1996
Deviations:
not specified
Qualifier:
according to guideline
Guideline:
ISO DIS 9439 (Ultimate Aerobic Biodegradability - Method by Analysis of Released Carbon Dioxide)
Deviations:
not specified
GLP compliance:
not specified
Remarks:
Study performed according to ISO/IEC 17025; however, no statement of GLP compliance was found in the test report.
Oxygen conditions:
aerobic
Inoculum or test system:
activated sludge (adaptation not specified)
Details on inoculum:
- Source of inoculum/activated sludge (e.g. location, sampling depth, contamination history, procedure): Klagshamn waste water plant, Sweden.
- Laboratory culture: not specified
- Method of cultivation: not specified
- Storage conditions: not specified
- Storage length: not specified
- Preparation of inoculum for exposure: not specified
- Pre-treatment: not specified
- Concentration of sludge: 5.9 g suspended solids/L to 8.9 g/L
- Initial cell/biomass concentration:
- Water filtered: not specified. Deionized water
- Type and size of filter used, if any:
Duration of test (contact time):
28 d
Initial conc.:
85 mg/L
Parameter followed for biodegradation estimation:
DOC removal
Parameter followed for biodegradation estimation:
TOC removal
Details on study design:
TEST CONDITIONS
- Composition of medium: 5.1 ml of inoculum, 85 ml (1g/L) test substance, deionized water, anhydrous potassium dihydrogenphosphate, anhydrous potassium monohydrogenphosphate, sodium monohydrogenphosphate dihydrate, ammonium chloride, magnesium sulphate, anhydrous calcium chloride, iron (III) chloride hexahydrate, concentrated hydrochloric acid, sodium acetate, mercury (II) chloride.
4 L of the test medium was prepared and 2 L was added in duplicate to 2 L flasks

- Test temperature: a room with temperature 20-25°C
- pH: 7.4
- pH adjusted: yes
- Suspended solids concentration: 5.9 g/L
- Continuous darkness:diffused light
- Other:


TEST SYSTEM
- Number of culture flasks/concentration: 2 flasks of test medium, 1 flask of control medium, 1 flask of inhibition medium, 1 flask sterile medium, 2 flasks blank medium
- Method used to create aerobic conditions: air was bubbled from an air pump through a long glass tube which ended at the bottom of the flask.

- Measuring equipment: Not reported
- Test performed in closed vessels due to significant volatility of test substance: Not reported
- Test performed in open system: Not reported
- Details of trap for CO2 and volatile organics if used: Not reported


SAMPLING

- Sterility check if applicable: filter for DOC samples was Scheicher and Schuell, boiled 3 times in deionized water to purify the filters from soluble carbon. Each boiling period lasted for at least one hour.



CONTROL AND BLANK SYSTEM
- Inoculum blank: For 1 L blank medium the following solutions were added
Anhydrous potassium dihydrogenphosphate (KH2PO4), Anhydrous potassium monohydrogenphosphate (K2HPO4), Sodium monohydrogenphosphate dihydrate (Na2HPO4. 2 H2O), Ammonium chloride (NH4Cl), Magnesium sulphate (MgSO4) Anhydrous calcium chloride (CaCl2), Iron(III) chloride hexahydrate (FeCl3 . 6 H2O) Concentrated Hydrochloric acid,
- 5.1 ml inoculum , deionized water.


- Abiotic sterile control: Anhydrous potassium dihydrogenphosphate (KH2PO4), Anhydrous potassium monohydrogenphosphate (K2HPO4), Sodium monohydrogenphosphate dihydrate (Na2HPO4. 2 H2O), Ammonium chloride (NH4Cl), Magnesium sulphate (MgS04) Anhydrous calcium chloride (CaCl2), Iron(III) chloride hexahydrate (FeCl3 . 6 H2O)
Concentrated Hydrochloric acid, Mercury(II) chloride (HgCl2), Di-Pentaerythritol
Deionized water (H2O)

- Toxicity control: Anhydrous potassium dihydrogenphosphate (KH2PO4), Anhydrous potassium monohydrogenphosphate (K2HP04), Sodium monohydrogenphosphate dihydrate
(Na2HPO4. 2 H2O), Ammonium chloride (NH4Cl), Magnesium sulphate (MgSO4)
Anhydrous calcium chloride (CaCl2), Iron(III) chloride hexahydrate (FeCl3 . 6 H2O)
Concentrated Hydrochloric acid, Sodium acetate (NaCH3COO), Di-Pentaerythritol
Deionized water (H2O), Inoculum

Reference substance:
acetic acid, sodium salt
Parameter:
% degradation (DOC removal)
Value:
7
Sampling time:
28 d
Parameter:
% degradation (DOC removal)
Value:
16
Sampling time:
35 d
Parameter:
% degradation (TOC removal)
Value:
1
Sampling time:
28 d
Details on results:
After the 28 day test period removal of DOC was less than 10% for both replicates. At the 2 additional time points (35 and 42 days) DOC removal did not progress beyond 21%.
Results with reference substance:
Removal of the reference substance was greater than 90% after 14 days. This fulfills the validity criteria for this study.

The validity criteria for this study were met and the test is considered valid, this confirms the suitability of the inoculum. Removal of the test substance was less then 10% after 28 days.

Validity criteria fulfilled:
yes
Interpretation of results:
under test conditions no biodegradation observed
Conclusions:
Di-Pentaerythritol cannot be regarded as readily biodegradable.
Executive summary:

According to SS-EN ISO 7827:1996/OECD 301 A the test is valid if the degradation of the reference compound (sodium acetate) after 14 days is more than 70 %. In this case the degradation is more than 90 % after 14 days. The quantity of DOC that is degraded in the toxicity control corresponds to the part that comes from the reference compound. This shows that the test article was not inhibitory to the inoculum. The removal of 5% DOC in the sterile medium indicates a small degree of abiotic removal.

 

According to OECD guidelines for testing chemicals a test compound is regarded as easily biodegradable if the loss of DOC within 28 days is greater than 70 %. The pass value has to be reached in a 10-day window within the 28-day period of the test. The 10-day window begins when the degree of biodegradation has reached 10% DOC and must end before day 28 of the test. This criterion is not reached for the test article and Di-Pentaerythritol can not be regarded as readily biodegradable. A plateau at approximately 16% is reached after 35 days.

 

Given this result Di-Pentaerythritol cannot be considered to be readily removed in the STP.

Description of key information

The substance, di-pentaerythritol, is not readily biodegradable or inherently biodegradable..

Key value for chemical safety assessment

Additional information

The potential for biodegradation of di-pentaerythritol was determined in a study according to SS-EN ISO 7827:1996/OECD 301 A. The quantity of test substance DOC that was degraded in this study was less than 20%. According to OECD guidelines for testing chemicals, a test compound is regarded as easily biodegradable if the loss of DOC within 28 days is greater than 70 %. This criterion was not reached for the test article and, as such, Di-Pentaerythritol cannot be regarded as readily biodegradable.


In a further supportive study the aerobic biodegradability of di-pentaerythritol was investigated in a study with methodology closely corresponding to OECD 301 D. The results of this study suggested that little biodegradation took place during the test period.


 


Additionally, the inherent biodegradability of Dipentaerythritol was assessed over a 28 day period by the Modified Zahn-Wellens Test. The test was designed to comply with the requirements of OECD (1992) Guideline 302B.  Dipentaerythritol was introduced to the test system at an addition rate of 400 mg DOC/L (Dissolved Organic Carbon/L). The reference item (aniline) was introduced to the test system at 400 mg DOC/L. The reference substance (aniline) was degraded by 99.5 % within 28 days. The test item is not considered to be inhibitory to the microbial inoculum as 55.2% biodegradation was observed in the toxicity control bioreactor (containing 400 mg DOC/L of test and reference items) on Day 28. Dipentaerythritol did not show signs of inherent biodegradability under the conditions of the test as the test item biodegraded by 25.0% during the 28 day test. According to the guideline, Dipentaerythritol is not inherently biodegradable.