Hexadecan-1-ol

Biodegradation in water: screening tests

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1996-10-14 to 1996-11-13

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

Test conducted according to the appropriate OECD test guideline, and in compliance with GLP.

Qualifier:

according to guideline

Guideline:

OECD Guideline 301 B (Ready Biodegradability: CO2 Evolution Test)

Deviations:

no

GLP compliance:

yes

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge, domestic (adaptation not specified)

Details on inoculum:

- Source of inoculum/activated sludge (e.g. location, sampling depth, contamination history, procedure): Severn Trent Water Plc STP at Belper, Derbyshire, UK

- Storage conditions: Maintained on continuous aeration upon receipt.

- Pretreatment: A sample of activated sewage sludge was washed three times by settlement and resuspension in culture medium to remove any excessive amounts of dissolved organic carbon.

Duration of test (contact time):

29 d

Initial conc.:

25.5 mg/L

Based on:

test mat.

Parameter followed for biodegradation estimation:

CO2 evolution

Details on study design:

TEST CONDITIONS
- Composition of medium: 76.5mg of test material dispersed in inoculated culture medium to total volume of 3 litres

- Test temperature: 21 C

- Aeration of dilution water: Approximately 24 hours prior to addition of the test and standard materials the vessels were filled with 2400ml of culture medium and 30ml of inoculum and aerated overnight

- Suspended solids concentration: 30 mg/l

- Continuous darkness: yes


TEST SYSTEM
- Culturing apparatus: 5 litre glass culture vessels each containing 3 litres of solution

- Number of culture flasks/concentration:Standard (sodium benzoate) in duplicate, 10mgC/l; test material in duplicate, 20 mgC/l

- Method used to create aerobic conditions: The culture vessels were sealed and C02-free air was bubbled through the solution at a rate of approximately 70 ml/min and stirred continuously by magnetic stirrer.

- Details of trap for CO2 and volatile organics if used: Two 500 ml Dreschel bottles containing 350 ml of 0.05M NaOH. The C02 absorbing solutions were prepared using purified de-gassed water.


SAMPLING
- Sampling method and frequency: 2ml samples taken from the first CO2 absorber vessel on days 0,1,2,3,8,10,12,14,16,18.20,22,24,27,28 and 29. The second absorber vessel was sampled on days 0 and 29. On day 28 1ml of conc hydrochloric acid was added to each vessel to drive off any inorganic carbonates formed. The vessels were resealed, aerated overnight and the final samples taken from both absorber vessels on day 29.

- Sample storage before analysis: Samples analysed immediately for C02.



CONTROL AND BLANK SYSTEM
- Inoculum blank: Control (inoculated culture medium) in duplicate

- Toxicity control: Preliminary investigational work was carried out to assess any toxic effect of the test material on sewage sludge microorganisms (method based on OECD 209 ASRI test). The test period was increased from 3 hours to 5 days in order to determine whether any long term toxic effects of the material occurred. No inhibition was found at the concentration employed in the test.

Reference substance:

benzoic acid, sodium salt

Value:

62

Sampling time:

28 d

Interpretation of results:

readily biodegradable, but failing 10-day window

Conclusions:

The substance was found to be readily biodegradable, but not meeting the ten day window, in a reliable study conducted according to an appropriate test protocol, and in compliance with GLP.

Reference 2

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

The study was conducted according to an appropriate OECD test guideline and in compliance with GLP.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 301 B (Ready Biodegradability: CO2 Evolution Test)

Deviations:

not specified

GLP compliance:

yes

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge, domestic (adaptation not specified)

Details on inoculum:

- Source of inoculum/activated sludge (e.g. location, sampling depth, contamination history, procedure): Activated sludge collected from Downingtown Regional water pollution control centre (DRWPCC)

- Preparation of inoculum for exposure: The sludge was passed through a 2mm sieve and the total suspended solids (TSS) level was determined.

- Concentration of sludge: 2500 Mg/l

Duration of test (contact time):

31 d

Initial conc.:

1 000 µg/L

Based on:

test mat.

Parameter followed for biodegradation estimation:

CO2 evolution

Details on study design:

TEST CONDITIONS
- Test temperature: 20.6 - 22.5 C

- Aeration of dilution water: The flasks were placed on a rotary platform shaker and aerated continuously with a CO2-free air

- Continuous darkness: no


TEST SYSTEM
- Method used to create aerobic conditions: Continuous aeration with CO2-free air

- Test performed in open system: Yes

Reference substance:

benzoic acid, sodium salt

Test performance:

No Data

Parameter:

% degradation (CO2 evolution)

Value:

82.4

Sampling time:

28 d

Details on results:

10 day: 73.7%
14 day: 77.2%
21 day: 80.9%
28 day: 82.4%

Results with reference substance:

The biodegradation value of 82.1% was obtained for the reference substance (Benzoic acid)

10 day: 74%
14 day: 76.6%
21 day: 80%
28 day: 82.1%

Table 1: Biodegradation results for Test Hexadecanol (X0526.02) and Benzoic acid (X0429.06)

				Computer regression analysis constants
TSIN	Flask No	Conc. (µg/l)	Final %T^14CO2	Asymptote %T^14CO2	Rate (Day^-1)	Lag (Days)
X0526.02 (X0526.03R)	21	1000	86.0	75.6±3.0	1.9±1.1	0.0±0.2
	22	1000	82.4	75.0±2.8	2.0±1.2	0.0±0.2
X0429.06 9X0429.07R)	25	1000	88.5	78.5±2.8	1.8±0.9	0.0±0.2
	26	1000	81.8	73.1±2.4	1.6±0.7	0.0±0.2

Validity criteria fulfilled:

not specified

Interpretation of results:

readily biodegradable

Conclusions:

A ready biodegradation values of 82.4% was obtained for hexadecanol using an appropriate test procedure. The result is considered reliable.

Reference 3

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1993-06-08 to 1993-07-26

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Remarks:

The study was conducted according to a similar OECD test guideline and in compliance with GLP but validity criteria not specified

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 301 B (Ready Biodegradability: CO2 Evolution Test)

Deviations:

not specified

GLP compliance:

yes

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge, non-adapted

Details on inoculum:

- Source of inoculum/activated sludge (e.g. location, sampling depth, contamination history, procedure): Activated sludge collected from Downingtown Regional water pollution control centre (DRWPCC)

- Concentration of sludge: 2500 Mg/l

Duration of test (contact time):

33 d

Initial conc.:

20 mg/L

Based on:

test mat.

Parameter followed for biodegradation estimation:

CO2 evolution

Details on study design:

TEST CONDITIONS
- Composition of medium: The test medium was modified BOD water containing the following standard reagent solutions litre of water: 1mL magnessiumsulfate solution, 1mL calcium chloride solution, 2mL phosphate buffer solution, 4mL ferric chloride solution, 1mL of a 4% (w/v) solution of (NH4)2SO4

- Test temperature: 20.9 - 22.1 C

- Aeration of dilution water: Yes

- Continuous darkness: No


TEST SYSTEM

- Method used to create aerobic conditions: The test flasks were placed on rotary shaker, connected to scrubbing train and aerated overnight to purge the system of background CO2.

SAMPLING
- Sampling frequency: Daily for the first 10 days, 5 days interval thereafter

Reference substance:

benzoic acid, sodium salt

Parameter:

% degradation (CO2 evolution)

Value:

80.7

Sampling time:

33 d

Table 1: Result

				Computer regression analysis constants
TSIN	Conc. (mg active/L)	Final %TCO2	Final SOC (mg/L)	Asymptote %TCO2	Rate (Day^-1)	Lag (Days)
X0526.02 X0526.02	20 20	87.1 74.3	0.9 1.1	93.2±3.0 75.2±1.3	0.10±0.01 0.15±0.01	2.3±0.3 2.6±0.2
Control Control	- 20	- -	0.6 0.7	- -	- -	- -

Validity criteria fulfilled:

not specified

Interpretation of results:

readily biodegradable

Conclusions:

A ready biodegradation value of 80.7% was obtained for hexadecanol using an appropriate test procedure. The result is considered reliable.

Reference 4

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2009-03-17 to 2009-04-15

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

The study was conducted according to an appropriate OECD test guideline. It was not compliant with GLP.

Qualifier:

according to guideline

Guideline:

OECD Guideline 301 B (Ready Biodegradability: CO2 Evolution Test)

GLP compliance:

no

Remarks:

At the time of the study, this lab was in the process of attaining formal GLP status and did not hold certification. The work was conducted in accordance with GLP-principles (personal communication, 2010) and to high quality standards.

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): Fairfield Wastewater Treatment Plant, Fairfield, Ohio

- Preparation of media: The media was prepared one day prior to test initiation. The media consisted of the following reagents (1ml/l) in high quality deionised water: magnesium sulfate (2.25%), calcium chloride (2.75%), ferric chloride (0.025%) and phosphate buffer (10 ml/l). The phosphate buffer solution consisted of potassium dihydrogen phosphate (8.5 g/l), dipotassium hydrogen phosphate (21.75 g/l), disodium hydrogen phosphate dihydrate (33.4 g/l) and ammonium chloride (0.5 g/l).

- Preparation of inoculum for exposure: Activated sludge solids centrifuged for 20 minutes at 3000rpm and the supernatant decanted. Solids resuspended in media and homogenised in a blender for 1 minute. The solids were washed a second time as descripbed above and the TSS (total suspended solids) measured. Sufficient inoculum was added to the media to obtain a solids concentration of 15 mg/l. This mixture was adjusted to pH7 and aerated overnight with CO2-free air.

- Concentration of sludge: 15 mg solids/l.

Duration of test (contact time):

28 d

Initial conc.:

15.3 mg/L

Parameter followed for biodegradation estimation:

CO2 evolution

Details on study design:

TEST CONDITIONS
- Composition of medium: test material, sludge inoculum and phosphate buffered media

- Test temperature: 22 C




TEST SYSTEM
- Culturing apparatus: 1 litre bottles

- Number of culture flasks/concentration: 15.3 mg/l. Two replicates.


SAMPLING
- Sampling frequency: 12h

- Sampling method: Conductivity probe immersed in 1% NaOH to measure production of CO2.

CONTROL AND BLANK SYSTEM
- Inoculum blank: Yes. Six replicates
- Reference substance: Sodium Benzoate. Three replicates

Key result

Parameter:

% degradation (CO2 evolution)

Value:

82.4

St. dev.:

5.3

Sampling time:

28 d

Details on results:

The final SOC levels ranged from 0.3 to 0.4 mg/l, and were <0.1 mg/L for reference substance.

Table 1: Degradation kinetics

Type of suspension	% degradation at sampling time (days)
	0	1	2	3	6	8	10	14	16	20	22	24	27	28
Test sample (mean of 2 replicates)	0	0	0	5.36	46.10	60.02	66.83	75.88	78.45	81.19	81.32	82.02	82.25	82.37
Reference substance (mean of 3 replicates)	0	0	26.03	41.45	68.48	76.51	79.90	81.88	82.14	82.10	82.19	81.76	81.42	81.69

 

Validity criteria fulfilled:

yes

Interpretation of results:

readily biodegradable

Conclusions:

A reliable study conducted according to an appropriate test protocol determined the substance to be readily biodegradable.

Reference 5

Endpoint:

biodegradation in water: screening test, other

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Remarks:

This information was obtained from the public IUCLID 2000 CD-ROM. Further review of the original source, to reassess the reliability, would not alter the overall conclusions concerning this end point.

Oxygen conditions:

aerobic

Inoculum or test system:

sewage, predominantly domestic (adaptation not specified)

Initial conc.:

0.05 mg/L

Based on:

test mat.

Parameter:

% degradation (CO2 evolution)

Value:

28

Sampling time:

5 d

Messgroessen: Konzentration der Testsubstanz, CO2-Entwicklung radioaktiv markierte Testsubstanz.

Validity criteria fulfilled:

not specified

Conclusions:

The information reported is insufficient to assess the validity of this study. Therefore the study is not assignable.

Reference 6

Endpoint:

biodegradation in water: screening test, other

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Remarks:

This information was obtained from the public IUCLID 2000 CD-ROM. Further review of the original source, to reassess the reliability, would not alter the overall conclusions concerning this end point.

Oxygen conditions:

aerobic

Inoculum or test system:

sewage, predominantly domestic (adaptation not specified)

Initial conc.:

50 µg/L

Based on:

test mat.

Value:

28

Sampling time:

5 d

Messgroesse: 14CO2-Entwicklung

Validity criteria fulfilled:

not specified

Conclusions:

The information reported is insufficient to assess the validity of this study. Therefore the study is not assignable.

Reference 7

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

The test failed to meet one of the validity criteria of the test guideline.

Principles of method if other than guideline:

Method: other: ISO 10708 (BODIS) and RDA Blok Test

GLP compliance:

yes

Oxygen conditions:

aerobic

Inoculum or test system:

other: activated sludge, predominantly domestic

Duration of test (contact time):

28 d

Initial conc.:

100 mg/L

Based on:

COD

Reference substance:

acetic acid, sodium salt

Value:

76

Sampling time:

28 d

Details on results:

Kinetic of control substance:
7 days = 81%
14 days = 95%
21 days = 97%
28 days = 98%

The test substance attained 76% degradation over the test period. However, the 60% pass level was not reached within the 10 day window, therefore it cannot be considered readily biodegradable. However, significant degradation was observed therefore the substance is considered to be inherently biodegradable.

Kinetic of test substance (in %):
= 40 after 7 day(s)
= 59 after 14 day(s)
= 67 after 21 day(s)
= 76 after 28 day(s)

The validity criteria were fulfilled: 

(1) degradation rate of the reference substance had reached a level of 60% within 14 days

(2) total oxygen uptake in blanks after the first week was lower than 3 mg O2 and lower than 1 mg O2 in the following weeks (3) residual concentration of O2 in test bottles did not fall below 0.5 mg/l.

However, the parallel assays did not fall within the acceptable 20% range. On day 28, the % degradation for the three replicates was 71%, 91% and 

67%.

Interpretation of results:

readily biodegradable, but failing 10-day window

Reference 8

Endpoint:

biodegradation in water: screening test, other

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Remarks:

This information was obtained from the public IUCLID 2000 CD-ROM. Further review of the original source, to reassess the reliability, would not alter the overall conclusions concerning this end point.

Oxygen conditions:

aerobic

Inoculum or test system:

other: other bacteria: Marines Sediment (adaptiert)

Initial conc.:

1.7 mmol/L

Based on:

test mat.

Value:

96

Sampling time:

42 d

Details on results:

Kinetic of test substance (in %):
= ... 62 after 25 day(s)

Messgroesse: substanzspezifische Analytik (IR-Spektroskopie)

Werte aus graphischer Darstellung entnommen

Validity criteria fulfilled:

not specified

Conclusions:

The information reported is insufficient to assess the validity of this study. Therefore the study is not assignable.

Reference 9

Endpoint:

biodegradation in water: screening test, other

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Remarks:

Well-documented scientific study.

Principles of method if other than guideline:

Method: other

Oxygen conditions:

anaerobic

Inoculum or test system:

other: municipal sewage digester sludge fortified with activated sludge

Duration of test (contact time):

28 d

Initial conc.:

1 mg/L

Based on:

test mat.

Value:

90.1

Sampling time:

28 d

Details on results:

Total gas production for hexadecanol was 90.1% (69.1% as 14CO2 and 21.0% as 14CH4). Approximately 9% of the starting material remained with the solids and 0.5% remained in solution.

The method involves a batch test system with a domestic wastewater treatment sludge inoculum. The evolution of
radiolabeled carbon dioxide and methane are monitored. The test temperature is 35 degrees C. Starting solids levels of
the test sludge ranged between 24 and 29 g/L. 
C14-hexadecanol (labeled in the first carbon) was obtained from Sigma Chemical in St. Louis, MO (purity >98%) and
hexadecanol was obtained from American Tokyo Kasei Inc. in Portland, OR. The final ratio of C14 -CO2 to C14 -CH4 in
the gas produced was 3.3 to 1. The mechanism of hexadecanol biodegradation would be catabolized by beta oxidation to
form acetate. Hexadecanol degradation exhibited first-order kinetics.

Reference 10

Endpoint:

biodegradation in water: screening test, other

Remarks:

(anaerobic)

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Remarks:

Well-documented scientific study.

Principles of method if other than guideline:

Method: other

GLP compliance:

not specified

Oxygen conditions:

anaerobic

Inoculum or test system:

other: activated sludge from municipal sewage digester

Duration of test (contact time):

28 d

Initial conc.:

10 mg/L

Based on:

test mat.

Value:

97

Sampling time:

28 d

Details on results:

Total gas production for the fatty alcohol was 97.1% (25.1% as 14CH4 and 72% as 14CO2). Approximately 4% of the starting material remained in the sludge and approximately 0.5% was in the supernatant.

The publication describes in detail the test system and method used to evaluate the anaerobic biodegradability of
several materials including 14C-cetyl alcohol obtained from Amersham-Buchler in Germany.

Reference 11

Endpoint:

biodegradation in water: screening test, other

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Remarks:

This information was obtained from the public IUCLID 2000 CD-ROM. Further review of the original source, to reassess the reliability, would not alter the overall conclusions concerning this end point.

Principles of method if other than guideline:

Method: other: BSB-Bestimmung nach AFNOR-Richtlinie NF T90/103 (1969)

Value:

0

Sampling time:

5 d

Abbaugrad: 0 % (Sauerstoffmangel durch Bildung eines Oberflaechenfilms?)
Einsatzkonzentration: nicht angegeben
Inokulum: nicht angegeben

Validity criteria fulfilled:

not specified

Conclusions:

The information reported is insufficient to assess the validity of this study. Therefore the study is not assignable.

Reference 12

Endpoint:

biodegradation in water: screening test, other

Adequacy of study:

supporting study

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Remarks:

This information was obtained from the public IUCLID 2000 CD-ROM. Further review of the original source, to reassess the reliability, would not alter the overall conclusions concerning this end point. A source of higher reliability is available.

Oxygen conditions:

aerobic

Inoculum or test system:

sewage, predominantly domestic (adaptation not specified)

Initial conc.:

50 µg/L

Based on:

test mat.

Parameter:

not specified

Value:

37

Sampling time:

5 d

Messgroesse: 14CO2-Entwicklung

Validity criteria fulfilled:

not specified

Conclusions:

The information reported is insufficient to assess the validity of this study. Therefore the study is not assignable.

Reference 13

Endpoint:

biodegradation in water: screening test, other

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Remarks:

This information was obtained from the public IUCLID 2000 CD-ROM. Further review of the original source, to reassess the reliability, would not alter the overall conclusions concerning this end point.

Principles of method if other than guideline:

Method: other: Warburg-Respirometer

Inoculum or test system:

activated sludge, non-adapted

Initial conc.:

500 mg/L

Based on:

test mat.

Value:

0

Sampling time:

1 d

Abbauversuche mit drei Belebtschlaemmen unterschiedlicher Herkunft der Stoff wirkte auf jeden der getesteten Belebtschlaemme
toxisch (Sauerstoffmangel durch Bildung eines Oberflaechenfilms?)

Validity criteria fulfilled:

not specified

Conclusions:

The study demonstrated the substance having toxic effects on each of the three activated sludge tested, probably due to an oxygen deficiency by formation of a surface film. The information reported is insufficient to assess the validity of this study. Therefore the study is not assignable.

Reference 14

Endpoint:

biodegradation in water: screening test, other

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Remarks:

This information was obtained from the public IUCLID 2000 CD-ROM. Further review of the original source, to reassess the reliability, would not alter the overall conclusions concerning this end point.

Inoculum or test system:

sewage, predominantly domestic (adaptation not specified)

Initial conc.:

73 mg/L

Based on:

test mat.

Value:

14.3

Sampling time:

20 d

durchgehender Oberflaechenfilm (begrenzte Bioverfuegbarkeit) Messparameter: Gewichtsverlust des eingesetzten Hexadecanols
nach Methanolextraktion.

Validity criteria fulfilled:

not specified

Conclusions:

The substance appears to have limited bioavailability but loss of the substance weight have been reported. The information reported is insufficient to assess the validity of this study. Therefore the study is not assignable.

Reference 15

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Remarks:

The information reported is insufficient to assess the validity of this study.

Principles of method if other than guideline:

Method: other: US EPA OPPTS 835.3100 Aerobic Aquatic Biodegradation Test

GLP compliance:

not specified

Oxygen conditions:

aerobic

Inoculum or test system:

other: no information provided on inoculum

Duration of test (contact time):

31 d

Initial conc.:

20 mg/L

Based on:

test mat.

Reference substance:

other: Sodium benzoate

Value:

61

Sampling time:

31 d

Details on results:

Kinetic of control substance:
4 days = 47.1%
10 days = 58.1%
17 days = 60.5%
24 days = 61.2%
31 days = 62.2%

The test substance attained <60% degradation over the test period therefore it cannot be considered readily biodegradable.

Kinetic of test substance (in %):
= 27 after 4 day(s)
= 47 after 10 day(s)
= 59 after 17 day(s)
= 60 after 24 day(s)
= 61 after 31 day(s)

There is no information given on the validity criteria.

Validity criteria fulfilled:

not specified

Remarks:

The information reported is insufficient to assess the validity of this study.

Interpretation of results:

inherently biodegradable

Conclusions:

This test followed the method set out in US EPA OPPTS 835.3100 Aerobic Aquatic Biodegradation Test (which corresponds to OECD 301B Modified Sturm Test) with one exception: after the samples were added, dichloromethane (30ml) was used to dissolve the non water-soluble alcohols. When the alcohol was dissolved the solvent was evaporated leaving an alcohol film on the bottom of the flask. This was done to increase the bioavailability of the alcohol.
The substance appears inherently biodegradable but the information reported is insufficient to assess the validity of this study. Therefore the study is not assignable.

Reference 16

Endpoint:

biodegradation in water: screening test, other

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Remarks:

This information was obtained from the public IUCLID 2000 CD-ROM. Further review of the original source, to reassess the reliability, would not alter the overall conclusions concerning this end point.

Inoculum or test system:

other: other bacteria: Pseudomonas sp. (adaptiert)

Initial conc.:

800 µmol/L

Based on:

test mat.

Value:

ca. 66

Sampling time:

2 d

Alkohole (C10 - C18) als Gemisch geprueft; Einzel-Abbauraten aus GC-Peaks bestimmt; Abbau-Werte aus Graphik ermittelt

Conclusions:

The information reported is insufficient to assess the validity of this study. Therefore the study is not assignable.

Description of key information

Readily biodegradable: 82.4% (CO2 removal) in 28 d (OECD 301B; GLP) and 82.4% (CO2 removal) in 28 d (OECD 301B; non-GLP)

Key value for chemical safety assessment

Biodegradation in water:

readily biodegradable

Additional information

A reliable study (Federle, 1993) conducted according to an appropriate test protocol (OECD 301B) and according to GLP determined the test substance Hexadecan-1-ol to be readily biodegradable (82.4% after 28 days), meeting the 10 days window.

This result is consistent with another reliable study (Federle, 2009) conducted according to an appropriate test protocol (OECD 301B), but not conducted according to GLP, determined samples of pure linear hexadecan-1-ol to be readily biodegradable (82.4% CO2 evolution in 28 days), meeting the ten-day window. Trichloromethane was used as a solubilising agent in this study. The solvent was then evaporated under a gentle stream of N2 gas to deposit the test materials as a film on the walls of the vessels.

This study (Federle, 2009), using a methodology with appropriate loading method for the low solubility of the substances, was carried out with a range of linear saturated alcohols from four carbon chain length (C4) to twenty-two carbon chain length (C22).

These results are significant and fit for purpose even though the study was not conducted to GLP. The study gave results of 76.1% (C4), 77.7% (C6), 77.9% (C8), 74.6% (C10), 69.0% (C12), 82.2% (C14), 82.4% (C16), 95.6% (C18), 88.4% (C20) and 87.9% (C22) in 28 days. All were readily biodegradable, meeting the ten-day window. The finding is consistent with very high levels of removal in other types of test systems.

Therefore both studies are used as Weight of Evidence.

Other supporting studies are also available :

- A study conducted according to OECD 301B reports the substance to be readily biodegradable (62% after 28 days at 17 mg/L) (Mead, 1997)

- A study conducted according to a comparable OECD 301B test guideline reports the substance to be readily biodegradable (80.7% after 33 days) (Federle, 1993b)

- A summary report from a BODIS test reports the substance to be readily biodegradable (76% in 33 days, but failing the 10 days window) (Henkel, 1992)

Various other results from non-assignable reliability studies are also available. Some of them (Freitag 1979, 1982, 1985) are showing very limited biodegradation rates. However, the information reported is insufficient to assess the validity of those studies.

It is quite normal to observe some inter-laboratory variation in screening studies, particularly for substances where solubility limits may be a factor in degradation rates under the conditions of the testing. Due to the very diluted nature of the inoculum and its limited size, it may sometime happen that no degradation-competent microorganisms are present in a particular inoculum.

In the case where multiple reliable studies exist showing a range of extent of biodegradation in the course of standard tests, the normal approach is to base the interpretation on the higher degradation results, this is in line with ECHA guidance on information requirements and chemical safety assessment. An important piece of additional evidence to consider is the availability of ready biodegradation data from a series of tests conducted at the same laboratory at the same time, to examine degradability throughout the series of linear alcohols from C4-C22. Whilst at the time of the study by Federle (2009), the laboratory was not GLP-certified, the data are reliable and consistent throughout the homologous series.

Discussion of trends in the Category of C6-24 linear and essentially-linear aliphatic alcohols:

Many biodegradation assays have been carried out on this family of alcohols. Studies generated on single carbon chain length alcohols for tests that conform most closely to ready test biodegradability methods (OECD 301 series) show that alcohols with chain lengths up to C22 are readily biodegradable. In all cases the inoculum was not acclimated. Older reliable data suggest that chain lengths above C18 are not readily biodegradable, however those studies used loading techniques which, while in general still reliable, did not make allowance for the reduced bioavailability caused by the low water solubility of these longest chain substances. Where the substances are introduced into the test vessels by coating onto the flask, very rapid biodegradation was confirmed at all chain lengths tested.

In the older supporting tests, alcohols with chain lengths up to C18 are readily biodegradable. At carbon chain lengths ≤ 14, most tests showed that pass levels for ready biodegradation were reached within the 10 day window. Chain lengths of C16-18 achieved ready test pass levels, but not within the 10 day window. The one test on a single carbon chain length greater than C18 (using docosanol) showed degradation of 37%.

Tests which allowed adaptation are considered to have significant methodological deficiencies in terms of REACH requirements for the present purpose, and are accordingly considered to be Klimisch reliability 3: Invalid. However these also consistently demonstrate extensive biodegradability. Aliphatic alcohols occur naturally in the environment and environmental organisms will be acclimated.

Reliable studies for decanol and tetradecanol that show low levels of degradation are considered to be unexplained outliers. It is usual in the interpretation of such data to take the highest levels of degradation as the key study.

P&G (2009) conducted ready biodegradation screening tests on even-numbered saturated single chain length alcohols (C6-C22) using an appropriate test method (OECD 301B). Although, the test was not conducted in compliance with GLP, the study was found to be consistent with other available data, reliable and acceptable for environmental assessment. All tests substances were found to behave in a similar way. The substances were found to be readily biodegradable meeting the ten day window after a brief lag period. A separate test using the same methodology has confirmed the ready biodegradability result, meeting the ten-day window, at the upper end of the carbon number range (docosan-1-ol) in a GLP-compliant study (Flach, 2012).

Some variability is seen in the ultimate percentage degradation over the course of the study (see Table 1 below). It is quite normal to observe some inter-laboratory variation in screening studies, particularly for substances where solubility limits may be a factor in degradation rates under the conditions of the testing. As discussed above, due to the very diluted nature of the inoculum and its limited size, it may sometime happen that no degradation-competent microorganisms are present in a particular inoculum. This is evidenced by the variable mineralisation levels seen for standard reference substances under the conditions of OECD 301. In the case where multiple reliable studies exist showing a range of extent of biodegradation in the course of standard tests, the normal approach is to base the interpretation on the higher degradation results, this is in line with ECHA guidance on information requirements and chemical safety assessment, and consistent with the availability of ready biodegradation data from a series of tests conducted at the same laboratory at the same time, to examine degradability throughout the series of linear alcohols from C6-C22. Whilst at the time of the study (P&G, 2009), the laboratory was not GLP-certified, the data are reliable and consistent throughout the homologous series. In this study (P&G, 2009) and all other chain lengths studied were found to be readily biodegradable.

Biodegradation under anaerobic conditions

The anaerobic biodegradability of a range of chain lengths within the category has been investigated (C6 and C16 alcohols, 2 studies;and C16-18 and C18 unsaturated alcohols, 2 studies). All test substances were anaerobically degradable. Results from available studies are presented in Table 2 below.

Biodegradation by algae

Rapid degradation in water is indicated by the difficulties encountered in aquatic toxicity tests (chronic Daphnia reproduction) for long chain aliphatic alcohols (Section 6.1.4). Alcohols in the range C10-C15 were found to be rapidly removed from the test medium. This was attributed to metabolism by algae present as a food source in tests, and in later stages of the 21-day tests to bacterial degradation by microbes adsorbed onto the carapace of the test daphnids, despite daily cleaning of the animals.

Natural occurrence

It is important for context to note the findings from studies in the EU and US which consistently show that anthropogenic alcohols in the environment are minimal compared to the level of natural occurrence. Using stable isotope signatures of fatty alcohols in a wide variety of household products and in environmental matrices sampled from river catchments in the United States and United Kingdom, Mudge et al. (2012) estimated that 1% or less of fatty alcohols in rivers are from waste water treatment plant (WWTP) effluents, 15% is from in situ production (by algae and bacteria), and 84% is of terrestrial origin. Further, the fatty alcohols discharged from the WWTP are not the original fatty alcohols found in the influent. While the compounds might have the same chain lengths, they have different stable isotopic signatures (Mudgeet al., 2012).

In conclusion, the environmental impact of these studies is that it has confirmed that the fatty alcohols entering a sewage treatment plant (as influent) are partly derived from detergents, but these are not the same alcohols as those in the effluent which arise from in-situ bacterial synthesis. In turn, the fatty alcohols found in the sediments near the outfall of the WWTP are derived from natural synthesis and are not the same alcohols as those in the effluent.

Table: Ready biodegradation data on single constituent alcohols

CAS	Chemical Name	Comment	Method	Result % degradation	Result 10 day window	Reliability	Reference
111-27-3	1-Hexanol		301B	77.7% in 28 days at 17 mg/L	69.8%	2	Federle 2009
111-27-3	1-Hexanol		OECD 301-D	77% in 30 days at 2 mg/L 61% in 30 days at 5 mg/L	>60% in 14 days	2	Richterich, 2002a
111-27-3	1-Hexanol		Non-standard	- half life of 8.7 hours	-	2	Yonezawa and Urushigawa 1979
111-87-5	1-Octanol		301B	77.9% in 28 days at 18.8 mg/L	79.2%	2	Federle 2009
111-87-5	1-Octanol		ISO ring test (CO2 headspace biodegr. test)	92% in 28 days at 20 mg/L	>60%	2	Procter & Gamble, 1996
111-87-5	1-Octanol		OECD 301-B	59 % in 29 days at 10 mgC/L	-	2	Huntingdon Life Sciences Ltd. 1996a
111-87-5	1-Octanol		Non-standard	- half life of 1.9 hours	-	2	Yonezawa and Urushigawa 1979
112-30-1	1-Decanol			74.6% in 28 days at 15.1 mg/L	68.6%	2	Federle 2009
112-30-1	1-Decanol		301-D	88% in 30 days at 2 mg/L	>60%	2	Richterich, 2002c
112-30-1	1-Decanol		301-B	29 % after 29 day(s) at 10 mg/L COD	-	2	Huntingdon Life Sciences Ltd. 1996b
112-53-8	1-Dodecanol		301B	69% in 28 days at 15.4 mg/L	63%	2	Federle 2009
112-53-8	1-Dodecanol	Supporting	301-D	79% in 28 days at 2 mg/L	>60% in 14 days	1	Werner, 1993
112-72-1	1-Tetradecanol		301B	82.2% in 28 days at 15.9 mg/L	77.2%	2	Federle 2009
112-72-1	1-Tetradecanol		BODIS ~ISO 10708	92% in 28 days at 100 mg/L COD	>60%	1	Henkel, 1992d
112-72-1	1-Tetradecanol		301-B	28 % after 28 day(s) at 25.4 mg/L	-	1	Mead 1997b
36653-82-4	1-Hexadecanol		301B	82.4% in 28 days at 15.3 mg/L	75.2%	2	Federle 2009
36653-82-4	1-Hexadecanol		301B	62% after 28 days at 17.1 mg/L	<60%	1	Mead, 1997c
36653-82-4	1-Hexadecanol		BODIS	76 % after 28 day(s) at 100 mg/L COD	<60% after 14 d	2	Henkel KGaA 1992a
112-92-5	1-Octadecanol		301B	95.6% in 28 days at 14.5 mg/L	90.2%	2	Federle 2009
112-92-5	1-Octadecanol	Supporting	301D	38% in 29 days at 5 mg/L 69% in 29 days at 2 mg/L	<60%	1	Henkel, 1992f
629-96-9	1-Eicosanol		301B	88.4% in 28 days at 15.6 mg/L	83.4%	2	Federle 2009
661-19-8	1-Docosanol		301B	87.5% in 28 days at 20 mg/L	75.6%	1	Flach, 2012
661-19-8	1-Docosanol		301B	87.9% in 28 days at 15.3 mg/L	83%	2	Federle 2009
661-19-8	1-Docosanol		301B	37% after 28 days at 12.4 mg/L	<60%	1	Mead, 2000

Anaerobic degradation of alcohols

CAS	Chemical name	Comment	Method	Source of sludge	Concentration of test substance	Duration	% degradation at end of test	Reliability	Reference
111-87-5	1-Octanol		Serum bottle, gas production + GC analysis	1oor 2odigesters	50µg/ml	8 weeks	>75%	2	Sheltonand Tiedje, 1984
36653-82-4	1-Hexadecanol		Batch test using14C labelled test material	Municipal digester sludge fortified with activated sludge	1 mg/L	28 days	90%	2	Nuck and Federle, 1996
36653-82-4	1-Hexadecanol		Batch test using14C labelled test material	Municipal sewage digester	10 mg/L	28 days	97%	2	Steber and Wierich, 1987
68002-94-8	Alcohols, C16-18 and C18 unsaturated	Supporting	ECETOC screening test	Municipal sewage digester	50 mg/L	8 weeks	89%	2	Steberet al. 1995

A study by Rorije et al. (1998) on structural requirements for anaerobic biodegradation of organic chemicals is relevant. The study used a computer-automated structure evaluation program (MCASE) to analyse rates of aquatic anaerobic biodegradation of a set of diverse organic compounds, and developed a predictive model. Primary alcohols were one of the most important fragments linked to biodegradability (biophore). The authors discuss how the presence of a biophore indicates a possible site of attack for microbes to follow a metabolic pathway for anaerobic biodegradation.

Biodegradation in STP-simulation tests

Other recent data on ethoxylated alcohols also suggest that the rate of degradation could be higher than usually assigned to readily-biodegradable substances. In an OECD 303A study of the fate of alcohol ethoxylate homologues in a laboratory continuous activated sludge unit (Wind,et al., 2006) useful data about the properties and environmental exposures of alcohols are presented, although the paper describes mainly the properties of alcohol ethoxylates (AE). The waste water organisms were exposed principally to ethoxylates, but the alcohols would be generated by the degradation of the ethoxylates. The test substance comprised a 2:1 mixture of two commercial alcohol ethoxylate surfactants with chain lengths of C12-C15 (odd and even numbered) and C16-C18 (even numbered), respectively. The test substance was dosed at a concentration of 4 mg/L in the influent.

 

Results are shown in Table 3 below:

Removal of alcohols during an activated sludge test on alcohol ethoxylates.

Alcohol	Conc in effluent ng/L	Conc in sludge µg/g	%removal
C12	18	0.6	98.6
C13	21	0.7	99.5
C14	5.5	0	99.6
C15	2.9	1.1	99.8
C16	1.6	0.01	99.5
C18	58	0.7	99.1
Total	130	2	99.4

 

This shows that most of the alcohol which does not degrade (itself a small amount) was found in the solids in recovery at the end of the study. This study is important in that it indicates that the extent of removal of alcohols is high, from an exposure route that can realistically be anticipated based on the known life cycle.

References:

EU Commission, DGIII, Study on the possible problems for the aquatic environment related to surfactants in detergents, WRc, EC 4294, February, 1997

Flach, F., 2012. Biodegradability in the CO2-evolution test according to OECD 301b (July 1992). Hydrotox laboratory, report number 737, company study number 8571, Sasol, 2 May 2012.

Mudge, S.M, Deleo, P.C., Dyer, S.D. (2012). Quantifying the anthropogenic fraction of fatty alcohols in a terrestrial environment. Environmental Toxicology and Chemistry, Vol. 31, No. 6, pp. 1209–1222.

Nuck, B.A. and Federle, T.W. 1996. Batch test for assessing the mineralization of 14C-radiolabeled compounds under realistic anaerobic conditions. Environ. Sci.. 30:12, 3597-3603.

Rorije E, Peunenburg WJGM, Klopman G (1998) Structural requirements for anaerobic biodegradation of organic chemicals: A fragment model analysis. Environmental Toxicology and Chemistry, Vol. 17, No. 10, pp. 1943 -1950.

Shelton, D.R. and Tiedje, J.M. 1984. General method for determining anaerobic biodegradation potential. Applied and Environmental Microbiology 850-857.

Steber, J., Herold, C.P. and limia, J.M. 1995. Comparative evaluation of anaerobic biodegradability of hydrocarbons and fatty derivatives currently used as drilling fluids. Chemosphere 31:4, 3105-3118.

Steber, J. and Wierich, P. 1987. The anaerobic degradation of detergent range fatty alcohol ethoxylates. Studies with 14C-labelled model surfactants. Water Research. 21:6, 661-667.

Wind, T., R.J. Stephenson, C.V. Eadsforth, A. Sherren, R. Toy. (2006) Determination of the fate of alcohol ethoxylate homologues in a laboratory continuous activated sludge unit. Ecotox and Environ Safety, 64: 42-60.

Federle (2009). Ready Biodegradability Test, The Procter and Gamble Co., Study number65522, 27thApril 2009

Richterich, K. 2002a. 1-Hexanol: Ultimate biodegradability in the closed bottle test.Final report R 0200259.

Yonezawa, Y. and Urushigawa, Y. 1979. Chemico-biological interactions in biological purification systems. V. Relation between biodegradation rate constants of aliphatic alcohols by activated sludge and their partition coefficients in a 1-octanol-water system. Chemosphere 3:139-142.

Procter & Gamble. 1996. Final report: ISO ring test CO2 headspace biodegradation test. Study ECM ETS 554/02.

Huntingdon Life Sciences Ltd. (HLS). 1996a. Report No. 96/KAS217/0325.

Richterich, K. 2002c.Final report R 0200257.

Huntingdon Life Sciences Ltd.(HLS).1996b. Report No. 96/KAS223/0327.

Richterich. 1993. 1-Dodecanol: Aerobic biodegradation: Closed bottle test. Biological Research and Product Safety/Ecology: Unpublished results; test substance registration no. SAT 910724, Henkel KGaA; Report No. RE 920247 (With English summary report no. R9901416)

Henkel KGaA.1992d.  Report No. 920026 (test substance registration no. SAT 910723, test run no. 118).5 Marz 1992.

Mead, C. 1997b. Safepharm Laboratories, SPL Project Number 140/598.

Mead, C. 1997c. Safepharm Laboratories, SPL Project Number 140/543.

Henkel KGaA. 1992a.Biological Research and Product Safety/Ecology: Report No. RE 920102; test substance registration No. SAT 910721, test run No. 120.26 Juni 1992.

Henkel KGaA.1992f. Report No.RE920246, 18 December 1992.

Mead, C. 2000. Safepharm Laboratories, SPL Project Number 140/1002.