Orange, sweet, ext.

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:

2 June 1992 - 30 June 1992

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study without detailed documentation

Remarks:

Performed under GLP, according to international guideline, test substance defined. Not all test details reported

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
not relevant

Oxygen conditions:

aerobic

Inoculum or test system:

other: secondary effluent from an unacclimated activated sludge plant

Details on inoculum:

Secondary effluent from an unacclimated activated sludge plant at URL North.
Inoculum used was 10% by volume of the effluent, filtered through a Whatman filter paper (541) to remove coarse particluate matter.

Duration of test (contact time):

28 d

Initial conc.:

10 mg/L

Based on:

DOC

Parameter followed for biodegradation estimation:

CO2 evolution

Details on study design:

Medium as proposed in 1988 OECD RIng Test on BIodegradation with deviation: Feriic chloride stock solution contained 0.25 g not 0.2 g. The EDTA stock solution contained 0.4 g and not 0.5 g.
100 ml medium in 160 ml vessels (hypovials)
Analysis of the headspace gas and liquid medium for CO2/DIC was performed on days 3,7,10,14,16,21,24,28, using a Ionics 555 Inorganic Carbon Analyser.

Preliminary study:

not relevant

Test performance:

No data on reference substance. However, the inoculum activity was proven by the ready biodegradation of the test substance

Key result

Parameter:

% degradation (CO2 evolution)

Value:

60.5

Sampling time:

14 d

Key result

Parameter:

% degradation (CO2 evolution)

Value:

72

Sampling time:

28 d

Remarks on result:

other: 95% confidence 69.3 - 74.8 %

Details on results:

The biodegradation curve is attached. It is concluded that Orange oil florida is classified as readily (meeting the 10d-window) and ultimately biodegradable.

Results with reference substance:

no information

Validity criteria fulfilled:

not specified

Interpretation of results:

readily biodegradable

Conclusions:

The test substance is readily biodegradable.

Executive summary:

The substance 'Orange terpenes' (purity 96% limonene) was assessed in a test for ready biodegradation according to OECD TG 301B (CO2 production) under GLP. After 28 days the biodegradation was 72 % (95% confidence interval 69.3 - 74.8%). Thus the test substance is classified as readily biodegradable. The report does not present all test details and therefore the study is rated reliability 2.

Reference 2

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

31 August 1990 - 28 September 1990

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study without detailed documentation

Remarks:

Performed under GLP, according to international guideline, test substance defined. Not all test details reported

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
not relevant

Oxygen conditions:

aerobic

Inoculum or test system:

other: secondary effluent from an unacclimated activated sludge plant

Details on inoculum:

Secondary effluent from an unacclimated activated sludge plant at URL North.
Inoculum used was 10% by volume of the effluent, filtered through a Whatman filter paper (541) to remove coarse particluate matter.

Duration of test (contact time):

28 d

Initial conc.:

10 mg/L

Based on:

DOC

Parameter followed for biodegradation estimation:

CO2 evolution

Details on study design:

Medium as proposed in 1988 OECD RIng Test on BIodegradation with deviation: Feriic chloride stock solution contained 0.25 g not 0.2 g. The EDTA stock solution contained 0.4 g and not 0.5 g.
100 ml medium in 160 ml vessels (hypovials)
Analysis of the headspace gas and liquid medium for CO2/DIC was performed on days 4,7,14,18,21,25,28, using a Ionics 555 Inorganic Carbon Analyser.

Preliminary study:

not relevant

Test performance:

No data on reference substance. However, the inoculum activity was proven by the ready biodegradation of the test substance

Key result

Parameter:

% degradation (CO2 evolution)

Value:

62

Sampling time:

14 d

Key result

Parameter:

% degradation (CO2 evolution)

Value:

83.4

Sampling time:

28 d

Remarks on result:

other: 95% confidence 76.4 - 90.4 %

Details on results:

The biodegradation curve is attached. It is concluded that Orange oil florida is classified as readily (meeting the 10d-window) and ultimately biodegradable.

Results with reference substance:

no information

Validity criteria fulfilled:

not specified

Interpretation of results:

readily biodegradable

Conclusions:

The test substance is readily biodegradable.

Executive summary:

Orange oil was assessed in a test for ready biodegradation according to OECD TG 301B (CO2 production) under GLP. After 28 days the biodegradation was 83 % (95% confidence interval 76.4 - 90.4%). Thus the test substance is classified as readily biodegradable. The report does not present all test details and therefore the study is rated reliability 2.

Reference 3

Endpoint:

biodegradation in water: inherent biodegradability

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

30/11/2007 - 07/01/2008

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

performed under GLP

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to other study

Qualifier:

according to guideline

Guideline:

OECD Guideline 302 C (Inherent Biodegradability: Modified MITI Test (II))

Deviations:

no

Principles of method if other than guideline:

Not relevant

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
Not relevant

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge, domestic, non-adapted

Duration of test (contact time):

28 d

Initial conc.:

31.6 - 35.7 mg/L

Based on:

test mat.

Parameter followed for biodegradation estimation:

O2 consumption

Reference substance:

benzoic acid, sodium salt

Remarks:

Sigma-Aldrich , lot nr 02720

Parameter:

% degradation (O2 consumption)

Value:

75

Sampling time:

28 d

Details on results:

After 28d a mean degradation value of 75% of the test item was measured. An abiotical degradation of the test item can be excluded as there was no oxygen uptake within the abiotic control. The calculated "toxic effect" of the test item towards microorganisms at the concentration tested can be
neglected as the oxygen uptake within the test solution with the test item was higher than within that of the toxicity control. This special behaviour can be explained by the so-called diauxic effect (in the presence of a second carbon and energy source not always both components are degraded simultaneously as much as within the individual test solutions with the test or control item alone). As there was a degradation >60% of the test item, there is evidence of an inherent biodegradability.

Results with reference substance:

The control item sodium benzoate was degraded 90% within 28d.

Summary of results of testing biodegradability according to OECD 302 C

Time (d)	% Degradation
	TS 1	TS 2	TS 3	TS, Mean value	Tox-control	Sodium Benzoate
0	0.0	0..0	0.0	0	0.0	0.0
0.8	3.8	4.2	3.3	4	15.6	6.3
2.0	7.5	10.7	9.4	9	20.2	36.2
2.9	22.5	25.8	17.9	22	20.9	47.6
3.9	27.9	36.6	24.6	30	26.2	54.4
4.9	30.9	45.2	29.6	35	33.8	58.1
5.9	33.6	49.0	32.7	38	38.0	61.6
6.9	36.4	51.2	35.5	41	40.4	65.9
7.8	38.4	52.4	37.3	43	41.5	69.0
8.8	40.9	54.3	39.8	45	42.5	73.8
9.9	43.8	56.7	42.8	48	44.0	79.7
10.7	45.5	58.3	44.2	49	44.9	82.7
11.8	47.5	60.6	46.1	51	45.8	85.1
12.9	49.5	62.2	48.1	53	47.2	86.9
13.9	51.0	63.9	49.9	55	48.4	88.0
14.8	52.4	65.2	51.9	56	49.6	89.3
16.0	53.5	67.4	55.0	59	51.3	90.2
17.0	54.1	69.9	57.6	61	52.5	90.6
17.9	55.2	72.7	59.9	63	53.6	90.6
19.0	56.4	75.4	61.4	64	55.1	90.8
20.0	57.4	77.7	62.4	66	56.6	90.6
21.0	58.9	79.8	64.0	68	58.4	90.8
22.0	59.8	80.7	64.6	68	59.3	90.8
23.0	60.6	81.7	65.7	69	60.5	90.8
24.0	61.7	83.5	66.8	71	61.3	90.4
25.0	62.8	85.4	68.2	72	61.9	90.4
26.0	63.4	86.7	69.1	73	62.6	90.4
27.0	64.1	88.1	69.8	74	62.8	90.6
28.0	64.6	89.7	71.0	75	63.4	90.2

Validity criteria fulfilled:

yes

Interpretation of results:

inherently biodegradable

Conclusions:

Regarding the results obtained within this study the test item may be considered "inherently biodegradable".

Executive summary:

"Orangenol WMB" was tested for biodegradability according to 'MITI-[II]-Test' (OECD Guideline 302C using a mixture of two natural inocula). The test was performed under GLP. The degradation process was followed by the oxygen uptake in the test solutions. After 28d a mean degradation value of 75% of the test item was measured. An abiotical degradation of the test item can be excluded as there was no oxygen uptake within the abiotoc control. The calculated "toxic effecf' of the test item towards microorganisms at the concentration tested can be neglected as the oxygen uptake within the test solution with the test item was higher than within that of the

toxicity control. This special behaviour can be explained by the so-called diauxic effect (in the presence of a second carbon and energy source not always both components are degraded simultaneously as much as within the individual test solutions with the test or control item alone). As there was a degradation >60% of the test item, there is evidence of an inherent biodegradability. Regarding the results obtained within this study the test item may be considered "inherently biodegradable". The control item sodium benzoate was degraded 90% within 28d. The threshold of ~60% was met within 6d of incubation. Therefore, the test is considered valid.

Reference 4

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

other: constituent approach was followed.

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Orange Oil is a Natural Complex Substance (NCS). WIth 96% of the constituents of Orange oil known, this NCS can be considered as a well defined NCS. If the fraction of readily biodegradable constituents of the NCS exceeds 75%, the NCS itself is readily biodegradable as well.

Justification for type of information:

The conclusion for this NCS is based on the constituent approach as explained in the 'principles of the method' section

Principles of method if other than guideline:

Assessment of the Ready biodegradability of the known constituents; addition of the percentage of readily biodegradable constituents in the mixture gives the fraction of readily biodegradable substances. If this fraction exceeds 75%, the NCS is classified as 'readily biodegradable'.

GLP compliance:

no

Oxygen conditions:

aerobic

Inoculum or test system:

other: the results originate from multiple test set-ups

Duration of test (contact time):

28 d

Key result

Parameter:

other:

Value:

>= 60

Sampling time:

28 d

Remarks on result:

other: based on > 75% of the constituents being readily biodegradable

Details on results:

Details for all constituents are summarised in window "Any other information on results" and in attached document

Assessment of the biodegradability of ORANGE OIL:

Each of the identified constituents, totalling 96% of the composition, is readily biodegradable, see underlying data in table below.

Constituent	Results of Ready biodegradation
D-limonene C10H16	READILY BIODEGRADABLE: - Headspace CO2: 71.4% (Quest 1993). The biodegradability of perfume ingredients in the sealed vessel test. - MITI (I) / OECD 301C, improved for volatile substance: Limonene 14d - BOD: 41, 81, 98% Direct analysis: GC: 100%; TOC 50, 73, 81% (MITI 1980 http://www.safe.nite.go.jp/sitemap/db_map_e.html) - Readily biodegradable (Misra, G.; Pavlostathis, S. G.; Perdue, E. M.; Araujo, R. Applied Microbiology and Biotechnology (1996), 45(6), 831-838.)
Linalool	READILY BIODEGRADABLE: - OECD 301C: 80% (Givaudan 1991) - OECD Modified 301B, sealed CO2: 97.1% (QUEST 1994)                               - Readily biodegradable (Misra, G.; Pavlostathis, S. G.; Perdue, E. M.; Araujo, R. Applied Microbiology and Biotechnology (1996), 45(6), 831-838.)  - MITI (I) / OECD 301C: 28d - BOD: 90% Direct analysis: TOC 99% GC: 100%; (MITI 1996 http://www.safe.nite.go.jp/sitemap/db_map_e.html)
Octanal C8H16O	READILY BIODEGRADABLE: Read across from decanal   supported by prediction from EPISuite/ BIOWIN v4.10 (March 2009): readily biodegradable - yes
Decanal C10H20O	READILY BIODEGRADABLE: - OECD 301D Closed Bottle test: BOD 66% (14d) AND - OECD 310 Carbon Dioxide Headspace test: CO2 production 72% (14d), > 80% (28d) (RIFM/Givaudan/Akzo Nobel: A. Lapczynski, D. Salvito, K. Jenner, R. Geerts, C. Van Ginkel: Ready biodegradability testing of volatile poorly water soluble fragrances; Possible pitfalls. Poster SETAC North America 29th Ann. Meeting Tampa, Fl, November 2008, http://rifm.org/doc/RIFM%20SETAC%20Tampa%20Poster%20112008.pdf)
Myrcene C10H16O	READILY BIODEGRADABLE: - OECD 310 Carbon Dioxide Headspace test: CO2 production: 73% (Van Ginkel 2009) - MITI (I) / OECD 301C, improved for volatile substance: 14d - BOD: 86% Direct analysis: GC: 99%; TOC: 97% (MITI 1987: http://www.safe.nite.go.jp/sitemap/db_map_e.html)
alpha-Pinene C10H16	READILY BIODEGRADABLE: - OECD301B, sealed vessels: 62% (28d), 70% (42d) (Astra Zeneca, 2001, ECB PBT#84 http://ecb.jrc.ec.europa.eu/documents/PBT_EVALUATION/PBT_sum084_CAS_91770-80-8.pdf).                                                 - MITI (I) / OECD 301C: 28d - BOD: 91, 90, 95% Direct analysis: GC: 100, 100, 100% (MITI 2007: http://www.safe.nite.go.jp/sitemap/db_map_e.html)
alpha-Terpineol C10H18O	READILY BIODEGRADABLE: - OECD 310 Carbon Dioxide Headspace test, CO2 production: 80% degradation for Isomeric terpineol mixtures [CAS 8000-41-7] e.g. mixture containing 86% alpha, 11% gamma & 1% beta (Van Ginkel 2008).   Primary degradation and complete mineralisation demonstrated using soil inoculum under aerobic conditions (Misra, G.; Pavlostathis, S. G.; Perdue, E. M.; Araujo, R. Applied Microbiology and Biotechnology (1996), 45(6), 831-838.)
Citral C10H16O	READILY BIODEGRADABLE: - Modified 301B, sealed vessels CO2: 92.1% in 28 d (Quest 1994) - MITI (I) / OECD 301C,improved for volatile substance (Citral is mixture of neral and geranial): 28d - BOD: 92% Direct analysis: TOC: 78%; GC: 100% The test substance formed 6-Methyl-5-hepten-2-one (2-2480) and Geranic acid in (Water + Test Substance) system. (MITI 1991: http://www.safe.nite.go.jp/sitemap/db_map_e.html)   Supported by prediction from EPISuite/ BIOWIN v4.10 (March 2009): readily biodegradable - yes
Citronellal C10H18O	OECD 301F: 61% (28d), 51% in 10d-window (Givaudan 1994)   Supported by prediction from EPISuite/ BIOWIN v4.10 (March 2009): readily biodegradable - yes
	Fraction of NCS READILY BIODEGRADABLE

 The complete asssessment is added attached to this study record. Studies mentioned by Company Name are included in the RIFM database.

 

Validity criteria fulfilled:

not applicable

Interpretation of results:

readily biodegradable

Conclusions:

Orange oil is considered as readily biodegradable.

Executive summary:

Orange oil is a Natural Complex Substance (NCS) for which 96% of the constituents is identified. Orange oil consists for 80% of d-limonene and a range of minor constituents. An assessment of the biodegradability of Orange oil was made based on the degradability of its constituents. The underlying assumption is that if the constituents of the NCS are readily biodegradable, the NCS itself is readily biodegradable as well.

With the exception of one minor constituent, for all constituents the available data are obtained in tests on the ready biodegradability. For one minor constituent the ready biodegradability is based on a structurally related substance.

The cut-off level for ready biodegradability of the NCS is derived from the approach used for the classification as R53 in the Preparations Directive 1999/45/EC and the CLP Regulation 1272/2008/EC. If more then 25% of a mixture is not readily biodegrable, the mixture is considered as not ready biodegradable. This implies that if more then 75% of the fraction is biodegradable, the mixture (in this case the Natural Complex Substance Orange oil) is to be considered ready biodegradable.

All known constituents, comprising 96% of the composition of this NCS, are in the category of ready biodegradable substances. Therefore, Orange oil is considered readily biodegradable.

Description of key information

All known constituents of Orange oil, comprising 96% of the composition of this NCS, are in the category of readily biodegradable substances. Therefore, Orange oil is considered readily biodegradable. 

Key value for chemical safety assessment

Biodegradation in water:

readily biodegradable

Additional information

Orange oil is a Natural Complex Substance (NCS) typically consisting for 80% of d-limonene and a range of minor constituents. Therefore the asssessment of the biodegradability of Orange oil is based on its constituents. The assumption is that if the constituents of the NCS are readily biodegradable, the NCS itself is readily biodegradable as well. All known constituents, comprising 96% of the composition of this NCS, are in the category of ready biodegradable substances. Therefore, Orange oil is considered ready biodegradable.

This finding is supported by the results of two studies on the ready biodegradation of Orange oil florida and on Orange terpenes (King 1993, 1992) showing a mineralisation (CO2 production) of 83 and 72% after 28 days , respectively (Rel. 2 due to concise reporting). Moreover a high degree of biodegradation found in an inherent biodegradability study with Orange oil (Orangenöl, Lebertz, 2008) carried out according to OECD TG 302 C (MITI [II]) with a reliability of 1. This test revealed a biodegradation percentage of 75% (O2 consumption) in 28 days.