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EC number: 227-813-5
CAS number: 5989-27-5
% biodegradation of D-Limonene
The ready biodegradability of D-Limonene
(purity 95%) was assessed in a GLP study conducted according to OECD TG
310 (CO2 production in a sealed vesssel, formerly under OECD 301B).
After 28 days, the biodegradation was 71.4% (95% confidence interval
68.3 - 74.5%). Thus the test substance is readily biodegradable.
Due to the wide range of degradation results
with other test substances, the viability of the inoculum is
demonstrated and the lack of a reference substance does not invalidate
the test results. The report does not present all test details and
therefore the study is rated with a reliability of 2.
Lag period (h)
Maximum degradation rate (mg l-1h-1)
Normalized degradation rate (h-1)
Unacclimated soil A extract
Acclimated (from CSR1)
a: Four hydrocarbons terpenes tested
b: Three hydrocarbons terpenes tested
NM: not measured because of lack of accurate
In a ready biodegradation study, d-limonene
was tested at concentrations of 0.5-3 mg/L. Both forest-soil extract and
enriched cultures were used as inocula for the experiments conducted
first without microbial acclimation (experiment 1), then with prior
microbial acclimation to the test material (experiment 2A). The
degradation of the test material was assessed by the determination of
the biomass, concentration of the test material and headspace CO2. The
test treatments and control (sodium azide, 2.5 g/L) were measured in
The normalized degradation rate in
experiments 2A and 2B were 0.076 and 0.034/h, respectively. The
normalized degradation rate was not measured in experiment 1 because of
lack of accurate biomass data. The maximum degradation rate in
experiments 1, 2A and 2B were 0.044, 0.40 and 0.137 mg/L/h,
respectively. The lag period in experiments 1, 2A and 2B were 180, 15
and 23 h, respectively. The total mass of the test material in the
control reactor remained unchanged throughout the incubation period.
d-Limonene was readily degraded by mixed
cultures derived from forest soils.
In a ready biodegradation study, d-limonene
was tested at concentrations of 0.6 mg/tube and the inoculum was 20%
(w/v) soil-slurry. The degradation of the test material in test
treatments, inoculum blank (soil-slurry samples without any test
material) and azide-amended (2.5 g/L) soil-slurry controls were assessed
by the determination of the biomass, residue test material and headspace
The observed volumetric biodegradation rate
and soil-normalised biodegradation rate were determined to be 0.38
mg/L/h and 1.9 µg/g/h.
d-Limonene was readily degraded by
indigenous soil microorganisms.
a ready biodegradation study performed according to OECD Guideline 301
C, dl-limonene was tested at concentrations of 100 ppm and the inoculum
was activated sludge (30 ppm). The degradation of the test material was
assessed by the determination of the oxygen consumption.
100 ppm test concentration, 41-98% of degradation by biochemical oxygen
demand was reached in 14 days.
dl-limonene was readily biodegradable.
Dissolved oxygen concentrations (mg/L) in the closed bottles.
Oxygen concentration (mg/L)
Ocs: Mineral nutrient solution
without test material but with inoculum and silica gel
Ot: Mineral nutrient solution
with test material (2.0 mg/L), silica gel, and inoculum
Oc: Mineral nutrient solution
with only inoculum
Oa: Mineral nutrient solution
with sodium acetate (6.7 mg/L) and with inoculum
Oxygen consumption (mg/L) and the percentages biodegradation of the test
substance, Dipentene (BOD/ThOD) and sodium acetate (BOD/ThOD) in the
Closed Bottle test.
Oxygen consumption (mg/L)
In a ready biodegradation study performed
according to OECD Guideline 301 D and GLP, dipentene multiconstituent
was tested at concentrations of 2 mg/L and the inoculum was activated
sewage sludge, domestic. The degradation of the test material was
assessed by the determination of the oxygen consumption. The test
treatments, inoculum blank, and reference (sodium acetate) were measured
At 2 mg/L test concentration, greater than
60% of biodegradation was reached on Day 14. Hence, the test material
met the 14-day window requirement for ready biodegradability. On Day 28,
the biodegradation was 80%.
The reference material, sodium acetate,
reached greater than 60% of biodegradation on Day 6. Dipentene
multiconstituent was considered to be non-toxic to micro-organisms as
inhibition of the endogenous respiration of the inoculum was not
detected during the test. Hence, it met the validity criteria for
reference material and toxicity control. The endogenous respiration at
Day 28 was 1.0 mg/L and oxygen concentration during the test was greater
than 0.5 mg/L.
Therefore, dipentene multiconstituent was
d-limonene is readily biodegradable according to the criteria of the OECD 301 guideline.
ready biodegradability of d-limonene
was assessed in a GLP study conducted according to OECD TG 310 (CO2
production in a sealed vesssel, formerly under OECD 301B). The result
was confirmed in two other biodegradation studies conducted with the
dl-Limonene and dipentene multiconstituent
were also found to be readily biodegradable according to OECD 301
criteria, confirming the results obtained with d-limonene.
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