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EC number: 227-813-5
CAS number: 5989-27-5
Table 1: DNA damage measured by the Comet
assay in isolated rat kidney cells 3–6 or 22–26 hours after a single
administration of d-limonene at dose levels of 1000 and 2000 mg/kg bw
Sampling time (h)
Significant difference (Mann–Whitney U-test)
as compared with the vehicle control; ***p < 0.001.
Olive Tail Moment median value
In an in vivo comet assay, groups of
4 OFA Sprague-Dawley male rats were administered a single oral dose of
d-limonene in 0.5% CMC by gavage at dose levels of 0, 1000 and 2000
mg/kg bw. After an exposure period of 3-6 or 22-26 hours, treated
animals were sacrificed and the kidney cells were isolated and the
prepared slides were scanned to determine mean Olive Tail Moment (OTM)
median value in 150 cells per animal using the method described by Tice
et al (2000). A preliminary range-finding test has also been conducted
using 4 males rats/dose and animals were observed at least 2 days for
any clinical signs of toxicity and any mortalities in order to determine
the maximum tolerated dose (MTD).
Positive control (streptozotocin, 20 mg/kg
bw) caused a clear increase in the mean OTM median value. d-Limonene
showed no substantial increase in the mean OTM median value.
Therefore, d-limonene is not considered as
mutagenic in Comet assay on isolated kidney cells and does not need to
be classified according to Directive 67/548/EEC and CLP Regulation (EC) No
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