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EC number: 204-617-8 | CAS number: 123-31-9
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Genetic toxicity: in vitro
Administrative data
- Endpoint:
- in vitro gene mutation study in mammalian cells
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- comparable to guideline study
Data source
Referenceopen allclose all
- Reference Type:
- publication
- Title:
- Responses of the L1578Y tk+/tk- mouse lymphoma cell forward mutation assay. II. 18 coded chemicals.
- Author:
- McGregor DB, Brown A, Cattanach P, Edwards I, McBride D, Caspary WJ
- Year:
- 1 988
- Bibliographic source:
- Environ Molec Mutagen 11, 91 - 118
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 1 989
- Report date:
- 1989
Materials and methods
Test guideline
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
- GLP compliance:
- no
- Type of assay:
- mammalian cell gene mutation assay
Test material
- Reference substance name:
- Hydroquinone
- EC Number:
- 204-617-8
- EC Name:
- Hydroquinone
- Cas Number:
- 123-31-9
- Molecular formula:
- C6H6O2
- IUPAC Name:
- hydroquinone
- Details on test material:
- - Name of test material (as cited in study report): hydroquinone
Constituent 1
Method
- Target gene:
- Thymidine kinase locus
Species / strain
- Species / strain / cell type:
- mouse lymphoma L5178Y cells
- Details on mammalian cell type (if applicable):
- - Type and identity of media: Fischer's medium with 5% heat-inactivated horse serum
- Properly maintained: yes
- Periodically checked for Mycoplasma contamination: yes
- Periodically "cleansed" against high spontaneous background: yes
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 mix from Aroclor 1254 induced rat liver
- Test concentrations with justification for top dose:
- - S9: 1.56, 3.12, 6.25, 12.5, 25, 50 µg/mL
+ S9: 0.652, 1.25, 2.5, 5, 10 µg/mL - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: DMSO (+ S9), methanol (- S9)
Controls
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: - S9: EMS; + S9: EMS or 3-methylcholanthrene
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium
DURATION
- Exposure duration: 4 h
- Expression time (cells in growth medium): 2 d
- Selection time (if incubation with a selection agent): 11-14 d
- Fixation time (start of exposure up to fixation or harvest of cells): 13-16 d
SELECTION AGENT (mutation assays): trifluorothymidine
NUMBER OF REPLICATIONS: 2, two independent assays
DETERMINATION OF CYTOTOXICITY
- Method: relative total growth - Evaluation criteria:
- Positive response: dose-related trend and statistically significant response at one of the three highest doses
- Statistics:
- Dose trend test (Barlow et al., 1972); variance analysis (pairwise comparisons) with P < 0.05
Results and discussion
Test results
- Species / strain:
- mouse lymphoma L5178Y cells
- Metabolic activation:
- with and without
- Genotoxicity:
- positive
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Vehicle controls validity:
- valid
- Positive controls validity:
- valid
Any other information on results incl. tables
Table: Results of the mouse lymphoma cell mutation assay without and with metabolic activation
Metabolic activation / trial |
Test concentration |
Cloning efficiency (%) |
Relative total growth (%) |
Mutant fraction |
Average mutant fraction |
- S9 / trial 1 |
0 |
47 |
68 |
73 |
74 |
35 |
57 |
96 |
|||
48 |
164 |
62 |
|||
60 |
110 |
65 |
|||
1.562 |
41 |
45 |
200 |
157* |
|
42 |
108 |
113 |
|||
3.125 |
25 |
23 |
454 |
333* |
|
42 |
50 |
213 |
|||
6.25 |
17 |
7 |
857 |
624* |
|
25 |
13 |
391 |
|||
12.5 |
TOX |
||||
TOX |
|||||
Positive control |
47 |
101 |
214 |
197* |
|
45 |
43 |
181 |
|||
- S9 / trial 2 |
0 |
76 |
96 |
19 |
22 |
61 |
100 |
28 |
|||
93 |
98 |
18 |
|||
85 |
106 |
24 |
|||
3.125 |
13 |
5 |
565 |
419* |
|
57 |
30 |
274 |
|||
6.25 |
48 |
13 |
542 |
542* |
|
37 |
9 |
541 |
|||
12.5 |
27 |
7 |
871 |
857* |
|
25 |
7 |
843 |
|||
25 |
28 |
6 |
863 |
774* |
|
30 |
7 |
685 |
|||
50 |
TOX |
||||
TOX |
|||||
Positive control |
120 |
114 |
108 |
108 |
|
- S9 / trial 3 |
0 |
93 |
114 |
19 |
38 |
62 |
109 |
47 |
|||
52 |
90 |
42 |
|||
90 |
87 |
45 |
|||
0.625 |
81 |
68 |
47 |
49 |
|
102 |
71 |
51 |
|||
1.25 |
36 |
22 |
164 |
179* |
|
27 |
16 |
195 |
|||
2.5 |
15 |
7 |
607 |
624* |
|
12 |
4 |
642 |
|||
5 |
TOX |
||||
12 |
1 |
1915 |
|||
10 |
TOX |
||||
TOX |
|||||
Positive control |
69 |
66 |
299 |
306* |
|
93 |
58 |
313 |
|||
+ S9 / trial 1 |
0 |
91 |
99 |
65 |
70 |
70 |
102 |
67 |
|||
68 |
98 |
70 |
|||
79 |
100 |
77 |
|||
0.625 |
93 |
116 |
67 |
72 |
|
90 |
133 |
78 |
|||
1.25 |
81 |
105 |
69 |
78 |
|
72 |
99 |
86 |
|||
2.5 |
75 |
81 |
138 |
134* |
|
98 |
93 |
130 |
|||
5 |
61 |
20 |
338 |
328* |
|
70 |
18 |
318 |
|||
10 |
43 |
7 |
485 |
464* |
|
44 |
7 |
582 |
|||
Positive control |
55 |
57 |
423 |
451* |
|
45 |
47 |
480 |
* Statistically significant increase compared to vehicle controls, P < 0.05
Applicant's summary and conclusion
- Conclusions:
- Positive with and without metabolic activation.
In the mouse lymphoma assay for induction of trifluorothymidine resistance in L5178Y/TK cells, HQ was positive at doses of 1.25 µg/mL and higher in the absence of S9, and at 2.5 µg/mL and higher in the presence of S9. - Executive summary:
Hydroquinone was tested in a mouse lymphoma mutation assay for induction of trifluorothymidine resistance in L5178Y/TK cells with a test protocol similar to OECD Guideline 476. Test concentrations of HQ ranged from 0.625-50 µg/mL without S9 mix and 0.625-10 µg/mL with S9 mix. Treatment time was 4 h, followed by a 2-day expression period and a 11 to 14-day selection period in the presence of TFT. Cytotoxicity was indicated by relative total growth. The lowest effective dose with a significantly increased mutant fraction was 1.25 µg/mL without S9 (relative total growth 32%), and 2.5 µg/mL with S9 (relative total growth 87%). The mutant fraction was statistically significantly and dose-dependantly increased from these concentrations up. HQ was positive in the mouse lymphoma assay both in the presence and absence of S9.
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