Registration Dossier
Registration Dossier
Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.
The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.
Use of this information is subject to copyright laws and may require the permission of the owner of the information, as described in the ECHA Legal Notice.
EC number: 283-919-1 | CAS number: 84775-95-1 Extractives and their physically modified derivatives such as tinctures, concretes, absolutes, essential oils, oleoresins, terpenes, terpene-free fractions, distillates, residues, etc., obtained from Krameria triandra, Krameriaceae.
Genetic toxicity: in vitro
Administrative data
- Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 05.07.2018 - 02.08.2018
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Justification for type of information:
- The study was performed in compliance with the Principle of Good Laboratory Practice, confirmed by Statement of GLP Compliance.
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 018
- Report date:
- 2018
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Principles of method if other than guideline:
- None known
- GLP compliance:
- yes (incl. QA statement)
- Type of assay:
- bacterial reverse mutation assay
Test material
- Reference substance name:
- Rhatany, Krameria triandra, ext.
- EC Number:
- 283-919-1
- EC Name:
- Rhatany, Krameria triandra, ext.
- Cas Number:
- 84775-95-1
- Molecular formula:
- not available
- IUPAC Name:
- Extractives and their physically modified derivatives such as tinctures, concretes, absolutes, essential oils, oleoresins, terpenes, terpene-free fractions, distillates, residues, etc., obtained from Krameria triandra, Krameriaceae.
- Test material form:
- liquid: viscous
1
- Specific details on test material used for the study:
- Name Krameria triandra extract obtained from Rhatany root by hydroalcoholic extraction
Batch no. PES180014
Appearance reddish brown viscous substance
Composition Krameria triandra extract obtained from Rhatany root by hydroalcoholic extraction
Purity not applicable, UVCB
Homogeneity inhomogeneous, warm up to about 60°C and stir
Expiry date Jan. 2019
Storage Fridge (2 - 8 °C)
Method
- Target gene:
- S. typh. TA 98: hisD
S. typh. TA 100: hisG
S. typh. TA 102: hisG
S. typh. TA 97a: hisC
S. typh. TA 1535: hisG
Species / strainopen allclose all
- Species / strain / cell type:
- S. typhimurium, other: TA97a
- Additional strain / cell type characteristics:
- DNA polymerase A deficient
- Species / strain / cell type:
- S. typhimurium TA 98
- Additional strain / cell type characteristics:
- DNA polymerase A deficient
- Species / strain / cell type:
- S. typhimurium TA 100
- Additional strain / cell type characteristics:
- DNA polymerase A deficient
- Species / strain / cell type:
- S. typhimurium TA 102
- Additional strain / cell type characteristics:
- DNA polymerase A deficient
- Species / strain / cell type:
- S. typhimurium TA 1535
- Additional strain / cell type characteristics:
- DNA polymerase A deficient
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9-Mix
- Vehicle / solvent:
- In a non-GLP pre-test, the solubility of the test item was tested in a concentration of 50 g/L in dimethyl sulfoxide (DMSO).
The test item is sufficiently soluble in DMSO, only.
Based on the non-GLP pre-test, DMSO was chosen as vehicle, because the test item was sufficiently soluble, and this solvent does not have any effects on the viability of the bacteria or the number of spontaneous revertants in the tested concentrations.
Controlsopen allclose all
- Positive controls:
- yes
- Positive control substance:
- other: 4-Nitro-1,2-phenylene diamine, C6H7N3O2; CAS-No.: 99-56-9
- Remarks:
- Concentration per plate: 20 µg Solvent DMSO Strains: TA97a, TA98 and TA102 Metabolic activation: none
- Positive controls:
- yes
- Positive control substance:
- sodium azide
- Remarks:
- Concentration per plate: 1 µg Solvent: H2O Strains: TA100 and TA1535 Metabolic activation: none
- Positive controls:
- yes
- Positive control substance:
- other: 2-Amino-anthracene, C14H11N; CAS-No.: 613-13-8
- Remarks:
- Concentration per plate: 1 µg Solvent DMSO Strains: TA97a, TA100, TA102 and TA1535. Metabolic activation: S9
- Positive controls:
- yes
- Positive control substance:
- benzo(a)pyrene
- Remarks:
- Concentration per plate: 20 µg Solvent DMSO Strain: TA98 Metabolic activation: S9
- Details on test system and experimental conditions:
- Different media and solutions were prepared preliminary (exact production dates are doc-umented in the raw data).
On the day of the test, the bacteria cultures were checked for growth visually. The incubation chambers were heated to 37 ±1 °C. The water bath was turned to 43 ±1 °C. The table surface was disinfected.
The S9 mix was freshly prepared and stored at 0 °C.
Date of treatment 18. Jul. 2018
Concentrations tested 5000 / 1500 / 500 / 150 / 50 µg/plate
Incubation time 48 h
Incubation temperature 37 ±1 °C
Tested strains TA97a, TA98, TA100, TA102, TA1535 with and without metabolic activation
Method plate incorporation method - Evaluation criteria:
- Evaluation
The colonies were counted visually and the numbers were recorded. A validated spread-sheet software (Microsoft Excel®) was used to calculate mean values and standard deviations of each treatment, solvent control and positive control.
The mean values and standard deviations of each threefold determination was calculated as well as the increase factor f(l) of revertant induction (mean revertants divided by mean spontaneous revertants) of the test item solutions and the positive controls. Additionally, the absolute number of revertants (Rev. Abs.) (mean revertants less mean spontaneous revertants) was given.
A substance is considered to have mutagenic potential, if a reproducible increase of revertant colonies per plate exceeding an increase factor of 2 in at least one strain can be observed. A concentration-related increase over the range tested is also taken as a sign of mutagenic activity.
Results and discussion
Test resultsopen allclose all
- Species / strain:
- S. typhimurium, other: TA97a
- Remarks:
- Experiment 1a
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- True negative controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 98
- Remarks:
- Experiment 1a
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- True negative controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 100
- Remarks:
- Experiment 1a
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- True negative controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 102
- Remarks:
- Experiment 1a
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- True negative controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 1535
- Remarks:
- Experiment 1a
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- True negative controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium, other: 97a
- Remarks:
- Experiment 1b
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- True negative controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 98
- Remarks:
- Experiment 1b
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- True negative controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 100
- Remarks:
- Experiment 1b
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- True negative controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 102
- Remarks:
- Experiment 1b
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- True negative controls validity:
- valid
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 1535
- Remarks:
- Experiment 1b
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- True negative controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium, other: 97a
- Remarks:
- Experiment 2
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- True negative controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 98
- Remarks:
- Experiment 2
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- True negative controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 100
- Remarks:
- Experiment 2
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- True negative controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 102
- Remarks:
- Experiment 2
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- True negative controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 1535
- Remarks:
- Experiment 2
- Metabolic activation:
- without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- True negative controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 1535
- Remarks:
- Experiment 2
- Metabolic activation:
- with
- Genotoxicity:
- positive
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- True negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- Experiment 1a:
Mutagenicity:
No increase of the number of revertant colonies in the treatments with and without meta-bolic activation could be observed. No concentration-related increase over the tested range was found. Therefore, the test item is stated as not mutagenic under the test conditions in this experiment.
To verify this result, a further experiment was performed with lower concentrations.
Experiment 1b:
Mutagenicity:
No increase of the number of revertant colonies in the treatments with and without metabolic activation could be observed. No concentration-related increase over the tested range was found. Therefore, the test item is stated as not mutagenic under the test conditions in this experiment.
Experiment 2:
Mutagenicity:
An increase of the number of revertant colonies in the treatments with metabolic activation could be observed in the bacteria strain TA1535 in the two highest concentrations (5000 and 2500 µg/plate). A concentration-related increase over the tested range was found. Therefore, the test item is stated as mutagenic under the test conditions of this experiment.
Any other information on results incl. tables
Mean Revertants Experiment 1a
Strain |
TA97a |
TA98 |
TA100 |
TA102 |
TA1535 |
||||||
Induction |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
|
Demin. water |
Mean |
94 |
115 |
40 |
38 |
95 |
96 |
445 |
453 |
11 |
13 |
sd |
15.2 |
27.6 |
4.6 |
8.3 |
20.2 |
14.0 |
44.1 |
26.6 |
2.6 |
2.6 |
|
DMSO |
Mean |
102 |
92 |
50 |
36 |
104 |
102 |
465 |
429 |
11 |
11 |
sd |
11.0 |
21.5 |
1.2 |
4.5 |
18.3 |
9.9 |
48.9 |
32.6 |
1.5 |
4.9 |
|
Positive |
Mean |
748 |
503 |
166 |
174 |
379 |
1001 |
989 |
1189 |
221 |
275 |
sd |
78.8 |
16.2 |
21.6 |
29.5 |
12.9 |
0.0 |
196.3 |
184.6 |
25.9 |
5.0 |
|
f(I) |
7.33 |
5.47 |
3.32 |
4.83 |
3.99 |
9.81 |
2.13 |
2.77 |
20.09 |
25.00 |
|
5000 µg/plate |
Mean |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
0 |
sd |
0.0 |
0.0 |
0.0 |
0.0 |
0.0 |
0.0 |
0.0 |
0.0 |
0.0 |
0.0 |
|
f(I) |
0.00 |
0.00 |
0.00 |
0.00 |
0.00 |
0.00 |
0.00 |
0.00 |
0.00 |
0.00 |
|
1500 µg/plate |
Mean |
0 |
35 |
0 |
0 |
0 |
48 |
38 |
55 |
13 |
11 |
sd |
0.0 |
12.5 |
0.0 |
0.0 |
0.0 |
11.6 |
4.6 |
8.3 |
3.8 |
2.5 |
|
f(I) |
0.00 |
0.38 |
0.00 |
0.00 |
0.00 |
0.47 |
0.08 |
0.13 |
1.18 |
1.00 |
|
500 µg/plate |
Mean |
0 |
107 |
41 |
44 |
12 |
105 |
397 |
461 |
13 |
11 |
sd |
0.0 |
19.4 |
4.5 |
2.6 |
3.5 |
16.0 |
60.0 |
44.1 |
1.7 |
3.1 |
|
f(I) |
0.00 |
1.16 |
0.82 |
1.22 |
0.12 |
1.03 |
0.85 |
1.07 |
1.18 |
1.00 |
|
150 µg/plate |
Mean |
83 |
108 |
39 |
37 |
80 |
112 |
339 |
491 |
9 |
10 |
sd |
7.2 |
12.5 |
1.7 |
4.2 |
7.8 |
25.6 |
30.3 |
12.2 |
1.5 |
1.0 |
|
f(I) |
0.81 |
1.17 |
0.78 |
1.03 |
0.77 |
1.10 |
0.73 |
1.14 |
0.82 |
0.91 |
|
50 µg/plate |
Mean |
82 |
96 |
48 |
46 |
103 |
120 |
409 |
501 |
15 |
10 |
sd |
12.5 |
14.3 |
1.7 |
11.2 |
15.0 |
21.2 |
71.1 |
23.1 |
4.0 |
0.6 |
|
f(I) |
0.80 |
1.04 |
0.96 |
1.28 |
0.99 |
1.18 |
0.88 |
1.17 |
1.36 |
0.91 |
|
1001 colonies per plate means the bacteria growth was too strong for counting
f(I) = increase factor, calculation see chapter7.4, page24
* Different positive controls were used, see chapter6.3, page14
Mean Revertants Experiment 1b
Strain |
TA97a |
TA98 |
TA100 |
TA102 |
TA1535 |
||||||
Induction |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
|
Demin. water |
Mean |
117 |
123 |
41 |
53 |
100 |
102 |
247 |
305 |
14 |
15 |
sd |
8.1 |
11.4 |
9.3 |
2.1 |
11.0 |
5.8 |
30.0 |
22.0 |
4.0 |
3.5 |
|
DMSO |
Mean |
139 |
109 |
47 |
44 |
94 |
117 |
303 |
269 |
16 |
15 |
sd |
19.6 |
10.3 |
0.6 |
4.7 |
9.9 |
11.5 |
66.0 |
15.1 |
2.6 |
2.1 |
|
Positive |
Mean |
593 |
1001 |
341 |
381 |
624 |
1001 |
1469 |
1477 |
455 |
377 |
sd |
155.0 |
0.0 |
37.8 |
72.6 |
108.2 |
0.0 |
66.6 |
95.4 |
148.3 |
66.5 |
|
f(I) |
4.27 |
9.18 |
7.26 |
8.66 |
6.24 |
8.56 |
4.85 |
5.49 |
32.50 |
25.13 |
|
5000 µg/plate |
Mean |
n.d. |
n.d. |
n.d. |
n.d. |
n.d. |
n.d. |
n.d. |
n.d. |
11 |
7 |
sd |
n.d. |
n.d. |
n.d. |
n.d. |
n.d. |
n.d. |
n.d. |
n.d. |
0.6 |
1.5 |
|
f(I) |
n.d. |
n.d. |
n.d. |
n.d. |
n.d. |
n.d. |
n.d. |
n.d. |
0.69 |
0.47 |
|
1500 µg/plate |
Mean |
n.d. |
11 |
15 |
14 |
n.d. |
31 |
47 |
51 |
14 |
12 |
sd |
n.d. |
2.0 |
3.2 |
0.0 |
n.d. |
8.5 |
6.6 |
5.0 |
2.3 |
1.5 |
|
f(I) |
n.d. |
0.10 |
0.32 |
0.32 |
n.d. |
0.26 |
0.16 |
0.19 |
0.88 |
0.80 |
|
500 µg/plate |
Mean |
96 |
94 |
41 |
46 |
46 |
88 |
269 |
276 |
14 |
16 |
sd |
14.4 |
6.0 |
8.1 |
14.6 |
7.6 |
11.2 |
23.1 |
55.0 |
1.2 |
2.6 |
|
f(I) |
0.69 |
0.86 |
0.87 |
1.05 |
0.49 |
0.75 |
0.89 |
1.03 |
0.88 |
1.07 |
|
150 µg/plate |
Mean |
114 |
102 |
43 |
47 |
115 |
82 |
228 |
284 |
14 |
17 |
sd |
7.5 |
16.1 |
7.6 |
11.4 |
4.4 |
10.4 |
59.7 |
4.0 |
3.5 |
2.6 |
|
f(I) |
0.82 |
0.94 |
0.91 |
1.07 |
1.22 |
0.70 |
0.75 |
1.06 |
0.88 |
1.13 |
|
50 µg/plate |
Mean |
120 |
103 |
48 |
36 |
113 |
99 |
265 |
289 |
19 |
14 |
sd |
8.5 |
16.5 |
6.0 |
2.0 |
6.7 |
27.0 |
58.3 |
55.2 |
3.8 |
3.5 |
|
f(I) |
0.86 |
0.94 |
1.02 |
0.82 |
1.20 |
0.85 |
0.87 |
1.07 |
1.19 |
0.93 |
|
15 µg/plate |
Mean |
123 |
132 |
44 |
40 |
93 |
128 |
276 |
288 |
16 |
15 |
sd |
4.0 |
11.0 |
6.1 |
7.0 |
13.8 |
17.8 |
26.2 |
36.7 |
2.1 |
2.3 |
|
f(I) |
0.88 |
1.21 |
0.94 |
0.91 |
0.99 |
1.09 |
0.91 |
1.07 |
1.00 |
1.00 |
|
5 µg/plate |
Mean |
103 |
117 |
49 |
44 |
97 |
125 |
287 |
301 |
n.d. |
n.d. |
sd |
17.2 |
22.3 |
6.4 |
3.1 |
8.7 |
13.8 |
34.5 |
26.6 |
n.d. |
n.d. |
|
f(I) |
0.74 |
1.07 |
1.04 |
1.00 |
1.03 |
1.07 |
0.95 |
1.12 |
n.d. |
n.d. |
|
n.d. = not determined, due to the toxicity effect
1001 colonies per plate means the bacteria growth was too strong for counting
f(I) = increase factor, calculation see chapter7.4, page24
* Different positive controls were used, see chapter6.3, page14
Mean Revertants Experiment 2
Strain |
TA1535 |
||
Induction |
-S9 |
+S9 |
|
Demin. water |
Mean |
15 |
17 |
sd |
2.0 |
3.5 |
|
DMSO |
Mean |
16 |
17 |
sd |
1.5 |
2.3 |
|
Positive |
Mean |
317 |
269 |
sd |
37.8 |
84.8 |
|
f(I) |
21.13 |
15.82 |
|
5000 µg/plate |
Mean |
17 |
137 |
sd |
4.2 |
7.6 |
|
f(I) |
1.06 |
8.06 |
|
2500 µg/plate |
Mean |
15 |
40 |
sd |
1.2 |
9.2 |
|
f(I) |
0.94 |
2.35 |
|
1250 µg/plate |
Mean |
13 |
15 |
sd |
2.5 |
1.2 |
|
f(I) |
0.81 |
0.88 |
|
625 µg/plate |
Mean |
15 |
15 |
sd |
3.6 |
3.2 |
|
f(I) |
0.94 |
0.88 |
|
313 µg/plate |
Mean |
13 |
14 |
sd |
1.5 |
2.5 |
|
f(I) |
0.81 |
0.82 |
|
156 µg/plate |
Mean |
14 |
13 |
sd |
1.2 |
2.6 |
|
f(I) |
0.88 |
0.76 |
|
78 µg/plate |
Mean |
14 |
13 |
sd |
3.1 |
3.2 |
|
f(I) |
0.88 |
0.76 |
|
f(I) = increase factor, calculation see chapter7.4, page24* Different positive controls were used, see chapter6.3, page14
Applicant's summary and conclusion
- Conclusions:
- The test item Krameria triandra extract obtained from Rhatany root by hydroalcoholic extraction showed an increase in the number of revertants in the bacteria strain TA1535 in in the second experiment in the treatment with metabolic activation (two highest concentrations).
Nearly all negative and all strain-specific positive control values were within the laboratory historical control data ranges indicating that the test conditions were adequate and that the metabolic activation system functioned properly.
Based on the results of this study it is concluded that Krameria triandra extract obtained from Rhatany root by hydroalcoholic extraction is mutagenic in the Salmonella typhimurium strain TA1535 in the presence of metabolic activation under the experimental conditions in this study. - Executive summary:
The bacterial reverse mutation test uses amino-acid requiring strains of Salmonella typhimurium to detect point mutations, which involve substitution, addition or deletion of one or a few DNA base pairs. The principle of this bacterial reverse mutation test is that it detects mutations which revert mutations present in the test strains and restore the functional capability of the bacteria to synthesize an essential amino acid.
The bacterial reverse mutation test is commonly employed as an initial screen for genotoxic activity and, in particular, for point mutation-inducing activity.
Principle of the test method: Suspensions of bacterial cells are exposed to the test substance in the presence and in the absence of an exogenous metabolic activation system. In the plate incorporation method, these suspensions are mixed with an overlay agar and plated immediately onto minimal medium. In the pre-incubation method, the treatment mixture is incubated and then mixed with an overlay agar before plating onto minimal medium. For both techniques, after 2 days of incubation, revertant colonies are counted and compared to the number of spontaneous revertant colonies on solvent control plates.
This study was performed in order to evaluate the mutagenic potential of Krameria triandra extract obtained from Rhatany root by hydroalcoholic extraction in the Bacterial Reverse Mutation Test using five strains of Salmonella typhimurium.
Three valid experiments were performed.
The study procedures described in this report were based on the most recent OECD and EC guidelines.
The test item Krameria triandra extract obtained from Rhatany root by hydroalcoholic extraction was tested in the Salmonella typhimurium reverse mutation assay with five strains of Salmonella typhimurium (TA97a, TA98, TA100, TA102 and TA1535).
The test was performed in three experiments in the presence and absence of metabolic activation, with +S9 standing for presence of metabolic activation, and –S9 standing for absence of metabolic activation.
Experiment 1a:
In the first experiment, the test item (dissolved in DMSO) was tested up to concentrations of 5000 μg/plate in the absence and presence of S9-mix in the strains TA97a, TA98, TA100, TA102 and TA1535 using the plate incorporation method.
The test item showed no precipitates on the plates at any of the concentrations.
The test item showed signs of toxicity towards the following bacteria strains (in the absence resp. in the presence of metabolic activation) in the following concentrations:
• TA97a without metabolic activation: 5000 μg/plate (no bacteria growth), 1500 μg/plate (no bacteria growth) and 500 μg/plate (no bacteria growth)
• TA97a with metabolic activation: 5000 μg/plate (no bacteria growth), 1500 μg/plate (decrease in the number of revertants)
• TA98 with and without metabolic activation: 5000 μg/plate (no bacteria growth), 1500 μg/plate (no bacteria growth)
• TA100 without metabolic activation: 5000 μg/plate (no bacteria growth), 1500 μg/plate (no bacteria growth) and 500 μg/plate (decrease in the number of revertants)
• TA100 with metabolic activation: 5000 μg/plate (no bacteria growth), 1500 μg/plate (decrease in the number of revertants)
• TA102 with and without metabolic activation: 5000 μg/plate (no bacteria growth), 1500 μg/plate (decrease in the number of revertants)
• TA1535 with and without metabolic activation:5000 μg/plate (no bacteria growth)
The bacterial background lawn was not reduced at any of the concentrations.
The results of this experiment showed that none of the tested concentrations showed a significant increase in the number of revertants in all tested strains, in the presence and the absence of metabolic activation.
Experiment 1b:
Based on the toxicity results of the experiment 1a, the test item was tested up to different concentrations in the absence and presence of S9-mix in all five bacteria strains using plate incorporation method.
The test item showed no precipitates on the plates at any of the concentrations.
The test item showed signs of toxicity towards the following bacteria strains (in the absence resp. in the presence of metabolic activation) in the following concentrations:
• TA97a without metabolic activation: 500 μg/plate (decrease in the number of revertants)
• TA97a with metabolic activation: 1500 μg/plate (decrease in the number of revertants)
• TA98 with and without metabolic activation: 1500 μg/plate (decrease in the number of revertants)
• TA100 without metabolic activation: 500 μg/plate (decrease in the number of revertants)
• TA100 with metabolic activation: 1500 μg/plate (decrease in the number of revertants)
• TA102 with and without metabolic activation: 1500 μg/plate (decrease in the number of revertants)
• TA1535 with and without metabolic activation:5000 μg/plate (decrease in the number of revertants)
The bacterial background lawn was not reduced at any of the concentrations.
The results of this experiments showed that the test item caused no increase in the number of revertants in all bacteria strains compared to the solvent control, in both the absence and presence of metabolic activation. The test item did not induce a dose-related increase in the number of revertants colonies in all strains, in the presence and absence of metabolic activation.
Experiment 2:
Based on the results of experiment 1a and 1b, the test item was tested up to the resp. concentrations of experiment 1b in the absence and presence of S9-mix in all bacteria strains using the pre-incubation method.
The test item showed no precipitates on the plates at any of the concentrations.
The test item showed signs of toxicity towards the following bacteria strains (in the absence resp. in the presence of metabolic activation) in the following concentrations:
• TA97a without metabolic activation: 500 μg/plate (no bacteria growth)
• TA97a with metabolic activation: 1500 μg/plate (no bacteria growth)
• TA98 with and without metabolic activation: 1500 μg/plate (no bacteria growth)
• TA100 without metabolic activation: 500 μg/plate (no bacteria growth)
• TA100 with metabolic activation: 1500 μg/plate (no bacteria growth)
• TA102 with and without metabolic activation: 1500 μg/plate (no bacteria growth)
No bacterial background lawn was present at the concentration 1500 μg/plate (bacteria strain TA97a and TA100 with metabolic activation) and 500 μg/plate (bacteria strain TA97a and TA100 without metabolic activation)
At the other concentrations, the bacterial background lawn was not reduced at any of the concentrations.
The results of this experiments showed that the test item caused an increase in the number of revertants in the bacteria strain TA1535 compared to the solvent control, in the presence of metabolic activation in the two highest concentrations (5000 and 2500 μg/plate). The test item induced a dose-related increase in the number of revertants colonies in this bacteria strain, in the presence of metabolic activation.
Based on the results of this study it is concluded that Krameria triandra extract obtained from Rhatany root by hydroalcoholic extraction is mutagenic in the Salmonella typhimurium strain TA1535 in the presence of metabolic activation under the experimental conditions in this study.
Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice.
Reproduction or further distribution of this information may be subject to copyright protection. Use of the information without obtaining the permission from the owner(s) of the respective information might violate the rights of the owner.
