N-(N6-trifluoroacetyl-L-lysyl)-L-proline

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

An Ames test conducted according to OECD TG 471 it is concluded that Epsilon-Trifluoracetyl-L-lysyl-L-proline did not show mutagenic activity in Salmonella typhimurium TA 1535, TA 1537, TA 1538, TA 98 or TA 100 either in the absence or in the presence of the S-9 mix, under the conditions employed in this evaluation.

Link to relevant study records

Reference

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1990-06-22 to 1990-07-17

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

1983

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Target gene:

His locus

Species / strain / cell type:

other: S. typhimurium TA 1535, TA 1537, TA 1538, TA 98 and TA 100

Metabolic activation:

with and without

Metabolic activation system:

Due to migration, the value was transferred to one of the current document's attachments

Test concentrations with justification for top dose:

Concentration range in the preliminary test (with metabolic activation) 0, 5.0, 50.0, 500.0, 5000.0, 50000.0 µg/plate
Concentration range in the preliminary test (without metabolic activation) 0, 5.0, 50.0, 500.0, 5000.0, 50000.0 µg/plate
Concentration range in the main test (with metabolic activation): 0, 620, 1850, 5560, 16670, 50000 µg/plate
Concentration range in the main test (without metabolic activation): 0, 620, 1850, 5560, 16670, 50000 µg/plate

Vehicle / solvent:

Water

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

2-nitrofluorene

sodium azide

other: 2-aminoanthracene

Details on test system and experimental conditions:

NUMBER OF REPLICATIONS:
- Number of cultures per concentration: triplicate
- Number of independent experiments: 2

METHOD OF TREATMENT/ EXPOSURE:
- Cell density at seeding (if applicable):
Experiment 1:
TA1535: 3.5E+08 cells/mL
TA1537: 1.5E+08 cells/mL
TA1538: 1.9E+08 cells/mL
TA98: 2.5E+08 cells/mL
TA100: 4.6E+08 cells/mL
Experiment 2:
TA1535: 1.2E+08 cells/mL
TA1537: 4.5E+07 cells/mL
TA1538: 2.5E+08 cells/mL
TA98: 1.8E+08 cells/mL
TA100: 1.8E+08 cells/mL
- Test substance added in agar (plate incorporation)

TREATMENT AND HARVEST SCHEDULE:
- Exposure duration/duration of treatment: 72h

METHODS FOR MEASUREMENT OF CYTOTOXICITY
- Method, e.g.: background growth inhibition

METHODS FOR MEASUREMENTS OF GENOTOXICIY
A positive response in the assay system is taken to be a two-fold or greater increase in the mean number of revertant colonies appearing in the test plates over and above the background spontaneous reversion rate observed with the vehicle, together with evidence of a dose-response.

Rationale for test conditions:

As recommended by the guideline

Evaluation criteria:

A positive response in the assay system is taken to be a two-fold or greater increase in the mean number of revertant colonies appearing in the test plates over and above the background spontaneous reversion rate observed with the vehicle, together with evidence of a dose-response.

Key result

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

(> 50000 µg/plate)

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Key result

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

(> 50000 µg/plate)

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Key result

Species / strain:

S. typhimurium TA 1538

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

(> 50000 µg/plate)

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Key result

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

(> 50000 µg/plate)

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

> 50000 µg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

RANGE-FINDING/SCREENING STUDIES (if applicable):
A preliminary test was performed at the following concentrations:
Concentration range in the preliminary test (with metabolic activation) 0, 5.0, 50.0, 500.0, 5000.0, 50000.0 µg/plate
Concentration range in the preliminary test (without metabolic activation) 0, 5.0, 50.0, 500.0, 5000.0, 50000.0 µg/plate
the test substance was not toxic for all strains at all dose levels, thus the highest concentration in the main test was 50000 µg/plate

STUDY RESULTS
- Concurrent vehicle negative and positive control data
Experiment 1:
negative control
-S9: TA1535: 21 ± 2; TA1537: 18 ± 5; TA1538: 27 ± 7; TA98: 48 ± 8; TA100: 78 ± 18
+S9:TA1535: 24 ± 3; TA1537: 23 ± 3; TA1538: 60 ± 10; TA98: 68 ± 3; TA100: 82 ± 3
positive control:
-S9: TA1535: 691 ± 84; TA1537: 1361 ± 310; TA1538: 1367 ± 60; TA98: 857 ± 56; TA100: 839 ± 26
+S9:TA1535: 226 ± 26; TA1537: 216 ± 42; TA1538: 812 ± 17; TA98: 669 ± 47; TA100: 610 ± 46
Experiment 2:
negative control
-S9: TA1535: 66 ± 6; TA1537: 11 ± 4; TA1538: 26 ± 5; TA98: 51 ± 7; TA100: 170 ± 5
+S9:TA1535: 17 ± 1; TA1537: 17 ± 5; TA1538: 47 ± 3; TA98: 72 ± 20; TA100: 149 ± 8
positive control:
-S9: TA1535: 734 ± 113; TA1537: 1820 ± 202; TA1538: 1033 ± 27; TA98: 627 ± 74; TA100: 719 ± 123
+S9:TA1535: 213 ± 25; TA1537: 327 ± 120; TA1538: 643 ± 24; TA98: 420 ± 26; TA100: 544 ± 41

For all test methods and criteria for data analysis and interpretation:
- Statistical analysis; p-value if any : No statistical analysis was reported
Ames test:
- Signs of toxicity yes at > 50000µg/plate
- Individual plate counts :
Experiment 1:
TA1535: 3.5E+08 cells/mL
TA1537: 1.5E+08 cells/mL
TA1538:1.9E+08 cells/mL
TA98: 2.5E+08 cells/mL
TA100: 4.6E+08 cells/mL
Experiment 2:
TA1535: 1.2E+08 cells/mL
TA1537: 4.5E+07 cells/mL
TA1538:2.5E+08 cells/mL
TA98: 1.8E+08 cells/mL
TA100: 1.8E+08 cells/mL
- Mean number of revertant colonies per plate and standard deviation
Experiment 1:
TA1535: 18
TA1537: 15
TA1538:24
TA98: 37
TA100: 89
Experiment 2:
TA1535: 33
TA1537: 18
TA1538:24
TA98: 25
TA100: 182
HISTORICAL CONTROL DATA (with ranges, means and standard deviation, and 95% control limits for the distribution as well as the number of data) not reported

Remarks on result:

other:

Experiments 1: Results

Dose µg/plate		TA1535		TA1537		TA1538		TA98		TA100
		- S9	+ S9	- S9	+ S9	- S9	+ S9	- S9	+ S9	- S9	+ S9
0		22	21	12	21	20	58	42	67	79	80
		22	26	19	21	27	52	45	71	95	82
		18	26	22	27	34	71	57	65	60	85
	Mean	21	24	18	23	27	60	48	68	78	82
	SD	2	3	5	3	7	10	8	3	18	3
620		19	22	17	22	26	62	49	64	67	71
		22	26	17	24	22	63	38	73	85	80
		21	29	21	19	32	56	40	67	80	89
	Mean	21	26	18	22	27	60	42	68	77	80
	SD	2	4	2	3	5	4	6	5	9	9
1850		22	19	18	25	36	81	38	68	103	103
		21	24	19	27	43	70	33	73	106	97
		24	18	22	21	40	61	50	64	107	120
	Mean	22	20	20	24	40	71	40	68	105	107
	SD	2	3	2	3	4	10	9	5	2	12
5560		24	22	25	27	28	44	35	82	105	95
		17	22	19	30	31	69	50	76	74	85
		21	29	19	27	30	65	51	72	80	83
	Mean	21	24	21	28	30	59	45	77	86	88
	SD	4	4	3	2	2	13	9	5	16	6
16670		24	15	9	19	16	55	34	96	72	89
		17	17	13	29	22	57	44	111	77	105
		29	19	14	30	27	64	36	82	86	93
	Mean	23	17	12	26	22	59	38	96	78	96
	SD	6	2	3	6	6	5	5	15	7	8
50000		18	14	9	19	27	42	14	52	127	128
		11	18	10	13	24	39	20	63	123	115
		19	13	17	17	19	36	22	39	100	116
	Mean	16	15	16	16	23	39	19	51	117	120
	SD	4	3	3	3	4	3	4	12	15	7
Pos. C.		613	223	025	245	1436	826	908	624	840	567
		780	201	635	235	1338	793	797	664	813	659
		679	253	422	168	1326	816	865	718	865	605
	Mean	691	226	361	216	1367	812	857	669	839	610
	SD	84	26	310	42	60	17	56	47	26	46

Experiment 2: Results:

Dose µg/plate		TA1535		TA1537		TA1538		TA98		TA100
		- S9	+ S9	- S9	+ S9	- S9	+ S9	- S9	+ S9	- S9	+ S9
0		28	17	12	17	22	43	43	94	174	157
		17	17	7	12	24	48	57	64	165	148
		20	18	14	22	22	49	54	57	172	141
	Mean	22	17	11	17	26	47	51	72	170	149
	SD	6	1	4	5	5	3	7	20	5	8
620		25	19	13	21	20	43	39	62	174	184
		37	14	16	8	22	53	48	61	159	165
		36	16	23	22	32	54	48	78	168	150
	Mean	33	16	17	17	25	50	45	67	167	166
	SD	7	3	5	8	6	6	5	10	8	17
1850		33	33	12	15	25	57	53	76	177	160
		29	28	12	21	36	57	39	89	212	177
		30	25	12	20	30	61	40	73	182	191
	Mean	31	29	12	19	30	58	44	79	190	176
	SD	2	4	0	3	6	2	8	9	19	16
5560		17	23	18	22	27	54	34	74	182	191
		34	25	13	20	27	49	39	65	190	209
		25	16	24	23	34	47	33	73	172	194
	Mean	25	21	18	22	29	50	35	71	181	198
	SD	9	5	6	2	4	4	3	5	9	10
16670		11	21	16	16	15	54	56	73	168	158
		30	22	12	16	21	53	48	57	162	169
		26	21	18	18	21	44	52	80	180	156
	Mean	22	21	15	17	19	50	52	70	170	161
	SD	10	1	3	1	3	6	4	12	9	7
50000		21	20	12	26	15	41	25	65	146	172
		11	16	2	18	37	35	26	43	156	170
		19	12	15	15	36	43	24	49	154	163
	Mean	17	16	10	20	29	40	25	52	152	168
	SD	5	4	7	6	12	4	1	11	5	5
Pos. C.		604	190	1590	440	1002	627	542	450	818	586
		801	240	1905	201	1051	631	674	404	581	543
		798	210	1966	341	1045	670	665	406	758	504
	Mean	734	213	1820	327	1033	643	627	420	719	544
	SD	113	25	202	120	27	24	74	26	123	41

Conclusions:

From the above findings from a study conducted according to OECD guideline 471 (1983) it is concluded that Epsilon-Trifluoracetyl-L-lysyl-L-prolin did not show mutagenic activity in Salmonella typhimurium TA 1535, TA 1537, TA 1538, TA 98 or TA 100 either in the absence or in the presence of the S-9 mix, under the conditions employed in this evaluation. Thus, the test item does not need to be classified as mutagenic according to Regulation (EC) No 1272/2008 (CLP) and the Globally Harmonized System for Classification and Labelling of Chemicals (GHS).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Genetic toxicity in vivo

Description of key information

In an in vivo mouse micronucleus study conducted according to EU Method B.12 (1992) , ε-Trifluoroacetyl-L-Iysyl-L-proline did not induce any chromosome mutations in mice by damage to the chromosomes or the mitotic apparatus at 24- or 48-hour intervals alter the animals had received a single intraperitoneal dose of 2150 mg/kg bw. Therefore, ε-TrifluoroacetyI-L-lysyl-L-proline is considered non-mutagenic in the reported in the vivo mouse micronucleus test.

Link to relevant study records

Reference

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1994-06-14 to 1995-05-18

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EU Method B.12 (Mutagenicity - In Vivo Mammalian Erythrocyte Micronucleus Test)

Version / remarks:

1992

Deviations:

no

GLP compliance:

yes

Type of assay:

micronucleus assay

Species:

mouse

Strain:

NMRI

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: 6 weeks
- Weight at study initiation: males : 27 - 33 g females: 24 - 28 g
- Assigned to test groups randomly: yes, under following basis:using computer generated random numbers.
- Fasting period before study: 16h before start of the experiments
- Housing: individually in Macrolon cages, type II with Animal bedding softwood granulation HW 300/500W supplied by Jelu-Werk, J. Ehrler, D-73494 Rosenberg
- Diet (e.g. ad libitum): Standard diet ad libitum, ssniff® M, "Special diet for Mice", supplied by SSNIFF Spezialdiaten GmbH, D-59494 Soest
- Water (e.g. ad libitum):Water ad Iibitum in drinking-water quality from the Stadtwerke Halle/Westfalen was provided after filtration by a Pall Seal- kleen Filter (Pall Sealkleen Filter with candle, type SLK 7002 ARP, supplied by Pall Filtrationstechnik GmbH, D-63303 Dreieich/Sprendlingen). Water was provided in drinking-bottles.
- Acclimation period: 6 days under test conditions with veterinary care.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.5 - 22.0
- Humidity (%): 58 - 63
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

intraperitoneal

Vehicle:

- Vehicle(s)/solvent(s) used: water
- Concentration of test material in vehicle: 100 mg/mL
- Amount of vehicle (if gavage or dermal): 21.5 mL/kg b.w. i.p.

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:
An orientating study of the acute toxicity of epsilon-Trifluoroacetyl-L-Iysyl-L-proline in mice after single intraperitoneal administration with the test substance batch used in the present experiment was performed. In this study, solely the maximum dose to administer intraperitoneally according to the guideline (2150 mg/kg body weight) caused first signs of toxicity. Therefore, this dose was used in the micronucleus test. Administration Volume: 21.5 mL/kg b.w.

Duration of treatment / exposure:

At 24- and 48-hour intervals after treatment, 6 mice each per sex of the negative control group and 7 mice each per sex of the test material group were sacrificed to prepare bone marrow smears. All animals of the positive control group were sacrificed 24 hours after treatment.

Frequency of treatment:

once

Dose / conc.:

2 150 mg/kg bw/day (nominal)

Remarks:

as recommended by the guideline

No. of animals per sex per dose:

Control group: 12 males and 12 females
Positive Control group: 6 males and 6 females
Test group 14 males and 14 females

Control animals:

yes

Positive control(s):

cyclophosphamide
- Justification for choice of positive control(s): according to guideline
- Route of administration: i.p.
- Doses / concentrations:31.6 mg/kg bw

Tissues and cell types examined:

bone marrow

Details of tissue and slide preparation:

CRITERIA FOR DOSE SELECTION: according to guideline
TREATMENT AND SAMPLING TIMES ( in addition to information in specific fields): Three groups of mice, the negative control group containing 12 males and 12 females, the positive control group containing 6 males and 6 females, and the test material group with 14 males and 14 females each received a single intraperitoneal administration (all groups: diet withdrawal: 16 h before treatment).
Group 1, the negative control, received physiological saline solution (0.9%).
Group 2, the test material group, received 2150 mg/kg body weight (males and females) of the test material. The test material was given as a freshly prepared solution in aqua ad iniectabilia.
Group 3, the positive control, received Cyclophosphamide(31.6 mg/kg b.w.) dissolved in physiological saline solution (0.9%).
The animals were observed for up to 7.5 hours after administration for the occurrence of toxicity symptoms; on days 2 and 3 only once daily in the morning. At 24- and 48-hour intervals after treatment, 6 mice each per sex of the negative control group and 7 mice each per sex of the test material group were sacrificed to prepare bone marrow smears. All animals of the positive control group were sacrificed 24 hours after treatment. After sacrifice , in group 2 animals the abdominal cavity was opened for optical control of the resorption of the test material solution.
DETAILS OF SLIDE PREPARATION: All mice were sacrificed by CO2 overdose. Both femurs were removed from each mouse and the bone marrow cells flushed into a labelled centrifuge tube with approximately 1.5 mL of fetal calf serum (Art. No. 210463, Boehringer Mannheim GmbH, D-68305 Mannheim). The tubes were centrifuged at approx. 180 x g for 5 minutes (Eppendorf Zentrifuge 5415, Eppendorf- Netheler-Hinz GmbH, D-22331 Hamburg), after which the supernatant serum was discarded and the bone marrow cells suspended upon a thin layer of serum. A small drop of the marrow serum suspension was smeared on a slide, which was identified by study number, animal number, species, sex, and date of preparation, and allowed to dry overnight. At least two slides per animal were prepared. The following day, the smears were stained using the panoptic stain method developed by PAPPENHEIM. Before evaluation, the slides were coded separately per sex. So they could not be assigned to animals of a specific group or sampling time during the microscopical examination.
METHOD OF ANALYSIS: For each sampling time the bone marrow smears from the first 5 animals per sex and group were used for evaluation. One slide per animal was examined. The remaining smears of each sex and group per interval were evaluated if macroscopical examination of the first smears revealed technical imperfections which precluded accurate microscopical analysis. From each animal one thousand polychromatic erythrocytes (PCE) were scored under the microscope (magnification 650- 1000 x, C. ZEISS, D-73447 Oberkochen) for the incidence of polychromatic erythrocytes with micronuclei. The ratio of polychromatic to normochromatic erythrocytes (PCE/NCE) was calculated, based on 1000 erythrocytes (PCE + NCE) scored per slide, as a measure of the toxic efficacy of the test material.

Evaluation criteria:

If a test material produced neither a statistically significant and reproducible positive response nor a dose related statistically significant response at any one of the test points compared to the negative control group, it is considered non-mutagenic in this system (significance level: 5%; p < 0.05).

Statistics:

The frequencies of polychromatic erythrocytes with micronuclei of the test material group and of the positive control group were compared with those of the negative control group at each sampling time (positive control only at 24 hours). A POISSON test (6) was applied. The data from each treatment group for each sex as well as for both sexes combined were compared with the respective negative control group data by means of a software developed by the Biometrical Department (FB) of ASTA Medica AG using a VAX 4000-400 computer (Digital Equipment Corp., D-81927 München). The values of the first 5 animals of each sex and sampling time were used for statistical evaluation.

Key result

Sex:

male/female

Genotoxicity:

negative

Toxicity:

no effects

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

RESULTS OF DEFINITIVE STUDY
- Induction of micronuclei (for Micronucleus assay): negative
- Ratio of PCE/NCE (for Micronucleus assay): please refer to any other information on results incl. tables.
- PCEs with MN:
Negative control:
24h application: Males: mean: 1.8 (0.84) n=5, females: mean: 1.2 (0.45) n=5
48h application:Males: mean: 2.2 (0.84) n=5, females: mean: 1.0 (1.00) n=5
Test item:
24h application: Males: mean: 1.6 (1.14) n=5, females: mean: 2.2 (1.64) n=5
48h application:Males: mean: 2.6 (1.67) n=5, females: mean: 2.4 (1.67) n=5
Positive control:
24h application: Males: mean: 29.0 (11.68) n=5, females: mean: 11.4 (5.18) n=5
- Statistical evaluation: please refer to any other information on results incl. tables.

Statistical Evaluation of the Data (POISSON Test)

	24h		48h
No. Of PCEs with MN	m	f	m	f
verum	8	11	13	12
negative control	9	6	11	5
test statistic F	0.800	1.5714	1.0833	2.000
degrees of freedom	20,16	14,22	24,26	12,24
p-value	0.685	0.166	0.419	0.072
No. of PCEs with MN
positiv control	145	57
negativ control	9	6
test statistic F	14.5000	8.1429
degrees of freedom	20,290	14,114
p-value	0.000	0.000

Results (Negative control)

24 h application				48 h application
Animal No.	PCEs with MN		Ration PCE/NCE	Animal No.	PCEs with MN		Ration PCE/NCE
	N	%			N	%
Males
1	3	0.3	1.95	7	3	0.3	2.55
2	2	0.2	2.38	8	2	0.2	2.36
3	1	0.1	2.60	9	3	0.3	1.98
4	1	0.1	2.29	10	2	0.2	1.74
5	2	0.2	2.60	11	2	0.2	1.81
Mean	1.8				2.2
SD	0.84				0.84
N	5				5
Females
13	1	0.1	2.56	19	2	0.2	1.95
14	1	0.1	2.77	20	1	0.1	1.91
15	2	0.2	1.84	21	0	0	2.08
16	1	0.1	2.45	22	0	0	2.62
17	1	0.1	2.34	23	2	0.2	3.18
Mean	1.2				1.0
SD	0.45				1.00
N	5				5
Mean
M+F	1.5				1.6
SD	0.45				1.07
N	10				10

Results Test item

24 h application				48 h application
Animal No.	PCEs with MN		Ration PCE/NCE	Animal No.	PCEs with MN		Ration PCE/NCE
	N	%			N	%
Males
101	2	0.2	2.17	108	4	0.4	2.14
102	2	0.2	1.44	109	0	0	1.70
103	1	0.1	2.26	110	2	0.2	2.64
104	0	0	2.16	111	4	0.4	2.57
105	3	0.3	2.25	112	3	0.3	2.95
Mean	1.6				2.6
SD	1.14				1.67
N	5				5
Females
115	1	0.1	1.24	122	1	0.1	2.50
116	2	0.2	1.60	123	3	0.3	1.97
117	2	0.2	3.46	124	5	0.5	2.27
118	5	0.5	1.82	125	2	0.2	1.46
119	1	0.1	1.78	126	1	0.1	2.42
Mean	2.2				2.4
SD	1.64				1.67
N	5				5
Mean
M+F	1.9				2.5
SD	13.37				1.58
N	10				10

Results Positive control

24 h application
Animal No.	PCEs with MN		Ration PCE/NCE
	N	%
Males
401	49	4.9	3.65
402	29	2.9	4.99
403	20	2.0	3.52
404	25	2.5	2.41
405	22	2.2	3.57
Mean	29.0
SD	11.68
N	5
Females
407	9	0.9	1.98
408	5	0.5	1.95
409	11	1.1	1.65
410	13	1.3	2.03
411	19	1.9	3.20
Mean	11.4
SD	5.18
N	5
Mean
M+F	20.2
SD	12.59
N	10

Conclusions:

In a study conducted according to EU Method B.12 (1992) , ε-Trifluoroacetyl-L-Iysyl-L-proline induced no chromosome mutations in mice by damage to the chromosomes or the mitotic apparatus at 24- or 48-hour intervals alter the animals had received a single intraperitoneal dose of 2150 mg/kg body weight. Therefore, ε-TrifluoroacetyI-L-lysyl-L-proline is considered non-mutagenic in the reported in vivo mouse micronucleus test and does not need to be classified according to Regualtion (EC)No 1272/2008 (CLP) and the Globally Harmonized System for Classification and Labelling of Chemicals (GHS).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Justification for classification or non-classification

In an in vivo mouse micronucleus study conducted according to EU Method B.12 (1992) , ε-Trifluoroacetyl-L-Iysyl-L-proline did not induce any chromosome mutations in mice by damage to the chromosomes or the mitotic apparatus at 24- or 48-hour intervals alter the animals had received a single intraperitoneal dose of 2150 mg/kg bw. Therefore, ε-TrifluoroacetyI-L-lysyl-L-proline is considered non-mutagenic in the reported in the vivo mouse micronucleus test. Thus, it does not need to be classified according to Regualtion (EC)No 1272/2008 (CLP) and the Globally Harmonized System for Classification and Labelling of Chemicals (GHS).

An Ames test conducted according to OECD TG 471 it is concluded that Epsilon-Trifluoracetyl-L-lysyl-L-proline did not show mutagenic activity in Salmonella typhimurium TA 1535, TA 1537, TA 1538, TA 98 or TA 100 either in the absence or in the presence of the S-9 mix, under the conditions employed in this evaluation. Thus, the test item does not need to be classified as mutagenic according to Regulation (EC) No 1272/2008 (CLP) and the Globally Harmonized System for Classification and Labelling of Chemicals (GHS).