Bis(ethyl acetoacetato-O1',O3)bis(propan-2-olato)titanium

Genetic toxicity: in vitro

Administrative data

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:

January 11, 1978 - May 19, 1978

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Even though study pre-dates GLP and current guideline, the study method is comparable to OECD 471 (Bacterial reverse mutation assay) with one deviation of not using TA 102 bacterial strain.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1978

Report date:

1978

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

Recommended TA102 bacterial strain was not used in the assay.

Principles of method if other than guideline:

Suspension of Salmonella Typhimurium cells were exposed to the test substance in the presence and in the absence of an exogenous metbolic activation system. These suspensions were mixed with with an overlay agar and plated immediately onto minimal medium. After two days of incubation, revertant colonies are counted and compared to the number of spontaneous revertant colonies on solvent control plates. Study was performed according to reference Ames, B. N., McCann, J. and Yamasaki, E. (1975) Methods for Detecting Carcinogens and Mutagens with the Salmonella/Mammalian-Microsome Mutagenicity Test. Mutat. Res. 31, 347-364.

GLP compliance:

no

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Bis(ethyl acetoacetato-O1',O3)bis(propan-2-olato)titanium

EC Number:

248-697-2

EC Name:

Bis(ethyl acetoacetato-O1',O3)bis(propan-2-olato)titanium

Cas Number:

27858-32-8

Molecular formula:

C18H32O8Ti

IUPAC Name:

Bis(ethylacetoacetato‐O1',O3")bis(propan‐2‐olato)titanium

Details on test material:

- Name of test material (as cited in study report): Haskell No. 11,939-02; Titanic acid, Diacetoacetyl-diisopropyl; Tyzor DC

Method

Target gene:

histidine locus in selected strains

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Species / strain 2

Species / strain / cell type:

S. typhimurium TA 1538

Metabolic activation:

with and without

Metabolic activation system:

rat-liver homogenate activation system (S-9)

Test concentrations with justification for top dose:

1000, 3000, 5000, 7000 and 10000 µg test substance/plate with and without metabolic activation

Vehicle / solvent:

acetone

Controls

Controls 1

Untreated negative controls:

yes

Remarks:

culture medium without solvent

Negative solvent / vehicle controls:

yes

Remarks:

acetone

Positive controls:

yes

Positive control substance:

other: 2-aminoanthracene

Remarks:

used for assay with metabolic activation

Controls 2

Untreated negative controls:

yes

Remarks:

test plate without glucose-6-phosphate and NAPDH

Remarks:

used for assay with metabolic activation

Controls 3

Negative solvent / vehicle controls:

yes

Remarks:

acetone

Positive controls:

yes

Positive control substance:

2-nitrofluorene

Remarks:

used for assay without metabolic activation

Controls 4

Untreated negative controls:

yes

Remarks:

culture medium without solvent

Positive controls:

yes

Positive control substance:

9-aminoacridine

Remarks:

used for assay without metabolic activation

Controls 5

Positive controls:

yes

Positive control substance:

other: N-Methyl-n'-nitro-N-nitrosoguanidine

Remarks:

used for assay without metabolic activation

Details on test system and experimental conditions:

Five histidine-requiring strains of Salmonella typhimurium were used in the mutagen assays.
Strains TA 1535 and TA 100 were used to detect base-pair substitution mutations, whereas strains TA 1537, T 1538 and TA 98 were used to detect frame-shift mutations. Two different experiments were performed to confirm the results.

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Vehicle controls validity:

valid

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Vehicle controls validity:

valid

Positive controls validity:

valid

Test results 3

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Vehicle controls validity:

valid

Positive controls validity:

valid

Test results 4

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Vehicle controls validity:

valid

Positive controls validity:

valid

Test results 5

Species / strain:

S. typhimurium TA 1538

Metabolic activation:

with and without

Genotoxicity:

negative

Vehicle controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Trial 1

Mutagenic activity of titanic acid, diacetoacetyl-diisopropyl with metabolic activation
	Histidine^+Revertants per plate**
Test item / μg/plate	TA 1535	TA 1537	TA 1538	TA 98	TA 100
NSC	22	5	32	40	168
Acetone	20	13	32	39	187
Tyzor DC / 10000 *	20	13	14	20	147
Tyzor DC / 1000	19	10	37	41	175
Tyzor DC / 3000	15	5	31	28	162
Tyzor DC / 5000	18	8	24	21	147
Tyzor DC / 7000	21	7	24	27	154
Tyzor DC / 10000	21	11	19	20	133
2AA / 5	-	-	-	-	1514
2AA / 10	555		1820	1889	-
2AA/1 00	-	493	-	-	-

NSC - Nonsolvent control

Acetone – solvent control

TYZOR DC – titanic acid, diacetoacetyl-diisopropyl

2AA – 2-aminoanthracene (positive control)

* - Negative control plate without glucose-6 -phophate and NADP

** - Average number of revertants from 2 plates

Mutagenic activity of titanic acid, diacetoacetyl-diisopropyl without metabolic activation
	Histidine^+Revertants per plate**
Test item / μg/plate	TA 1535	TA 1537	TA 1538	TA 98	TA 100
NSC	35	8	14	24	159
Acetone	28	5	25	26	159
Tyzor DC / 1000	34	7	20	23	140
Tyzor DC / 3000	22	8	17	25	116
Tyzor DC / 5000	20	6	16	24	119
Tyzor DC / 7000	22	4	13	20	104
Tyzor DC / 10000	15	4	14	19	103
MNNG / 2	1727	-	-	-	2031
9AAc / 50	-	386	-	-	-
2NF / 25	-	-	1861	1958	-

NSC - Nonsolvent control

Acetone – solvent control

TYZOR DC –titanic acid, diacetoacetyl-diisopropyl

MNG – N-Methyl-N´-nitro-N-nitrosoguanidine (positive control)

9AAc - 9- Aminoacridine (positive control)

2NF - 2 -Nitrofluorene

** - Average number of revertants from 2 plates

Trial 2

Mutagenic activity of titanic acid, diacetoacetyl-diisopropyl with metabolic activation
	Histidine^+Revertants per plate**
Test item / μg/plate	TA 1535	TA 1537	TA 1538	TA 98	TA 100
NSC	22	15	32	47	180
Acetone	28	10	36	40	198
Tyzor DC / 10000 *	22	9	25	31	170
Tyzor DC / 1000	17	9	32	51	210
Tyzor DC / 3000	16	10	28	57	180
Tyzor DC / 5000	15	5	36	44	203
Tyzor DC / 7000	15	7	34	42	178
Tyzor DC / 10000	18	4	29	33	177
2AA / 5	-	-	-	-	1012
2AA / 10	473		1844	2102	-
2AA/1 00	-	426	-	-	-

NSC - Nonsolvent control

Acetone – solvent control

TYZOR DC –titanic acid, diacetoacetyl-diisopropyl

2AA – 2-aminoanthracene (positive control)

* - Negative control plate without glucose-6 -phophate and NADP

** - Average number of revertants from 2 plates

Mutagenic activity of titanic acid, diacetoacetyl-diisopropyl without metabolic activation
	Histidine^+Revertants per plate**
Test item / μg/plate	TA 1535	TA 1537	TA 1538	TA 98	TA 100
NSC	23	10	12	34	162
Acetone	24	8	11	15	147
Tyzor DC / 1000	25	8	12	12	133
Tyzor DC / 3000	20	8	12	12	133
Tyzor DC / 5000	17	4	8	13	16
Tyzor DC / 7000	12	10	9	14	96
Tyzor DC / 10000	10	8	8	13	81
MNNG / 2	1300	-	-	-	1548
9AAc / 50	-	534	-	-	-
2NF / 25	-	-	1416	1949	-

NSC - Nonsolvent control

Acetone – solvent control

TYZOR DC –titanic acid, diacetoacetyl-diisopropyl

MNG – N-Methyl-N´-nitro-N-nitrosoguanidine (positive control)

9AAc - 9- Aminoacridine (positive control)

2NF - 2 -Nitrofluorene

** - Average number of revertants from 2 plates

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative without metabolic activation
negative with metabolic activation

Genetic toxicity in vitro of Bis(ethyl acetoacetato-O1',O3)bis(propan-2-olato)titanium was evaluated using five histidine-requiring strains of Salmonella Typhimurium. Test substance was not mutagenic in this bacterial reverse mutagenetic test.

Executive summary:

Bis(ethyl acetoacetato-O1',O3)bis(propan-2-olato)titanium was tested in Salmonella typhimurium strains TA 1535, TA 1537, TA 1538, TA 98 and TA 100 in concentrations up to 10 000 µg per plate. The compound was not mutagenic in the microbial assays either in the presence or absence of a liver microsomal system, i.e., it did not induce a significant increase over the spontaneous mutation frequency.

This study was regarded reliable with one restriction; the study does not include full range of recommended strains. However, the study performed is comparable to OECD 471 guideline. Thus, the result of this study is used as a key study in hazard assessment.