Genetic Toxicology - Mammalian Cell Mutagenicity

Study Information
Study Attribute	Value
CEBS Accession Number:	002-01385-0002-0000-1
Chemical Name:	3,3',5,5'-Tetramethylbenzidine
CASRN:	54827-17-7
NTP Study Type:	Genetic Toxicology - Mammalian Cell Mutagenicity
NTP Study ID:	509405
Study Result:	Negative

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Table Instructions and Notes:
Studies are indicated to be "Nontoxic" when the observed reductions in relative total growth did not indicate that the maximum tolerated dose was achieved.

Nonactivation Trial: 1 Experiment Call: Positive
		Conc. ug/mL	Cloning Efficiency	Relative Total Growth	Mutant Colonies	Mutant Frequency	Avg Mutant Frequency
Vehicle Control	Dimethyl Sulfoxide	0	81	96	41	17	22
			77	96	67	29
			82#	106	53	22
			71	102	43	20
Test Chemical	3,3',5,5'-Tetramethylbenzidine	12.5	68#	115	28.5	14	14
			88#	146	34.5	13	14
		25	75	126	40	18	12
			85#	144	16.5	6	12
		50	84	68	39	16	19
			79	80	51	22	19
		75	82	81	78	32	24
			92	86	44	16	24
		100	44	9	91	69	63*
			52	15	91	58	63*
		125	49	31	78	53	42*
			70	55	64	31	42*
		150	39	2	132	113	83*
			61	12	99	54	83*
		200

Positive Control	Ethyl Methane Sulfonate	500	20	12	607	1012	1000*
Positive Control			17	8	514	988	1000*

Nonactivation Trial: 2 Experiment Call: Negative
		Conc. ug/mL	Cloning Efficiency	Relative Total Growth	Mutant Colonies	Mutant Frequency	Avg Mutant Frequency
Vehicle Control	Dimethyl Sulfoxide	0	105	105	41	13	21
			76	99	49	22
			96	105	69	24
			93	91	68	24
Test Chemical	3,3',5,5'-Tetramethylbenzidine	50	97	68	61	21	20
			90	82	39	14
			62	54	47	25
		75	89#	46	42	16	25
			86	32	83	32
			69	28	54	26
		100	56	6	110	65	52*
			45	3	86	63
			89	30	71	27
		125	94	15	85	30	25
			67	15	47	23
			50	4	34	23
		150	72	16	58	27	23
			108	17	60	19
			42	4	29	23
		200	83	12	73	29	25
			72	12	64	30
			102	25	45	15
Positive Control	Ethyl Methane Sulfonate	500	11	4	474	1394	1126*
			16	7	438	942
			11	4	344	1042

Nonactivation Trial: 3 Experiment Call: Equivocal
		Conc. ug/mL	Cloning Efficiency	Relative Total Growth	Mutant Colonies	Mutant Frequency	Avg Mutant Frequency
Vehicle Control	Dimethyl Sulfoxide	0	94	92	128	45	35
			107	100	117	37
			119	99	103	29
			105	109	88	28
Test Chemical	3,3',5,5'-Tetramethylbenzidine	50	101	82	103	34	33
			101	78	83	27
			94	84	107	38
		75	87	46	155	60	62*
			91	54	121	44
			95	32	234	83
		100	67	15	166	83	60*
			101	33	161	53
			101	37	130	43
		125	101	22	171	56	51*
			98	20	121	41
			86	11	145	56
		150	52	9	106	68	56*
			79	18	97	41
			82	17	144	58
		200	75#	9	104	46	39
			87#	12	85	33	39
Positive Control	Ethyl Methane Sulfonate	500	7r	1	333	1514	966*
			35	8	530	510
			11	2	462	1422

Nonactivation Trial: 4 Experiment Call: Negative
		Conc. ug/mL	Cloning Efficiency	Relative Total Growth	Mutant Colonies	Mutant Frequency	Avg Mutant Frequency
Vehicle Control	Dimethyl Sulfoxide	0	63#	95	156	83	76
			71#	102	154	73
			66#	102	143	73
Test Chemical	3,3',5,5'-Tetramethylbenzidine	40	90	86	187	69	61
			70	105	131	62
			91	133	139	51
		60	71	74	149	70	72
			70	76	169	80
			96	99	191	67
		80	81	48	143	59	60
			74	42	154	69
			98	59	154	53
		100	73	27	189	86	78
			94#	26	226	80
			75	36	155	69
		120	78#	12	134	57	67
			86	27	187	72
			77	14	168	73
		150	86	13	180	70	84
			65#	9	181	93
			67#	8	179	89
Positive Control	Ethyl Methane Sulfonate	250	33	33	844	853	684*
			42#	43	794	630
			48#	44	820	569

Non-Induced S9 Trial: 1 Experiment Call: Negative
		Conc. ug/mL	Cloning Efficiency	Relative Total Growth	Mutant Colonies	Mutant Frequency	Avg Mutant Frequency
Vehicle Control	Dimethyl Sulfoxide	0	97	98	265	91	80
			113	101	269	79
			109#	89	249	76
			115	112	256	74
Test Chemical	3,3',5,5'-Tetramethylbenzidine	50	84	84	223	89	89
			81	60	217	89	89
		100	74#	34	210	95	101
			77#	34	166	72
			55	17	225	136
		200	84	39	234	93	83
			72	31	179	83
			80	23	174	72
		300	39	6	141	122	119*
			61	12	222	122
			82	19	276	113
Positive Control	3-Methylcholanthrene	7	31#	7	363	390	393*
			36#	12	382.5	359
			33	20	422	428

Non-Induced S9 Trial: 2 Experiment Call:
		Conc. ug/mL	Cloning Efficiency	Relative Total Growth	Mutant Colonies	Mutant Frequency	Avg Mutant Frequency
Vehicle Control	Dimethyl Sulfoxide	0	82#	89	105	43	32
			93	106	101	36
			89	88	60	22
			94	117	80	28
Test Chemical	3,3',5,5'-Tetramethylbenzidine	100	88	73	99	37	36
			82	53	99	40
			97	81	91	31
		150	92	65	82	30	33
			97	48	105	36
			89	54	93	35
		200	96	55	92	32	35
			107	45	118	37
			77	14	82	35
		300	83	53	66	26	39
			88	48	97	37
			73	4	118	54
		400	78	32	91	39	37
			104	28	129	41
			113	27	102	30
Positive Control	3-Methylcholanthrene	3.5	94	61	423	151	145*
			91	52	447	163
			80	53	292	121

Induced S9 Trial: 1 Experiment Call: Negative
		Conc. ug/mL	Cloning Efficiency	Relative Total Growth	Mutant Colonies	Mutant Frequency	Avg Mutant Frequency
Vehicle Control	Dimethyl Sulfoxide	0	84	113	106	42	47
			97	110	104	36
			72	92	117	54
			76	85	127	56
Test Chemical	3,3',5,5'-Tetramethylbenzidine	12.5	77#	86	132	57	48
			102	100	125	41
			95	118	134	47
		25	86	103	123	48	44
			101	99	125	41	44
		50	101	153	122	40	52
			69	77	103	50
			83#	73	160.5	65
		100	96	86	100	35	33
			87	45	93	36
			90	57	76	28
		200	65	7	140	72	47
			61	36	52	28
			52	31	63	40
		400	31r	4	59	64
Positive Control	3-Methylcholanthrene	5					330*
			7r	1	78	355
			15	2	145	330

Induced S9 Trial: 2 Experiment Call: Negative
		Conc. ug/mL	Cloning Efficiency	Relative Total Growth	Mutant Colonies	Mutant Frequency	Avg Mutant Frequency
Vehicle Control	Dimethyl Sulfoxide	0	107	111	97	30	37
			93	85	90	32
			97	99	112	38
			103	105	140	45
Test Chemical	3,3',5,5'-Tetramethylbenzidine	12.5	115	71	133	39	34
			101#	92	90	30
			79	56	82	35
		25	62	64	133	71	58*
			86	56	150	58
			77	59	99	43
		50	95	52	113	40	38
			106	43	128	40
			110	68	114	35
		100	79	40	57	24	24
			125r	39	69	18
			86	31	62	24
		200	56	14	115	69	44
			40	5	24	20
			78	17	104	45
		400	21#	1	20	32	55*
			64	8	142	74
			80	13	142	59
Positive Control	3-Methylcholanthrene	5	22	4	245	374	314*
			19	3	169	305
			33#	5	260	263

Footnotes:
Asterisks(*) indicate significant responses.
r = rejected value due to quality control criteria
# = reduced sample size because of the loss of one culture dish due to contamination