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Study Information
Study Attribute Value
CEBS Accession Number: 002-01465-0015-0000-4
Chemical Name: 4,4'-Methylenebis(2-chloroaniline)
CASRN: 101-14-4
NTP Study Type: Genetic Toxicology - Mammalian Cell Mutagenicity
NTP Study ID: G10679
Study Result: Positive


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: Negative
  Conc.
ug/mL 		Cloning
Efficiency 		Relative Total
Growth 		Mutant Colonies Mutant Frequency Avg Mutant Frequency
Vehicle Control Dimethyl Sulfoxide 
 0         
 121r 161 87 24
    113 124 88 26
Test Chemical 4,4'-Methylenebis(2-chloroaniline) 
5         
 98# 68 85.5 29 30
    95 46 90 32
  
10         
 77 41 104 45 36
    93 50 74 26
  
20         
 82# 57 102 42 36
    86 54 78 30
  
40         
 62 24 86 46 42
    65 19 75 38
  
60         
 81 10 71 29 32
    78 8 83 36
  
80         
   
Positive Control Ethyl Methane Sulfonate 
500         
 51 32 751 488 444*
    62 32 750 401
Negative Control Negative (Not Specified) 
 0         
 79 89 65 27 33*
    78 87 91 39

Nonactivation Trial: 2 Experiment Call: No Call
  Conc.
ug/mL 		Cloning
Efficiency 		Relative Total
Growth 		Mutant Colonies Mutant Frequency Avg Mutant Frequency
Vehicle Control Dimethyl Sulfoxide 
 0         
 99 89 48 16 16
    107# 104 53 16
Test Chemical 4,4'-Methylenebis(2-chloroaniline) 
5         
 86 82 77 30 29*
    62 54 51 27
  
10         
 85# 64 66 26 19
    94 88 36 13
  
20         
 112# 96 55.5 16 20
    99# 83 67.5 23
  
40         
 94# 69 72 25 23
    101# 63 64 21
  
60         
 114# 57 67 20 22
    95 58 70 25
  
80         
   
Positive Control Ethyl Methane Sulfonate 
500         
 46 23 809 588 583*
    40 31 700 579
Negative Control Negative (Not Specified) 
 0         
 108 118 59 18 24*
    102 89 92 30

Nonactivation Trial: 3 Experiment Call: No Call
  Conc.
ug/mL 		Cloning
Efficiency 		Relative Total
Growth 		Mutant Colonies Mutant Frequency Avg Mutant Frequency
Vehicle Control Dimethyl Sulfoxide 
 0         
 78 101 71 30 33
    123r 118 101 27
    111 99 122 37
Test Chemical 4,4'-Methylenebis(2-chloroaniline) 
12.5       
 111 145 46 14 22
    102 133 95 31
  
25         
 117 112 65 19 20
    108# 126 72 22
  
50         
 104 86 71 23 22
    109 89 70 21
  
70         
 121r 61 113 31
    118 65 84 24
  
75         
 120 53 129 36
  
80         
 117 34 131 37 36
    111 67 117 35
Positive Control Ethyl Methane Sulfonate 
500         
 66 31 1255 634 683*
    51 37 1120 732

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         
 95 90 32 11 12
    86 102 23 9
    102 115 52 17
    94 93 35 12
Test Chemical 4,4'-Methylenebis(2-chloroaniline) 
5         
 86 98 31 12 10
    75 79 22 10
    91 98 26 10
  
10         
 103 92 35 11 12
    69 65 25 12
    92 67 38 14
  
20         
 85 79 31 12 12
    97 84 32 11
    94 86 39 14
  
40         
 77 50 46 20 17
    96 70 35 12
    81 54 43 18
  
60         
 78 19 25 11 10
    101 22 22 7
    103 61 36 12
  
80         
 86 10 27 10 14
    91 8 46 17
   
  
100         
   
   
Positive Control Ethyl Methane Sulfonate 
500         
 31# 16 821 876 900*
    40# 24 919 771
    27 15 864 1054

Nonactivation Trial: 5 Experiment Call: Negative
  Conc.
ug/mL 		Cloning
Efficiency 		Relative Total
Growth 		Mutant Colonies Mutant Frequency Avg Mutant Frequency
Vehicle Control Dimethyl Sulfoxide 
 0         
 83 94 182 73 65
    90 87 188 70
    72# 100 135 62
    98 119 160 55
Test Chemical 4,4'-Methylenebis(2-chloroaniline) 
5         
 111 114 160 48 51
    94 99 127 45
    93 112 166 59
  
10         
 91 101 177 65 57
    71 95 104 49
    90 91 153 57
  
20         
 99 110 153 51 60
    65 76 133 69
    88 105 161 61
  
40         
 117 79 182 52 60
    94 84 160 57
    79 74 169 71
  
60         
 100 46 173 58 85
    81 8 260 107
    92 20 245 89
  
80         
   
   
   
Positive Control Ethyl Methane Sulfonate 
250         
 50# 35 862 575 604*
    46 30 872 634

Induced S9 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         
 109 106 59 18 18
    91 89 51 19
Test Chemical 4,4'-Methylenebis(2-chloroaniline) 
5         
 80 31 156 65 68*
    67 34 143 71
  
10         
 66 47 183 93 92*
    75# 35 207 92
  
20         
 87 36 113 43 63*
    64 39 161 83
  
40         
 89 40 152 57 60*
    86 42 161 62
  
60         
 88 13 164 62 69*
    69 23 157 76
  
80         
 85 13 169 66
   
  
100         
   
Positive Control 3-Methylcholanthrene 
5         
 44 14 538 411 350*
    56 28 482 289
Negative Control Negative (Not Specified) 
 0         
 81 121 57 24 24*
    89# 84 66 25

Induced S9 Trial: 2 Experiment Call: Positive
  Conc.
ug/mL 		Cloning
Efficiency 		Relative Total
Growth 		Mutant Colonies Mutant Frequency Avg Mutant Frequency
Vehicle Control Dimethyl Sulfoxide 
 0         
 72 116 82 38 44
    73 83 111 51
Test Chemical 4,4'-Methylenebis(2-chloroaniline) 
5         
 79 112 72 30 37
    85 96 111 43
  
10         
 81 55 107 44 49
    72 48 114 53
  
20         
 76 22 148 65 62
    81 23 146 60
  
40         
 40 4 170 142 124*
    51 5 164 106
  
60         
   
Positive Control 3-Methylcholanthrene 
5         
 30# 8 315 348 422*
    10 2 151 495
Negative Control Negative (Not Specified) 
 0         
 82 101 78 32 34*
    104 100 115 37

Induced S9 Trial: 4 Experiment Call: Positive
  Conc.
ug/mL 		Cloning
Efficiency 		Relative Total
Growth 		Mutant Colonies Mutant Frequency Avg Mutant Frequency
Vehicle Control Dimethyl Sulfoxide 
 0         
 65 75 83 43 38
    74 96 82 37
    78 107 90 38
    69 122 71 34
Test Chemical 4,4'-Methylenebis(2-chloroaniline) 
5         
 43# 46 103 80 64*
    57 75 106 62
    65 54 100 51
  
10         
 63 41 151 80 72*
    58 46 112 64
    71 50 151 71
  
20         
 80 27 249 104 107*
    64 26 230 120
    70 34 205 98
  
30         
 50 20 232 156 142*
    57 18 214 125
    61 21 267 146
  
40         
 60 16 217 121 142*
    54 16 227 141
    50 15 244 163
  
60         
    70 8 222 106
   
  
80         
   
Positive Control 3-Methylcholanthrene 
5         
 44 33 505 381 438*
    36 28 529 490
    37 21 495 442

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