Document wDar3D62x8n0j7n8N0rLVMKxd
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Copies to:
AR226-2859
E. 1. du Font de NemourS and Company
HaskelI Laboratory for Toxicology and Industrial Medicine
HASKELL LABORATORY REPORT NO. 622-77
a+erial Tested: l^^--------f
aterlal Submitted By: H^^HBHBHqf' ^S'snic Chemicals Dept.,
Jackson Laboratory
"
Haskel I No. 11,308
Sample Ready for Testing: 6
IN THE SAIM019ELLA/WCROSOWE ASSAY
aterials and Methods: Five his+idlne-requlring s+rains of Salmonella typhimupiwn were used In the n trains TA 1535 and TA 100 are used to detect base-pair substitution mutations', whereas strains TA 155 ^ 98 are used to detect frame-shift mutations. (Ames, McCann and YamasakI, Mutation Research 3^ (197
The tests were performed In the presence and absence of a rat-liver homogena+e activation syste
?sence of metabolic activation, O.I ml of a solution of the test compound and approximately 108 bact ml of top agar (0.6? agar, 0.6? NaCI, 0.05 mM L-his+ldine, 0.05 mM blo+in). The solution was mixed jrface of a Davis minimal agar plate. The metabolic activation sys+een involved the addition of 0.5 m i the chemical-top agar solftion. The S-9 mi:.: contains per- ml: 0.5 ml of the 9,000 X g sipernatant iver, 8 mM MgCfz, 55 mM KCI, 5 mM glucose-6-; >osphate, 4 mM NADP and 100 1 sodium phosphate (pH 7.4 is added directly to the top agar Immediate )efore It was poured over the minimal agar plate.
Prior to testing for mu+agen i c I+y, the t-ompound
ie general toxici+y range for an strains.
Is tested for toxicl+y. Routinely, TA 1555 is
Appropriate controls were included for each strain. In -the .lonactlvated system these controls ga+lve, or solvent control, and positive controls. A second negative control (-S-9 control) Is Inc .say +o measure any activity of the compound In the absence of the S-9 ac+Iva+or mixture.
All plates were Incubated at 57C for 48 hours.
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-2-
Resul+s: Tables I and II.
HIHB^as Summary:
tested In Salmonella typhimuri-tim strains TA 1555, TA 1537, TA 1538, TA 98 and
tra+ions up to 10,000 ug per pe+ri plate. The compound was not mu+agenic in the microbial assays eithe
absence of a liver microsoniat system. I.e., It did not Induce a significant Increase over the spontaneo
quency.
JFR-.DFK.-ms
Da-t-e: August iq,
IQ77
Report by:
Approved by:
David F. Krahn Chief, Molecular Biology S
TABLE (
MUTAGENIC ACTIVITY OF ------------IN SALMONELLA TYPHIMURIUM
STRAINS TA 1535, TA 1537, TA 1538, TA 96 AND TA 100 WITH "ETABOLIC ACTIVATION
HZO 11,508
-S-9*
2AA
Compound Added
lig/pla+e
10,000
ti
2000
11
400^
ii
6000
ii
8000
ii
10,000
ii
5
10
11
100
(i
TA 1535
20
7
20
16 11 12 11
369
H20
11,308
2AA
*
**
^ Distilled water (solvent control)
-^--------ftl
= 2-Amlnoan+hracene (positive control) = Test plate without S-9 and activalors = Average of two plates
His+idine4 Rever+ants Pei- Plate**
TA 1537
TA 1538
TA 98
20
17
24
10
6
16
17
20
29
13
16
32
14
19
28
20
18
24
16
19
28
1870
2058
235
*
TABLE
PI|N MUTAGENIC ACTIVITY
O F ^ f STRAINS TA 1535, TA 1537, TA
TA
SALMONELLA TYPBIIWRIUM
1538,
98, AND TA 100 WITHOUT METARni IP An-n/flTi <,.
,., ,
.
.
--
w
l -' 1 V f^ | | \JH
H20 11,308
MNNG 9AAc 2NF
Compound Added
.
2000 4000 6000 u8u0u0u0 10.000
2
50 25
yg/pla+e
"
"
"
"
"
"
"
"
TA | K^il.t
15
17 15 10 10 5
3086
His+ldir^ Revertants Per Pl
i-f"\ 1 ^3--5--7
TA 1538
T
22
10
19
12
15
12
7
11
12
8
13
5
1364
1651
23
HzO
11,308
MNNG 9AAc 2NF
*
= Distilled water (solvent con+ro1) =i----------k|
=
N-Me+hyl-N'-ni+ro-N-nI+rosoguanidinei (positive control) = 9-AmInoacrldlne (positive control) = 2-NI+rofluorene (positive control) = Average of two plates