Document YD7j1MbdeL7Z5w3N7vXMYxVMk
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AR226-2860
E. 1. du Font de Ne'nours and Company
Haskell Laboratory for Toxicology and Industrial Medicine
HASKELL LABORATORY REPORT NO. 645-77
Material Tested:)^------------ V
jimHIfHMHK Material Submitted By:
Iprganic Chemicals Dept.,
Jackson Laboratory -
Haskel I No. 11,556
Sample Ready for Testing:
OFlUHBj MUTAGENIC ACTIVITY
IN THE SALMOmLA/MICROSOME ASSAY
Katericis and Methods: Five his+idine-requ I ring strains of Salmonella typkimurium were used in th Strains TA 1555 and TA 100 are used +o detect base-pair substitution mutations, whereas strains TA TA 98 are used to detect frame-shift mutations. (Ames, McCann and Yamasaki, Mutation Research 5J_ (
The tests were performed in the pres nee and absence of a rat-liver homogena+e activation sy
absence of ma+abol ic activation, O.I ml of a solution of the test compound and approximately 108 l^ 2 ml of top agar (.0.6% agar, 0.6^-NaCI, 0.05 inM L-his+idlne, 0.05 mM bio+ln). The solution was nii:surfocp oi a Davis minimal agar plate. The metabolic activation system involved the addition of 0. to the chemical-top agar solution. The S-9 mix contains per ml: 0.5 ml of the 9,000 X g supernalo liver, 5 rr,M MgClz, 55 mM KCI, 5 mM glucose-6-phospha+e, 4 mM NADP and lOO.mM sodium phosphate (pi I 7
was adc'3d directly to the top agar immediately before it was po'-red over the minimal.agar pidte.
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the
compound
is tested for toxici+y .
TA 1555 is routin
Appropriate controls were included for each strain. In the nonac+ivated system these contr negative, or solvent control, and positive controls. A second negative control (-S-9 control) is essay to measure any activity of the compound in the absence of the S-9 ac+iva+or mixture.
.n ,l,te, .er. locub.ted .1 37-C for <8 hours.
C.mp, S-t
-2-
Results: Tables I. and 11.
f^^^Bl Sun-Tiary: IMl^^^^yas tested in Salmonella typhimuriwi s+rains TA 1535, TA 1557, TA 1538, TA yS>
tra+ionsup +o 10,000 \ig per pe+rl pla+e. The conpound was not mu+agenic in the nii.crobial asss-.'s
or absence of a liver microsomal system, i.e., it did not Induce a significant increase over tne
frequency.
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Da+e: A..g^- iq,
|Q77
Report by:
lLitw /
ames ^'i^uss ir E'i
Approved by:
/^y/jr/ r\/\/\^>^'^
David ,-. Kr
Chief, Mol;ecular Biology
Company SanHlzed
TABLE I
MUTAGEN 1C ACTIVITY OFfl----------iS^AIIMNOSELLA rypsjMu, ^IUM
STRAINS TA 1535 , TA 1557. TA 1558. TA 98 'AND 1rA 100 WITH METABO LIC ACTriVATlON
HzO 11,336
-S.9*
2AA
Compound Added
10,000 2,000 4,000 6,000 8,000 10,000
5
IJig/pla+e
iii in 11 n i.i n
10
n
100
n
tA IS3S
16
50 17 18 16
17 16
414
H20 11,336
2AA
3 01s+111 ad water (solvent control)
"UBM)
* 2-Amlnoanthracene (positive control)
" Test plat without S'9 and act 1valors
Average of two pintes
HIstldIne+ Revertants F'er Plat
TA 1557 ----W]p38--------IS
16
226
31
11
7
21
11
22
27
8
23
29
7
23
18
10
26
35
12
22
38
2015
192
520
Company SMiRlxe
TABLE
^fi--ri| MUTAGENIC ACTIVITY <
IN SALMONELLA TTPB IWRIVM
^STRAINS TA 1535 . TA 1557. TA 1;53fl', TA" 98 1WD TA I'OO WtTHOUT METABOLIC ACTI^/AT
Compound Adde d
H^O 11.336
MNNG 9AAc 2NF
2.000 4,000 6,000 8.000 10,000
2
Hg/plate
ii ii ii
ii ii
50
ii
25
n
----tA Tn?3S
1 His>t5ld3l7n--e---1---Revertants
tA t538
PCir
23
18
10
26 27 31 22 26 1897
11 14 10
8 12
1213
16 18
9 11 11
1950
H20
" . ^ MNN6
9AAc 2NF
DIs+lllod later (solvent c ;ontrol)
-ffl--------n
= N-Methyl-N'-nltro-N-nI+rosoguiBnIdlne " 9-Amlnoacrldlne (positive control) 2-Nltrofluoriane (positive con+ro1) * Average of two plates
(positive
control)