# llllllllllllllllllllllllllllllllllllllllllllllllllllllllllll 

# • OTS0523259 

# N One Source. One Search. One Solution. 

# MUTAGENICITY OF TWO LOTS OF FM-100 LOT 53 AND RESIDUE OF LOT 3322 IN THE ABSENCE AND PRESENCE OF METABOLIC ACTIVATION WITH TEST DATA AND COVER LETTER

08 MAR 1990

U.S. Department of Commerce 

### National Technical Information Service

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## For rnore information visit our website: 

# www.ntis.gov 

# /'S':t' U.S. DEPARTMENT OF COMMERCE 

# . l!J'-

**~** Technology Administration 

# \ ·r / National Technical Information Service .,..."' Springfield, VA 22161

### CODING FORM FOR SRC INDEXING REVISED 10115166 OTS0523259

I'IUTRGENICI"TY, OF !TWO LOTS OF FM-100 LOT 53 AND RESIDUE OF LOT 3322 IN THE ABSENCE AND PRESENCE OF METABOLIC ACTIVATION WITH TEST DATA AND coVER LETTER ':

**HEXABR~YCLODODECANE** (3194-55-6)**Chem3~al** Corporation


### March 8, 1990 Document Processing Center (TS-790) Room L-100, Office of Toxic substances Environmental Protection Agency 401 M Street, SW Washington, DC 20460 Attn: S(d) Health 

### & Safety Reporting (Notification;Reporting) Dear Sir: 

# Contains Nc CBi,

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## ·' 0 2 

# - ::;; 

### ::! :;;.; 

### w ..

**~** 

### _,

**:..~ :~**  

### -; ::.3 N :;:! 

### In response to the TSCA Section S(d) rule published in the December 12, 1989 Federal Register, the enclosed information is being sub- mitted on the following substances: 2,4,6-Tribromophenol Tetrabromophthalic anhydride Hexabromocyclododecane 1,2-Bis(tribromophenoxy)ethane Decabromodipheriyl ether Pentabromodiphenyl ether octabromodiphenyl ether

**~** 

### CAS Number llB-79-6 632-79-1 3194-55-6 37853-59-1 1163-19-5 '32534-81-9

32536-52-0 The studies or1 the first four compounds are submitted on behalf of the following manufacturing facility: Great Lakes Chemical Corporation Route 7, Box 529 El Dorado, AR 71730 T",1e studies on the last three compounds are submitted on behalf 

## Of the following manufacturing facility: Great Lakes Chemical Corporation Highway 15 South, P.O.BOX 1878 El Dorado, AR 71731-1878
0 D

## ' There are no known additives to these compounds. Pentabromodi- phenyl .ether consists of hexa-, penta-, tetra-cbromodiphenyl ether. Octabromodlphenyl ether consists of nona-, octa-, hepta-:, and hexa- bromodiphenyl ether. We are not aware of any ongoing studies with any of the listed compounds. If you have any questions concerning this submission, please call me at (317.) 497-6391. Sincerely,



## -:u..L~t1£.J/[ **j.(</"~** Dennis 

### L. McFadden, Ph.D. Product Sa:fety Coordinator DLM:sb:030!)90 Enclosures

### TEST DATA 2.4,6-Tribromophenol Acute toxicity studies in rats and rabbits Primary skin irritation study in albino rabbits Eye irritation study in albino rabbits Acute inhalation toxicity in the albino rat Acute dermal toxicity in male and female albino rabbits Acute oral tcixieity (LD50 ) 

### study in albino rats Dermal sensitization study in the albino guinea pig Bioaccumulation study in fat tissue with 2,4,6-tribromophenol 48-Hour dynamic aquatic toxicity study with 2,4,6-tribromophenol in Daphnia , Four-day statio aquatic t.oxieity studies with 2, 4, 6-i::.ribromophenol in rainbow trout and bluegills . , · Screening test for microbiological inhibition with 2,4,6- tribromophenol (' . Acute dust inhalation toxicity study with 2,4,6-ti:"ibromophenol, lot number 3106 in rats 28-0ay subacute dermal toxicity study with 2,4,6-tribromophenol in albino rabbits' (10/27/77) 28-Day subacute dermal toxicity study wth 2,4,6-tribromophenol in albino rabbits (12/12/77) Pilot teratology study in rats Mutagenicity evaluation of 2, 4, 6-tribromophenol, lot number 3287, in the Ames Salmonella/Mi,crosome Plate Test Acute oral toxicity (LD5

### .,J study in rats · The bioaceumulation of 2,4,6-tribromophenol in bluegill sunfish 2,4,6-Tribromophenol biodegradation (9/30/75) 2,4,6-Tribromophenol (TBP) biodegradation (6/i/76) Pharmacokinetics study of 2,4,6-tribromophenol in rats Photolysis of 2,4,6-tribromophenol Partition coefficient of several flame retardants and industrial chemicals Water solubility of severa1 flame retardants and industrial chemicals ...•·,., .. Photolysis of 2, 4, 6-trib;romophenol in aqueous solution 

## '•:•o

### Tetrabromophthalic Anhydride Acute oral study Acute dermal study Acute toxicity studies ox tetrabromophthalic anhydride Primary ..skin**irritation~**

### study in albino rabbits Eye irritation study in. •albino rabbits Acute inhalation toxicity in the albino rat Dermal sensitization s:t;udy in 

### the albino guinea pig Acute inhalation toxicity in the albino rat after 1'pyrolysis (3/12/75) 

### Acute inhalation toxicity in the albino rat after pyrolysis (3/26/75) . 

### 28-Day dermal toxicity study in rabbits**2l~Day** 

### inhalation toxicity in rats Hum'an repeated insult patch test with Firemaster PHT4 Mutagenicity evaluation of 859-74-4 Pilot teratology study in rats The acute toxicity of Firemaster PHT4 to the water flea, papl}hia magna 

### Straus 

### The acute toxicity of Firema!<ter PHT4, lot nunilier 6332::..8 

### to the rainbow trout Salmo gairdneri Richardson The acute toxicity of Firemaster PHT4, lot number 6332-B 

### to**the·blu~gill** 

### sunfish Lepomis macrohirus Rafinesque Acute oral toxicity sttluy:' in mice V•, 

### Comparative inhalation toxicity tests Evaluation of the inhalation toxicity of combt,•,stion prod(icts of sel,ected polYJiiers II c 

### The bioaccumulation of tetrabromophthalic**anhydrid~,in** 

### the bluegill sunfish ·· . ·· .., \:,< 

### Persistence of PHT-4 in so.il at 1 

### and 10 

### !"g/g Photolysis of PHT-4 Effects of PHT-4 on soil microflora at l and 10 

### 1"9/9 Pharmacokinetics of PHT-4 in rats ' Partition coefficient of several flame retardants and industrial chenica!s ' Water solubility of several flame retardants,; and industrial chemicals ·· ::.:;::,

### ijexabromociyclododecane /.Hexabromodyclodcidecane: 28-day feeding trials with rats and 90-day feeding trials with rats ·v,rn vitro microbiological mutgenicity studies of four Ciba- Geigy Corporation compounds Pilot cataractogenic study in chicks Cataractogenic study in chicks Mutagenicity evaluation of 421-32B · Acute toxicity' studies in rabbits ad rats Mutagenicity of two lots of FM-100, lot 53 and residue of lot 3322 in the absence and presence of metabolic activatl.on ·· ·· . The acute toxicity of HBCD lot number 990-17 to. the bluegill sunfish Leoomis macrohirus Rafinesque · Partition coefficier.t of dicarnba, endrin, VEL 3510 and several industrial chemicals and flame retardants Water solubility of several industrial chemicals, flame retardants.and a herbicide VEL 3510 Hydrolysis of Firemaster 100 HBCD - biodegradabil'ity study 0

### 1,2-Bisftribrornophenoxylethane

**~water** 

### solubility of several flame retardants and industrial chemicals

**~Photolysis** 

### of Firemaster 680 -Pharmcokinetic study of FM-680 -Rate of hydrolysis studies -Partition coefficient of MC-680, chlorendic anhydride and chlorendic acid -28-Day rat feeding study -Acute oral. toxicity study in the albino rat -Acute dermal LD

## 50 (1 level)

**~Acute** 

### inhalation toxicity in the albino rat -Acute oral toxicity study in beagle dogs

**~cute** 

### toxicity

**(T~ )**  

### study in the bluegill -Acute toxicity (LD50 ) 

### study in the rainbow trout -Acute heated vapor inhalation toxicity study in rats -Modified Draize multiple insult test in male humans -28-Day dermal toxicity study in rabbits

**~21-nay** 

### inhalation toKicity in rats ··

**·~cute** 

### inhalation toxicity in the albino rat after pyrolysis (ABS resin;FM-680/SbzO)) Acute inhalation toxicity in the albLno rat after pyrolysis (ABS resinjsb2 03 ) 

### -14-Day range fnding study in rats '-.28-Day toxicity study in=-rats 'Biodegradation study with "c-tagged MC-680 '-Biodegradability study with "c-:tagged MC-680 'Primary skin irritation test with mother

**li~1or** 

### 680

. process, code no. 7-29-76-B in albino rabbits -skin sensitization test with mother liquor 680 process in albino guinea pigs

**~0-Day** 

### subacute OJ:al toxtcity study with Firemaster 680 in albino rats (Jan. 6, 1977)

**~90-Day** 

### subacute oral toxicity study with Firemaster 680 in albino rats (Mar. 10, 1977) '-Mutagenicity evaluation of 859-74-·5 -Test report (method for testing the of chemical substances in fish in fish) final report degree of accumulation bod.¥) (Bioaccurnulation

**~Acute** 

### inhalation toxicity study in the albino rat after pyrolysis (ABS

**resin/Sb~a,Jcontaining** 

### 680) Pilot teratology study in rats · '-Teratology study in 

### rats " "Tissue residue accumulation/depletion study with Firemaster 680 in albino rats ·

### pecabromodiphenyl ether \Acute toxicity.,_,studies., ....in.,.,rats"-a.nd-':ra!Jbits 

### \Mutagenicity evaluation of compound 364-5A '-28-Day toxicity study in'' rats (6/12/74) f· 

### '28-Day toxicity study in rats (6/24/76)

**~B-Day** 

### toxicity study in rats (3/2/77) "-Mutagenicity evaluation of compound 277-·10 Pentabromodiphenyl oxide 

## .,

ii

**~utagenicity** 

### evaluation of compound 345-76A final report.

'The acute oral toxicity of pentabromotl.iphenyl ether to rats • \Acute toxicity studies in rats and rabbits \28-Day toxicity study in ·rats 'The bioaccumulation of compound s-512 by carp

**~30-Day** 

### dietary study in rats with pentabromodiphenyl oxide including recovery periods of 6, 12, and 24 weeks '-90-Day dietary study in rats with pentabromodiphenyl. oxide {DE-7:1,) Octabromodiphenyl oxide 

# /j The



# bioacc~mulation of compound S-511 by carp

**~cute** 

### toxicity studies in rats and rabbits v15Ubacute inhalation toxicity study in rats

**~~utagenicity** 

### evaluation of compound 345-79A final report j;[n vitro sister chromatid exchange in Chinese hamster ovary cells with octabromodiphenyl ether (DE-79)



### J,unschedule~ J?NA syt!.thesis assay, compound DE-79 •28-Day

**tox~c~ty** 

### study in rats '13-Week feeding study in rats 'A range-finding teratology study in rats with DE-79



## " 

# '

## ,, 

# J,.!iuJ.tuaJ, 

# a 1

# o . n s·r .l!tz/;aA.LZ.f.Miet,, 

# .91ti: · • " _ . 1610 FRONTAGE ROAD •

## "onfartt~ 

# ru.;.;**oft~~** NORTHsRoox.•LLINOis 6D062 ... EP_,.-oTS 

# · ··**li~IIIUitil\111** DODb3bll42T 

# ,**z~~Doocio~b~**

LOT 53 .Al'ID 

## fie x.t**6r-o»u>c)'cf,.d~lec.t£** 

## c.;+<:;.q:/;:. 

## 31'11-ST"-..{: '•. 

### REPORT TO , VELSICOL

# CHEM!C~L**~OR!?ORATIOi** }..!UTAGEl'l'ICITY OF TWO LOTS OF FM-100, RESIDUE 

### OF LOT 332:2 L"i 

### THE ABSENCE .AJ."'D PRESENCE 0!' 

### METABOLIC ACTIVATION P.O. NO. 01515 

### AUGUST 26, 1977 IBT NO. 8536-10908 

## "

### ,, 

# !/

**.!J~** BfO -TEST


## ~.ie"-. 

## !Jnc. I!
·;.

Dr. D •i\M• WhitaC:::::e 1810 S:RONTAGE ROAD NORTHSROOK,

**ILLJNO~S** 60062

Velsicol Chemical 

### Corporation 

## ., " 

## " • ?.::GULA.TORY OIVISI0!'4 

## SEP 13 J97( 

# R!="C;;:_IVED
<\ 

## '

3H East Ohio Street 'chicago. 

### illinois 

### 606U 

## f/ef.A l•r o 

## "<<ocylo.U/et1-r)/e. 

### Dear Dr. Whita=e 

# C"-As'\3/7"?- ;Si.:. (, Re: 

### IBT No. 8536-10908"- Mutagecicit-;;-o£ TwooLots of

**F~I-100,** 

### Lot 53 and Residue of 

### Lot 33Z2 i.r'the Absence a."lc! l?::-ese:1ce 

### o£ Metabolic Acti\•.:tio::1.- P.O. No 01515 We are subx::itrlng herewit.'l ou:r 

### re,i.SeC. pages 

### prePared in 0

**co::...~ec:ioil** 

### wi_th

**t.~e** above study.
l' 

### A:IP/cl

.;:. 

## ' /)

**~.** V( 

# p.

**:.Y'~** A. 

### J. t'!"tsque ?::-esic!er:.t



## corm <· 

### TIAL 

## '

docur.:ent cnr.t:!= s 

### t:

**.1:~;:-::.td flltrJK:'l~:-~a**  u 

## w ich is the pr1"lH1'/ . : 

### ttJ··::i t 

### i•::r..-::.1 

### .C rporati!ln.ll m!!z.!'!:.: :;::::-:.. .:

**~ :·~**  **··~** "<t•; 

### r-· .

thout

**r~t:ro::!c~f:cr**.. **~;;:...-.,** '.!t;

**.~.:.:...~**.. i.••

**·~** ,;..,Ji.

;ved 

### c: nEJ

**l!~· .:\:.~,.;**  ..

**~** 

## ..1810 FRONTAGE ilOAO
**No~iHaROOK.ILLINOJS** 6ooo2 0

**~ugust.** 

### 26, 1977

**~!r.** 

### D.. **H~ ~1itacor**  

### Velsicol Chemical Co=?c:ation 341 East Ohio St=eet 

### Chicago, Illinois 60611 

## . . 

### Dear Mr. 

### Witacor Re: IB'l' No. 8536-10908 - **Hu~a<;e::licity** 

### of 'I'Wo Lots o:!: E'M-100, Lot 53 anC ResiCue 

### oi Lot 3322

**i~** 

### the

**Abse~ce a~d**  

### Presence of Met:bolic **Ac~ivation** 

### - 

### P.O. No 01515 we are **sub~tting he~ewi~h**  

### our laboratory 'report prepa=ed in connection 

### with the above study.


### AJE'/fd Very truly "YOtJ:s,



# /?/lflvt~

**?-~** ..

## ~ ( 

### ::":isque 

### ?.resiC.ent



# ,,**~**



### '":t~-=··-~~::d 

## a J

## c 

# -t: 5 

## r ..

**~** I "

**..::=;;,;,.~:k;4** ;•r.::. 

### "

BE?ORT TO v"ELSICOL CE::O.fiCAL CORPORAT!ON

**.l\.fli'TAG2~'ICITY** 

### OF T'\'10 LOTS OF 

### FM-100, LOT 53 Ac'l!l 

### 3.ESlDu"E OF LOT 3322 

### n; Ti-::E 

### A5SENCE 

## :-.1>m ??..ES2NCE OF :O.!ETABOUC ACTIVATION 

### P.O. NO. 

### 01515 

### AOGOST 26, 1977 IBT 

### NO. 

### 8536-10908 I. btroduc:tion

**~!utagenic:** 

### assays we:-e requested fo!" :'M-100 Lot 

### No. 53 (Fi:re!:laster-, white powder) and FM-100 Residue Lot No. 3322 (::t=::.a'ble lic;_cid) o::ith and .,;."ithout :::letaboiic ac;_, .. :a:ion 2.S !:"a: live:- ::::tic:-osortas. The concen.:raticns of the st:.bjec: sa=plas testeC were

**H-~:.eC.** 

### by thei::- 

### respective salu.bilities b ,;.; ..... **e:_~y1sc.Ea=dde~**

(j 

### "

## ' I. (r



## ' 

# z Tee nr-100 Lot 53 (25 to 250 !-'g/10 

### Jlil a:oc the FM-100 Resicue, Lot 3322 (50 to 1000

**~g/10 ~1)**  in

**t..~e** abse::.ce of rat !.ive::- ciC"osoces

**we~e** :::.ct r::.t::age:".i=. Analysis of va.:-ia.."'lce a:'lC =:ultiple co::pa:i.sor:.s of these

**da~a** C.emof..sZated that the levels of significa:1ce obtained in the statiStical analyses were consistently

**attribut~ble** to

**nit:"osogua~..i.Cine** teSted as the positive 

### control. L"l the presence of rat liver mic-osoces. mutants were

**evi~i!!e::.ced** in the 2-aminofluorene positive cont:;ol for each of the 5 tester- strains. Only TA-1535 shOwed I:lt:.tations related li.."le""::-ly to dosage, significant at the 1% 0 level (analysis of va.-i.a...,ce; Appendix, Table lliC) for FM-100, residue Lot



### ~322. That FM-100 Lot 53



### evidenced,"~ mutagenic effects in the



### pr~sence of metabolic activation may 

# be related entirely 

### to the concenh"ations of this proC.uct assayed. The highest concent::"ation of EM-100 Lot 53 that could be dissolved b Ci::::e,;-,ylsclioxicie W2S 250 ll;;/10 

## Jll. 

## ' The occur:::-e:c.ce a£ sig:1llica..'"!t :nu:age:ric

**e~ects** with TA-1535

**fo~** FM-100 Residue Lot 3322 2.!"e .fndicative of base pair substitt:.tions. \ .Report pre?a=eC.


### Director of Data. Ve::i.i'ied by 

### Quality Assurance Unit Date: 8/16/77 cl 

### .. ', . ..:;

### 3
l. :::-::ce:::.:.:::e ?:.:.!:i!is!:eC 

## •.
adjusted C.ov..-nw·a!"C.. A linear dosage respo::.se is desirable 

### fo:- t..1e Ce!l_'":i:io::. of ::lt!tage:ric e!:ects. 3. Teste:- Sr:-a.:.:::.s: Stai::.s TA-98, TA-100, TA-1535, TA-1537 =C: TA-1533 are usecl i:c. t.'-J.e p!"esence of :-at live!" 

### :::ieoso=.es. I!l

**~e** absence cf :::leRbolic ac=.... ·a.tio::. the last strain, TA..;l538, is not included in the assay. Eacl-: test species is 4:. Co'C.:l-:.s of a positive co::.i::'Ol, a solven: c::::l=.t:-ol 

### a:::.C. S?O::.ta::eous :reve::-sion are obtaL'""!eC. fo:- ea:::l:.

**teste~** s-.::-a!n.

a.

**~!ale** :-ats - 200 - 300 gc each (ChZ!'!es

**!live~ s~~ai:1,**  Cha.::-les 

### B.ive::- 5!"eeC::.g La:,., Inc,)

**b~** Si...'"'lgle L"'l.jection of Aroc:lo:r 124.2 (diluted i:t corn oil 

### to a r:onc. 

### of ZOO 

### :::lg/cl) LP. at a rate of 500 ::tg/kg

**.fiv~ ci.~j:i**  

### be!o:-e sac:-:=.ce e.
**D:-~bg** water giv·en 

## !£ libit'..t.'"'n &

**P~i:!a** Labora

### tor:;.· Chow 

### until 12 hou:s beiore

**sa~.iice.** 

### d. 0::. fifth C.ay of i::duc!ion,

**:r-a~** head and then decapitated. 

### stu."l.ned by a blow to the 

## co•l-

**-::~:r:.,.** 

### Th

**dQ:t:rne~t** 

### • ... ·• · ·• .. wh ch is tr.a rop:rtJ' Cot. cratinn. It 

### ::t h=

**~ec~::. ~!!d**  **:·~:** -:;, 

### With 

### ut u:pr"d ..

### t:cn, t:;::n =:t,:. ;;:: tn\'cl d

**~ ~~~~~**  c.;..:.o. ::.;.;:;;;:1.

**~**

### a. All 

### st.e?S**a=~** 

### at 0-4°C 

### cs:i=;; cold, sterile solu::!o=s 

### a:1C.**;:U.S~aze.** 4.


### b.



# ,, Livers placed i.J::. beal-ce:::-s con:a:.:l!:lg' 0.15 J..Lg: K:Cl (ap?:-ox. 1 :::ll/1>"' of wet liver) for 

### weig"-i::q;. 

### c:. ititer weisl:i!:.g, llve:!:'s a:e t:-a::.sfe!"!'ed to a**bea..~e::-** containing 0.15 !-lg KCJ (3 cl/g::::: wet l:!,ve:-) oi:o.ceci wit.':t sterile scissors, and ho:mogeni:eC in a Potter Elvejha.::n apparatus with a Teflon ?estle. d. 

### Hoo.og=ate c=t:::"-'cgee £::::: 10 :::,utes at 9000 : g (5700 

### :::?=)
g. Qcic..1<ly f:-oz= and 

### sto:!:'eci at -BO"C. 3..**?'!"e::~a:"ation** of S-9 M.;:;t; 

### S-9 (0.0'!-0.1 ol) 

### used 50"11=1 

## \i ""' MgClz 8!-!.M Kc.l**~3 't~-'vf**  Gl:.:cose**-6~1:.c~phate** 

### NADP 4flM SaCit= ?hosp!:ate Bt:.:.::e:- :;::E 7.**~** 

### 100!-D-i 

### S-9 M!:t is .:. esi::.ly**;:::-e?<!.=~C. e~::h**  clay, 

### .5.l::er s.:e....-:..l:.:ec!, 

### a=.C. ke?t Oil 

### iee be.fo::-e a::.C. 

### C.t:...-:...::.; \:.Se. 

### C.**Ml.::~::a.~e:esis** 

### Assaws o::. ?l:a.::es Top Aga:::o: 0**~** 

### 6% Di:::o Agar 

### 0.5% NaCl Au:ocla-.reC. !..::! 

### !.00 

### :::11 vol::=:tes a:::.C. ke?t c;.t 

### rcoc te::.?e:-a:-...:::-e 

### beio:!:'e use.-" '·' 

## u . 1!inimal-glucose agar mei:!ium in
**Vog~l-:Sor-""J.e:-** )..[eCit!::l E 

### 1. 5% Difco



### Ag~J:. 2, 0% Glucose 2. Mutaeenesis Assav P:.-ocedu:re ,·r;rtth 

## s-q ML""<: a; 

## " Acded in order to 

### Z ml !:!Olten top agar at 

### 45 'C l. -:-.0,: 1 

## m:I of 

### overnight nutrient bl"oth cultt.J;e of ,bactei-i.al tester strain. 

# z. Sample _to be tested - 

### lOf!l. ::;;> 

### .3. 

### 0.5 ml o£ the S-9 •mix. 4. S-9 mb: should not be left at 45 

### 'C fo:: more than 

### a few seconds . b. The contents a!"e mb<ed by rotati."'1g the tube between the pabs. c. The contents of the tube are then poc:ed on 

### :::li... **~al** 

### glucose aga:- plates. 

### d. U::ifor:n dist:-!bc.:ion of the 

### top

**aga~** is ac::Or::plished by gently tilt::.!lg and rotating the unco,re'!"eC. plite. and 

### e. ther:. sat:i.;.g down to ba:den. Mbd ... 

### 'P'!g, pou..."'"i!lg, a.""!cl **ciistribt:ti..~g** should ta.lte 

### seconds. D. Mu.t:aqer-ic Assav Without Ma:n::nalia.-.i **M!ecso::~.es** 1. 

### a. Sa."'Ile as Section C, 1. E.. Statistical Co::toutation 

# '
'· 

### 5

All data are subjected to analys;s of varia."'1ce and the si'gni.iii:ant ''· (
**a~:.:e!"e:lcies** 

### are fw:.t.."le:- de.:;:1ed 

### by Tukey 1

### s ou!tiple coz:npa:t"iso:t"'..s. 

### Tests e.'"(:ceedng

**t.~e** 

### para.r:J.eters a£ spontaneo·u.s **reve~sio!'l** 

### for eacl; teste 

### st:-air::. a!"e reg2.!:'cied. as !:lutZ.ge:lic pro'\'idec! the· counts obtai:led with 

### the test 

## mat2~i~·:i:.is . . 

### . **~·~· signiii~.?·nUy**  **dj~J!'ere.!tt,** 

### statisti.t;:a!ly, frO!:J, 

### those6 IV .

**R.est!l~** The data obtained for assays with 2-'"ld without oetabolic actiir:ation are 

## ,.. sl!.!':l:na:i:ized in Tables I andIII. The cutat:.on counts obtaineC in the. absence 

## d 0 of rat liver mic:rosomes are given i.."'l Table II; those obtained in t-1-te prese:1ce of rat liver r:::ticrosot:leS are tabulated in Table IV. All statistical cocputat:ions pe:-fo:r.ned 2.!"'e presente.d in the Appendi::< and data t"elevant to giire!l tests (tabie number and teste::- strain) are so-referenced.**-=--=--~** i 

## · .. Sa..":!ole Firemaster

**~** TABLE I Triplicated Cou::>ts of l.!utager.ic Effects P:ooduced by 

### Fi:o=aste:- 100 Lot 

### 53, and 

### FM 100 Residue No. 

### 3.322 for Fott:!" Genotypes of SaL""nonella. tvchimu::-iUI:'l in the Absence of Rat Liver Mic:rosomes 7
Triplicated Counts and Computed Ave!'ages (Nearest Whole NUI:tbe:-) o£ 4 

### Test Strains Cone. 

### (IJ.<:!/10 ul.) 

### TA-98 TA-100 TA-1535: 

### TA-153i 100 Lot 

### 53 250 16 121 10 6 

### 15 130 6 

### 3 

### 15 80 

### 5 6 Average 

## IT ITii 

## T 5 

### 100 14. II 

### 106 12 6 23 128 8 5 

# )I 17 116 13 

### 6 Aveliage 

## iB .117 

## IT 6 ! 

### 50 26 98 11 

### 9 21 

### 119 14 3 

### 27 117 8 

### 5 Ave!"age 

## IS 

# m IT 

## 6 ?" 

# _, 20 112 

### 12

**~** 

### 20 12:1 

### 10

**~** 

### 13 122 

## ...2. 5 Ave!"age 

### 18 118 

### 10 4 FM 

### 100 Residue No. 3-??

**~--** 

### 1000 26 

### 128 23 

### 3 20 162 15 5 

## ll 128 20 

## 1 Ave:-age 

### 23 139 

### 19 

# "' .;::,} 

### 500 19 114 

### 11i 7 

### 18 118 13 

### 3 20 108 10 3 Ave:·age 

### 19 113 

## IT • 

# .. 100 18 113 

### 6 16 106 5 19 109 3 Ave!"age 

### 18 

# i:ii9 5 50 25 13 

### 14 Ave!"age: 

# IT I I

### "

**::.~** 

## ( 8 

### TABLE I continued Tri;:olicated

**Colll\~S** 

### of Mutagenic Effects Produced 

### by Fire::1aste:r 100 Lot 53, 

### and FM 100 Residue No. 3322 for Four Genotypes of Salmonella tvohimurium in 

### the Absence of Rat Liver Microsornes Triolica.ted

**Co~i'>.ts** 

### and Comouted Averages



### (N~arest Whole N=ber) o£.4 Test Strains 

### Colic. S ,_,.,nle (U<Z/10 

### pl.) TA-98 TA-100 TA-1535 TA-153i MNNG** 10 48 7103 7208 

### 42 60 7205 5541 25 55 6956 5549 

### 23 Average 

# -.,

**~-** 

### 7088 6099 30 

### 2 20 2403 2567 10 29 

### 2891 2669 

### 13

**~** 

### 3083 2478 

### 18 Average 

### 24 279Z 2571 

## 14 DMSO*** 20 127 6 

### 7 21 10& 

### 6 7 

# ' 18 

### 113 l3 

### 5 Average 

### 20 

## liS 8 

## 6 Spontaneous • ?..eversion

**~** 

### 17 134 

### 16 6 25 

### 101 9 

### 5 23 .100 

# _s 7 

### Average 22 

### 112 10 

## 6 .. 

# ·- **

**~fNNG:** 

### N,...:cethyl-N-r:.i:b:'o-N-nit:-osoguanidine

**!~at** 

### #"08047, AlC=ich Che:::ri.cal 

### Co. *** DMSO: Dimethylsulfoxide, Lot illOF27A 

### Matheson, Coleman & 

### Bell

**Mf~.**

**~** 

### ..

### 9 TABLE. II Nu:nbe::- of 1luta."lt Color.!.es **Computed~** F:-om

**~** Averaged Triplicated Counts of

**Feu~** Teste:- St:ains 

### oi Salmonella h-ohi::::=iu:n Ex;:>osec! tp F1! 

### 100 J::ot 53 and

**~M** 

### 100 Residue Lot 3322 i!! Absence of

**~** Rat Liver !v!ic:-osomesc- 

### .4

**~~:::.** ?=- \) l :;-::. ....

**~** Cone. Counts 

### oi
**:\·luta..~ts.** Each 

### oi 4 Tester Sa:nole FM lOO Lot 53 FM 

### 100 Residue Lot 3322 MNNG** DMSO*** **(~g/10** 

### 250 100

50 

### 25 1000

500 100

50 10 2 ul) 

### TA-98 N.M.

**)(*~=~=** N.M. N.M. N.M. N.M. N.M. N.M. N.M. 

### 32 N.M. 

### N.M. TA-100 

### TA-153o 

### N.M. N.M. N.M. N.M. N.M. N.M. N.M. .N.M.



### . **·~·rc** N.M. 

# N.M.
- N.M.

**:N.~{.** 

### N.M.-!" N.M. N. 

### ;.,,r. N.M. 

### 6976 6089 

### 2680 2561 N.M. N.M. St!";tins 

### TA-1537

N.M. N.M:;: N.M. N.1L

**N.~L** N.M. N.M. 

## N.M.

24

**r~LM.** 

### ·:-;.M. 

## c: 

# * Cot:lputed as Cp-Cs 

### in w};.J.:::._'-1 Cp is the

**co~"'lt** obtair..ed of a given genotype

deter:ni::t.ed :for the proC.uct mL"'lus. Cs, the coun:!:: of tl:.e nt:=.ber of reve:-:ants

**occu...~~.g** spo:1t.aneous!y. The **sig-•:;ca.~ce** o.f t..-.,ese d"ffere::.C:es are evalt.:.atet; in term of the analysis of va.ria..'"'l.ce a.."'ld m1.:.lo:iple comparisons of the data. ** MN!i'<G: N-methyl-N-:Utro-N-::-.i::::-osoguaniCine Lot 

### #0804i, A1cirich Che:nl.c:al'Co. 

# *** DMSO: Dicet.:"lylsulio:cide, Lot #10F27A Mat..l-teson# Cole::J.an & :Cell M·fg. 

#   N .M. == 

### Non-Mutagenic

# . 1x;,·e'':id. BI 0 • T E S T

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### lU 

## '

**'.~!"** ''·· {!

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### 100

Lot 53 'TABLE III Tri?licated Counts of Mutagenic EffeCts Produced 

### by 

### Fi:t"e!:laste:t" 100 Lot 53, and FM 100 Residue No. 3322 ior Five Ge:1ot;rpes of

**~nella tv":)h~mu~iu..~**  in the P::-esence of Rat Liver Microsomes 

# "'·

# .,, Triplicated Cou.'"'!ts a.."ld .Averagtis·'-Co!:::::n.:teC. to 

### Nearest Whole Number EaCh 

### oi 5



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### TA-100 TA-1535 TA-1537 

### TA-1538 < 

### 250 

# so 114 

### 20 13 19

**~··** 4l 

### 121 20 9 24 J.:i 

### ill 

### lB. ...2. 

### .ll Average 

### 42 123 23 10 21 ... I 

# -

# -· 100 44 127 

### 16 9 

### 29

**~4'0** 

### 122 

### 19 8 27 

## ;u ill l3 

### ll J£. c 

### Average (: 

### 40 lZ1 16 9 

### 30 50 

### 46 1Z3 17 15 

### 24 4:4: 

### 120 24 15 27 

### 48 

## ...2.2. lJl.

**~** 

## 6.2. Average 

### 46 113 20 12 27 25 

### 46 129 30 

### 6 27 42 

### 119 17 10 21 

## £ !.Qi 24 20

**~** Aver2.ge 

## ...

# - 118 24 

### 12 25 

# _, FM 100 Residue No. 3322 1000 47 

### 172 79 8 19 43 

### 147 71 

### 15 22

**~** 

## ill. 63 

## il 25 Average 

### 43 

### 152 

## ,, 71 11 22 

### 500 48 

### 135 43 8 

### 28 43 121 38 6 

### 26 

### £1 ill d" 

## .2. 22 ..2. 

### .Average 

### 47 133 

### 42 8 

### 25 100 34 

### 124 

### 21 12 19 36 

### 116

**30~)** 

### 10 19 

# .... 134 . 42 

## ..1 18 -=2 

### Average 

### 39 125 31 10 19 

# ,, 50 37 126 22 34 

### 102 42 

### 116 

### Average 

### 38 

### 115

### TABLE ill 

### continued 

### '.0 Triplicated" 

### <;punts of Mutagenic E:iects Produced by 

### Fi.:-e:::aster lOG<Lot 

### 53; and FM 

### 100 Residue No, 

### 3322**~** .... . G \\**~** c:: 1 11 t..: · • Ior !:: 1ve enor-es o: ...**a~::1cne**....a tv:Ja!mu:-lwn 1n 

# the**Pre~)~nce** 

### of Rat Li,ter 

### Mic::-osomes ll

Tri;:llicated Counts and Averages Co:::':luted 

### to Cone.

### Ne~est Whole Nu.a""Dber 

### Each o£ 5

### '!'est~ Sb"a.ins Sarnnle 

### (g,;/10 ul.) 

### TA-98 TA-100 TA-1535 TA-1537 TA-1538**2-~-~nofiuorene 
(/ 

### DMSO** * Spontaneous Reve:-sion 

### 10 Average "5 

### Average Average AYerage 

### 2259 1998 2076 2111 1387 1275 1187 

## I2"83 49 

### 48 49 

# 49 49 

### 50 47 49 **Aldrich Chemical Co, Lot No, 032547 

### 1087 39 46 

### 1039 34 48 1087 39 46 

### 107l 37 47

736 39 3l 

### '789 28 39 854 23 29 793 30 

# rr 133 26 14 123 17 12 134 19 

### 14 

## Do 

## 21 13 127 18 17 129 

### 21 15 137 15 

# ..1. ill 18 14 

# "** DMSO: Di:nethylsulfo::cic!e, Lot**~10F27A** 

### Matheson, Coleman & 

### Bell Mfg,

") 

### 2608 2611 2661 2627 1835 1800 1720 1785



### "'33



### 24 28 28 25 20 33 

### 26 

## "' ""

### ."J~ticl 

## ll. I 0 • TEST **.!~.1e4** 

## J.C. 12 TABLE IV Nu=J.ber of Mutant Colonies Coc:::n.:.ted* FrOm Averaged Triplicated. Cou...""tts of :E"ive T.ester Strains of 

### ~Salmonella tvchim.\U"it:.r.:l Exp'oseci to 

### FM 100 Lot 53 and FM 100 Residue Lot 3322 in P:--esence of Rat Liver Microsomes Cone. l\:!utation Counts, each of 5 Teste::- St:"ai."':.s

### Saoole --"'( U"'2'-'l...:l"'O-'ul=·I..)**_..::Tc:.A:..-.~9 B** 

### _ _;;T:;:.A:..-..::1.::.0

### O::__,T"-A:..-...:l;::5.::.3S;:__..::T=A -153

### 7 T.>.

**~** 

### 1538 FM 

### 100 

### Lot 53 FM 100 Residue, 

### No. 3322 2-

### ..A.I:li..'"'l.oiluorene** DMSO*** 

### 250 100

50 

### 25 1000 500 100 50 

### 10

5 '.·, )! 

### ' N 

### .M.  

### N.M. N.M. N.M. N.M. N.M. N.M. N.M. 2062 1234 

### N.¥. N.M. N.M. N.M. N.M. N.M. N.M. N.M. N.M.

940 662 

### N.M. N.M. N.M. N.M. N.M. N.M. N.M. N.M. N.M. N.M. N.M. N.M. N.M.

53 

### N.M. N.M. 24 

### N.M. N.M. 

### 13 

### N.M. N.M.

6 

### N.M. N.M. 19 33 2601 

### 12 

### 19 li59 

### N.M. N.M.

** N.~L** 

# * Cocputed as Cp-·Cs in which C- is the count:;:obtainf!cl o£ a given geno-cype deter:nined fo!"' the product mi.r...u.s Cs, the coun.t 

### oi the number of revertants·
**occu~** .... 

### ,.g **spon't2-~eollsly**

**~** The sig::i:ica.."'lce of these ,.::H.:e.:-ences are· evaluated 

### in te..-m of the 2-'"lalysis of \'"2.:"ia..'"'lce and multiple ccmp2.:."isor...s oi the data.. 

# ** Aldrich Che=ical Co., Lot No. 03Z547 

### **"' DMSO: Dbet.,.ylsulio:cide, Lot f,!lOF27A 

### Matheson, Colema., & 

### Bell Mig, 

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## http://www.ntis.gov. 

# • NTIS Ensuring Permanent, Easy Access to U.S. Government Information Assets¢OF "o



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### US. DEPARTMENT OF COMMERCE Technology Administronon Nononol Technicollnformonon Service Springfield, VA 22161 (703) 605·6000

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### BIN: M30 06-27-07 INVOICE: 1661132 SHIPTO: 1*979351 PAYMENT: CSH*V