## -- 

# RECEIV'EB ' JuanB PerezlDClUSEPAlUS TO NCIC HPV@EPA 

# OPPT 

# CBtC
1 111 612006 08:25 AM CC 

# 2006 

# OEC 

# - I 1 

# An 7: 

# 32 bcc Subject Fw: HPV submission



### ----- Forwarded by JuanB PerezlDCIUSEPAIUS on 11/16/2006 08:25 AM 

## ---- "StafFord, Fran (FM)" <FMStafFord@dow.com> TO NClC OPPTQEPA, Rtk Chem@EPA 1 111 512006 05:48 PM "Cosse, Pam (P)" <PCosse@dow.com>, "Deford, Connie cc (CL)" cCLDeford@dow.com>, "Stott, William (WT)" <WStott@dow.com> Subject HPV submission



### Attached please find the HPV submission test plan and dossier for Tris(hydroxymethy1)aminomethane 

### (77-86-1). A 

### hard copy and CD were sent via Fed Ex yesterday, (Tuesday, 14 November) with an arrival to EPA by the morning of today, Wednesday, 15 November. However, according to Fed 

### Ex there were weather conditions and a missed courier pick up which did not allow for a Wednesday delivery of the test plan and dossier to EPA. We were not informed that Fed Ex did not deliver the package until after 5:00 pm today (Wednesday, 15 November). We apologize for this unforeseen delay of FedEx not being able to do the next day delivery 

## . We made every attempt to make the November 15th submission day. If you have any questions please contact Pam Cosse 989-638-9660 or Dr. William Stott 989-636-8203.

<<Dossier Tris(hydroxymethy1)aminomethane 

### CAS# 77-86-l.rtf>> <<Test Plan Tris(hydroxymethy1)aminomethane 

### CAS# 77-86-l.doc>> Thank you. Pamela F. Cosse Toxicology and Environmental Research and Consulting Bldg 1803 > 

### The Dow Chemical Company > 

### Midland, MI 48674 phone 989-638-9660

Fran Stafford TERC 1803 Bldg., Midland, MI 48764 989-636-3010 E-Mail: fstafford@dow.com

Dossier Tris[hydroxymethyl]arninomethane CAS# 77-86-1

. rtf Test Plan Tris[hydroxymethyl]aminomethane CAS # 77-86-1 .doc

### AMINO ALCOHOL

HPV Challenge Program

TEST PLAN For 

### Tris(hydroxymethyl)aminomethane (77-86-1)

Submitted t6the U.S. Environmental Protection Agency Under the High Production Volume (HPV) Chemicals Challenge Program

The Dow Chemical Company

Midland, Michigan, 48674

November 14,2006-- 

# I I .'.: -:3 

## 1 .': j L -: 3 

# -1 1: :. .L --

;l -;3 

## I , -

, :

. /,[O&&d:4 

### TABLE OF CONTENTS '. ,' , '.

., ,, ,! 

# 1 ,I--=--

....... -: 4

. ... '.*

\ 

## .-__A : .' ::j -:7 

## J

, ' ., 7 ' . . 

### I. INTRODUCTION



### ..................................................................................................... .. :!.:.; .!-:7 ,

. . .

). ,,1

..... 

### II. IDENTIFICATION OF THE ANALOGUES..



### ....................................................................

, . - -. -. - -. . ,: ,- : : '.
........ . / 

### III. JUSTIFICATION FOR ANALOGUE DESIGNATION.



### ..................................................... '-
,, ,, ,

... '.',r 

### IV. DEVELOPMENT OF ROBUST SUMMARIES AND STUDY SCORlNG CRITERIA


### ... ...... ! .: :; :; DQka6Bd: 8 ;. : 

### . : 

# -7 V. 

### TEST ENDPOINT RESULTS FOR ANALOGUES.



### . ..................................

. ....

F. PHOTODEGRADATION 

### ................................................................................................................................... 

# " -.;. --'-:9-9 

## I G. IN WATER (HYDROLYSIS)


### .......................................................................................................... .. .. STA~~ITY 

### H. ENVIRONMENTAL


### ..................................................................................................................... 

### 3 -".

~ . ''L~:lo

-- - 

### I TRANSPOR T. 

### -- I. BIODEGRADATION


### ........................................................................................................................................ 

## I J. ACUTE FISH TOXICITY .% 

### .............................................................. . .. ............................................................ K. AQUATIC INVERTEBRAT ES L. AQUATIC PLANTS


### .......................................................................................................................................

EYE IRRITATIO SKIN SENSITIZATION..


### ......................................................................................................... REPEATED-DOSE


### .. ...................................................................................................................... TOXICITY GENETIC TOXICITY: AND CHROMOSOME


### ... ......................................... ... GENE MUTATIONS ABERRATION S CARCINOGENICI TY



### . ..................................................................................................................................... REPRODUCTIVE



### . ......................................................................................................................... TO XICIT Y DEVELOPMENTAL


### . ................................................................................................................. .... TOXICI TY.
,::'",:',,L 

### CONCLUSIONS



### . ......................................................................................................... 

# !$;i7::,c,:,,:r -: I, ':-,,.,.3 ,.,,, !,+. 

## ...... 

# 1 i!,,."., ,,. , ,:.::.::. 

### :,:1 -: 12 

# II. INTROD UCTION Th e D ow C hemi c al Co mpany h a s co mmitted to spo nsor TRI S A M IN O in th e EPA H i gh Pr o duction Volu me Ch emical Progr am. II. IDEN T I FIC A TION OF T H E SU BSTAN C E For mo r e th a n5 0 y e ar s, T ri s ( h y dr o xyme thyl )a mino me th a n e (T RI S AMI N O) C A S#7 7

­ 86

- 1 , tec hnic al l y desc ribe d a s 2

- Ami n o - 2 ( H y dr ox yme th y l )

- 1 , 3

- Pr o pane di o l , ha s bee n
u s ed inapp l ic ation s rang i ng from ph arm a ce u ticals, biolo gical buff e rs, andtr ansp o rt of l i v e aqu ati c speci e s, to emu l sify ing ag e n t s incos m eti c s. TRIS AMINO is widely know n and used as a th er ap e utic bio l o gical bu f f er, pre d omin antly in p atients suf fering a d e creas e in bloo d pHdue to di ab ete s or a wid e ar r a yof re spir ato ry di s eases and
con di ti ons. It is al so use d clin i cal ly as a drug fo r th e trea tment o f me t abo li c acidos is
w here it is i njected intr ave n ous ly or vi a oth e r rou tes. I tis liste d in both th e U

. S. Ph arm a c o p e i aandth e N ati on al Formularyof drug s fo r th er ap e u ti c drug ap p li c ation s . TRIS AMIN O is admi n i s tered in hum an p ati en ts p ri m ari l y v i a th e in tr av en ous route
re g ard less o f th e cau s e o f th e acidos is. It h a s b een ri gorous ly ev alu ated in numerous clin i c a l exp e rime nt s w i than imal s andhu mans, a n d has consisten tly sh own a low i n c id e n c e of lo ng-te rm t oxi colog i c a le ffe ct s at e f f ectiv e dos es withre l ati v elyfewan d mi nor si de-effec t s. T RIS AMINO i s a l so use d i nthe an ti -i nflammatory Kete rolac ® to imp r o v e drug s o lu b i li ty . I nth i s f o rm , itha s bee nap pr ove d f o r

u s e i ntr eati ng a c u te p o st-op e r ati v e p ainvi a an intr amus cu lar in jectio n . Kete ro l a c ® h a s be enwi de l y use d due to its ou tsta n di ng p ain con tro l propertie s, andis an ond ep end e n c y- formin g
al te rn ativ e t o opi ates w i th a low occ urren c e of s i d e -ef f e c ts. T RIS AMIN O is also l i s te d in th e CTFAIntern ation al Cosme tic I ngredie n t Diction ary(p age 62 8, en try name of Tro me th ami ne), re fl ectin g its us e as an ingredi e n tin v ari ous co sm eti c fo rmulations.
B e cause of i t s low a qu ati c toxi city, TRIS AMIN O is also an eff e ctiv e buf f e r u s ed to
re gulate th e pHof th e wate r during the tr a n sp o rt of liv e a qu a ti c sp eci es. TRIS AMINO i s also wid e l y u s ed a s a b u ff e r , solubi li ze r a n d n e utr a li z i ng ag e nt in co sm eti c cream s, solu tion s , an d lotions, miner a l o il a n dpar a f f in w ax emuls i ons, po lish e s an dclea n i n g comp ound s. Finally, TRIS AMINO is also of ten use das abu ffer in v ario us bio l ogical te st sy stem s a n d li f e s c i e nce ap pl i cati ons su c ha s ce ll cu lture me di a.

3II I. JUSTIFIC A T ION FORT H E US E OF SU RROGA T E SUBSTANCES TO SU PPORT THE SU BMISSION FOR TRIS AMINO Table 1 : I d entificatio n of the s u rro ga te s ubsta nces CA S No. Chemical Acronym Che m ical Name 000077-8 6 -1 T R IS A M IN O, TH AM , TA, TRI S , TR OM ET H A M I NE Tris (hydroxy methyl) amin ome thane (2-Ami no-2

- hydro xymeth y l

- 1, 3-propanedi ol) 000 115-6 9 -5 AM PD 2-Amino-2-methyl-1,3-propanediol 000124-6 8 -5 AMP, AMP-95 2-Ami no-2-me thyl-1-propanol Acomm o nm ech an ismof a c ti on , b a sed on stru c tur a l and ch e mical simi l arity , is on e o f th e b a s e s th at th e EPA h a s p r ovid e d u n d er th e H P Vprog ram to use stru ctur al ly-s imil ar
su bs tanc es as d atasurr o gate s (U.S. EPA , 199 9). We p ropos e to utilize sur rog ate ch emicals to fill g aps in our H P V d a ta b a s e fo r TRIS AM I N O.
TRIS AMIN O (2-A mino-2

- hydrox y m e thy lprop anedi ol) and th e p rop osed surrog ate comp ound s 2 -Amino -2- me thy l -1,3- pro p a n e dio l (A MPD) an d 2-A mino

- 2 -meth yl-1

­ p rop anol (AMP) p ossess an id entical mo lec u lar “bac k bon e” s tru cture and commo n f unc ti ona l g r ou p s , f o rming a c o nti n uu m f r om tri o l t o d i ol to mo n o - a lc o h o l , re sp ectiv ely

. Th e s e stru c ture s are sho w nin Fig u re 1. TRIS AMINO , AM PD a n d AM P allc o n t ain a p rim ary amin e con n e c ted to a terti ary c arbo n , with on e or m o re termin al primary alcoho l group s Figu re 1. R e pre s e n ta ti ve Stru ctu re s o f th e Su rr og ate Su b s tance ( s) Formatted: Left, Indent: First line: 36

pt

NH 2

OH O H

O H CH 3

N H 2 OH O H N H 2

CH 3 OH C H 3

TRIS AMIN O AMPD AMP

Table 2: Ch e mical Prop erties of TR I S AM I N O and of th e Surrog ate Substan ces
Test Ch emi c a l CA S No. Molecular Wt. g/mol Melti n g Po i n t Boil ing Po i n t Va p o r P r essure Partitio n C o ef ficient Water Solubility

4( o C) ( o C) (hPa) Log K ow(g/L ) TRI S AM INO 77-86

- 1 121.14 171-1 72 21 9-2 20 @ 10 mm H g 3.04x10 -6-2.31 800

AMPD 115-6 9 -5 105.14 1 10 15 1-1 52 @ 10 mm H g 8. 71x1 0 -3 <-3. 8 2 500

AMP 124-6 8 -5 89

. 14 30.5 165.5 @ 7 60 mm H g 4.5.3x10 -1 -0.63 Mi s c ible all proporti on s

C o ns i s te nt wi thth e ge nera l stru ctu r a l an d c h e mic al si mi lari tie s of T RI S AMI N O, A MPD and AMP, studies hav e d e mon s tr ate d a s i mil ar p attern of phy si c a l ch emistry , env i ro nme n tal f ate, and ec o l ogical an d mammaliantoxicity profiles. Th e comp ound s
are so li d c rysta ll i n e ma sse s i nth e p ure or ne atstate , an d p o s s e s s c o mmong e ne ral ph y s ic al/c hemi ca l pr ope rtie s . B e c au s e of th ei r stru c tu re , a l l the su bstanc es ar e h ig h l y
so l ub le inwate r , ha ve ve ryl o w vapo r p re ssu re s, po sse ss r e lativ e l y lo w parti ti o n
co e f f i ci ent s (log K ow ’s), and s i milar disso c iati on c onsta nt s, maki ng th e mli ke l y t o
re m ai n dis s o lv e d in th e wate r comp artme n t upo n th e e v en t of an envi ro nm ent al re l e a s e,
w h ere b i o d e gr ad a ti on is ultim atel y exp e cte d

. M acK ay Le v e lIII fu g a city mod e l i n g p r edi c ts th at TRIS AMI N O and th e surrog ate subst a n c es w i l l te n d to p artition pr e d omi n at el y t o wate r Gon s i o r ( 2 00 6)

. T h e re is al so a l o w po te ntia l t o b i oa cc u mu late in a qua tic o rg ani sms ba se d o n l o w l og K owvalu es . Th e h igh wate r solub i l ity an d
n e g l ig ible v ap o r pre s sure o f all thre e substa n c es suppo rt th e low estim a ted Henry ’ s Law Con s tants (4.54x 10 -8 and 6

. 48x 1 0 -10Pa m 3 /mol for TRI S AMINO and AM P , re sp ectiv ely , a n d 8.6 7 x1 0 -1 3 atm

- m 3 /mole for AMPD). De let

e d: , S

.

De

let

e d:

T h e dataon f i sh , a quati c i n ve rte b rate , an d al g a l toxicity, in additi on to th e use of TRIS AM INO a s abu ff er for l i v e f i sh tr an sport, indi cate a p attern o f low to xi ci ty for this ch emi c al a n d th e sur r o g ate subs t a n c e s . Th e s e m ate ri als are o f lo w to xi city to aqu ati c
o rg a n i sms w ith EC50s greater th an 1 00 mg/L.
Mamma l ian t o x ici ty stu die s ha ve di sp la y e d si mi lar re sul t s . Th e ora l L D 50valu e fo r TRI S AM INO is 5 500 mg/kg in th e mo use, and its surrogates r ang e from 2 150 to greater th an

55 000 mg/kg in th e r a t an d m ouse . TR IS AM IN O w a s no n-irritating to ey e s wh e n a 40 % aqu e ou s solutio n w a s ap p l ie d to th e e y es of r abb its (p H 10 .4 for 0.1 M a qu e ou s solu tio n). In
con tr a st, 95 % AM P in w a te r w a s s e v e re ly ir ri tati ng to th e e y es, p re s um ably du e to th e
se ve re ly a lka l i n e p Ho f th e te stso l u tio nu s e d ( p H 1 1

. 3 f o r

0. 1M a q ue ou s so l u ti o n) ; howe v e r , mo r e n e utral c o s metic f o rmula ti ons co n tai ni n g l o we r c onc en tra ti o n s o f AMP ar e o n l y mi ni ma ll y irri tati ng. T h ere i s no se ns itiz ation data a v ai lable for T R IS AMINO; howe v er, b a s e don th e follo wing d ata, TRI S AMINO is not exp e cted to be a sensitizer. Labor atory
an i mal te stsample s of AMP di d not cause a l le rgic skin reactions when te sted ingu in ea p i gs fo llowin g to pi ca lo r in tr adermal a dmi nistrati on. I n patc h tests w ith human s , AMP and
cosmetic formulation s contain i ng e ither AMP o rAMPD w e re ne g ativ e for derm al
se ns iti zati on. Re pe ate d

- dos e mammal i a nto x i city s tudies condu c ted on TRI S AMIN O and th e tw o surrogate c h e mic al s i ndicate th a t th e comp ound s are g e n e r a lly well- t oler ate dat
con c en tr atio ns as h i gh as 5 00 mg/kg/d a y v i aIVinfusion f o r TRIS AMIN O and in g e stion of
up to 3200ppm in th e ro den tdiet (25 0-7 50 mg/kg/d ay fo rr ats and mice, e s tim ated). A
nu mb e r o f huma n c li n i c a l trial s o f th e I Vi n f u si on o f TR I S AM I NO ha ve a ls o be e n
suc c e s s f ullycon du c te d. In a l l studi e s, th e onlyt arget ti ssu e , w h e n obs erv e dat al l, h a s been the li ve r with AMP. Hu ma n c l i n ica lstudie s withKe t e rola c® (amaj o r c o mpone n t of whi c h is TR I S AMI N O) h a ve su g g e s te d th atp a tie n t s with de c re a s e dl i v er f unc tionn o t be giv e n th e dru g o v e r e x te nde d tr eatme n tperi o d s b a se dup onc h an ge s i n s e ve ral c li n i c a l c h e mi s try p ar a me ter s . Ing e stion o f relativ e ly high d o sa g e s o f AMP has cau sed liv e r histop atholo gical ch ang e s in rats a n ddog s. Th e m o st sign if i can t t oxi colo g i calac tiv ity h a s been af e tot o xi c effect of AMP w h e nin gested at re lativ e lyhighlev e ls by pregnan t r ats. Subsequ e n tdermal exp o sure to comp ar ab le do sag e s f ai l ed to e l i c i ta d e v e lopment a l eff e ct in r ats. Ov er all, th ere ha ve bee n no c o n s i s ten tly

- n ot e dob serv ati ons or treatmen t- rela ted finding s a m o ng th e
nu merous repeated -dos e mammalian toxicity s tudie s th at h a v ebee ncondu c ted ov er th e la s t 5 0 years on th e s e comp o unds th at w o uld indicate long-term sig nificant tox i city of eith er comp ound at ty pical hu m a n exp osure lev e ls. R e flectiv e of these findin g s is th e f a ct that bo thTR I S AMI N O an d AMP d i sp lay s i mi lar patte rn s of e x c reti o n f r o m th e bo d y , be ing pri m aril y e limi n ate du n ch an ge dviathe urine over a r e la ti ve l ysh ort pe ri o d o f ti me . Furthe r , n o ev id en ce of e i th e r dire ct re a c tiv i ty or metaboli sm t o rea c tive s p ec ies t o war dge ne tic

6m a te rial h a s b een ob serv e d . G e n e ti c t oxi ci ty studies condu c te d onth e TRI S AMIN O and th e
su rr o gate s ubs tanc e s i nth e pre s e n c e o r ab se nc e of mammal ianme tab o l ic e n z y me s have al l be e nnega ti v e . Fina l l y , TR IS AMI N O an d th e s urrogate c h e mic al s ha v e displaye d li ttle i f an y t oxici ty to hum a ns during th eir long history of u s e a s hu m a ndrugs an d/or inp e rs onal care p rodu cts an d c osmetic s. TRIS AMI N O has fo un d u s e as an I V dru g for the
m a n a g e me n t of acidos is in hum a n s fo r manyy e ars and th e tox i c i ty of AM PD a ndAMP ha ve bee n r e v i e w e db y th e C o sme tic I n g re d ie n t Re v i e w E x pertPa nel whic h c o ncl u ded th a t th es e m ate ri als are s a f e as us ed inc o smetic f ormula tion s up to 1% ( C I R, 1 990). Based upon the p r op ertie s , us e s an d toxic i ties of T RI S AM I N O , A M PD and AM P, the us e of AMPD and AMP as s u rr ogate subs tances to fulfill any data gaps for TRIS AM I NO is w a rranted
. IV. DEVELOPMENT OF ROB U ST SUMM ARIES A N D ST U DY SCORING CRITERIA Th e D o w C h emi c al Co mp any h a s ch ose n t o us e th e IUC L ID (Inte rn atio n a l U n iform Ch emicaL I nformation D a tab a se) fo rm at for p re p ar ation o f ro b u st summ aries for th e H P V p rogram . Becaus e ma n y of th e field s in th e IU C L ID d a tab a se prog r am are
o utsid e th e s c op e o f th e H P V p r o gram, these fi el ds are typ i call y l e ft b l ank in th e I U C L I D r o bu stsu mmary . Sc o ri n g of s tu d ie s fr om c o mpa n y f i le s o r fr o mth e li ter ature fo r reliability to fu lfill th e testing re quire ment for each e n dpoin tus e da sys te m simi lar t o th at pub li shed b y Kl i misch et al

. (19 97). Stu d ie s w e re g i ven a s c o r e of “1 ” if th e data c oul dbe c ons i d e re d v a l i dwithou tre stric ti on b a sedo nth e c o mp le te ne ss o f th e pr o t oc ol a nd a d e qu ate detail s in r e p o rtin g . Stu die s we re gi ve nasc or e of “2 ” i f th e data andstu dy d e sig n could b e con s id eredscien tif ically valid to ad dress th e e n dpoin tbut wi thre stric tio n s du e t o lac k of vari o u s tec hnic a l or re po rti n g d e tail s or de viati o n s fr o m
curre n t OEC D gu idelin es . Studie s w e re giv e n a s c ore of “ 3 ” if th eir c o ndu ct was no t accep t able and “ 4 ” if th ere wasn’t en o ugh inform atio n p resen t to assignare liability
rati n g . Ho we ver , astu dy re ce i v in g as c ore of “ 4 ” c oul d p r o v i d e s u pple me n tary i n form a tio n th at c o u ld b e us ed to ad dre ss th e en dp oint i na w e ig ht of ev id e n ce
e val uati oninth e ab se nce of othe r data

.
V. TES T ENDPOIN T RES U LT S FO R TRIS AMIN O and SU RR OGATE SUBS TA NC E S Asummary of th e d ata o n HPV/ SIDS e ndpoin t s for TRIS AMINO and s tru cturally similar a min o alcoh o ls can b e f o und in Table 3: Summary of th e E ndp oints

. Com m

ent [

P

FC

1]: Pam does thi s refe re n c e in clu de AMP D ?

De

let

e d: ¶ ¶

7Ev alu ati on o f this d at a l eads to th e c o nclu sio ns th at (1 ) a subs t a nti a l qu a n tityof d at a
curre n tly e x i s t t o a d equ ate lyrep resen t th e t oxico logica l and eco l ogical p r o f ile, (2) th ere is con c ur ren c e and similarity a m ong th e existing d ata fo r th e variou s HP V/SID S endp o i nts, ( 3 ) av a i l abl e d at a f r o m T RIS and th e stru c tur ally simi lar subst a n c es can b e
us e d t o a d e qu ate l yre p rese ntth e vario u s HPV/ SI DS e n dp o i nts th a tmay notha ve be e n sub j ected to th e same lev e l of te stin g, a n d (4) util i z atio n o f th ese d at asup p o rt th e
c o ncl u si o nthatno f u rthe r te sti n g i s n e e d e dfo r m o st of th e HPV/S I D S cate g o ri e s . Th e d at afo r ea ch o f th e HPV/ SID S endpoi nt s are di scu ssed inth e fol l o win g sectio n.
A . Melting Po i n t IUCL ID 2.1 : TRIS AMIN O is c ry s t al l i ne m a ss es inth e n e at o r p ure st ate w i th m e lting po i n t s o f 171

- 1 7 2 o C (M erck, 19 89). Th e d atasu gg e s t th at d ecompositionis un likely,
an d be cau s e the me l ti n g p o i nti s we l l

- doc u me ntedi n pe e r- re vie we d l ite ratur e an d pu b l ic dat aba ses , furth er testing for t h is endpo int w o uld not be p r odu c tive

.
B. Boil ing Po in t IUCL ID 2.2: T h e b o i li ng p o i ntf o r TR I S AMI N O i s we ll doc ume n t e d(Me rc k , 19 8 9 )

. No furth er tes ting is planne d forthis e ndpoint.
C . Va por Pr es sure IUCL ID 2.4: Th e v apo r p ressurefor TRIS AM I N O w a s estimate dby EPIW IN MPBPWIN pro gr a m, an d altho ugh estim a tio ns are not cons idered re liable f o r th is cate go ry, bo th th e exp e rime n t al or estim ated v alu e s for th e stru ctur ally s i mil ar amino
al co h o l s are ex tre mel y l o w at 3

. 0 x 1 0 -6(TRIS AM INO ) hPa at 20 -25 o C

. Th e vapor pr e s s ur e , f o r th e stru c tural l y re late d su bs tanc e s have bee nwe l l

- doc ume nte di n e i th e r
measured re p orts, pub lished literature o r c h emical h andbook s. Th is re su lt is con s is tent wi thth e ph ysic a l/c he mical nature of the ami n o a lc o h o l s an dsug g e st s averyl o w d e g r ee
o f

vol atil ity ; th ere is no furth e r tes t ing planned for this end p oint

.
D . Pa rtitio n Coefficient IUCL ID 2.5: Th e v alu e fo r TR IS A M IN O is –2 .31(Lho t ak, 19 96). Th e p artition
co eff i ci ent i s con s is te nt with h i gh w ate r so lubi li ty a n dwhi c hbydefi n itio n w ouldbe

8i ndi cativ e o f lowlog Kow v a lu es. Bas e d on th e s tru cture, an d th e low log Kowvalu es, no furth er testing of t h is endpoint is necessar y .

E. Water Solu bility
IUCL ID 2.6.1: Mea s ure ddatai n dicate th at T R I S AMI N O i s hi g h l y s o l uble to mi sc i b le in wate r. Measured aqu e ou s solubilitie s o f at least 800 g/l at 2 5 o C fo r TRIS A M IN O f urthe r d o c u me n t i ts very h i ghwate r so l u bi li ty (L

. T r oe ste r , pe r s o n a l c o mmu n i ca ti on, 2 006). Furtherm ore , TRIS AMIN O is sold as an aqu e ou s solutio n, and is formulated as p art of th e drug Kete ro l a c® to enh an ce it s solub i l ity

. Suff i c i e nt d at aexis t for th is endp o i nt to c h ar acteri ze wate r solub i l ity for th e T R IS AM IN O, such th at no fur th er te stingis needed

.
F. Photod eg ra dation IUCL ID 3.1.1: TRIS AM IN O do e s not absorb lig ht >290 nm , and th erefore direct p hoto l ysis is no t possib l e. I n dire ct ph o t olysis (hy droxy l r a di cals) is possib l e , h owev e r. Mo delin g h a s show n th at th e h a l f-li f eof TRIS AM INO in th e atm o sph e re is app r o x im ately 3

. 8 ho urs ( W est, 2 004 a ). Si nce vola ti li zati on is not ani mporta nt
env i ro nmental f ate p r o cess for th e surrog a te substan ces, te s ting f o r pho to lysis in th e atmo sp he re i s no tre le van t , a n d th e ref o re no further testing for t h is endp oint is
necess ary .
G. Stabili ty in W a ter ( H y d r oly sis ) IUCL ID 3.1.2: TRIS AM IN O do e s not possess mo lecu lar stru ctures th at c o n t a i n fu n c tio n a l g r o ups sub j ect to hydro l y s is un d e r n e u tr a l amb i ent con di tions ( Reaction Mec h an i s ms i nE nvi ronme n ta l Organic Che mistry, 1994). It i s c o mme rcia ll y a v ailable
a s ana que ou s mi xtu re , fu rthe r d o cume n ti n g h i ghs tab il ity in wate r , an dis re po rt e d stable for severa l years <200 o C. Th is te stin g en dpoin tis w e l l c h aracte ri zed, an d no additional testing is required.

H. Environmental Transp ort IUCL ID 3.3.1: En vironmental transpo rt d ata w e re obtain e d for th e TRIS AM I N O b y estim atio n using th e EPIW I NLevel II I Fug a city p r ogr am ( C an a dian Env i ronmen ta l

9Mo delin g C e n ter )

. M e asured w ater solubility d ata and th e p rev iou s ly

- rep orted v a lu e s fo r v ap or p re s sure w e re used as th e mod e l input. Th e p r o g r am also us edinput v a lu es foroc tanol/wate r p artition c o effic i ents , ai r-wat er c o e f fi cie n ts , me lti ng poi ntand
m o lecu l ar w e ig ht th at w ere e ith er c a l c ul ated b y oth e r prog r am s i nth e EPIWINsu i te or
us e d as re p o rte d i npee r-re v i e we d l i ter ature o r databa se s . Th e fug a ci tyma ss a m ou nt (p arti tion i ng ) v a l u e s o b t ain ed indi cate th at di stributio n in to th e air wou l d be n e g lig ible (<0. 1 % ), a s w ould b eexp e cted fo r prim ary amin o alcoho ls comp ound s w ith lo w vo l ati lityand hi gh sol ubil ityin wate r. Like w i se, th e distri bution t o sedime nt w a s estim ated to b e <0

. 1 %. Th e c a l c u l a te dm a ss am oun ts i nth e so i l
env i ro nment a l comp artmen t wa s <0.1 % ; th e pre di c te d m a ss am ou nts distri buted to
wa ter wa s 10 0

. 0 %, ahi ghle ve l th a t is c o nsi s te n t wi thth e kn own h i g h wate r so l u bil ity of

me mbe r s o f th i s catego ry ( G on si or , 2 0 0 6 )

. A ltho ugh th e v alu es o b t ain e d u s in g this mod e l sh ou ld no t b e reg ard ed as qua ntita tiv e , th e m odel results are con s iste ntwith th e know n p h y s ical and ch emical p roperties of TRIS AMIN O, i. e ., th e kn ownhighwate r solub i l ity an d n e gl igib l e vo l ati l ity. Fu r th er tes ting forthis endpoint is not n e eded.

I. Biod eg r ad a tion IUCL ID 3.5: Bio d e g r ad atio n is th e co nv ersion o f a ch e mi c a l b y mic r oorg anism s in th e envi ron me n t into i t s simpl e r c o mponents an d ultim ately to carbo n di ox id e and it s
o th e r cons ti tu en t m o le cu l e s. Ch emi c als are cl assifi ed as re a di ly biod eg r a d ab l e by th e O r gan i zatio n for Econo mic D e v e lo pment (O EC D) g uid el in es if th ere is a 7 0% d e g r ad atio n of disso l v e d org anic carb on w ith in a 10-d ay p e ri o d during a ty pica l 28 -d ay labor atory p roto c o l

. Th ere is a biodeg r ad atio nstu dy, O E C D 30 1D, fo r TRIS AM IN O w hic h indica tes th at it is n o t re adily biod e g rad ab l e (IWL, 199 0). Th ere is e v id e n ce b a s e d on s tudies with stru ctur ally

- re late d c o mpoun ds to su gges tth at TRIS AMIN O wo uld b i odeg r a d e un d e r f a vor ab le co nditions. Th ere is anOECD 3 02C study o n AMPD wh ic hsh o w ed 96 .7% bi o de g ra dati o n a f te r 22 day s . I ts ho uldth e re f o re be c ons i dere d in he r e ntl y b i od egr a d ab l e, whi c h in di cates th at i two uld n o t persis t ind e f i nite ly in th e env i ron me n t

10andth a t it w ould e x hib it similar biod eg r a d ationp o tentia l b a s e d on stru ctureb i od egr a d ab ility rela tio n s h i ps measur ed fo r similar c o mpoun ds ( W est, 20 04 b).
Th es e d at ai n di cate th at th ere is suf f i c i e nt info rm ation on th e bio d e gr ad ati o n po tenti a l of T R I S AMI N O an do f th e s urr o gate su bs tanc es, a n d j u sti f i e s th at no additional te stingis needed.

J. Ac ute F i sh T o x i ci ty IUCL ID 4.1: Th e LC 50 valu e of TRI S AM IN O f o r 29 sp ecies of fish w a s repo rted to be greate r th a n4 4 0 0 mg /Lwhe no b s e rve df o r a pe ri odo f 30 day s ; iti s useda s a b uff e ring ag en t for shipp i ng liv e fish at d ose lev e ls r a n ging 44 0-11 00 mg/L und er
ae rate d or static conditions (McF arland and Nor ris, 1958 ). AMP h a s b e e n ev alu ated in 9 6 -hourLC 50te sti ng with Lepom is m acro c hirus , w h e re th e L C 50 u nde r se mi- s tati c
con di tions was reporte d to b e appr oxima te l y 190 mg /L ( P are kh, 1 9 80a ). N o ad ditional te stingis requi red.

K . A q u a ti c I n v e r teb r a tes IUCL ID 4.2: B a sed on to xicity tren ds in fish, an d c o mp arable physical- c h e mical pr o p e rtie s, T R I S AMI N O i s e x pec ted t o be a s non - t o x ic t o a qu atic i nve rte b r ate s as A M P. A quati c in v e rtebr ate tox i ci ty dat aar e av ai l abl e fo r A M P onl y, an d th e au tho r s
re port 4 8 - & 96

- h our LC 50 v a lu es fo r C r ang on cra ngonto b e 179 mg/L ( Y o ung a nd Tapp, 1983 ). In Da phniamag na , AMP w a s re p o rted w i th a 48-hour EC 50of 19 3 mg/L ( P are kh, 1 9 80b )

. No a dditional te sting is re quired

.
L. Aqua tic Pla n ts IUCL ID 4.3: TRIS AMIN O w a s tested with Se lena strumCa pricornu tu m , and a N O EC of > 100 m g/L ina 96 -h our g row th r ate test (A d a m s , et al

. , 198 5). No
a dditional testing i s co nsider ed ne ce ssa r y.
M. Acute Oral To xicity IUCL ID 5.1.1: A c u t e or al tox i ci ty studi es w e re con ducte dwith TRIS A MIN O using a ga stric tube i n the Swi s s mo u s e an d W i star rat
. So l uti o n s of 2 0 % a n d 5 % we re gi ve n
v i ath e g a stri c tub e to r ats and mice resp ectiv e ly at d os e s o f 1 00 0, 20 00, and 3 000

11m g /kg a s a s o lu tion. Th e LD 50for r ats and mi ce are estim a ted to b e >300 0 mg/kg in solu tion. Th ere w a s n o toxicity n oted in ei th e r s pecies at th e top do se lev e ls, alth ough abu nd a nt urine ou tpu t w a s no te d fo r so me an imals ( G irouxa n d Beau laton , 1962 ). Additional testing is no t c o ns ider ed ne ce ssar y .
N. Acute Inhalation Toxicity IUCL ID 5.1.2: T h ere i s n o a v ai labl e d ataon T RI S AMI NO o r th e surr ogate su b s tance s fo rth is endp oint. Th e re is, h owev e r , alo w po tential for inh alation exposure b a s e d on know n use p atterns, phy sical st ate of th e pure m ate ri als, and v apo r p r es sure , a n d therefore testing is con s idered unnecessar y .
O. Acute D e rmal To xicity IUCL ID 5.1.3: TRIS AM IN O has b een e v a l u ate din th e r a t and m ouse vi a i ntr ad erm a l i n

jections, an d th e LD 50 for bo th sp ecie s e x ceed ed100 0 mg/kg bo dy w e ig ht , th e h i gh est d ose tested. A tth at do se le v e l, lesio n s w e renoted, bu t no oth e r sign s o f toxicity were
re ported (Girou x a n d B eau la to n, 19 6 2). AM P was ev alu atedin rabb its v i a a 24 -hour skinp atch te st, a n dth e LD 50was re p orted greater th an 20 00 mg/kg (Parekh, 1980 c). Th ere w e re no signs o f systemi c t o x i city , how e v e r th e tre atm ent site s w e rene croti c
w ithin2-3 d ays, an d re m ain ed so at stu dy termin ation. Treated group s ex hibited a los s in body weight ov er th e 14 -d a y post-treatme n t p e riod.
N o add i t i onal te sting fo r this e n dpo int is c on s id ered n e cessary , as rel i ab le datais
av ailable for TRIS AM I NO andth e surrog ate c o mp oundAMP.

P. Skin Irritation IUCL ID 5.2.1: Skin ir ri t atio n te st s fo llow i ng th e Drai ze M e th o d hav e b e enperfo rmed
o n TRIS AMINO , alth oughdocumen tation is n ot su f f icie nt fo r f u ll a s ses s ment (rel i ab ili ty4 ). It w a s fou nd to b e mod e r a te ly i rri t ati ng on ab r a d e d rabb it skin , but
re so lv e dwithin48 h ours (Baldwin, 1 96 1). Th ere was no n oticeab l e irri tationon
u n ab rad e d s kin sites. I nam o re re liable study, AM P was fou nd to b e irritating to
r abbits, w ithburr o w ing le s i ons no tedw h en ap plied to abr a d e dskin site s ( Machle et a l ., 1 940); th ere was mild ir rita tion noted wh e napp lied to un abr a d e d skin . Th e sev e rity of i rri t ation is dire ctly re l ate d to th e b a se s tre ng th of th e a mino al c oh ol. A t2 5°C th e p K a

12o f TRIS A M IN O is 8. 03, th e pK a o f AMPD is 8

. 76, and th e p K a of AM P is 9. 72. Since limited dataex ists fo rth e comp ound, but exists f o r th e surrog ate substan ces, additional testing is no t c o nsid ered necess a r y . Q. Ey e Irrita tion IUCL ID 5.2.2: Un d i lu tedT RI S AMINO wa s f o un d t o be e s se ntial l y no n-i rritati n g t o
r abbits, alth oug h documen tation is n o tsu fficie nt f o r fu ll a s ses s ment (reliability4). A noth e r test con du c te d on a 40 % solution of TRIS AMIN O in water h a s show n th is m a teri al to b e
n on-ir ri t a ti n g to r abb its ( Po w e r , 197 5)

. N o a ddi ti o n al te sti n g i s ne ce ssary.
R. Skin Sensitiz ation IUCL ID 5.3: While th e toxico logical/s af ety p rof ile of TR I S AMI NO h a s b e e ne x te nsi v e l y
es tab lish e d f o r m any endp o ints, esp e ci al ly tho s ea s so ci ated w ith i ts us e i nth e tr e a tme n t of
me tabo lic ac idosis, no labor atory stu dies o f skins e nsi tiz ati o n ha ve bee nr e p o rte df o r

th i s
m a te rial , an d th e ass e ssme n t fo r d e rm al s e nsi ti z ationh a s b e e n b a s e d on th e f a v o r abl e us e exp e ri en c e a s so ci ated w i th v ario us us e s , su c ha s app l i c ations wh e re it is an in gre di e nt in fo rmul a tio ns a s so c i ated w ith anti c i p ated d e rm al con t a c t. Ina d dition, stru c tur ally re l ate d
su rr o gate s AM PD an dAMP h a ve b e e nne gati ve in labor atory an imal a nd /or hum a n p atch te sts fo r sen s iti zati o n. AMP did n ot caus e all e rgi c skin re actio ns wh en te sted in guin e a p i gs fo llowin g to pi ca lo r in tr adermal a dmi nistrati on. I n patc h tests w ith human s , c o smeti c fo rmula tio ns c o nt a i n i n g eith er 0.2 2 % AMP or 0.5% AMPD or 0.07 5% A MPD w e re
n e g ativ e for dermal s e n s itization (Kocib a , 2003 ; CIR, 1 990). Bas e d upon ex te n s ive cl i n ica l use a nd s u rr oga te re l a t e d c h em ic al ac ti vit ies no fur therte stingi s recommende d.
S. R e p e ate d -D os e To x i city IUCL ID 5.4: T RIS AM INO was admi n i ste r ed t o vario u s sp eci es o f a n i m a l s and humans in mu ltipl e stu di e s and tri als prim arily vi a th e intr av e no us route, sin ce it is d e liv ere din th at m ann er t o hum a np ati en ts su ff erin g acido s is a n d re qu i rin g treatmen t. Th e test dur atio ns r ang ed from 1 0 to 99 days in r ats, mi ce , a n drabb it s. In all s tudi es,
re g ard less o f dur a tio n or species, th e ob serv a tio ns c on s isten tlynoted w e re lesions an d /
o r g a n gre n eat th e in f u sion site s, an d , in m a nycases, in creas es in wh ite b l ood cell Com m

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13cou n t s asso c i a ted w i th g ang re n e . Sp or a dic ki dn e y a nd liv er lesions w e re no ted in th e long er stu die s. B e cau s e th e stu di e s were g e n e rally condu cted at o n e dos e lev e l ( p lu s
con tro l ) co mp ar ab l e to hum an th er apeu ti c do s e s , no -obs e rv e d- a d v e rs e eff e c t l e v e l (N OAEL’s ) / low e st-obs erv e d- adv e r s e ef f e ct lev e l (LO A EL’ s ) are no t us eful to repo rt
. It is reason ab le to assu me, how ev er, th a t sin c e th e s tudies were c o ndu cted under v e ry l i ke lyhuman expo sure sce n arios, th e rel ati v e to xi city o f TRIS AMINO to hu ma n s i s l ow. Th e assump tion of low t oxi ci ty is a l so supp orte d by mu l tipl e cl ini c al tri als wi th bo thhea l th y an d ai li n g humanpati e n t s tre ate d withTR I S AMI N O atsimi lar d o se l e v e l s . In th ese c l in i cal tri als, th e m o st c o mm onobserv ati ons in clud ed a tr ansi e nt d e creas e in re sp ir ato ry rate , an in crease in urin e o utput, hyp erkalemia, a nd hypog ly cemia. In cas e s wh e re th e d o s e lev e l f ar ex ceed e d th e recomme n d e d th e r ap eutic dose, vomiting, sweatin g , hu nger , di zz in ess, and di arrh eao c c u rre din a small subs et o f th e test g roup. Th ere are no d o c u me n ted lo ng -te rm effects of TRIS AM INO treatme n t , a n d no serio us side- e f f e cts o n record th at are di re ctly attributed to tr eatmen t w i th th e comp ou nd.
Th e surrog ate ch e mica l AMP has b een mo re tho r o ughlyte s tedin mammalia nsp ecies
v i am ore tr adition al indu s tri al saf e ty andh a n d lin g t oxi ci ty te st th an TRIS AMINO

. The
re pe ate dd o s e t o x ic i ty of AMP ha s bee n we ll

- s tu die d an ddoc ume n te d

. T h e s e d ietary studi e s r a ng e d from e i g htwee k s to o ne y e ar in mu ltip l e sp eci e s. Th e p ri m ary tr e atm en t- re l ate d e f f ect noted h a s b een h istop ath o lo gi cal c h anges con s is ting of in c reased liv e r weights and vac u o l ationof th e liv er in r ats an d dog s ( P arekh , 198 1; 1 981b ; Grif f i n, 1 990). No fur th er te stingof TRIS AMINO is r eco mm e nde d.

T. G e n e ti c To xi ci ty : G e n e Muta tions and C h r o moso m e A b er ratio n s IUC L

ID 5. 5 a n d 5
. 6: TRIS AMI N O was neg ativ e in a g e ne mutation ass a yin Sacc romy ce s c e re v i sia e , an dne gati ve i nth e ba cte ria l ge ne mu tati o n a ssay wi th E. c o l i (Livtak and McEvoy, 19 90). No a dditional te sting is consid ere dn e cess ary .
U . Ca rcinog e n i c i ty IUCL ID 5.7: TRIS AM INO didnot in du ce tu mo rs w hen tested in m a le Syri a ngo lden ha mster s re ce i vi n g 0. 2 ml of ami xture of Tri s b uffe r an d 0

. 9% Na C l i n tratr a c hea ll y in t o the lungs weekly for lif e (Ketk ar, M. et al ., 1 979). Th e lo ng historyof saf e use o f TRIS

14AM INO a s ab i olog i c a lbuff e r in th e treatmen t of acid osis, i t s use in Ke te r ol a c®, an an tiinfla mm atory drug, and th e comp oun d’s us e in co smetic s o v e r many y e ars suggest th at TRIS is no t carc inog enic. R e p e ated dose t oxicity studies o f TRIS and th e s tru ctur ally
re lated primary amino alc o ho l, AM P, do not sho wany evid en ce o f a n y p re n e op l astic lesions, an d mutag e n i c itystu dies are all n e g ativ e ; ao n e -y ear studyof AM P indog s (Griffin , 1 990), indicate d no ev id en ce of carc ino ge nicity. T h ese datasugg e s t that the materia l s
a re no t c a rci nogen i c . No a dditionalte stingis c onsider ed n ece ssa r y.
V . Repro duc ti v eTo x i cit y IUCL ID 5.8.1: T h e re ha ve bee nn o re pr oducti o n stu d i e s c o ndu c te d wi thT RI S AMI N O. D a t aare av ail ab l e to indi rectly ev alu ate th e p o te nti a l for re pro duc tiv e eff e c ts f r ome x p o sure
v i achron i c studies th at includ e histo l og ical e x amination of gonad al tissues f o r e v id en ce of adv e rse eff e c t s. No adv e rse cli n i cal, histolo g i cal, or h e m ato l ogi cal e f f e ct s were no te d in
m ore th a n a dozen rep e ated -dos e tox i city stu dies condu cte d w ith AMP an d AMPD that
wo u l d i n d i cate t o xic i ty to the r e pr o duc ti ve or gans . L i ke wi se , the re h a ve b e e nno re ports of anyre produ ctiv e eff e cts in th e multip l e studies c o n ducte d on hu ma np a tients with TR I S AM INO. Ho wev e r, in a re cen tr at re p r odu ctiv e/d e v e lopme nt al s c re ening stu dy, th e H C l salt
o f AMP has been o b s erv e d to b e f e to to xic in r ats. A n OECD 421 stu dy w a s co ndu cted
u s in g AMP- H C l in wh ich m a le an d f e m a le CD rats were f e d die t s supp lying 0( c on tro l ), 1 00, 3 00, o r 1000mg/kg/day o f AM P-HCl ( Carn e y et al

. , 200 5). M a les were e xpos ed fo r at lea s t tw o weeks p rio r to bree din g an dc o n tinu i n g throu ghou t breeding f o r 37 days. The f e males w e re expo s e d fo r two week s prior to breedi ng, continu i ng throug h breeding (up to two w eeks ) , ge s t atio n ( three w e e k s), and lactatio n (four d a y s ). Ev id en ce of comp lete litte r
re so rp tion ( 10 0% po st-imp la n t ationloss) w a s se e nat 10 00 mg/kg/d ay, and sign ifican t
re so rp tions were s een at 300 mg/kg/day. Ef fect s asso ciated with , or s e condary to th e p o sti mpl ant atio nloss in creas e at300m g / k g/d a yin cl u d ed d e c rea sed li tter si z e , in crea sed p u p b ody weight , and d ecrease d g e statio n body weig ht a ndbody weightg a in

. Th e no-ob s e rv ed effect lev e l ( NOEL) for s yste mic to x i city in male s was 100 m g/kg/d a y , bu t cou l dnot b e
es tab lish e d f or f e m a l e s du e to l i v e r eff e c t s in th e l owdose gro up ; th e NOEL fo r
re produ ctiv e toxicity w a s 100mg/kg/day. Th ere w e re no treatmen t re lated effects on
re produ ctiv e per fo rm an c e in th e 100 m g/kg/d a y g roup. Th e N O EL for g e n e r a l toxicity in
ma les wa s 300 mg /kg /day , wh i l e the ge neral to xicityNO EL for f e m a les could not b e

15d e termin ed, based up on the p resen ce of v e ry sl i g ht mi c r o s co p i c l i v e r ef f e c t s. T h e N O E L f o r
re produ ctiv e eff e cts w a s cons id ered to be 100 m g/kg/d ay. Com m

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re produ ctiv e toxi city ass oci a ted withTRIS A M IN O, we beli ev e th at
re produ ction/d e v e lop ment a l tox i city d at a m a ye x ist. We p l an to explo re th is furth e r by filin g a Freedo mof Info rmationAct ( F OIA) requ est w ith th e US Food a ndDrug A d min i str ati on (FD A ) to dete rmin e if su c h te sting has b e en co ndu cted. In th e ev e n t th at we d o no t obtain this d ata , the nw e c ommit to condu c tin g an O E CD 4 21 re pro ductiv e an d de vel o p me n ta l scre e n i n g stu dy si milar t o th at c o ndu c te donAMP t o f u ll y e v al uate a ny Formatted: Not Highlight Formatted: Not Highlight Formatted: Not Highlight p ote n ti a l f e t otoxi city o f TRIS A M IN O inrats.
W. Dev e lop m ental To xi c i ty IUCL ID 5.82: D at aare not av a i l abl e for TRIS AM IN O. How e v e r, anOECD 4 14 GL P g u id el in e study onAMP is av a i l abl e. In th is study , female r ats w e re e xpo sed 6 hours d aily d e rmally to 0 , 30, 100, or 300mg AMP/kg/d a yfro m gestatio n d a ys (GD ) 6-20 ( C arn e y and T h o r sru d , 20 06 )

. Derm a l a d mi n i str ati o n of 3 0 0mg /k g/da y of AMP pr odu c e dsi gni f i c an t
e ffe ct s at th e te st s ite , as ev id en ced b y scabbing andm oder ateto s e v e re s c aling

. Th e d e rm al fin ding o f slig ht s c aling at 30 and10 0 m g/kg/d a yw a s n ot co n s id ere d adv e rse, as th e
o b serv ation w a s tr a n si en t in n ature andrel ati v e lyl o w inin ci d e n c e. Th e re was n o e v id e n ce
o f te s tarti c l e re late d sy stemi c m ate rn al o rd e v e l opm ent al to xi ci ty at any dose l e v e l te sted. U nder th e con dition s of th is study, the N OAEL for mate rn al tox i city b a s e d on d e rm al eff e cts
w a s 10 0 mg/kg/d ay. Th e NO EL fo r d e v e lop me n tal to xicityw a s 3 00 mg/kg/d ay, th e hi g hest d ose l e v e l teste d. W hil e th is s tudy is beli ev ed to re p re s e n t a d e fin itiv e e v alu ati on o f p ote n ti a l d e velopmen t a ltoxi city fo r TRIS AMINO in addi ti o n to AMP, it is b e l i e v e d th at g i v e n th e ex te nsiv e u s e of T R IS AM INO a s a th er ap e u ti c ag ent d a t aex ist w ithin US FD A
arc h iv es to address th is en dpoint (FO IA re f e re n ce noted in R e p rodu ction Sectio n and b e low ). Sho uld n o d ata b e id entif i e d , a p ropos e d OECD 4 21 re p rodu ctiv e and d e v e lopmen t a l screen ing study wi llp r ovid e an ad dition al e v al uation o f an y p ote nti a l f or d e v e lopmen t a l toxi city TRIS AMIN O. Formatted: Not Highlight

16VI. CON C L U SIONS E v al uati on o f th e e x i s ti ng datale ads to th e c onc lusi ons that [1] a substantia l quan ti tyof d at ac urren tl y ex ist to ad e qu ate l y re p resent th e to xi colo g i cal a n d ec olog i c al scre en ing profile of the cate gory, [ 2 ] th e re is a c onc urrenc e a n d s i mi lari ty among the e x i s ti n g dataf o r

th e v ari ou s HPV/ SI DS e ndp o i nt s [ 3 ] a v ai lable dat a f r o m pr e v i ous l ys tu die d stru ctur allysimi l ar prim ary a mino al c o ho ls can b e us ed to ad e qu ate ly re p resent th e
m a j o rityo f the v ario us H P V/SI D S en dpoints fo r the compoun dth at w a s n ot sub j ected t o th e same l e v e l of te sti n g (withth e ex ceptionof acu te inh a l ation a n d d e v e lo pmen t a l t oxi ci ty), an d [ 4 ] utiliza tionof th ese datasupp ort th e con c lusion th a t no furth e r tes ting is n e e d e d to satis f y e n dp oints fo r H P V/SID S withth e e x ce p tio n of R e p ro ductiv e toxicity

. ( T able 4: Test Plan M atrix ). Ho wev e r, TRIS AMIN O has a lon g history o f safe us e inp h armaceutical p r odu cts in te nd e d f o r us e i n terna l ly in huma n s. W e be lie ve th a t re produ ction/d e v e lo pme nt al tox i city d at a mayexist , s o th e FOIAinqu iry with the FD A will d e te rmin e if su ch te s ting h a s be en con ducted. I f no dat ai s foun dwe wil l th e n
commit to c ondu cting an OECD 42 1 re p r o du c tive andd e v e lopme ntal s c reening stu dy. Furth e r, a p ropo sed O E CD 421 r e p ro du c tiv e/d e v e lo pmen ta l to xic i tyscre en ing test will p rovid e d ata onth e potenti al d e v e lo pme nt al tox i city o f TRIS AMIN O.

17VII. REFER E NCES

A dams, N., G ouldi ng, K.H. , a n d Do bbs, A

. J. ( 1 985). Tox i cityof Eight W ater -Solub l e Orga nic Chemic als to Se le na strum capric ornu tum: AStudyof Meth ods fo r C a lculatin g Tox i c V a lu es Using D i ffere n tGrow th Parame ter s . Arch. En viron. Con t a m . Tox i co l. 14 :333­ 34 5. B a ldwin , R

. ( 1 961). Tris (hydro xymethyl) amino me th an e PrimarySkin Ir ritatio n S tudies. Intern al co rre s po nd en ce o f Th e Dow C h emical C omp a n y. M ay 19, 1961

. C arn ey, E. W. and Th o r srud, B

. ( 2 0 06)

. AMP: D e rm al D e v e lopmen tal Toxicity S tu dyin CRL:CD (SD ) Rats. Th e Do w Ch emical Comp any R e p ort No: DR-039 9

- 3 940-0 01

. Formatted: Font: Italic De

let

e d:

Formatted: English (U.S.) Carn ey, E. W., Qu ast, J.F., a n dZab l otny, C. L. ( 200 5)

. 2- Meth y l

- 2

- A mi n o p r opa no l H ydroch l oride: A Repro du c tiv e / D e velopmen talToxi city S c re e n ing Test in CD R ats. De let

e d: o f th e Do w Ch e mic al Co mp a n y De

let

e d: Th e D o w Chemi c al Comp a n y Report No

. D R -0 038-3 1 86-001

. CI

R: Co sm etic Ingre die nt Review Board ( 19 90). A Fi nalRep ort on the Sa f

e t

yA s se ss me nt of Am inom eth y lpropan ol a n d Amin om eth y lpro pan e diol. J. Am

. Co ll. o f

T

ox i co l. 9:2 03-2 28

. G i roux, J. and B e au lato n, I. (1962 )

. Rec h erc h es Pharm a c o lo giqu es Sur Le TH AM. Soc ie te P h ar mac i e M o nt pe l l i e r. 2 1 :2 06-217

. Formatted: Normal, Indent: Left: 67.5 pt De

let

e d: ¶ De

let

e d: ¶

Formatted: Font: Italic De

let

e d: ¶ G onsior, S. (2 006). Prediction o f th e Env i r o nmen ta l Dis tri bution a n d Tr an sport B e tw een Env i ronmen ta l C o mpartments for Tris( h ydrox y me thy l)a min ome tha n e Us in g Le vel I an d Lev e l I II Fu gacity Mod e ls. Dr a f t R e p ort of T h e Dow Chemical Compan y. Grif f i n , T. ( 1990 ). A O n e- Y e ar Or al Tox i c i tyStudyof AMP in Do gs. Co ulston R e searc h In corpor ate d , W hite S an ds R e s e arch C e n ter, A l amog ordo, NM , U S A

. Amend e d bythe orig in al au th o ron Ap ril 20, 1993

. IW L (1990 )

. Institut für gew e rb lich e Wasserwirtsc h aft und Leftre inh altun g e . V. IW L– Auftr a gsnu mm e r: Lab 59 4

. Ke tk ar , M

. , Green, U ., S c h n e id er, P., andMohr, U. ( 1 979). In vestig ationo nth e carc in og enic b u rd e nby air pollu tion inma n . Can c er Le tters 6 : 2 79-284

. Klimis c h , H., A ndreae, M. and Tillmann, U . (1 997 ). A Sy s te m atic Ap pr oac hf o r E v a l uati n g De let

e d: t

Formatted: Font: Italic De

let

e d: ,

Formatted: Font: Italic
Formatted: English (U.S.) Formatted: English (U.S.) De

let

e d: De

let

e d:

18 th e Qua l ity o f E x peri me n t a l T o x i co lo gi ca l an d Ec ot ox ic olo g i c al Data
. Re gul

. Toxic o l

. Pha r m a c o l

. 25 :1-5. Koc i b a , R. ( 20 03). A s s e ssm ent o f th e Pote nti a l fo r Skin S e ns iti zation o f Stru c tur al ly- R el ated A na l ogs Repre s e n ted by 2-Amino-2- (Hy droxy methyl) -1,3-Propan e di ol, 2

- A mino -2-Methyl1-Prop a no l , a n d 2 -Amino-2-M e thyl- 1,3

- Pr o p a n e dio l

. Written commu nication

.Lho t ak, H. (19 96). Particip ation C o e fficie nt of V arious AN GU S Products (AMPD APD ). ANGUS Chemie GmbH. Liv t ak, K

. M

. and M c Ev oy, G

. K. (1 990). Ketoro l ac, an in jectab le n onn arco tic an alg e sic. Cl i nic alP h arma c y 9 : 921

- 9 3 5. Machle, W., Scott, E. W. and Treo n, J. F. (194 0). Th e Physio log i cal Resp onse o f Anim a l s to So me Simple Monon itr o p ar a ffin s an d to Certain D e ri v ativ e s of Th ese Co mp ound s. J. I n d

. H

y g . Toxico l. 22 : 3 15

- 3 32

. McF arland, W.N

. a n d N orris, K

. S. ( 1958 ). Th e C o ntro l of p HbyB u ffer s inFish Tr ansport. C a lifo rn iaFi sh and G a me. 2 9 1

- 310

. Formatted: Font: Italic De

let

e d:

De

let

e d:

.

De

let

e d:

Th e M e rc k I ndex (Ele v e nth Edition) (198 9)

. M e rc k & Co. , In c . Rahw ay, NJ , USA. Pare kh , C. (1 980a). A c ute Toxicity Ef f ects o f A M P-95 o n B l u e g i ll Sunf ish, N i nety-six Hour LC50. Repo rt D R -0 309

- 439 1-002 fo r th e I ntern ation a l Min e r a ls & C h emical Corpo ration, Ph arm a co l o g y L ab o r at o ri e s , A n i m alS c i e n c e R e s e archb y A n a l y ti c a l B i o C h emi s try Labor ato ries , Columb us, MO, U S A. Pare kh , C. (1 980b ). AMP-95 Acu te To x i c i tyEf f e cts on D ap hnia, 48 Hr LC50. R e p o rt #2 5561 for the Intern atio nal Min e rals & Ch e mi cal C o rp or ation , Pharm aco logy Labo ratorie s , A n im al S c i e nce R e s e ar c hbyAnaly ti c al Bio Ch emistry Lab o r atories, Colu mbus, MO, USA

. Pare kh , C. (1 980c). D e rm al Toxicity of P-18 26 (AMP). Report PLR-11 3 o f Pharm aco logy L ab o r at o ri e s A n i m al S c i en c e R e s e arch , T e rre H au te , I N , USA Formatted: Space Before: 12 pt De

let

e d: 1989.

Formatted: English (U.S.) Pare kh , C. (1 981). A Three Mon th In D i e t Tox i cityStudyo f 2-Amino-2 -methyl-1 -prop a n o l inRats. Report o f Bio/dynami cs In c, Ea s tM ill st on e, NJ , U S A

. Sp onsored by th e Inte rn atio n a lMin e r a ls & Ch emi c al C o rpor ation, T e r r e H aute , IN , U S A

. De let

e d: ¶ Po wer , C . ( 1 975). Ey e Ir ritatio n, Bio a ss a y Report #2 6775

. Pharmacolog y Lab orato ries R e s e arc h an d Dev e lo pmen t, Commerc ial Solv en ts Corpo r a tio n . May 21 , 1975

. R e actio n M ech an ism s inEnviron m e n tal O rg anic Ch e mistry (1 994). Richard A. Lars on and Formatted: Indent: Left: 67.5 pt Formatted: Font: (Default) Times New Roman, 12

pt Formatted: Font: (Default) Times New Roman, 12

pt Eric J. Web e r Lew i s Publish e r s , A nn Arbor, MI , USA

. Chap ter 2 "H ydro l y s is ", p a g e s 103­ 16 7

. U S EPA (19 99). Th e u s e o f stru cture- activity re lationsh i ps ( S AR) in th e H i gh Pro duction V o lume Ch emicals Ch alle n g e Prog ram. O PPT, EPA. W e st, R.J. (2004 a ). A re p o rt of Th e D owC h e mical Compa ny using Lev e l I mod e l v e r s io n 2. 11, O b tain e dfro m th e Can a di a nEn vironment al Mod e l i ng C e n tre, Tre nt Univ e r s ity, Peterbo roug h, Ontario, C a n a d a . West, R.J. ( 200 4b). A re p o rt of Th e D ow C h e mical Compa ny using BIOWINestim ation software

. De let

e d: ,

19Y oung , B.E. and Tap p, J

. F. (198 3). Th e Toxicity of AM P 95 to th e b ro w nshri mp (Cr a ng on crang on) an d plaice ( P leuron ec tes platessa)

. Rep o rt K 349/B f or th e C e ntr al Electricity G e n e r ating B o ar d, Man c h e ster, U K by Imperial Ch e mical In dustries, PLC.

20Table 3: Su mma ry o f th e Da ta
Formatted Table SURR OGA T E SUB S TA NC ES TRIS AMINO (77- 86-1 ) A M PD (1 15- 69-5 ) AMP ( 12 4 - 68- 5)

PHY S ICAL C H EMISTR Y Melting poi n t , ° C 171

- 1 72 110 30 .5 B o iling poin t , ° C 219

- 2 20 o C @ 13 3.3hPa 290 .9o C @ 101 3hPa 151

- 1 52 16 5 o C @ 10 13 hPa
Vapor Press ure @ 20 ° C 3

. 0x 10 -6hP a 8

. 71 x 10 -3 hPa 4.5.3x10 -1 hPa
Wate r So lub ili ty @ 25 o C 800 g/L 2500 g/L Mis cib l e a l l p rop or ti o n s Log Kow -2.3 1 <-3. 8 -0.63 Estim ated LogD (log Kow@ p H7) -2.2 2 -- -2.77 Density 0.93 2 g/cm 3 @ 2 0 o C -- - 0 .928 g/c m 3 @ 20 o C ENV I R O NME NTAL FA T E Biodegradation No t inh e ren tl y inh e re n tl y Not inh e re n tl y ; do es not b i od egra de Hydrol ys i s No t ex p ecte d No t ex p ecte d Not exp e c te d Photodegrada tion Di re ct p hotoly s i sn o t ex

pec ted. Di re ct p hotoly s i sn o t ex

pec ted. D i rect pho to lysis no t exp ec te d. Tran sport between Environ m e n tal Co mpartments: ( Fuga ci tyLev el I II M o d e l ) Defau lt a s sumptio n: 100 0 kg/hrreleased simu lt an e o u s ly into air, w ate r, a n d so il. <0.1% to air 68.6% to w a te r

31.4 %to soil <0.1% to sedim e n t -- - 0.07 % to a i r 73.9 %to water 2 6.0%to so il 0.03 % to sedimen t

EC OT O X I C IT Y Acute Tox i city to Fish ( LC 50 ) Non-toxi c to 2 9speci e s in con c e n tra tio ns up to 4400 mg/L. Used as a buffering ag e n t a t 440

­ 110 0 mg /L for shipp i ng li ve fish 96-hsta tic L C 50

in ze b r afish is >10, 000 mg/L Marin e L C 50 ( 96h )= 184 mg /L in Pl e uron e ctes plate s sa Fresh w a ter LC 50 (48h ) = 33 1mg/L in Formatted Table

Formatted Table

Formatted: Font color: Auto

Formatted Table

21Leu c iscus idus Acute Tox i city to Aqua tic Inv e rtebr ates (48hr LC 50 ) -- - -- - LC 50= 179 mg/L in C r ang on cra ngon EC 50 = 193 mg/L in D aph nia m agna Toxi city to Aqua tic Plan t s 7 2-hour g ro wth ra te NOEC > 20 0 mg /L; 96

­ h our growthrate NOEC > 100 mg/L in Selenas trum c a

p r icornu tum -- - 72 -hour EC 50= 520 mg/L in Scened esm us sp.

TO X I COLO GICAL DA T A Acute Tox i city (or a l) LD 50> 3000 mg/kg in rats ( 20%solu tion ) &

m i ce ( 5 %soluti on ) LD50 was 3500 - 44 00 mg /kg in m i ce LDLo=140 mg /kg in m i ce LDLo = 1500 mg /kg in rabbits LD 50= 2 900 + 140 mL/kg i n ra ts LD 50= 2 150 mL /kg in mice LDlo = 1000 -2000 mL /kg in ra bbit s

Acute Tox i city (d e rm a l) mg/kg LD 50> 1000 mg/kg in rats & mice -- - LD 50 > 2 000 mg/kg in rab bits Acute Tox i city (inh alation ) -- - -- - --- Acute Tox i city (oth er ro utes ) LD 50= 3 280-40 40 mg /kg in ra t s via tai l ve in LD 50 = 6000 mg/kg in rats v i atai lvein LD 50 = 6100 mg/kg in m ice vi a tail vein -- - LC 50= 325 mg /kg in m ice i.p. d o s e

Acute Skin Irri tation In rabbits, a mild irritant on a b r a de d skin, re so lv ed with i n 48 ho urs -- - Inra bbit s, a n irri tan t in ab rad e d (b urrowin g lesions ) or inta ct (d

e r

ma ti tis) skin

. Acute Ey e Irri tation In rabb it s, s a tu ra ted T

R IS AM IN O sol u tion (4 0%) is e s s e n t ia lly non-irrita tin g. -- - In rabbits, visio n was de stroy e d if ey es were flush ed or u n flush e d. Neat AMP is hig hly irri tat i n g li kel y du e to highpH. Se nsi tizati on -- - 0.1% n e g a tive on I n t r ac utane o u s te st Formatted Table

Formatted Table

22Gu i n e a Pi g an d a la te r pa tc h te st with g u i n e a pigs fou nd

AM P to b e an onse n s it iz e r. Rep e ated D o se Toxicity NO AEL = 5 00 mg/kg o f 0.3M THAM in S

p ra gue-Dawley rats for 20 da y sof in fusions. NO AEL > 0. 5mg/kg o f 0.3M T H

AM for 4 w e eks viaIV in rabbits 2 0-Day d e rm al rabb it – No e ffec ts LOA E L

< 25p p m v i a diet for 12 weeks in Sprag ue-D awley ra ts ( i ncrea s e d l i ver we i g hts) LOAEL ~100 0 ppm v i a die t f or

8 w e e k s i n Sprag ue-D awley ra ts (a lopecia& fo cal s k in ero s io ns) NO AEL > 3 200 ppm v ia di et fo r 8 we eks in CD

- 1 m i c e NO AE L = 25 pp m v i a diet for 13 weeks in do gs (clin chemis try & o r

g a n w e i g ht c h an ge s su ggestin g liver as ta r g e t or ga n) NO AEL > 1 10p pm via die t for 1 year in d ogs Ge ne tic T o xic ity Muta ti on Ba cte r ia l cell g e n e mu tationassay (b acte ri al stra ins)- nega tive Not muta g e nic in 4 stra ins o f

Sa lmonella, with o r w ith ou t metab o li c a c ti va tion A M E

S

Te st

– ne gat iv e Mam m alia n cell g e n e

m utation ass ay – ne gat iv e Ba cte r ia l Revers e M u t. A ssay – neg a tive Ge ne tic T o xic ity C hromosomal Aberrations -- - -- - Mous e Mic r onucleus A ss a y- ne gative Toxi city to R e p r o duc ti on -- - -- - A 1 ye a r di etar y dog stud y reveale d no gross o r histo path o lo gic effect on testes, uteri, or ovaries ( NO AEL > 110 ppm ) NO E L

fo r ge nera l Formatted Table

23toxi cit y in m a l e ra ts w a s 3 0 0 mg/ k

g/

day ; th e g en era l toxi ci t y NOE L for femal e rats cou ld not b e dete rm in e d , base d up on th e presen ce of ve ry slight mic ro sc opic liver e ffects. NOE L

for re

p roduc tive ef fe cts i s 10 0 mg /kg / d

ay. (OECD 42 1 stud y) Dev e lopmen ta l Toxicity -- - -- - NOAEL fo r m atern a l toxicity ba s e d on dermal effects w a s 1 00 mg/kg / da

y. The NOEL fo r d e velop m e n t a l toxicity was 300 mg /kg /da y, t h e h igh es t dos e le ve ltes te d

. (D eve lopm ent a l stud y inra ts OE CD414 )

24T a ble 4: T e s tPl an M a t r ix TR IS A M INO

(7 7 - 8 6 - 1 ) AM PD (11 5- 69-5) AM P ( 124- 68- 5 ) PHYSI C A L CHEM ISTRY Melting point, ° C 1 71-17 2 (m e a su red) A No D at a 11 0 (me a su re d)
No Data 30.5 (measured)

NoDa t a Bo il ing p o in t, ° C 219-2 20 (me a su re d) A No D at a 15 1-152 (m easure d)
No Data 165 ( m e asure d )
NoDa t a Vapor Press ure @ 2 0 ° C 3.0x10 -6hP a (c alc u late d)

Calc No D at a 8.714x10 -3 hP a ( c a l cu la ted ) Ca lc No D a ta 4

. 5.3x10 -1 hPa (calcu lated ) Calc No D at a W ate r Sol u b ili ty @ 25 o C 8 00 g/L (m e a su red) A No D at a 25 00 g/L (me a su re d)

Ex

p No Data Mis ci bl e al l p rop or ti o ns (measured)
NoDa t a Log Kow - 2

. 3 1( me as ure d) A No D at a <
- 3. 8(me a su re d)
NoD a t a -0.6 3 Calc No D ata E N VIRONME N TAL FAT E Bi odegradation Not Inherently(measure d ) A No D at a I nhe re n tly (m e a su re d)
NoD a t a Not inh e re n tl y ; do e s no t b i ode gra d e C a lc
No Data H ydrolysis Not exp e cte d NA No D at a Not ex pec ted NA No Data No t ex p ecte d NA No D at a Photodegradation D i rect pho to lysis no t exp ec ted

. NA No D at a Dire c t ph otoly s i snot ex

pe cte d

. NA No Data Di re ct p hotoly s i sn o t ex

pec ted. NA No D at a Tran sport b e twee n Environme n tal Co mpartments: ( F u gacity Level I II M o del ) Defau lt assu mp tion: 100 0 kg/hr re leased simult a n e ou s lyinto ai r, water, and s oil. <0

. 0 01 % to ai r 68.6 %to water

3 1.4%to soil <0 .1 % to se di me nt Calc No D at a --- 0 .07% to air 73.9% to w a te r 26.0 %to soil 0.03% to sedim e n t

Calc No D ata EC OT O X I C IT Y Acute To x i cityto Fi sh ( LC 50 ) No n-toxi c A No D at a LC 50 (9 6-h) static in ze b r afish is >10 ,000 mg/L LC 50 (4 8h ) i nB l ueg i ll sunfish
Y Acute To x i cityto A q u ati c I n v e rtebr ates (4 8hr LC 50 ) R R EC 50in Daph nia

ma gna Y Formatted: Font color: Black Formatted: Font color: Black

25Toxicity to A quatic Plants 72-h our gro w th ra te NOE C i n Se le n a s t r um c a

p ric or nutu m A No D at a --- 7 2 -hour EC 50in Scenede s mus sp.

No Data T O X I C O LOGICAL DATA Acute To x i city (o ral) LD 50in rats A No D at a LD 50in m i ce and ra ts LD 50 in mi ce a nd rats
N Acute To x i city (d erm a l) mg/kg LD 50in rats & m ice A N --- LD 50in rab bits
N Acute To x i city(inh a l ation) N A NA NA Ac ute Ski nIrrita ti on Irritan t A N --- Irrita nt N Acute Ey e Ir ritatio n N o n-ir rita ting

A No D at a --- Highly irrita tin g likely du e to high pH.
N Se nsi ti zati on No n-sen s it izer R N o n- se ns i tize r Non-s e n s it iz e r N R e pe ate dDo se T o xic ity 20 -d a

y IV inra ts 4

- w e ek IV inra bbits

R --- D i et ( 12 weeks) in ra ts No D at a D iet ( 8w e eks) in ra ts No Data Diet (8 we eks)in mice
No Data D i et ( 1 3 we eks) in do gs NoDat a Diet ( 1 year ) in do gs Y N, Y C arc i n oge nici ty Te s t Negative for tumors A N e g a t i ve

Ge ne tic T o xic ity - M uta tion Nega tive A N Ne gative A l lne gative
Y Ge ne tic T o xic ity C hromosomal Abe rrations R --- N e g ative

Y Toxicity to R e p rodu ctio n T e s t (D ep en de n t o n FOIA re su lt) --- O n e year di etar y dog stud y Y OECD421 in rats

Y Dev e lo pmen t a l To xi city T e s t (D ep en de n t o n FOIA re su lt) --- OECD414in rats Y

26Sy m b o l R T e s t Cal c A E xp NR N A
Y N NoDat a Legen d Descriptio n E ndpoint re qu i rement f ul f i ll e d using S tructurally rel at ed in f orm ati o n E ndpoint re qu i rements t o be fulf ille d with te sting E ndpoint re qu i r ement f ul f i l l e d based on calc u l ated dat a E ndpoint re qu i rement f ul f i ll e d w ith adequateexisti n g data E ndpoint re qu i rement f ul f i ll e d via experime ntation Not required per the OEC D S I DS guidan c e N o t applic abl e due t o phys i c al/c h e mical properties Yes (GLP ) No ( G L P ) N o data on GLP s t atus

27

# RECEIVED v- ! 

## OP6T 

# CErlC 2@6 

# OEC 

# - I 

# A!"! 

# 7: 

# 32 

# IUCLID Data Set

Existing Chemical 

### : ID: 77-86-1 CAS NO. : 77-86-1 Molecular Formula : C4HllN03 CAS Name : Tris (hydroxyrnethyl) aminomethane
Producer related part Company 

### : The Dow Chemical Company Creation date 

### : 11.11.2006 Substance related part Company : The Dow Chemical Company Creation date : 11.11.2006 Status Memo Printing date : 14.1 1.2006 Revision date Date of last update 

### : 14.1 1.2006 Number of pages 

### : 60 Chapter (profile) 

### : Chapter: 1,2,3,4, 5,6,7, 8, 10 Rellablllty (profile) 

### : Reliability: without reliability, 1,2, 3, 4 Flags (profile) 

### : Flags: without flag, confidential, non confidential, WGK (DE), TA-Luft (DE), Material Safety Dataset, Risk Assessment, Directive 67/548/EEC, SlDS

## 1. General Information Id 77-8 6 -1 Dat e 14.11.20 06

1.0.1 APPLIC ANT AN D C OMP ANY INF OR M ATIO N
Ty p e : manufa c turer Nam e : Do w Ch emic al, TERC Con t ac t pers on : Dr. William T. Stott Dat e : 27.08.20 06 Street : 1803 Buildi n g To w n : 4867 4 Midlan d, MI Coun try : United States Phone : Telefa x : Telex : Cede x : Email : Homepa ge : Source : Do w Ch emical, TERC Mi dland, MI 30.08.20 06
1.0.2 LOCA TION OF PR OD UCTION SITE, I M POR T ER OR FOR M ULATOR
Ty p e : manufa c turer Name of pla n t : Do w Ch emic al Street : To w n : Sterlington, Loui siana Coun try : United States Phone : Telefa x : Telex : Cede x : Email : Homepa ge : Source : Do w Ch emical, TERC Mi dland, MI Reliabilit y : (2) v alid with rest rictio ns Data from h a ndbo ok o rcol l ection of dat a. 13.11.20 06 (1) Ty p e : manufa c turer Name of pla n t : Do w Ch emic al Street : To w n : Ibbenbu ren Coun try : Germ any Phone : Telefa x : Telex : Cede x : Email : Homepa ge : Source : Do w Ch emical, TERC Mi dland, MI Reliabilit y : (2) v alid with rest rictio ns Data from h a ndbo ok o rcol l ection of dat a. 13.11.20 06 (1)

2 / 60

## 1. General Information Id 77-8 6 -1 Dat e 14.11.20 06

1.0.3 IDENTITY O F RECIPIENTS

1.0.4 DETAILS O N CATEG ORY / TEMPLATE

1.1.0 SUBST A NCE IDENTIFICATIO N

IUPAC Nam e : 2-Amino
- 2
- hy droxymethyl-1,3-p r op ane d iol Smiles Code : OC C( N) (C O) CO Molecular fo rmula : C4 H11 NO 3 Molecular w e ight Petrol class : : 121.14

Source : Do w Ch emical, TERC Mi dland, MI Reliabilit y : (2) v alid with rest rictio ns Data from h a ndbo ok o rcol l ection of dat a. 09.11.20 06 (2)
1.1.1 GENER A L S UBST A NCE INFO RM ATIO N

P u r i tyty p e : typical for ma rkete d su bsta nce Substa nce ty pe : orga nic Ph y s ical status : solid Purit y: Colour : White Odour : Odorl ess Reliabilit y : (2) v alid with rest rictio ns Data from h a ndbo ok o rcol l ection of dat a. 13.11.20 06 (3)
1.1.2 SPECTRA

T y pe of spectra : NMR Resul t : 13-Ca rbo n NMR sp ect r a o fTRIS were measured bet wee n 407 a n d 461 K. Source : Do w Ch emical, TERC Mi dland, MI Reliabilit y : (2) v alid with rest rictio ns Meets ge ne ra lly acce pted scientific stand ard s , well -do c ume nted an d accepta b le fo r asse ssment

. 09.11.20 06 (4)
1.2 SYNONYMS AND TRADE NAMES

Talatrol Source : Do w Ch emical, TERC Mi dland, MI Reliabilit y : (2) v alid with rest rictio ns Data from h a ndbo ok o rcol l ection of dat a. 19.03.20 04 (2) THAM Source : Do w Ch emical, TERC Mi dland, MI 3 / 60

## 1. General Information Id Dat e 77-8 6 -1 14.11.20 06

Reliabilit y 19.03.20 04 : (2) v alid with rest rictio ns Data from h a ndbo ok o rcol l ection of dat a. (2) Trimethlol aminomethan e Source Reliabilit y 19.03.20 04 : : Do w Ch emical, TERC Mi dland, MI (2) v alid with rest rictio ns Data from h a ndbo ok o rcol l ection of dat a. (2) TRIS Source Reliabilit y 19.03.20 04 : : Do w Ch emical, TERC Mi dland, MI (2) v alid with rest rictio ns Data from h a ndbo ok o rcol l ection of dat a. (2) TRIS AMINO ® Source Reliabilit y 10.11.20 06 : : Do w Ch emical, TERC Mi dland, MI (2) v alid with rest rictio ns Data from h a ndbo ok o rcol l ection of dat a. (2) Tris Buffer Source Reliabilit y 19.03.20 04 : : Do w Ch emical, TERC Mi dland, MI (2) v alid with rest rictio ns Data from h a ndbo ok o rcol l ection of dat a. (2) Tris(hy d roxy m e thy l )am i nom e tha n e Source Reliabilit y 19.03.20 04 : : Do w Ch emical, TERC Mi dland, MI (2) v alid with rest rictio ns Data from h a ndbo ok o rcol l ection of dat a. (2) Tris-steril Source Reliabilit y 19.03.20 04 : : Do w Ch emical, TERC Mi dland, MI (2) v alid with rest rictio ns Data from h a ndbo ok o rcol l ection of dat a. (2) Trisamine Source Reliabilit y 19.03.20 04 : : Do w Ch emical, TERC Mi dland, MI (2) v alid with rest rictio ns Data from h a ndbo ok o rcol l ection of dat a. (2) Trizma Source Reliabilit y 19.03.20 04 : : Do w Ch emical, TERC Mi dland, MI (2) v alid with rest rictio ns Data from h a ndbo ok o rcol l ection of dat a. (2) Trometamol 4 / 60

## 1. General Information Id 77-8 6 -1 Dat e 14.11.20 06

Source : Do w Ch emical, TERC Mi dland, MI Reliabilit y : (2) v alid with rest rictio ns Data from h a ndbo ok o rcol l ection of dat a. 19.03.20 04 (2) Trometh a ne Source : Do w Ch emical, TERC Mi dland, MI Reliabilit y : (2) v alid with rest rictio ns Data from h a ndbo ok o rcol l ection of dat a. 09.11.20 06 (2)
1.3 IMPURITIES

Purit y: typical for ma rkete d su bsta nce CAS -No : EC-No : EINECS- N a m e : Molecular fo rmula : Value : Rem a r k : The co mpo u n d is sol d as 1 00% cry s tal o ra 40% aqu e ous solutio n of tris(hydroxym ethyl)amin om ethane. Source : MSDS of the Do w Ch emical Comp any. 2003. Do w Ch emical, TERC Mi dland, MI Reliabilit y : (2) v alid with rest rictio ns Data from h a ndbo ok o rcol l ection of dat a. 09.11.20 06
1.4 ADDITIVES

P u r i tyty p e : typical for ma rkete d su bsta nce CAS -No : 7732 -18
- 5 EC-No : 231-791 -2 EINECS- N a m e : water Molecular fo rmula : H2O Value : c a . 40 % v/v Function o f additiv e : Solvent Source : Do w Ch emical, TERC Mi dland, MI Reliabilit y : (2) v alid with rest rictio ns Data from h a ndbo ok o rcol l ection of dat a. 09.11.20 06 (1)
1.5 TOTAL QUANTITY

1.6.1 LABE LLING

1.6.2 CLASSIFICATION

1.6.3 PACKAGING

5 / 60

## 1. General Information Id 77-8 6 -1 Dat e 14.11.20 06

1.7 USE PATTE RN

T y pe of use : use Category : other: Emulsif i er for cosmet ics, mine ral oi l and wax em ulsio n s, leath e r dre ssi ng s, textiles, clean ers, pharma ce u t icals, an d ch emical inte rm ediate buffer Source : Do w Ch emical, TERC Mi dland, MI Reliabilit y : (2) v alid with rest rictio ns Data from h a ndbo ok o rcol l ection of dat a. 10.11.20 06 (1) (2)
1.7.1 DETAILE D USE PATTERN

1.7.2 METHO D S O F MANUF A CTURE

Resul t : May be pre p a r ed by re du ction or cata lytic hydro gen ation of co rre sp ondin g nitro co mpo u nd. Prepa rati on by electrol yt ic redu ction : McMillan, US Patent 2,485,98 2 (1 949 TO COM M SOLVENT S CORP ), CA 44, 1836B (1 950) Source : Ha zardou s S ubsta nces Da ta Bank. Reliabilit y : (2) v alid with rest rictio ns Data from h a ndbo ok o rcol l ection of dat a. 13.11.20 06 (5)
1.8 REGU LA TO RY MEASU R ES

1.8.1 OCCUP ATIONAL EXPOS URE LIMIT VALUES

1.8.2 ACCEPTABLE RESIDUE S LEVELS

1.8.3 WATER POL LUTIO N

1.8.4 MA JOR ACCID EN THA ZAR D S

1.8.5 AIR POLL UT ION

1.8.6 LISTINGS E.G. CHEMICAL INVENTORIES

1.9.1 D EGRAD ATION /TR AN SFOR MA TION PR ODUC TS
Ty p e : degradatio n p r odu ct CAS -No : EC-No : EINECS- N a m e : 6 / 60

## 1. General Information Id 77-8 6 -1 Dat e 14.11.20 06

IUCLI D Cha p ter : Rem a r k : THAM is stab le at room te mperatur e fo r period s a s lo ng as 1 2 years. Source : Do w Ch emical, TERC Mi dland, MI Reliabilit y : (2) v alid with rest rictio ns Meets ge ne ra lly acce pted scientific stand ard s , well -do c ume nted an d accepta b le fo r asse ssment

. 09.11.20 06 (6)
1.9.2 COMPO N E N TS

1.10 SOURCE OF EXPOSURE

Source of e x posure : Huma n: expo sure by pro d u c tion Exposur e to the : Substan c e Source : Do w Ch emical, TERC Mi dland, MI Reliabilit y : (2) v alid with rest rictio ns Data from h a ndbo ok o rcol l ection of dat a. 09.11.20 06 (1) Source of e x posure : Huma n: expo sure thro ugh i n tende d use Exposur e to the : Substan c e Source : Do w Ch emical, TERC Mi dland, MI Reliabilit y : (2) v alid with rest rictio ns Data from h a ndbo ok o rcol l ection of dat a. 09.11.20 06 (1) Source of e x posure : Huma n: expo sure of the co nsum er/by s ta nder Exposur e to the : Substan c e Source : Do w Ch emical, TERC Mi dland, MI Reliabilit y : (2) v alid with rest rictio ns Data from h a ndbo ok o rcol l ection of dat a. 09.11.20 06 (1)
1.11 AD DITIO N A L REMA RKS

1.12 LAST LITERATURE SEARCH

T y pe of search : Internal an d External Chap ter s co v ered : 3, 4, 5 Date of se ar ch : 03.11.20 06 13.11.20 06
1.13 REVIEWS

7 / 60

## 2. Phy s ico-Chemical Data Id 77-8 6 -1 Dat e 14.11.20 06

2.1 MELTING P O INT

Value : Sublimation : Metho d : Year : GLP : Test s ubs ta nce : Source : Reliabilit y : 19.03.20 04 Value : Sublimation : Metho d : Year : GLP : Test s ubs ta nce : Source : Reliabilit y :

24.03.20 04
2.2 BOILING POINT

Value : Rem a r k : Source : Reliabilit y : 28.06.20 04 Value : Source : Reliabilit y :

24.03.20 04
2.3 DENSITY

2.3.1 GRANUL OM ETRY

2.4 VAPOUR PRESSURE

Value : Decomp ositi on : ca. 171 - 172 °C

1989 as prescribe d by 1.1 - 1.4 Do w Ch emical, TERC Mi dland, MI (2) v alid with rest rictio ns Data from h a ndbo ok o rcol l ection of dat a. (2) ca. 171 - 172 °C

1940 as prescribe d by 1.1 - 1.4 Do w Ch emical, TERC Mi dland, MI (2) v alid with rest rictio ns Meets ge ne ra lly acce pted scientific stand ard s , well -do c ume nted an d accepta b le fo r asse ssment

. (7)

ca. 219 - 220 °C at 101.33 hPa @ 10mm H g Do w Ch emical, TERC Mi dland, MI (2) v alid with rest rictio ns Data from h a ndbo ok o rcol l ection of dat a. (2) ca. 219 - 220 °C at Do w Ch emical, TERC Mi dland, MI (2) v alid with rest rictio ns Meets ge ne ra lly acce pted scientific stand ard s , well -do c ume nted an d accepta b le fo r asse ssment

. (7)

ca. .0000 03 hPa at 25 °C
8 / 60

## 2. Phy s ico-Chemical Data Id 77-8 6 -1 Dat e 14.11.20 06

Metho d : other (cal cul a ted) Year : 2004 GLP : Test s ubs ta nce : Source : Do w Ch emical, TERC Mi dland, MI Reliabilit y : (2) v alid with rest rictio ns Acce pted cal c ulatio n meth od. 28.06.20 04 (8)
2.5 PARTITIO N COEFFI CIEN T

Partition co e fficien t : oc tanol-water Log po w : = -2.3 1 at 20 °C pH v alue : Metho d : OECD Guide

- line 10 7 "Partition Coeffi ci ent (n-octa nol /water), Flasksha k in g Meth od" Year : 1996 GLP : no data Test s ubs ta nce : as prescribe d by 1.1 - 1.4 Conclu sion : The cal c ul ated partition coefficient is consi s tent with high water solubility and which by definition wou ld be indi cative of low log Kow value s
. Reliabilit y : (1) v alid with out rest rictio n Comp arable t o a guidelin e study. 13.11.20 06 (9) Partition co e fficien t : oc tanol-water Log po w : < = -3.8 at 20 °C pH v alue : Metho d : OECD Guide

- line 10 7 "Partition Coeffi ci ent (n-octa nol /water), Flasksha k in g Meth od" Year : 1997 GLP : no data Test s ubs ta nce : other TS:AM P D Test s ubs ta nce : AMPD (2 -Amino-2
- methyl -1,3-p rop ane d i ol) CAS# 00 0115 -69
- 5. Molecula r weig ht = 105. 14 13.11.20 06 (10 ) Partition co e fficien t : oc tanol-water Log po w : c a . -2.22 at °C pH v alue : = 7 Metho d : other (cal cul a ted) Year : 2004 GLP : Test s ubs ta nce : as prescribe d by 1.1 - 1.4 Source : Do w Ch emical, TERC Mi dland, MI Test c onditi on : Model Input Paramete rs: Data Tem perature (°C) = 2 5 Chemi c al Typ e = 1 Molecula r Ma ss (g/mol ) = 1 21.14 Wate r Solubil i ty (g/m3) = 5 50,000 Vapor Pressu re @ 25 ° C (Pa) = 3.0 x 10-4 Melting Point ( C) = 171.5 Estimated He nry's L a w Co nstant (H) (Pa m3/mol) = 6.7 x 10-8 Log Kow (Octanol-Wate rPartition Co efficient) = -2.22 / 1.38 Simulated Emissi on (kg ) = 100,0 00 9 / 60

## 2. Phy s ico-Chemical Data Id 77-8 6 -1 Dat e 14.11.20 06

Conclu sion : This mate rial has very hig h water solubili ty, very low vapor p r e s sure, and very low log Kow. In the abse n ce of ad vective and reactive proce s ses, these physi c al properties di ctate that the material will partition exclusively to the water compartment at equilibri um
. Reliabilit y : (2) v alid with rest rictio ns Acce pted cal c ulatio n meth od. 07.04.20 04 (11 )
2.6.1 SOLU BILITY IN DIFFERE NT MEDI A

Solubilit yin : Wate r Value : ca. 800 g/l at 25 °C pH v a lue : conce ntra t io n : at °C Tempera t ure effe cts : Examine different p ol. : pKa : 8.03 at 25 °C Des cription : Stable : Reliabilit y : (2) v alid with rest rictio ns Data from h a ndbo ok o rcol l ection of dat a. 13.11.20 06 (12 ) Solubilit yin : Wate r Value : ca. 550 othe r:mg/mL at 2 5 °C pH v a lue : ca. 10.4 conce ntra t io n : .1 other:Mola rat 25 °C Tempera t ure effe cts : Examine different p ol. : pKa : at 25 °C Des cription : Stable : Deg. produ c t : Metho d : Year : 1989 GLP : Test s ubs ta nce : as prescribe d by 1.1 - 1.4 Source : Do w Ch emical, TERC Mi dland, MI Reliabilit y : (2) v alid with rest rictio ns Data from h a ndbo ok o rcol l ection of dat a. 19.03.20 04 (2)
2.6.2 SURF A C E TENSION

2.7 FLAS H POINT

2.8 AUT O FL AM MABILITY

2.9 FLAMM ABIL ITY

10 / 60

## 2. Phy s ico-Chemical Data Id Dat e 77-8 6 -1 14.11.20 06

2.10 EXPLOSIVE PROPERTIE S

2.11 OXIDIZING P R OPERTIES

2.12 DISSOCIA TION C ONST A NT

Acid-base c onstant Metho d Year GLP Test s ubs ta nce : : : : : 8.21 1989

Source Reliabilit y 28.06.20 04 : : Do w Ch emical, TERC Mi dland, MI (2) v alid with rest rictio ns Data from h a ndbo ok o rcol l ection of dat a. (13 )
2.13 VISCOSITY

2.14 AD DITIO N A L REMA RKS

Memo Source Reliabilit y 13.11.20 06 :

: : pKa= 8.3 @ 20° C; 8.03 @ 25°C; 7.8 @ 37° C L. Troe ster (2 006) ANGUS data. (2) v alid with rest rictio ns Data from h a ndbo ok o rcol l ection of dat a.

11 / 60

## 3. Environmental Fate and Pathways Id 77-8 6 -1 Dat e 14.11.20 06

3.1.1 PHOTO D EG RADATION

3.1.2 STABILITY IN WATER
Ty p e t1/2 pH 4 t1/2 pH 7 t1/2 pH 9 Deg. produ c t Metho d : : : : : : abiotic at °C at °C at °C

Year : GLP : no data Test s ubs ta nce : as prescribe d by 1.1 - 1.4 Conclu sion : TRIS AMINO is stabl e for several yea r s at ambient temperature s a c cordi ng to ANGUS files

. Reliabilit y : (2) v alid with rest rictio ns Data from h a ndbo ok o rcol letion of data. 13.11.20 06 (14 )
3.1.3 STABILITY IN SOIL

3.2.1 MONITO RING DATA

3.2.2 FIELD STUDIES

3.3.1 TRANSPORT BETWEE N EN VIRON M ENTA L COM P ARTME NT S
Ty p e : fugacity model level III Media : water - air Air : % (Fugacity Model Level I ) Wa ter : 100 % (Fuga city Model Le vel I) Soil : % (Fugacity Model Level I ) Biota : % (Fugacity Model Level I I/III) Soil : % (Fugacity Model Level I I/III) Metho d : other: calcul a t ed Can adia n Environment al Modelin g Centre, Trent University, Peterbo r o ugh, Ontario, Can ada. Year : 2006 Resul t : Tris(hyd roxymethyl)amin o m ethane h a s high water so lubility, a low vapor pre s sure, and low log Kow. The sub s tan c e ha s a lo w potential for adsorptio n to soil or sedi m ents, and a lo w potential to volatilize fro m water or soil to the atmosphere. If released to air, the subst ance will react with hydroxyl radi cal s
. If released di re ctlyto water, the most proba bl e emission route ba se d o n physi cal propertie s an d use p a ttern s, most of the substa nce will rem a in in the water co mpartme n t and is expe cte d to be biode grad ed. If relea s e d to soil, the sub s tance is expe cted to be biod egra ded. Test c onditi on : Input Parame ters for L e vel I Model: Data Tem perature (°C) = 25 Chemi c al Typ e = Type 1 indicates che m ical can pa rtition into all 12 / 60

## 3. Environmental Fate and Pathways Id 77-8 6 -1 Dat e 14.11.20 06

environ menta l compa rtmen ts Molec ula r Ma ss (g/mol ) = 1 21 Wate r Solubil ity (g/m3) = 8

. 0 x 10^5 Vapor Pressu re @ 25 ° C (Pa) = 3.0 x 10^-4 Melting Point (°C) 171.5 Estimated He nry's L a w Co nstant (H) (Pa m3/mol)= 4. 54 x10^-8 Log Kow O c t anol-Wate rPartition Co efficient = -2.3 (Estimated va lue of Log D at pH 7.0 / Estimated value for neutral sp ecie s) Simulated Emissi on (kg ) = 100,0 0 0 (Level I Defa ult Value) Input para me t ers to Level I II Model: Data Tem perature (°C)=25 Default environmental tem peratu r e Chemi c al Typ e = Type 1 ind i cate s ch emical can p a rtitio n into all environ menta l compa rtmen ts Molec ula r Ma ss (g/mol )= 1 21 Wate r Solubil i ty (g/m3)= 8. 0 x 10^5 Vapor Pressu re @ 25 °C (Pa)= 3 x 10^-4 Melting Point (°C)= 171.5 H e nry' s La w C o ns ta n t( P a* m3/mo l e) = 4.5 4 x1 0 ^
- 8 Log Kow (Octanol-Wate rPartition Co efficient)= -2.3 Simulated Emissi on Rate (kg/h r )= 1,00 0 Simulated Environment Level III Default environment Rea c tion Half -lives (hr) Inp u t to Level III Model: Air (vapo r ph ase ) 4 *Wate r(no su sp. soli ds) 36 00 *Soil 7200 *Sediment 72 00 **Susp end ed Sediment 1.0 x 10^11 **Fish 1.0 x 1 0 ^11 **Aero s ol 1.0 x 10^11 *Half-live s extrapol ated ba sed on inh e re n tbiodegrada b ility classifi cat i on, according to Tech nical Gui dan ce Docu ment of the Europ ean Com m issi on. **Default value used in Lev el III m odel when reaction is expected to be negligible in this compartment. Reliabilit y : (2) v alid with rest rictio ns Acce pted cal c ulatio n meth od. 14.11.20 06 (15 ) Ty p e : fugacity model level III Media : water - air Air : 0 % (Fuga city Model Leve l I) Wa ter : 100 % (Fuga city Model Le vel I) Soil : 0 % (Fuga city Model Leve l I) Biota : % (Fugacity Model Level I I/III) Soil : % (Fugacity Model Level I I/III) Metho d : Year : 2002 Resul t : This mate rial has very hig h water solubili ty, very low vapor p r e s sure, and very low log Kow. These prop ertie s di ctate that the material h a s l o w potential to volatilize from water or soil to air, or adsorb to soil and sedim ents fro m the dissolved state. Wh en rele ased to water (the most likely emissio n scena rio ) , the materi al will remain di ssolved in wate r and is expecte d to be ultimately biodeg rad ed. If released to soil, the mate rial will be prim arily d i ssolved, and remai n mobi l e in, soil pore water (gro un dwate r). Source : Do w Ch emical, TERC Mi dland, MI Test c onditi on : Input Parame ters for L e vel I Model: Data Tem perature (°C) = 25 13 / 60

## 3. Environmental Fate and Pathways Id 77-8 6 -1 Dat e 14.11.20 06

Chemi c al Typ e = Type 1 indicates che m ical can pa rtition into all environ menta l compa rtmen ts Molecula r Ma ss (g/mol ) = 1 21.14 C Wate r Solubil i ty (g/m3) = 5 50,000 Vapor Pressu re @ 25 ° C (Pa) = 3.0 x 10-4 Melting Point ( C) 171.5 Estimated He nry's L aw Co nstant (H) (Pa m3/mol) = 6.7 x 10-8 Log Kow O c t anol-Wate rPartition Co efficient = -2.2 2 / 1.38 (Estimated value of Log D at pH 7.0 / Estimated val ue for neut ral spe c ie s) Simulated Emissi on (kg ) = 100,0 0 0 (Level I Defa ult Value) Estimated He nry's L a w Co nstant (H) = (Pa m3/mol) 6

. 7 x 10-8 Rea c tion Half -lives (hr.) Inp u t to Level III Model Air (vapo r ph ase ) 3.8 Wate r (no su sp. soli ds) 3600 Soil 7200 Sediment 7200 Suspe nde d Sediment 1.0 x 10^11 Fish 1.0 x 10^11 Aero sol 1.0 x 10^11 Reliabilit y : (2) v alid with rest rictio ns Acce ptable calcul ation me thod. 09.11.20 06 (16 )
3.3.2 DISTRIBUTI ON

3.4 MOD E OF DEGR ADA TION IN AC TU AL U SE

3.5 BIODE G RADATIO N
Ty p e : aero b ic Inoculum : Deg. produ c t : Metho d : OECD Guide-line 301 D " R eady Biode gradability: Closed Bottle Test" Year : 1990 GLP : no data Test s ubs ta nce : as prescribe d by 1.1 - 1.4 Conclu sion : TRIS AMINO is not rea d ily biode gra dabl e. Reliabilit y : (1) v alid with out rest rictio n Meets natio n al stand ard m ethod s (AFN OR/DI N ). 13.11.20 06 (17 )
3.6 BO D5, CO D OR B OD 5/C OD R A TIO

3.7 BIOA C CUM ULA T ION

3.8 AD DITIO N A L REMA RKS

14 / 60

## 4. Ecotoxicity Id 77-8 6 -1 Dat e 14.11.20 06

4.1 AC UTE/PR O LON GED TO XICITY TO FISH
Ty p e Species Exposur e pe riod Unit : : : : other: ae rate d and stati cwere both te ste d other:29 spe c ies were te sted

Metho d : Year : 1958 GLP : Test s ubs ta nce : as prescribe d by 1.1 - 1.4 Metho d : Fish were exposed to the buffer for 2 d a ys, then pla c ed in u n treat ed wate r and ob se rved for 30 days. Fre s h-wate r speci es were left in the buffer solutio ns for 9 days. The vessels were tested un der static an d aerated con d ition s se parately. Fish were monit o red fo r sig n s of toxicity. Resul t : Incre a ses in weig ht occurred in all sp eci e s. The p H d e clin e over th e 9 day fresh

- water te st wa s 0.02 p H units. One spe c ime n of chi c lid (Aq u id ens portale gren sis) die d du ring the third day, likely die to starvation sin c e none were fed until the fourth da y. There wa s no other mo rtality in any other fish throug hout th e study peri o d. The only ob servation s of toxicity noted were to spe c i mens of the o paleye (Girella nig r icans), whi c h e x ibit ed a marked inte rrupti on of opercul a r movement s a fter 5 hours in the TRIS buffer. Irregul ari t ies app eared in solutio ns ran ging from 5

- 2 0g TRIS pe r gallon water. The irregula r ities we re cha r a c teri ze d by two or three larg e ope rcula rbeat s, followed by a pau se lasting 5
- 10 seco nd s. Total beats varie d from 8-2 0 pe r minute. Th e upset in opercul a r rhythm did not seem to hav e an adverse effect upon th e fish. They appe ared normal in all other a s pe cts. Source : Do w Ch emical, TERC Mi dland, MI Test c onditi on : The followi ng marine spe c i es were te ste d: Heterodo ntus franci s ci Holo ce nt ris m i cro s t o m u s My ripri s tis m urdja n Pseud ope ne us bifa sci atus Amphipri on p ercula Lepid aploi s bi lunulatu s Iniis tius pavo Girella ni gri c a ns Kuhlia margin atus Hipp ocampu s punctul a tus H e pa tu s o livac e u s Hep a tus b a ri ene Microcanthi s strigatu s Zebrasoma flavescen s Z a n c lu s c a nes ce ns Het e r o st i c hu s rost r a t i s Lepto c ottus a r matu s Clinocottus analis Balistes vidu a The followi ng fresh water speci es were tested: Lebi stes reticulatus Molliene sia sp. Aequide ns p o rtaleg r en si s Cichla soma n i grofa sci atus Conclu sion : Test s indicate that 29 spe c ie s of fish ca n stand hi gh con c e ntration s of buffer (2 0g / gallon ) dissol ved in transp o rt wate r. Light dosa g e s (2 -5g / gallon ) are ad equate to sta b ilize p H du ri ng tran spo rt of fish, rega rd less of 15 / 60

## 4. Ecotoxicity Id 77-8 6 -1 Dat e 14.11.20 06
the pre s en ce of aeration (p rovided the weight of fish transporte d do es not excee d 25-50 g / gallon in aclosed sy st e m). The dataindicate that TRIS buffer is no ntoxic to the 29 spe c ie s teste d unde r these conditio ns. Buffered solu tions are stab le for up to three mo nths atnormal ai r temperature s
. Reliabilit y : (2) v alid with rest rictio ns Meets ge ne ra lly acce pted scientific stand ard s , well -do c ume nted an d accepta b le fo r asse ssment

. 09.11.20 06 (18 ) Ty p e Species Exposur e pe riod Unit : : : : st at ic Brachydani o reri o (Fi s h, fresh water) 96 hou r(s) mg/l LC50 : > 100 00 measu r ed/n omin al Metho d : OECD Guide

- line 20 3 "Fish, Acute Toxicity Test" Year : 1997 GLP : Test s ubs ta nce : other TS:AM P D Resul t : Hour LC50 (mg/L) 24 >10 000 48 >10 000 72 >10 000 96 >10 000 Test s ubs ta nce : AMPD: 2-Ami no-2
- methyl -1,3-p rop ane d iol Conclu sion : The acute LC50 96-hou r fo r Bra c hydani o reri o is > 1 0 ,000 mg/L for AMPD. Reliabilit y : (2) v alid with rest rictio ns Guideli ne stu dy with accep t able re stri ctions. 14.11.20 06 (19 ) Ty p e Species Exposur e pe riod Unit : : : : Leu ciscu s idu s (Fi s h, fre s h water) 96 hou r(s) mg/l LC50 : > 100 00 cal c ulate d Metho d : Year : 1990 GLP : Test s ubs ta nce : as prescribe d by 1.1 - 1.4 Source : IWL lab s 199 0 Conclu sion : The 96 -h L C 5 0 in golde n orfe is >10,0 00 mg/L. Reliabilit y : (2) v alid with rest rictio ns Data from h a ndbo ok o rcol l ection of dat a. 13.11.20 06 (20 )
4.2 AC UTE TOXI CITY TO AQ UATI C INVE RTEB R A TES
Ty p e : Species : Cra ngo n cran gon (Crusta c ea) Exposur e pe riod : hour(s) Unit : mg/l LC50 : = 179 mea s ured/n omin al Metho d : Year : 1983 GLP : no data Test s ubs ta nce : other TS: AMP
16 / 60

## 4. Ecotoxicity Id 77-8 6 -1 Dat e 14.11.20 06

Test s ubs ta nce : AMP-95 (2 -A mino-2-m e thyl-1-pro pan ol) Conclu sion : The 48 - & 96-hour L C 5 0 values for Cran gon cran gon i s 179 mg/L. Reliabilit y : (2) v alid with rest rictio ns 13.11.20 06 (21 ) Ty p e : Species : Dap hnia ma g na (Cru sta c e a) Exposur e pe riod : 48 hou r(s) Unit : mg/l EC50 : = 193 mea s ured/n omin al Metho d : Year : 1980 GLP : no data Test s ubs ta nce : other TS:AM P Test s ubs ta nce : AMP95 (2-A mino-2-m e thyl-1-pro pan ol) Conclu sion : In Daph nia m agna, AMP-9 5 wa s rep o rte d with a 48-h our EC50 of 193 mg/L. Reliabilit y : (2) v alid with rest rictio ns 13.11.20 06 (22 )
4.3 TOXICITY TO AQ UA TIC PLAN TS E.G. ALGAE
Species : Selena strum cap r icornutu m (Algae ) Endpoint : growth rate Exposur e pe riod : 4 day(s) Unit : μg/l NOEC : ca. 100 mea s ured/no mina l Metho d : Year : 1985 GLP : Test s ubs ta nce : as prescribe d by 1.1 - 1.4 Metho d : Dilution s of the actively-g ro wing cultures were added to tes tflas ks to give an app roxima te initial countof 1000 cell s per mL, and i n cu bated 2 4 hours. An appropri a te volume of test materi al wa s add ed to achieve the desi r ed con c e ntration , and the cult ure volum e made up to 10 0 m L with pa rticle-f ree deioni zed wat er. Cell co un ts we re mad e every 24 hou rs. The cell counts were used to cal c ulate the gro w th pa ram e ters to e s tab lish qu antitative toxic values. Source : Do w Ch emical, TERC Mi dland, MI Test c onditi on : Stock culture s of Selena strum we re mai n tained at 22 C und er con s tant illumination from 2 fluore scent tubes, on slop es of Oxo i d agar contai ning Bold's b a sal medium. About 7 days pr i o r to use, 2 conical flasks of 90mL particl e-free BBM were a s eptically ino c ulated with S elena stru m from sto ck cultures. The culture s were incu bated a t22C, and sh ake n at 175 rpm unde r co nsta nt illumination

. Sub-cultu r e s we re ma de into fresh m e dium 2 days pri or to i noculation of the testflasks, and incu bat ed und er ide n t ical conditions. Reliabilit y : (2) v alid with rest rictio ns Meets ge ne ra lly acce pted scientific stand ard s , well -do c ume nted an d accepta b le fo r asse ssment

. 09.11.20 06 (23 ) Species : Selena strum cap r icornutu m (Algae ) Endpoint : growth rate Exposur e pe riod : 96 hou r(s) Unit : μg/l NOEC : ca. 100 mea s ured/no mina l Metho d : 17 / 60

## 4. Ecotoxicity Id 77-8 6 -1 Dat e 14.11.20 06

Year : 1985 GLP : Test s ubs ta nce : as prescribe d by 1.1 - 1.4 Metho d : Dilution s of the actively-g ro wing cultures were added to tes tflas ks to give an app roxima te initial countof 1000 cell s per mL, and i n cu bated 2 4 hours. An appropri a te volume of test materi al wa s add ed to achieve the desi r ed con c e n tration , and the cult ure volum e made up to to 1 00mL with p a rticle
free deio n ize d water. Cell cou n ts were made eve ry 24 hours. The cell cou n ts were use d to cal c ul ate t he gro w th para meters to establish quantitative toxic value s
. Source : Do w Ch emical, TERC Mi dland, MI Test c onditi on : Stock culture s of Selena strum we re mai n tained at 22 C und er con s tant illumination from 2 fluore scent tubes, on slop es of Oxo i d agar contai ning Bold's b a sal medium. About 7 days pr i o r to use, 2 conical flasks of 90mL particl e-free BBM were a s eptically ino c ulated with S elena stru m from sto ck cultures. The culture s were incu bated a t22C, and sh ake n at 175 rpm unde r co nsta nt illumination

. Sub-cultu r e s we re ma de into fresh m e dium 2 days pri or to i noculation of the testflasks, and incu bat ed und er ide n t ical conditions. Reliabilit y : (2) v alid with rest rictio ns Meets ge ne ra lly acce pted scientific stand ard s , well -do c ume nted an d accepta b le fo r asse ssment

. 09.11.20 06 (23 )
4.4 TOXICITY TO MICR OO R G A N ISMS E.G. BA CTERI A
Ty p e : aquati c Species : Pseud omo na s putida (B act eria ) Exposur e pe riod : Unit : mg/l EC10 : ca. 334 47 measure d /nomi nal Metho d : other: Acute Bacteia Cell Multiplicatio n Inhibition Te st Year : 1990 GLP : yes Test s ubs ta nce : other TS: TRI S AMINO 40 % Metho d : A small amou nt of bacteria from a 7- day old sto ck cult ure of Pse ud omona s putida was in ocul ated in fluid nutrie n t medium in Erle nmeyer fla s ks. The prelimi nary culture s we re i n cu bated at 2 5 °C fo r 16-20 hours. Subse que ntly, the extinction of the m ono chromati c rad i ation at 436 nm for a 10 mm laye r of the bacte rial su sp en sio n wa s determ i ned by photo electri c measurement . On the basi s of the value s mea s u r ed, the final turbid ity value if the bacteri al su sp ensi on was a djuste d. Four p a rall el dilution serie s in 300 ml Erlenmeye rflasks stopp ere d with aluminiu m ca ps were p r e p a red from the formulated te st sub s tan c e stock solutio n and sterile Milli -Q water. Each flask co ntaine d 80 ml of liquid at the start. Each flask of three dilutio n seri ed to be i noculated, was mad e up to a final volume of 10 0 ml by addin g 5 ml of a stock sol u tion I, 5 ml of a stock solution II, and 10 ml of the prepare d bacterial suspension from the prelimi nary culture havin g a kno w n a d ju sted extinctio n value. The flasks of the dilution serie s that are not inoculate d , were m ade up to 100 ml by adding 5 ml of stock solution I, 5 ml of stock sol u tion II, and 10 ml of saline. Five culture flasks of the re feren c e seri e s we re ma de up with 80 ml of the 18 / 60

## 4. Ecotoxicity Id 77-8 6 -1 Dat e 14.11.20 06

methanol concentration, 5 ml stock solut ion I, 5 ml stock solution II, and 10 ml prep are d bacte rial solu tion. Ten control culture flasks with 80 ml sterile Milli-Q water, 5 ml stock solutio n I, 5 ml stock solut ion II and 10 ml prep are d bacte rial solu tion were also in clu ded. A ll f l asks we r e lef tat25°C f or 16- 20 ho u r s. S ubs equ ent ly , cell su spe nsi on s were hom ong enized an d t he extinction o fthe monoch r omatic radiatio n at 436 nm in a 10 mm layer in all seri es we re measured. Becau s e, afte r terminatio n of the test period, colo ratio n or turbi d ity occurre d in the dilution seri es for chemi c al-phy sical re aso ns, the an alogo us step s of dilution of the non -ino culate d serie s we re u s ed as p hotom etric blan k value s for turbi d ity fo the inoculate d dilution s se ries. Rem a r k : The stu dy pro c ed ure wa s b ase d on the followin g guid eline: Umweltbun de samt (UBA) Guideli n e s : wa sserg e fäh r dend er Stoffe, III Bestimmun g der a k uten B a kteri entoxizit ät, ad-ho c-A r beid s grupp e I (Obm ann Dr. Niemitz), LT WS, Nr. 10, Septembe r 19 79. Resul t : TRIS AMINO 40% was investigated for its ability to inhibit the cell multiplicatio n of the bacte ria spe c ie s Pseudom ona s p u tida. Cultures of Pseud omo na s putida b act eria were exp ose d to con c entration s ran ging from 432 to 8 85.6x10^3 T R IS AMINO 4 0 % per litre. Based o n the solubility of TRIS AMI N O 40% in water, a toxi c ity thres hold value of TRIS AMINO 40% of 33.4x10^3 mg/l for Pseud om o n a s putida coul d be determin ed. Conclu sion : The Asse ssm ent figure o rvalue for ba cte r ia toxicity is 1.5. Reliabilit y : (1) v alid with out rest rictio n GLP guideli n e study. 13.11.20 06 (24 ) Ty p e : aquati c Species : Pseud omo na s putida (B act eria ) Exposur e pe riod : Unit : mg/l EC10 : = 400 mea s ured/n omin al EC50 : = 160 0 measured/no mina l Metho d : DIN 38 412, p art8 Year : 1990 GLP : no data Test s ubs ta nce : other TS: TRI S AMINO 40 % Resul t : EC50 = 1 600 mg/l EC10 = 40 0 mg/l Test c onditi on : Con c e n tratio ns (g/l ): 10, 5, 2.5, 1.25, 0.625 and 0.3 1 5 Tempe r atu r e: 20-22 °C Reliabilit y : (2) v alid with rest rictio ns Meets natio n al stand ard m ethod s with a c ceptabl e re striction s
. 13.11.20 06 (25 )
4.5.1 CH RO NIC T O XICITY TO FISH

4.5.2 CH RO NIC T O XICITY TO AQ UATI C IN VERTEB RAT ES

4.6.1 TOXICITY TO SEDIMENT DWELLI NG ORGANISMS

19 / 60

## 4. Ecotoxicity Id Dat e 77-8 6 -1 14.11.20 06

20 / 60 4.6.2 TOXICITY TO TERREST RIAL PL ANT S

4.6.3 TOXICITY TO SOIL DWE LLING O R G ANISMS

4.6.4 TOX. TO OTHER NON MAMM. TERR. SPECIES

4.7 BIOLO G IC A L EFFECTS MONITO RIN G

4.8 BIOT RA NSF ORM ATIO N AN D KI NETICS

4.9 AD DITIO N A L REMA RKS

## 5. Toxicity Id 77-8 6 -1 Dat e 14.11.20 06

5.0 TOXICO KIN E TICS, META B O LISM A ND DISTRIB U TIO N

5.1.1 ACUTE O R AL TOXICITY
Ty p e Value : : LD5 0 ca. 500 0 ml/kg b w Species Strain : : mouse no data Sex : no data Number o fa nimals : 10 Vehicle : Dose s : 1000 0, 7000, 5000, 35 00 a nd 200 0 mg/kg bw Metho d : Year : 1955 GLP : no Test s ubs ta nce : as prescribe d by 1.1 - 1.4 Metho d : Ten mice per dose we re ad ministe r ed T R IS AMINO solution s via gavage at a con s tant vo lume of 0.05 ml/gm bw. Conclu sion : The oral LD5 0 in mice is 5 500 mg/kg. Reliabilit y : (2) v alid with rest rictio ns Meets natio n al stand ard m ethod s with a c ceptabl e re striction s
. 13.11.20 06 (26 ) Ty p e Value : : LD5 0 > 300 0 mg/kg bw Species Strain : : other: rat an d mice other: W ista r and Swi ss Sex : Number o fa nimals : Vehicle : Dose s : 1000, 20 00, a nd 300 0 mg/kg as a solutio n Metho d : Year : 1962 GLP : no data Test s ubs ta nce : as prescribe d by 1.1 - 1.4 Metho d : Solutions of 2 0 % and 5% o fTRIS AMIN O we re given via gastric int ubation to rats an d mice re sp ectivel yat doses of 1000, 20 00, a nd 300 0 mg/kg. Resul t : There wa s no toxicity noted in eit her sp e c ie s at the top dose levels, althoug h abu ndant uri ne o utput wa s not ed for some a nimals Conclu sion : The LD50 for rats an d mice are estim a te d to be >30 0 0 mg/kg in so lution. Reliabilit y 13.11.20 06 : (2) v alid with rest rictio ns (27 ) Ty p e Value : : LD5 0 ca. 335 0 mg/kg bw Species Strain : : mouse no data Sex : no data Number o fa nimals : Vehicle : Dose s : 7000, 50 00, 3 530, 250 0, an d 2000 mg/ k g bw Metho d : Year : 1955 GLP : no Test s ubs ta nce : other TS:AM P D 21 / 60

## 5. Toxicity Id 77-8 6 -1 Dat e 14.11.20 06

Test s ubs ta nce : AMPD: 2-Ami no-2
- methyl -1-p rop anol Reliabilit y : (2) v alid with rest rictio ns Meets natio n al stand ard m ethod s with a c ceptabl e re striction s
. 13.11.20 06 (26 )
5.1.2 ACUTE INHALATION T O XICITY

5.1.3 AC UTE DE R M AL TOXICI TY
Ty p e : LD5 0 Value : > 100 0 mg/kg bw Species : rat Strain : Sex : Number o fa nimals : 5 Vehicle : Dose s : 500 mg/kg (d elivered via 5 % solution ) Metho d : Year : 1962 GLP : Test s ubs ta nce : as prescribe d by 1.1 - 1.4 Metho d : Five rats were injecte d wit h 500 or 1 0 0 0 mg/kg (via 5% solution ) of THAM. Resul t : 500 mg/kg ca use d irritatio n at the injection site. 1000 mg/ k g caused the formation of lesi ons. There we re n o other ob se rvations note d

. Source : Do w Ch emical, TERC Mi dland, MI Reliabilit y : (2) v alid with rest rictio ns Meets ge ne ra lly acce pted scientific stand ard s , well -do c ume nted an d accepta b le fo r asse ssment

. 09.11.20 06 (27 ) Ty p e : LD5 0 Value : > 100 0 mg/kg bw Species : mouse Strain : Sex : Number o fa nimals : 5 Vehicle : Dose s : Metho d : Year : 1962 GLP : Test s ubs ta nce : as prescribe d by 1.1 - 1.4 Metho d : Mice were inj e cted with 50 0 or 100 0 mg /kg THAM (vi a 5% solution ). Resul t : 500 mg/kg ca use d irritatio n at the injection site. 1000 mg/ k g caused skin le sion s at the si te. There we re n o other ob se rv ations of toxicity noted. Source : Do w Ch emical, TERC Mi dland, MI Reliabilit y : (2) v alid with rest rictio ns Meets ge ne ra lly acce pted scientific stand ard s , well -do c ume nted an d accepta b le fo r asse ssment

. 22 / 60

## 5. Toxicity Id 77-8 6 -1 Dat e 14.11.20 06

09.11.20 06 (27 ) Ty p e Value : : LD5 0 > 200 0 mg/kg bw Species Strain : : rabbit no data Sex : male/female Number o fa nimals : 12 Vehicle : other: no vehi cle Dose s : 1000, 15 00, a nd 200 0 mg/kg Metho d : other: Pharm a col ogy Lab Protocol Year : 1980 GLP : yes Test s ubs ta nce : other TS:AM P Metho d : Each g r ou p o frabbits was treated with 1 000, 150 0, or 2000 mg of test material p er kg body wei gh t(mg/kg ). The desi r ed d ose wa s sp rea d over the prep ared abdomi nal ski n area (ab r ad ed or smooth as de sig nate d). The ski n wa s covered with a g auze and a s heet of impervious ru bbe ri zed cloth to prevent an y loss of the test materi al. The trun k wa s furthe r en cl ose d with a flexible wire scree n held in pla c e by tape. The animal s we re returned to in dividual cage s. After 24 hours dermally expos ed, the bi nding s an d p atche s were removed, the exposed area s ge ntly clea ned, an d observed fo r ski n irrita ncy. The animal s we re obse rved for anothe r 14 d a ys for any gross symptom s of toxicity. At th e end of the 1 4 day observ a tion peri od, the animal s were weig ed, sa crif iced, an d the orga ns exa m i ned for g r o ss patholo gy. Resul t : At the end of the 24 hou r e x posu r e pe rio d , the intact and abrade d treated ski n site s we re severely irri tated and bla ck in colo r. The site s be ca me necrotic withi n two to three days and re mained n e cro t ic for the 14 days. The treate d si tes had severe escha rform ation by the 14th day. The rabbit s in the three treatment grou ps lo st bodyweig ht over the 14 day ob servatio n perio d. The animal s in all treated g r ou p s sh owed no sign s of toxici ty or abno rmal p h a r ma colo gical behavio r. At necrop s y, all orga ns in all rabbits were grossly norm a l. The treated skin sites in all rab b its we re ne crotic. Test c onditi on : 12 Ra bbits weighin g 3.0 +/- 0.5 kg were divided into 3 group s of 4 e ach, and their a b d o men s we re shave d free o fhair. The skin of 2 rabbits were further p r ep ared by abra s io ns. The ab ra sion s were m ade 2-3 cm a part over the areaof exposure with a blunt h y podermic n e edle with out bleedi ng. Test s ubs ta nce : AMP:2-Amin o-2
- methyl-1-prop anol; CA S# 124-68-5; molecula r formula C4 H11 N O; m olecular weig ht 89.14 Conclu sion : The acute de rmal L D 50 for P-1826 for th e rabbit was >20 00 mg/kg. The test material wa s dermally nontoxic , but wa s a seve re skin irritant. Reliabilit y : (2) v alid with rest rictio ns Meets ge ne ra lly acce pted scientific stand ard s , well -do c ume nted an d accepta b le a s sessme nt. 13.11.20 06 (28 )
5.1.4 AC UTE TOXI CITY, OTHE R RO UTES
Ty p e : LD5 0 Value : ca. 335 0 ml/kg b w Species : mouse Strain : no data Sex : no data Number o fa nimals : Vehicle : Dose s : 4000, 36 00, 3 250, 250 0 an d 2000 mg/ k g bw Rou t e of ad min. : i.p. 23 / 60

## 5. Toxicity Id 77-8 6 -1 Dat e 14.11.20 06
Exposur e time Metho d : : 96 hou r(s) Year : 1955 GLP : no Test s ubs ta nce : as prescribe d by 1.1 - 1.4 Metho d : Ten mice per dose we re ad ministe r ed T R IS AMINO solution s via intrape riton a l at a con s tant volume of 0.015 ml/gm bw. Conclu sion : The acute LD50 i.p. of TRIS AMINO is 3 350 mg/kg b w
. Reliabilit y : (2) v alid with rest rictio ns Meets natio n al stand ard m ethod s with a c ceptabl e re striction s
. 13.11.20 06 (26 ) Ty p e Value : : LD5 0 ca. 16.5 other: mM TRIS/kg Species Strain : : mouse Sex : Number o fa nimals : 10 Vehicle : no data Dose s : Rou t e of ad min. : i.v. Exposur e time Metho d : : .5 minute(s) Year : 1961 GLP : Test s ubs ta nce : as prescribe d by 1.1 - 1.4 Metho d : IV injections of Tris ove r3 0 se con ds u s i ng un-neut rali zed a nd ne utralize d s o lutions of 0.3M Tris

. Resul t : The LD50 = 1 6 .5 mM Tris / kg Mice that we re given a leth al dose co nvulse d immedi ately before d eath. Source : Do w Ch emical, TERC Mi dland, MI Test c onditi on : Ten mice for each do se, in jected IV. Mice were o b se rved for 24 h ours and LD5 0 cal c ulat ed from the p e r ce nt of mice that died. Reliabilit y : (2) v alid with rest rictio ns Meets ge ne ra lly acce pted scientific stand ard s , well -do c ume nted an d accepta b le fo r asse ssment

. 09.11.20 06 (29 ) Ty p e Value : : LC5 0 3.28 - 4.04 o t her:g/kg Species Strain : : rat Sprag ue-Da wley Sex : male/female Number o fa nimals : 6 Vehicle : physiol. salin e Dose s Rou t e of ad min. : : 2.0, 2.5, 3.0, 3.5 g/kg of 0.6M THAM, 4.0 and 4.5 g/kg of 0.9M THAM. i.v. Exposur e time Metho d : : 1 minute(s) Year : 1965 GLP : Test s ubs ta nce : as prescribe d by 1.1 - 1.4 Metho d : Grou p 1 Males a nd fe males (3 rat s each sex) we re admi niste red variou s do se s of THAM via the tail vein. Th e dose levels of THAM ad ministe r ed were 2.0, 2.5, 3.0, 3.5 g/kg of 0.6M THAM , 4.0 and 4.5 g/kg of 0.9M THAM. Grou p 2 A replication of experime n ts co ndu cted with Group 1. Instead of admini s teri ng a single give n dose level of THAM, to all rats in on e unit in 24 / 60

## 5. Toxicity Id 77-8 6 -1 Dat e 14.11.20 06
one day, the different do sa ge levels were given at on e sitting to pa irs of rats, an d rep eated, on sub s eq uent day s, until all 36 animals in thi s grou p had be en u s e d

. Ultimately, 3 male and 3 female rats from Gro up 2 received T H AM at each of the do sag e le vels sh own for Gro up 1. Grou p 3 As a partial control (to rule out adverse effects of ra pi d infusio ns oflarge volumes of fluid), 4 pai rs o fmale and female rat s re ceived IV infusions, via the tail vein, of physiolo g ical salin e (0.9 g Na Cl per 1 00mL )

. The infusio n rates a nd vol umes of salin e were simil a r to the fluid infusio n rate s and volumes req u i red for e a ch dose level of THAM. In all cases, the rate of ad ministration o fthe test solu tions was e s t ablished individually b a se d ea ch rat ' s body weigh t. Each animal received th e test material su ch that they receive 0. 45g T H AM per kg b ody weig ht would be admini s tered in one minute

. Sterile equipment wa s u s ed throug ho ut the study, and ea ch anim a l wa s treate d with a sepa rate syringe an d ne edle. Animals that survived the i nfusio ns were held an d ob serve d for 2 h ours post-i nfusi on and were sa crificed a nd ne cro psi ed. Ne cro psi es we re like w ise perfo rmed o n anim a ls dying spo n taneo usly fo llowing treat ment. Specim en s from all org a n s and tissue s were preserv ed, embed de d, and staine d. All h i stopath o logi c and hist o c h e m ical meth od s we re pe rformed according to establi s h ed method s. Resul t : In mos tc a s e s , rats that did not s u rvive the post
- infusion peri od die d durin g the infusio n o rvery soo n afterwa r d. Tho s e anim a ls th at survived lo nger than 10 minut es follo wing treatment su rvi v ed the entire post-infu sio n period and re cove re d with no gro ssly-ob se rva ble ill effects. All 8 control animals survived the i n fusio n
. There we re n o signifi cant g r oss le si on s n o ted at necro psy for any ofthe animal s , with the exceptio n of the liver and kid neys. Pera cute toxic neph ro sis wa s noted con s i s tently in the kidn eys up to 2 hours po stinfusio n
. The lesion vari ed in that it was limited to a moderate deg ree to pykno s i s of the nucl e i of iso l ated se gm en ts of the renal tubular epith elium in rats infu se d with 2 and 2.5 g/kg T H AM dose s , and incre a sed in severity with the do se
. In higher do se level s , the lesio n s were cha r a c teri ze d by severe pykn o s is of the nu cl ei of sw oll en renal tubul ar e p ithelial cells of carrie d se gm ents of the co rtex. The cytopla s m of the affected cell s was coa gulate d , distinctly gra n u l ar, and inten s ely eo sino ph ilic. Lumen s of the affected tubul es were di ste nded with eo sino philic, am orph ou s tissu e debri s and secretion s
. Affected tubule s we re o b se rved adj a c ent to app arently norm al tubule s
. The inci de nce of le sion s incre ased wit h increa sing dose levels, and were note d in asimilar d o se-respon se p a ttern in ani mal s dying spo ntane ou sl y or at sch ed uled sacrifice. Lethargy wa s noted sp ora d i cally in rats a t3 -4 g/kg do se level s
. There were all note d with lesi on s of acute toxic hep atitis. The lesi on was cha r a c teri ze d by pyknosi s of the nuclei of the hepato c ytes an d clo udy swelling of th e cytopla s m of hepato c yt es. Although the lesi on s are thought to be related t o THAM admi n istratio n, they did not con s titute a con s i s tent cha r a c teri stic lesion a s did the pera c ute t oxic nep hro s i s
. Source : Do w Ch emical, TERC Mi dland, MI Test c onditi on : Eighty, 200-3 00g Spragu e-Da wley rats (males & fema les) we re ob served for 14 days p r ior to study st art for health evaluation. They were se g r egate d into 3 grou ps: 2 experime ntal grou ps of 3 6 animal s ea ch (1 8 male s and 18 females), and one co ntrol g r oup (4 male s and 4 female s). Conclu sion : The act ual L D 50 of T H AM given intrave nou sly wa s calcul ated to b e 3.5 g/kg +- 0.1 fo r Gro up 1, an d 3.6 g/kg +- 0.2 for Gro u p 2. Statistical evaluation cal c ulate d a ma ximum likelih ood e s timate of 3.55 and 3. 64 g/kg of body weig ht resp ectivel y , with a fiducial prob ability of 95%. The value of LD5 0 in gro u ps 1 an d 2 were expe cted to be 3.28-3. 83 g/kg a nd 3

. 28-4.04 g/k g , res p ec tively. A NOAEL wa s not ob serve d

. Obse rvations, tre a tmen t-relate d, were noted 25 / 60

## 5. Toxicity Id 77-8 6 -1 Dat e 14.11.20 06

at the lowe st dose level tested. Reliabilit y : (2) v alid with rest rictio ns Meets ge ne ra lly acce pted scientific stand ard s , well -do c ume nted an d accepta b le fo r asse ssment

. 09.11.20 06 (30 ) Ty p e Value : : LC5 0 ca. 600 0 mg/kg bw Species Strain : : rat Sex : Number o fa nimals : 10 Vehicle : Dose s : 100, 200, 40 0 , 500, 1000, 3000, 50 00, 6 000, 700 0 mg /kg (1 & 20% solut i o ns ) Rou t e of ad min. : i.v. Exposur e time Metho d : : Year : 1962 GLP : Test s ubs ta nce : as prescribe d by 1.1 - 1.4 Resul t : There we re n o notation s of any toxicity a tdose s lower than 3000 m g /kg. At 5000 mg/ k g, 30% mortality was n o ted, a t6000 mg/kg, 60% mortality was noted, and at 7000 mg/ k g, 70% mortality was n o ted. Source : Do w Ch emical, TERC Mi dland, MI Reliabilit y : (2) v alid with rest rictio ns Meets ge ne ra lly acce pted scientific stand ard s , well -do c ume nted an d accepta b le fo r asse ssment

. 09.11.20 06 (27 ) Ty p e Value : : LC5 0 ca. 610 0 mg/kg bw Species Strain : : mouse Sex : Number o fa nimals : 10 Vehicle : Dose s : 100, 200, 40 0 , 500, 1000, 2 000, 300 0, 50 00, 6000, 70 0 0 mg/kg (1% solut i o n ) Rou t e of ad min. : i.v. Exposur e time Metho d : : Year : 1962 GLP : Test s ubs ta nce : as prescribe d by 1.1 - 1.4 Resul t : There wa s no mortality noted at doses l e ss than 50 0 0 mg/kg. At 6000 mg/kg, 40% mortality wa s noted, and at 7000 mg/lg, 1 00% mortality was recorded. Animals expe rie n ce d muscle wea k n e ss a c comp anie d b y respiratory difficulty prior to death. Source : Do w Ch emical, TERC Mi dland, MI Reliabilit y : (2) v alid with rest rictio ns Meets ge ne ra lly acce pted scientific stand ard s , well -do c ume nted an d accepta b le fo r asse ssment

. 09.11.20 06 (27 ) Ty p e Value : : LC5 0 Species Strain : : rabbit Sex : 26 / 60

## 5. Toxicity Id 77-8 6 -1 Dat e 14.11.20 06

Number o fa nimals : 5 Vehicle : Dose s : 250, 500 mg/ k g (5% soluti on) Rou t e of ad min. : i.v. Exposur e time Metho d : : Year : 1962 GLP : Test s ubs ta nce : as prescribe d by 1.1 - 1.4 Metho d : Margi nal vein in the ear wa s used. Resul t : There wa s no treatment-rel ated mortality

. Change s in respi r ato ry rate and amplitude we re the only ob servatio ns n o t ed. Source : Do w Ch emical, TERC Mi dland, MI Reliabilit y : (2) v alid with rest rictio ns Meets ge ne ra lly acce pted scientific stand ard s , well -do c ume nted an d accepta b le fo r asse ssment

. 09.11.20 06 (27 ) Ty p e Value : : LC5 0 > 125 ml/kg bw Species Strain : : dog Sex : Number o fa nimals : 5 Vehicle : Dose s : 125 mg/kg (5 % solution ) Rou t e of ad min. : i.v. Exposur e time Metho d : : Year : 1962 GLP : Test s ubs ta nce : as prescribe d by 1.1 - 1.4 Metho d : Saphen ou s vein wa s u s ed. Resul t : Alteration s in respiratory ra te and amplit ude were the only observat i ons noted. Source : Do w Ch emical, TERC Mi dland, MI Reliabilit y : (2) v alid with rest rictio ns Meets ge ne ra lly acce pted scientific stand ard s , well -do c ume nted an d accepta b le fo r asse ssment

. 09.11.20 06 (27 ) Ty p e Value : : LD5 0 > 300 0 mg/kg bw Species Strain : : rat Wista r Sex : Number o fa nimals : 10 Vehicle : Dose s : Rou t e of ad min. : other: ga stri c tube Exposur e time Metho d : : Year : 1962 GLP : Test s ubs ta nce : as prescribe d by 1.1 - 1.4 Metho d : Do se s of 100 0 and 30 00 m g/kg were a d m iniste red bygastri ctube i n a 20% s o lution. Resul t : There we re n o sign s of acute toxicity noted. 27 / 60

## 5. Toxicity Id 77-8 6 -1 Dat e 14.11.20 06

Abunda nt uri ne output wa s re co rde d for som e anima ls. Source : Do w Ch emical, TERC Mi dland, MI Reliabilit y : (2) v alid with rest rictio ns Meets ge ne ra lly acce pted scientific stand ard s , well -do c ume nted an d accepta b le fo r asse ssment

. 09.11.20 06 (27 ) Ty p e Value : : LD5 0 > 300 0 mg/kg bw Species Strain : : mouse Swiss Sex : Number o fa nimals : 10 Vehicle : Dose s : Rou t e of ad min. : other: ga stri c tube Exposur e time Metho d : : Year : 1962 GLP : Test s ubs ta nce : as prescribe d by 1.1 - 1.4 Metho d : Do se s of 100 0, 2000, and 3000 mg/ k g were delive r e d by gastri ctube in a 5% solution. Resul t : There wa s no significant toxicity not ed, other than in creased uri ne o u tput. Source : Do w Ch emical, TERC Mi dland, MI Reliabilit y : (2) v alid with rest rictio ns Meets ge ne ra lly acce pted scientific stand ard s , well -do c ume nted an d accepta b le fo r asse ssment

. 09.11.20 06 (27 )
5.2.1 SKIN IRRITATION
Species Conc entrati on : : rabbit Exposur e Exposur e time Number o fa nimals : : : Vehicle : PDII : Resul t : moderately irritating Classific atio n : Metho d : Drai ze T e st Year : 1961 GLP : Test s ubs ta nce : as prescribe d by 1.1 - 1.4 Metho d : A saturate d solution (p H 1 0 .8), 25% sol u tion, and the crystallin e produ ct were tested o n intact and a b rad ed skin o frabbits. Resul t : The sat urate d solutio n an d the cry s talline pro du c t produ ced comp arabl e prima ry irritati on scores. T here wa s no notice able irri tation prod uced by any of the test concentratio n s on u nab ra ded skin. All sign s of irritat i on were complet e ly resolved in 48 hours. Solution Int a c t Abraded Total c ry s tals 0 0.83 0.4 saturated 0.16 0.83 0.5 25% s o ln 0 0.16 0.08 The test mate rial is con s ide r ed a mild irrit ant to the ski n. 28 / 60

## 5. Toxicity Id Dat e 77-8 6 -1 14.11.20 06

Source Reliabilit y 09.11.20 06 : : Do w Ch emical, TERC Mi dland, MI (4) n ot assign able Do cume ntation insufficientfor asse ssm ent. (31 ) Species Conc entrati on Exposur e Exposur e time Number o fa nimals Vehicle PDII Resul t Classific atio n Metho d Year GLP Test s ubs ta nce : : : : : : : : : : : : : rabbit

highly irritatin g irritating Drai ze T e st 1975 no other TS:AM P Source Test s ubs ta nce Reliabilit y 13.11.20 06 :

:

: ANGUS Che m ie Gmb H Ibbenb ure n Europ ean Co mmission - E urop ean Che m ical s Bure a u Ispra (VA). AMP: 2-Amino-2
- methyl-1-prop anol; Mol e cul a r form ul a C4 H11 N O; Molecula r wei ght 89.14 (4) n ot assign able Do cume ntation insufficientfor asse ssm ent. (32 )
5.2.2 EYE IRRITATION
Species Conc entrati on Dose Exposur e time C o mment Number o fa nimals Vehicle Resul t Classific atio n Metho d Year GLP Test s ubs ta nce : : : : : : : : : : : : : rabbit undiluted

6 none not irritating not irritating 1975 as prescribe d by 1.1 - 1.4 Resul t : Combi ned Av erag e Sco r e = 0/110

Source Reliabilit y 13.11.20 06 : : Clas s ific a tion = Non-irritating Do w Ch emical, TERC Mi dland, MI (4) n ot assign able Do cume ntation insufficientfor asse ssm ent. (33 ) Species Conc entrati on Dose Exposur e time C o mment Number o fa nimals Vehicle Resul t Classific atio n : : : : : : : : : rabbit 40 %

6 water not irritating not irritating 29 / 60

## 5. Toxicity Id Dat e 77-8 6 -1 14.11.20 06

Metho d Year GLP Test s ubs ta nce : : : : 1992 as prescribe d by 1.1 - 1.4 Source Reliabilit y 13.11.20 06 : : L. Troe ster (2 006). Person al comm uni cation. (2) v alid with rest rictio ns Data from h a ndbo ok o rcol l ection of dat a. (34 )
5.3 SENSITIZATION
Ty p e Species Conc entrati on

Number o fa nimals Vehicle Resul t Classific atio n Metho d Year GLP Test s ubs ta nce : : :

: : : : : : : : Intracut aneu s test guine a pig 1 st : Inductio n .1 % 2 nd : Chall enge .0 5 % 3 rd : Chall enge .0 1 % 30 water not sen s itizi n g not sen s itizi n g 1982 other TS:AM P Metho d : One group was treat ed wit h 0.05mL of 1 % P-1826 sol u tion, a negat ive control group wa s treated with sali ne, a nd a po sitive control group wa s treated with d i nitrochlo r obe nze ne (DNCB solubili zed i n alco hol an d made to volume with salin e). After 24 ho urs, sites we re cl ea ned an d scored for erythema a n d edema a c cording to Draize (Drai z e, JH, "Appraisal of the Safety of Chemical s in Foo d s, Drug s, an d Co smetics". Asso c. of Food and Dru g Official s of the United Stat es, p. 48, 1957). At 48 hours, the appli c ation was re peate d with ea ch gro up, and conti nued 2
- 3 time s per wee kuntil 10 appli c ation s were mad e
. Animals were allowe d a 2 wee k recovery pe ri od, and then chall enge d ata virgin site. The test and negative control anim a ls we re challe nged with 0.1 mL of 0.05% and 0.01% so lutions of P-1826. Positive and n egative co ntrol animal s we re also ch alle nged with 0.3% an d 0.03% DNCB solution. After 24 hou rs, they were de pilated, and thre e ho urs late r sco r ed for erythe ma and ed em a. Sites were sco red again at 48 h ours.

Resul t : Test mate rial is co nsi dered a sen s itize rif the challe ng e elicits ski n rea c tion s in alarge nu mbe rof test anim a ls when com pare d to the negative control. Duri ng the in ductio n pha se, the first in jection at 1% and second i n jectio n at 0.5% P-1826 indu ced n e crotic lesi on s, so the remai n i ng 8 inje ction s we re made with 0. 1% solution s. The 0.3% DNCB site s w e re ne cr otic for the entire 10 inj e ction s
. At challeng e with 0.05% a nd 0.01% P-1 826, one a n i mal in the test group sho w e d mild rea c tion s with 0.05%, but none of the ne gative control s chall enge d wi th P-1826 sh owe d any rea c tion s at 24 o r48 hou rs. In the repe at chall e nge, non e of the animal s in the test grou p sho w e d an y rea c tion s with P-1826, but of the negat ive control grou p, 4 animals at 0.05% and 1 animal at 0.0 1 % sho w ed skin re actio n s at 24 hours. A challen ge with the po sitive control in d u ce d skin re a c tion s in the positive control group. The 0.03% solutio n did n o t elicit any skin re actio n a t48 hours. 30 / 60

## 5. Toxicity Id 77-8 6 -1 Dat e 14.11.20 06

Test c onditi on : Thirty male g u inea pi gs (2 50-3 00g e a ch ) we re divide d into 3 grou p s of 10 each. The an imals' b a cks and flan ks we re sh aved fre e of hair. The guine a pigs were intradermally-inj ected with the solutio n s. Reliabilit y : (2) v alid with rest rictio ns Meets ge ne ra lly acce pted scientific stand ard s , well -do c ume nted an d accepta b le fo r asse ssment

. 14.11.20 06 (35 ) Ty p e Species Conc entrati on : : : Patc h-Tes t guine a pig 1 st : Inductio n 5 % 2 nd : Chall enge 2. 5 % 3 rd : Chall enge 5 % Number o fa nimals : 30 Vehicle : water Resul t : not sen s itizi n g Classific atio n : not sen s itizi n g Metho d : other: Buehl e r Year : 1982 GLP : Test s ubs ta nce : other TS: AMP Metho d : : One group was treat ed wit h 0.5mL of 10 % P-1826 sol u tion, a negative cont rol group wa s treated with salin e, and a positive control grou p wa s treated with dinitro c hl orob en zen e (DNCB sol ubil i zed in al co ho l and made to volu me with sali n e ). Afte r 24 hours, the patches were rem o ved, and site s were clea ned a n d score d at 24 and 48 h ou r s for e rythe ma and edema a c co rding to Draize (Drai z e, JH, "Appraisal of the Safety of Chemi c al s in Food s, Drug s, and Co sm et ics". Assoc. of Food and Drig Officials of th e United States, p. 48, 195 7). At 48 hours, the ap plication wa s rep eated with each group, and cont inued 2
- 3 tim es pe r we ek until 10 appli c ation s were mad e
. Animals were allowe d a 2 wee kre cove ry period, and then chal lenge d at a virgin site. The test and neg ative control animal s were ch allen ged with 0.5 m L of 2.5% and 5% solutio n s of P-18 26. Positive and ne gative control anim a ls we re al so challen ged wit h 0.03% DNCB solutio n
. Afte r 24 hou rs, the test materia l was clea ned away, depilat ed, and thre e ho urs late r sco r ed for erythe ma and ed em a. Sites were sco red again at 48 h ours. Test mate rial is co nsi dered a sen s itize rif the challe ng e elicits ski n rea c tion s in alarge nu mbe rof test anim a ls when com pare d to the negative control. Resul t : Duri ng the in ductio n , the 10% P-1826 solution was fo und to be mil d ly irritating to all animals in th e test gro up, so the remain ing 8 do se s d u ring the indu ction were mad e with a 5% so lution. The posi t ive control, DCNB, elicited a mild to strong rea c tion du ring t he 10 ap plica t ions. There wa s one death in the p o sitive co ntrol group, but was de emed n o t treatment-related via necrop s y. At challeng e with 2.5% an d 5% soluti on s of P-182 6, none of the a nimals in the test or po sitive control grou ps sho w ed any skin reactio n s at 2 4 hours, but the positi v e control a ni m als sho w ed mild ski n re actions at 48 ho urs. Nine of ten p o sitive co ntrol s wh en challe nged with DNCB at 24 hou rs sho w e d skin reactio n s, an d 7/10 sho w e d reactio n s at 48 hou rs. On ly 4/10 negative cont rols sho w e d reactio ns whe n chall enge d with DNCB at24 hours, and n o ne at 48 hou rs. Test c onditi on : Thirty male g u inea pi gs (2 50-3 00g e a ch ) we re divide d into 3 grou p s of 10 each. The an imals' b a cks and flan ks we re sh aved fre e of hair. The guine a pigs were top i cally treated with the sol u tions a pplied u nder a n occlu s ive patch. Conclu sion : 2-Amino
- 2
- m ethyl-1-pro pa nol wa s a no n-sen s itize ri n the topical 31 / 60

## 5. Toxicity Id 77-8 6 -1 Dat e 14.11.20 06

sen s itization test in guin ea pigs, un der th ese te st con d itions. Reliabilit y : (2) v alid with rest rictio ns Meets ge ne ra lly acce pted scientific stand ard s , well -do c ume nted an d accepta b le fo r asse ssment

. 14.11.20 06 (36 ) Ty p e Species Number o fa nimals : : : other human Vehicle : Resul t : Classific atio n : Metho d : Year : GLP : no data Test s ubs ta nce : other TS: AMPD and AMP Resul t : In patch test s with human s, cosm etic formulation s co ntaining eith e r0.22% AMP or 0.5% AMPD or 0.0 75% AMPD were neg ative for dermal sensitization. Test c onditi on : AMP The skin irrita tion potential of a cosm etic formulation containin g 0.2 2 % AMP-95 was examined u s i ng a sin g le in sult occlu s ive patch test on 15 paneli s ts. On e paneli s t formulation cont aining 0.2 2 % AMP-95 ha d a negligibl e pri mary skin irritation potentia l. AMPD A cosm etic fo rmulatio n co n t aining 0.07 3 % AMPD was tested for sen s itization potential in a grou p of30 h uman test su bject s usi ng a repe ated in su lt open patch test. The material was a ppl ied to the arm daily 4 days/wee kfor 2 we eks, a l ternating a r m s daily. In addition, an occl usive patch wa s wa s appli ed on t he first day ofthe test. After the 2-we ek appli c ation p e riod, the r e was a 2-we ek nontre atmentperiod. After this 2­ wee kpe riod, the test subj e c ts received a reap plicatio n of the formulation of the formulatio n to the test site along with an occlu s ive patch at an a d jacent site. The orig inal patch, ch alleng e patch , and open ch alleng e test si tes were re ad at 24, 48, and 9 6 hour. No re action s were observed in a n y of the test subj e c ts. The form ulation contai ning 0.07 3% AMPD wa s n e ither a prima ry irritan t, nor a fatiguing age nt, nor a sen s itize r , and the form ulation wa s safe u n d e r the conditi ons of the stu d y. A modified re peated in sult patch te st ofa co smetic fo rmulatio n co n t aining 0.5% AMPD wa s perfo rme d on a pan el of 39 wome n and 20 me n. The test material, 0.5 ml, was ap pli ed to a semio pen pat ch on the arm of ea ch paneli s t every Monday, Tuesd a y, Wedn esd a y, and Thursday for t w o wee k s. The patch sites were g r ad ed a pproxim ately 24 h after ap plicatio n. In addition, a clo s ed p a tch wa s appli ed to each pan eli s t on the first day fo the study and on the day of challe nge. No patch es we re appli ed for 2 wee k s after the indu ction pha se. On M onday follo wing the nontre atment perio d, chall e nge pat che s were appli ed to the original test site and an adja c ent site; the second close d patch p a tch was al so applied at thi s time. The ch allen g e sites were grad ed 1, 2, and 4 day s after appli c atio n. Slight erythema wa s noted at on e adja c ent ap plic atin site at each of the g r adin g times, but it was cl ear whet her the s e rea c tion s occu rred in the pan elist. The form ulation co ntainin g 0.5% AMPD wa s not a se nsitizer un de r the c o nditions fo the tes t. Reliabilit y : (2) v alid with rest rictio ns Data from h a ndbo ok o rcol l ection of dat a 14.11.20 06 (37 ) (38 )

32 / 60

## 5. Toxicity Id 77-8 6 -1 Dat e 14.11.20 06

5.4 REPEATED DOSE TOXICITY
Ty p e : Sub-chroni c Species : rabbit Sex : Strain : Rou t e of ad min. : i.v. Exposur e pe riod : 1-99 d ays Freque nc y o ftrea tm. : once daily Post exp osu re period : Dose s : 5-10 0 mL of 0.3M Tri s per kg bodywe ig ht, given at pH 7.4 and at pH 5.5 Con t rol grou p : Resul t : Treatm ent-rel ated mortalitywas first not ed a few day s after sta rt of treatment. Result s of the study indi cate that neutralized Tri s is le ss toxic. Toxic sympto ms noted in cl uded a norexia , bloody urin e, hindleg p a ralysis, and irreg ular respiration. Gro s s ob serv ations at ne cropsy in clud ed abno rmally re d lung s, necrosi s at t he po int of infusion , bleach ed liver, darken ed spl een, bloate d stoma c h, and lesion s on th e heart an d ki dney. Histol ogi cal e x amination s of the organ s were neg ative. Blood sa mpli ng followi ng treatme nts wit h TRIS indica ted that the levels increa sed u p on su cce ssiv e treatment s, and re main ed high the follo wing day. Result s indicate an a c cumulatio n of Tris at 25m L/kg (0. 3 M), the top dose evaluat ed for this pa rameter, but n o t at dose s lo wer. Blood sa mpli ng followi ng treatme nt with Tris (0.3M n eutrali zed a n d nonneutrali ze d) could not e s ta blish a di re ct relation shi p b etwee n Tri s con c e n tration s and gl ucose con c e n trati ons in the bl o od due to lo st sampli ng dat a. The gluco s e con c entrations, ho weve r, drop ped sig n ificantly durin g the infusio n s, but re turned to no rmal or ab ove norm a l followi ng the end of the infusio ns (Tri s-i ndu ced hy p o g lycemia p e rsiste d long er than the Tris-ne u trali z ed). Tri s and Tris -Ne utrali z ed b oth ca u s ed tra nsi ent hypoglycemia

. Blood analysis on extra c ted blood (Tri s add ed to bl ood dropl ets at varying levels) also d e te rmi n ed that there wa s no del eteriou s effect on eryt hro c ytes. Uri nalysi s (u rine colle cted via Foley cathete r measured ev ery hou r for 7 hours followi ng start of inf u sio n ) reveal ed that the amount ofTris excrete d in the urine reached a ma ximum at the end of infusio n , and drop ped rapi dly after infusion stop ped. Only a small quantity of chlori de was excreted. With Tri s pH 5.5, a large ramount of chl o ride than with Tri s was ex crete d

. At the end of the 7 hours, 44% of the infuse d Tris wa s foun d in the urine, while with T r i s pH 5.5, 77 % was foun d. Cau s e s of local necro sis a r ound the infu sion site we re investigated usin g Tris p H 5.5 a nd 7.4. Usin g injecte d Try pan dye, the irritation cau s ed by the solutio n s wa s evaluated byobse rving th e amount of e xtravasated d y e. Neutralization of the Tris re duced the ir ritation, sug g e s ting that the pH of the Tris i s the proba ble cau s e of the de rmal irritation. Source : Do w Ch emical, TERC Mi dland, MI Test c onditi on : Two o rthree rabbit s for group, inje cted IV over 3-6 h ours for 1

- 99 con s e c utive d ays (de pen di ng on the en d point bein g measure d) at adose s rangi ng 5
- 10 0 mL of 0.3M Tris p e r kg bo dyweig ht. The doses were given at pH 7.4 and at pH 5.5 to investigat e diff eren ce s rel ated stri ctly to pH. Conclu sion : 1. Acute IV toxicity of 0.3M Tris is 55 mL/kg. Neither ne utralization of addition of gl uco s e o rNa Cl decrea s e s the toxicity. 2. Maximum su bch r oni c no n-lethal do se of0.3M Tris in rabbits i s 75­ 90mL/kg over 5 hours. Ne utralizati on a ppea rs to de crea se the toxicity. 3. If high doses of Tris are giv en frequently, Tris will accumulate. 4. Injection of Tris ca uses hyp oglycemi a du ring infu sion and afterwa r d

. Tris n eutrali zed ca uses hy poglycemia o n ly during inf u sio n
. There is no dire ct relatio n s hip b etwe en Tris an d glu c ose con c e ntrations in the blood. 33 / 60

## 5. Toxicity Id 77-8 6 -1 Dat e 14.11.20 06

5. Dermal irritation is likely ca use d by pH, and not toxici ty per se of Tris. Reliabilit y : (2) v alid with rest rictio ns Meets ge ne ra lly acce pted scientific stand ard s , well -do c ume nted an d accepta b le fo r asse ssment

. 09.11.20 06 (29 ) Ty p e Species Sex : : : Sub-a c ute rabbit Strain : Rou t e of ad min. : infusion Exposur e pe riod Freque nc yo ftrea tm. Post exp osu re period Dose s : : : : 10 days once daily

Con t rol grou p Metho d : : Year : 1961 GLP : Test s ubs ta nce : as prescribe d by 1.1 - 1.4 Resul t : No treatm ent-related m o rtal ity was noted in either spe c ies. There were no toxic sympto ms, and a hi stological st ud y of the organ s wa s ne gative. Source : Do w Ch emical, TERC Mi dland, MI Test c onditi on : Thre e of each spe c ie s for each do se, in jected IV over 30 se con ds f or 10 con s e c utive d a ys at a dosa ge of 50 mL a nd 10 mL of 0

. 155M Tri s pe r kg bodyweight. Reliabilit y : (2) v alid with rest rictio ns Meets ge ne ra lly acce pted scientific stand ard s , well -do c ume nted an d accepta b le fo r asse ssment

. 09.11.20 06 (29 ) Ty p e Species Sex : : : Sub-chroni c rat male/female Strain : Sprag ue-Da wley Rou t e of ad min. : oral feed Exposur e pe riod Freque nc yo ftrea tm. Post exp osu re period Dose s : : : : 3 months
0, 25, 150, 25 0, 2500 ppm i n feed Con t rol grou p LOAEL : : yes, con c u r re nt vehicle = 250 0 ppm Metho d : Year : 1981 GLP : Test s ubs ta nce : other TS:AM P Rem a r k : Re sults & me thods se ction s refe ren c e d appe ndix dat a, which is no t provide d in Dow' s co py of the rep ort. As s u ch, tes tmaterial intak e (mg/kg/day) could not be e s timated. Test s ubs ta nce : AMP-95/HCl 66.1ai (L ot 10 9-1 ) Reliabilit y : (2) v alid with rest rictio ns Meets ge ne ra lly acce pted scientific stand ard s , well -do c ume nted an d accepta b le fo r asse ssment

. 13.11.20 06 (39 ) Ty p e Species Sex : : : Sub-a c ute rat Strain : Sprag ue-Da wley 34 / 60

## 5. Toxicity Id 77-8 6 -1 Dat e 14.11.20 06

Rou t e of ad min. : infusion Exposur e pe riod Freque nc yo ftrea tm. Post exp osu re period Dose s : : : : 20 days 1 infusio n dail y 24 hou r or 7 day recovery perio d witho u tinfusion s pri or to ne cro psy 0.3M THAM (0.5, 1.5g/kg T H AM: 10 or 2 0 infusio n s) Con t rol grou p NO AEL : : yes, con c u r re nt vehicle ca. 500 mg/ k g Metho d : Year : 1965 GLP : Test s ubs ta nce : as prescribe d by 1.1 - 1.4 Metho d : Grou p 1 Twelve rats (6 each sex) were given 2 0 daily infusions via the tail vein of 0.3M THAM at 0.5 g/kg ad ministe r ed at 0.45g/kg/min. Grou p 2 Twelve rats (6 each sex) were given 2 0 daily infusions via the tail vein of 0.3M THAM at 1.5 g/kg ad ministe r ed a s a single inj e ction in a 1-2 minute perio d. Grou p 3 Twelve rats (6 each sex) were given 1 0 daily infusions via the tail vein of 0.3M THAM at 0.5 g/kg ad ministe r ed at 0.45g/kg/min. On the 11th day, the infusio n s were discontin ue d and IP injections were co mmen c ed at the same d o se le vel. The IP in jection s were given daily for 10 days a s a singl e injectio n in a 1-2 min u te pe riod. Grou p 4 As c ont rols , 4 rats (2 each s e x) rece ived 20 daily IV in fusion s, via the tail vein, of a solution co ntaini ng 9g NaCl a nd 0.37g KCl per liter of wa ter. The rate of injecti on wa s 1mL/ kg/min. Grou p 5 As c ont rols , 4 rats (2 each s e x) receiv ed 20 daily IP in jection s of a solution contai ning 9 g NaCl a nd 0.3 7g KCl pe r liter of wate r. The rate of inj e ction wa s 1mL/kg/min. Half of the su rviving rats ofGroup 1 & 2 and of the rat s of the remai n ing grou ps we re observed fo r 24 hou rs afte r the 20th infu sion a nd were sa crificed an d necrop s ie d. The remaini ng rats of gro ups 1 & 2 we re observed a n addition al 7 d a ys po st-inf u s ion b efore a sched uled n e c ro psy. At necro psy, tissue an d org an sp ecim en s we re ta ken, preserve d, pro c e s sed, a nd evaluate d accordi ng to establi s h ed hi stopath o logi c and histo c hemi c al methods. Resul t : There we re n o finding s not ed in any ani mal in Gro up 1. Group 1 & 3 animal s expe rien ced d ry g angrene at th e sites of the tail injection s
. Approximatel y half of the rats in Group 2 & 3 were n o ted with a m ild inflammation of various p a rts of the visce ral pe ritone u m , or fat necrosi s and hemo rrh age of the serosa of various p a rts of the stom ach, inte stine , and periton eum. No gross le si ons were not ed in any co n t rol animal. Microsco pic e x amination oftissue s re vealed a chro ni c cell ulites at the injectio n site s, and pera c ut e toxic neph ro si s of the kid neys (5/6 rats of Grou p 1 ne cropsi ed 24 h o u rs afte r inje ction, but not seen in anim a l s allowed the 7 day recove ryperiod )

. In group 2, all rats ne cro p si ed at 24 hours an d 5/6 rats in the 7-day reco very grou p presen ted simila rly. Source : Do w Ch emical, TERC Mi dland, MI Test c onditi on : Sprag ue-Da wley, 200-30 0g , rats (19 m al es & 19 femal es) we re ob served for 14 days p r ior to study st art for health evaluation. They were se g r egate d into 5 grou ps. Conclu sion : NOAEL = 0.5 g /kg of 0.3M THAM Reliabilit y : (2) v alid with rest rictio ns Meets ge ne ra lly acce pted scientific stand ard s , well -do c ume nted an d accepta b le fo r asse ssment

. 09.11.20 06 (30 ) 35 / 60

## 5. Toxicity Id 77-8 6 -1 Dat e 14.11.20 06
Ty p e Species Sex : : : Sub-a c ute mouse Strain : Rou t e of ad min. : infusion Exposur e pe riod Freque nc yo ftrea tm. Post exp osu re period Dose s : : : : 10 days once daily

Con t rol grou p Metho d : : Year : 1961 GLP : Test s ubs ta nce : as prescribe d by 1.1 - 1.4 Resul t : No treatm ent-related m o rtal ity was noted in either spe c ies. There were no toxic sympto ms, and a hi stological st ud y of the organ s wa s ne gative. Source : Do w Ch emical, TERC Mi dland, MI Test c onditi on : Thre e animal s for ea ch do se, inje cted IV over 30 se cond s for 10 con s e c utive d a ys at a dosa ge of 50 mL a nd 10 mL of 0

. 155M Tri s pe r kg bodyweight. Reliabilit y : (2) v alid with rest rictio ns Meets ge ne ra lly acce pted scientific stand ard s , well -do c ume nted an d accepta b le fo r asse ssment

. 09.11.20 06 (29 ) Ty p e Species Sex : : : Sub-a c ute rabbit male/female Strain : Ne w Zeala nd white Rou t e of ad min. : i.v. Exposur e pe riod Freque nc yo ftrea tm. Post exp osu re period Dose s : : : : 4 wee k s, 5 days per wee k once daily 24 hou rs o r2 0 day recovery period s 0.3M THAM at 0.5g/kg at a rate of 0.5 mL/min. Con t rol grou p : yes, con c u r re nt vehicle Metho d : Grou p 1 Each ani mal i n the first gro up wa s given 20 daily infusi ons (5 days p er wee kfor 4 weeks) of 0.3M THAM at 0.5g/kg at a rate of 0.5 mL/min. Grou p 2 Animals were given 20 dail yIV infusions of normal sal ine to whi c h 5 m Eq of KCl were a dded p e r liter. The dosage and rate of a d minist ration were equal in volu me and time, on a kg b odyweig ht basi s , to the first gro up. Two mal es a nd 2 female s from ea ch group were sa crificed a nd ne cro psi ed within 24 h o u r s of the com p letion of the last treatme nt. The remaini ng animal s in ea ch group we re held for an addition al 20 days po st-inf usio n prio r to necro psy. Bodywe ight, food and water inta ke, and urin e out put were re co rde d daily, urinal ysis wee k ly. Re ctal tempe r ature wa s re corded twice d aily du ring the p r e- and po st-infu s ion p erio ds, and imme diat ely before a nd h ourly until temperature s returne d to no rmal after all i n fusio n s given over th e 20 day treat ment peri od. Urin alysi s (p H, albumin, gl uco s e, ben zidine te st for hemoglo b in on centrif uged sedim e nt and su pern a te, and microsco pic e x amination fo r red a nd white blood cell s and cast s) was perfo rmed o n the first urine voided after infusio ns d uri ng the 20 day treatment pe riod on the first day of infusions, an d twice a wee kfor t he remai ning 3 wee k s. Two days a wee kdurin g the infusio n peri od, a postinfusio n urin a ry THAM excretion wa s me asu r ed o n 2 males a nd 2 female s in each group. Heart blood s p ec im ens were collec t ed for BSP retention tests on ce d u r ing the p r e- and po st-i nfu s ion wee k s a nd duri ng the se con d wee kof the infusio n peri o d. Blood spe c imen s were colle cted fro m an ear 36 / 60

## 5. Toxicity Id 77-8 6 -1 Dat e 14.11.20 06
vein for the following te sts, once duri ng the pre
- an d p ost-infu s io n p eriod s and twi c e a week d u rin g the infusio n s: total serum proteins, A/G ra tios, serum bilirubi n, ceph alin flocculatio n , and se rum tra n sami na se. Also, blood spe c im ens were coll ected at the same time for hemogl obin a nd hemato c rit de terminatio ns, and for white blood cell, re d blood cell, differential, a nd platelet co unts. At necro psy, spe c ime n s from all org a n s and tissue s were extra c te d, pre s e rved, an d examined g r ossly and mi cro s copi cally. Histochemi c al examination s were p e rfo r med on liv er and ki dney specim en s fixed in formalin -calci um for the followin g : alkalin e pho sph a tase, acid ph osp hatase, esterase, pep tidase, DP N diaph ora s e, a nd PPN diap hora s e. Resul t : Food an d wat er co nsumpti on and b ody tempe r ature was not effecte d by treatment in a n y group, no r was water di ure s is p r o d u c ed. Body weight fluctuated through out stud y in all animals , inclu d ing control, but notin any treatment -rel a ted pattern. Seven of 8 rabbits receiving THAM ha d inflammatory lesio ns of the external ea r. The lesi on s carri ed from swelling and re dne ss to dry gang re ne and e r o s io n. Wee k ly analy s e s on blo od sampl es we re norm al for the followi ng para meters: total serum proteins, A/G ra tio, serum bili rubin, ceph ali n flocculation, serum transa minase, RB C, differential counts, hem og lobin, microhem ato c rit, and plate l et count s. Wh ite blood cell count s in excess of 13,000 were see n in 5/8 ra bbits re ceivin g THAM. In all ca se s, ele v ated WBC coin cid ed with the o bse rvation s o fdry gangren e in the external ear. No si gnificantfindings were noted in an y of the para m eters tested during urinaly s is. No gross le si ons were not ed in co ntrol animal s at ne cro psy, ho we ver 2/4 t e strab b it s n e cr op sied 2 0 day s po st -inf usio n pre s e nted gro s sly visible infarct s in the kidney s. No gro ss le sio ns were noted in any other organ or tissu e in any other ani mal. In 7/8 test animals in which there we re g r oss le sion s o fthe ear, there were microsco pic l esio ns of chr onic cellulite s and ne cro s i s at sites of injection in the sub c uta n eou s tissue s of the ear. The gro s s kid n ey lesion s in the 2 animal s we re confirm ed mi cro s copi cally. They were a l so foun d to have chroni c interstitial nephritis. Infiltrati ons of lymphocytes were see n in tissue se ction s of the liver and ki dney of 3 add it ional test ra bbits. The inf iltrations were se en in animal s that were allo wed to recove r 20 days po st-inf usio n, as well as tho s e sacrifice d immediat ely followin g the treatme nt seri es. Pera cute toxic nep hro s i s was ob se rved i n 1 rabbit, wh ich al so presented urolithia s i s
. Lesio ns of pe racute to xic n ephrosi s were not observe d in any of the other 7 rabbits re cei v ing THAM. Source : Do w Ch emical, TERC Mi dland, MI Test c onditi on : Ne w Zeala nd rabbits (16 a dult, 3-4kg) were u s ed. After a 2 we ek observation p eriod, the ani mals were se greg ated into 2 grou ps com posed of 4 male and 4 female ani mals pe r grou p. Conclu sion : Other tha n the necrotizi ng effects at the injectio n site, and tran sie n t body temperature cha nge s, the IV adminis trat ion of up to 20 daily doses of 0.5g/kg of 0.3 M THAM pro duces n o re a d ily detected deleteri o u s effects in mature rabbit s
. Reliabilit y : (2) v alid with rest rictio ns Meets ge ne ra lly acce pted scientific stand ard s , well -do c ume nted an d accepta b le fo r asse ssment

. 09.11.20 06 (30 ) Ty p e Species Sex : : : Chroni c dog male/female Strain : Beagle Rou t e of ad min. : oral feed Exposur e pe riod Freque nc yo ftrea tm. Post exp osu re period Dose s : : : : 1 year Contin uou s 0, 1.1, 11, 110 ppm 37 / 60

## 5. Toxicity Id 77-8 6 -1 Dat e 14.11.20 06

Con t rol grou p NO AEL Metho d : : : yes, con c u r re nt vehicle > 110 ppm Year : 1990 GLP : Test s ubs ta nce : other TS: AM P-H Cl (47.1% AMP) Rem a r k : Test mate ria con s um ption. l inta ke i s esti mated ba sed on we ek 3 6 b ody weig hts a nd food

Males Control es ti (mg/k g /day) 0 mated 1.1ppm 11ppm 110p pm .031 0.31 2.98

Female s
es timated (mg/ k g /day) Control 0 1.1ppm .029 11ppm 0.31 110p pm 2.55 Test s ubs ta nce : 2-ami no-2-m e thyl-1-pro pa nol su pplied by Angus Ch emical Co, Northb roo k , IL. Supplied as an a que ou s sol u tion of AMP-HCl. Concentratio n of AMP in the solutio n was 47.1%. Conclu sion : Based o n the findings u n d e r these stud y condition s, there i s no effect at any dose level on gen eral appe ara n ce, behavio r, bod y weight, food con s um ption, ophthalmo scopic exam s, clinical chemi s try, hematolo g y, orga n wei ght s, or tissue hi stopath o logy. Based on th e absen ce of statistically and biolo g icall ysignifica nt finding s in do se-respon se p a tterns, the No-Ob s e rved Effect Level for AM P in the diets of Beagle do gs i n greate rthan 110 ppm. Reliabilit y : (2) v alid with rest rictio ns Meets ge ne ra lly acce pted scientific stand ard s , well -do c ume nted an d accepta b le fo r asse ssment

. 13.11.20 06 (40 )
5.5 GENETIC TO XICITY ‘IN VITRO‘
Ty p e : Gene mutatio n in Saccha ro myces ce revi siae Sy stem of te sting : Test c once n tratio n : C y cotoxic concen tr. : Metab olic ac tiv a tion : Resul t : negative Metho d : Year : 1990 GLP : Test s ubs ta nce : other TS Resul t : No evide n ce of ketorolac tromethami ne-i ndu ce mutag ene sis in in vi tro Saccha romyces cerevi siae [Syntex Laboratori e s, Inc. Tora dol IM (ketorola c tromethami n e ) pre s cribi ng i n formatio n, produ ct mon o g r aph a nd formulary facts. Palo Alto, CA; 1990] Source : Do w Ch emical, TERC Mi dland, MI Test s ubs ta nce : Ketorola c is commeri cally available a s the trometh a mine salt. Ketorola c tromethami n e is comm erci ally av ailable as a ra ce mic mixture. 38 / 60

## 5. Toxicity Id 77-8 6 -1 Dat e 14.11.20 06

Reliabilit y : (4) n ot assign able Do cume ntation insufficientfor asse ssm ent. 13.11.20 06 (41 ) Ty p e Sy stem of te sting Test c once n tratio n : : : Bacteri a l gen e mutation assay E.coli C y cotoxic concen tr. Metab olic ac tiv a tion : : Resul t : negative Metho d : Year : 1990 GLP : Test s ubs ta nce : as prescribe d by 1.1 - 1.4 Resul t : No evide n ce of ketorolac tromethami ne-i ndu ce mutag ene sis in in vi tro Esch eri c hi a coli [Syntex La borato r ie s, Inc. Tora dol IM (keto r ol ac tromethami n e ) pre s cribi ng i n formatio n, produ ct mon o g r aph a nd formulary facts. Palo Alto, CA; 1990] Source : Do w Ch emical, TERC Mi dland, MI Test s ubs ta nce : Ketorola c is commeri cally available a s the trometh a mine salt. Ketorola c tromethami n e is comm erci ally av ailable as a ra ce mic mixture. Reliabilit y : (4) n ot assign able Do cume ntation insufficientfor asse ssm ent. 13.11.20 06 (41 ) Ty p e Sy stem of te sting Test c once n tratio n : : : other: Me cha nism of nu cle olysis in DMA Streptomyce s lividans C y cotoxic concen tr. Metab olic ac tiv a tion : : Resul t : positive Metho d : Year : 1992 GLP : Test s ubs ta nce : as prescribe d by 1.1 - 1.4 Metho d : The autho rs investigate d reactivity us in g a test syste m of electro p horeti c activation of electroph oresis buffe r conp onent s and a combi nation of the activated sa mples with S.lividans pla s mid DNA. After termin ation of the rea c tion, DNA sample s were a s sayed for dou ble
- strand cl eavag e
. The ass a y involved gel elec trophores is of the DNA samples in HEPES buffer, comp one nts of which are non-re active with the DNA modificatio n s, Southern tran sfer, an d hybridizatio n. Resul t : The autho rs concl ude that two finctio nal grou ps of T R IS are involve d in stran d scissio n
. Initial electroph oretic a c tivation at the anod e is ab solutely requi re d for the formatio n of a TRIS derivative, a presumptive oxygencente r ed radi cal spe c ie s which can b e scaven ged by thioure a
. This s p ec ies reac t s with the DNA modific a tions in the firs t ins t anc e
. As a result, the lesio n s a r e then su sce p tible to further attack, re sulting in stra n d cleava ge. The authors infe r that it is the amine g r ou p in particula r which confe r s o n T R IS its nucle olytic activity. Source : Do w Ch emical, TERC Mi dland, MI Reliabilit y : (2) v alid with rest rictio ns Meets ge ne ra lly acce pted scientific stand ard s , well -do c ume nted an d accepta b le fo r asse ssment

. 09.11.20 06 (42 ) Ty p e Sy stem of te sting Test c once n tratio n : : : Salmonella ty phimu rium re verse m u tatio n assay
C y cotoxic concen tr. : 39 / 60

## 5. Toxicity Id 77-8 6 -1 Dat e 14.11.20 06

Metab olic ac tiv a tion : Resul t : negative Metho d : Year : 1990 GLP : Test s ubs ta nce : as prescribe d by 1.1 - 1.4 Resul t : No evide n ce of ketorolac tromethami ne-i ndu ce mutag ene sis in in vi tro Salmonella ty phimu rium [Syntex Laborat orie s, Inc. Toradol IM (ketorolac tromethami n e ) pre s cribi ng i n formatio n, produ ct mon o g r aph a nd formulary facts. Palo Alto, CA; 1990] Source : Do w Ch emical, TERC Mi dland, MI Test s ubs ta nce : Ketorola c is commeri cally available a s the trometh a mine salt. Ketorola c tromethami n e is comm erci ally av ailable as a ra ce mic mixture. Reliabilit y : (4) n ot assign able Do cume ntation insufficientfor asse ssm ent. 13.11.20 06 (41 )
5.6 GENETIC TOXICITY ‘IN VI VO‘
Ty p e : Mic r onuc l eus as say Species : mouse Sex : Strain : Rou t e of ad min. : Exposur e pe riod : Dose s : Resul t : negative Metho d : Year : 1990 GLP : Test s ubs ta nce : as prescribe d by 1.1 - 1.4 Resul t : There wa s no evidence of mutageni citywhe n the micronu cle us a s say wa s use d to test for ch rom o so me bre a ks in vivo mice that had re ceived ketorola c tro m ethamin e [Syntex Laboratorie s, Inc. Torad o l IM (ket orola c tromethami n e ) pre s cribi ng i n formatio n. Palo Alto, CA; 1990] Source : Do w Ch emical, TERC Mi dland, MI Test s ubs ta nce : Ketorola c is commeri cally available a s the trometh a mine salt. Ketorola c tromethami n e is comm erci ally av ailable as a ra ce mic mixture. Reliabilit y : (4) n ot assign able Do cume ntation insufficientfor asse ssm ent. 13.11.20 06 (41 )
5.7 CARCINOGENICITY
Species : dog Sex : male/female Strain : Beagle Rou t e of ad min. : oral feed Exposur e pe riod : 1 year Freque nc y o ftrea tm. : contin uou s Post exp osu re period : Dose s : 0, 1.1, 11, 110 ppm Resul t : negative Con t rol grou p : yes, con c u r re nt vehicle Metho d : Year : 1990 40 / 60

## 5. Toxicity Id Dat e 77-8 6 -1 14.11.20 06

GLP Test s ubs ta nce : : other TS: AMP-HCl (47.1% AMP) Rem a r k : Test mate rial intake i s esti mated ba sed on we ek 3 6 b ody weig hts a nd food con s um ption. Males Control 1.1ppm 11ppm 110p pm es timated (mg/k g /day) 0

.031 0.31 2.98

Female s

Test s ubs ta nce
Conclu sion

Reliabilit y

13.11.20 06 :

:

: es timated (mg/ k g /day) Control 0 1.1ppm .029 11ppm 0.31 110p pm 2.55 2-ami no-2-m e thyl-1-pro pa nol su pplied by Angus Ch emical Co, Northb roo k , IL. Supplied as an a que ou s sol u tion of AMP-HCl. Concentratio n of AMP in the solutio n was 47.1%. Based o n the findings u n d e r these stud y condition s, there i s no effect at any dose level on gen eral appe ara n ce, behavio r, bod y weight, food con s um ption, ophthalmo scopic exam s, clinical chemi s try, hematolo g y, orga n wei ght s, or tissue hi stopath o logy. Based on th e absen ce of statistically and biolo g icall ysignifica nt finding s in do se-respon se p a tterns, the No-Ob s e rved Effect Level for AM P in the diets of Beagle do gs i n greate rthan 110 ppm. (2) v alid with rest rictio ns Meets ge ne ra lly acce pted scientific stand ard s , well -do c ume nted an d accepta b le fo r asse ssment

. (40 ) Species Sex Strain Rou t e of ad min. Exposur e pe riod Freque nc yo ftrea tm. Post exp osu re period Dose s Resul t Con t rol grou p Metho d Year GLP Test s ubs ta nce : : : : : : : : : : : : : : Syrian ham ster male other: intrat ra che al

negative yes, con c u r re nt vehicle 1979 no data other TS: TRI S AMINO an d 0.9% NaCl Metho d : Thirty 8-wee k old male Syrian gold en ha msters were use d in the control grou p. The a n imals re ceiv ed a pellete d diet and wate r ad libitium. Wee k ly intratra ch eal i n stillation s of a 0.2 ml mixture of TRIS b u ffer and 0.9 % NaCl were pe rform ed for the life of the hamste r
.

Resul t : Animals were obse rved for life or sa crifi c ed wh en mo ri bund. Body weights, averag e su rvi v al time and tumor in cid e n c e for the respirato ry tract (tra che a , larynx and lu ngs) we re a n a lyzed. Com p lete ne cro p sies were p e rf orme d and the orga ns fixed in a 10% buffere d formalin. TRIS AMINO did not indu ce tumors wh e n tested in m a le Syrian gol den hamst e r s. 41 / 60

## 5. Toxicity Id 77-8 6 -1 Dat e 14.11.20 06

13.11.20 06 (43 )
5.8.1 TOXICITY TO FERTILITY
Ty p e : other Species : rat Sex : male/female Strain : other:CD Rou t e of ad min. : oral feed Exposur e pe riod : Males we re e x pose d for at least two wee ks p r io r to bre eding & thro u ghout bree ding fo r 37 days. Fe males we re e x pose d for 2 wee k s prio r to bree ding, con t inuing thro ug h bree ding (u p to 2 wee k s), gestation (3 wee k s), & lactation (4 days). Freque nc y o ftrea tm. : Premating e x posur e peri od Male : At least two wee k s prio r to bree ding. Female : Two wee ks p r ior to b r eedi ng Dura tion of test : No. of gen e r a tion : studie s Dose s : 100, 300, or 1000 mg/ k g/d a y Con t rol grou p : yes, con c u r re nt no treatme nt Metho d : OECD Guide

- line 42 1 Year : 2005 GLP : yes Test s ubs ta nce : other TS:AM P Metho d : Male and fem a le CD rats were fed diet s sup p lying 0 (control), 10 0, 300, or 1000 mg/ k g/d a y of AMP-HCL. Males were expo se d for at least two wee k s prio r to bree d i ng and conti nuing throug h out bree ding f or 37 day s. The females we re expose d for two wee ks p r i or to bre edin g, continui ng throug h bree ding (up to two wee k s), gestation (th r ee wee k s), a nd lactatio n (four days). Effects on go nad al function, mati ng beh avior, con c e p tion, developm ent of the con c ep tus, partu riti o n , litter size, pup survival, sex, pup body wei ght, pup gross external m o rp hol ogical alterati ons, an d path o logy of adult gona ds were a s se ssed. Resul t : Incre a ses in absolute an d relative liv er weig hts, acco mpanie d by a very slight de gre e of microva c u o lizatio n of peripo rtal he pa tocytes, with or without vacuo lization of he patocyte s we re noted in m a les. Femal e s in all treatment g r o ups exhi bited similar hi stop athologi cal ch ange s in the l i ver, but in the absence of an organ wei ght chang e. AMP had no effe ct on mating pe rformance or con c eptio n, but caused ma rke d , dose
- relate d increa se s in post-i m pla nta tion loss (e m b ryo re so rptio n ). At the high dose level, all 12 pregn ant femal e s sho w ed ev iden ce of co mplete litter reso rption (100% p o st-i mplantation l o ss), while at 300 mg/kg/da y, post-impla n tation loss wa s 70% (vs. 10% in control s ). Effects a s soci ate d with, or seconda ry to the post-im plantation lo ss increa se at 300 mg/kg/da y include d decrea s e d litter si ze, incre ase d pup b od yweight, and decrea s ed g estation body wei ght and bo dy wei ght gain. The r e we re n o treatment rel a ted effects on rep r odu ctive pe rforma nc e in the 100 mg/kg/day grou p. Test s ubs ta nce : Chemi c al Na me: 2-Amino
- 2-methyl prop anol hydrochl oride salt Molecular Formula: C4H11NO Molecula r We ight: 89.14 Synonyms: AMP, AMP-HCL Conclu sion : The NOEL for gene ral toxi city in males wa s 300 mg/ k g/day, whil e the gene ral toxici ty for females could n o t be determi ned, b a se d on the pre s en ce of very slight mi crosco pic live reffects. The NOEL for rep r od uctive effects was consi dered to be 100 mg/ k g /day. Reliabilit y : (1) v alid with out rest rictio n GLP guideli n e study 42 / 60

## 5. Toxicity Id 77-8 6 -1 Dat e 14.11.20 06

13.11.20 06 (44 )
5.8.2 DEVELOPMENTAL TOXICITY/TERATOGENI CITY
Species : rat Sex : female Strain : Rou t e of ad min. : derm al Exposur e pe riod : 6 hours daily day from ge station days (G D) 6

- 20 Freque nc y o ftrea tm. : daily Dura tion of test : from ge statio n days (GD) 6-20 Dose s : 30, 100, or 3 00 mg AMP/kg/day Con t rol grou p : yes NO AEL m ate rnal tox. : = 100 mg/kg bw other : NOELfor : = 300 mg/kg bw dev e lopmental Tox. Resul t : There wa s no evidence of test arti cle rel a ted syste m ic maternal o r developm ent al toxicity at a n y dose level tested. Metho d : OECD Guide

- line 41 4 "Teratogeni city" Year : 2006 GLP : yes Test s ubs ta nce : other TS:AM P Metho d : Female rats were expo se d 6 hours dail ydermally to 0, 30, 100, or 300 mg AMP/kg/day from ge station days (G D) 6
- 20 (Carney a nd Tho r srud, 2006 ). Rats were sa crificed on G D 21, and su bjecte d to a pathologi cal examination i n clu d ing exa m ination of t he external tissue s an d all orifice s , stoma c h, liver, kidney s, an d uterine findi ngs. The first four pre s um ed preg nant fem ales fro m ea ch dose gro up were sel ecte d for blood co llection to evaluate systemic expo sure followi ng dermal a d mi nistratio n of AMP on the last day o fdosing (GD 20). Resul t : Dermal admi n istratio n of 300 mg/kg/d a y of AMP produce d sig n ifica n t effects at the test site, as e v idenced by scab bing a nd mode rate to severe scaling. The derm a l findin g of slight sca ling at 30 and 100 mg/kg/d a y was not con s id ere d adverse, as the obs e rvation wa s tran si ent in nature and relatively low in incid e n c e. There wa s no evidence of test arti cle rel a ted systemi cmat e rnal o rdevel opmental toxi city at any dose level teste d

. Analyse s of blood sample s confirm ed sy stemic exposure to AMP in a dose
respon sive m anne r, althou gh the st udy wa s not de sig ned to qua ntify percent ab so rption. Test s ubs ta nce : CAS Numb er: 124-68-5 IUPAC Name: 2-Amino-2-m ethyl-1 -p ropa nol Molecular Formula: C4H11NO Molecula r We ight: 89.14 Synonyms: AMP, AMP-95 Conclu sion : Und e r the co ndition s of this study, the NOAEL for maternal toxicity based on dermal effects wa s 100 mg/kg/day. The NOEL fo r developm ent al toxicity was 3 00 mg/kg/d a y, the highest dose level tested. Reliabilit y : (1) v alid with out rest rictio n GLP study. 13.11.20 06 (45 )
5.8.3 TOXICITY TO REPR ODUC TION , OTHER STUD IES

5.9 SPECIFIC INVESTIGATIONS
Endpoint : other: De rmal Absorbtion 43 / 60

## 5. Toxicity Id 77-8 6 -1 Dat e 14.11.20 06

Stud y descr. in chapter Referen ce : : Wepi erre, J. and Noel-Hu d so n, M.S. Ty p e Species Sex : : : other: hum an skin Strain : Rou t e of ad min. : 100 uL a ppli c ations to the ski n. No. of animals : Vehicle : Exposur e pe riod Freque nc yo ftrea tm. Dose s : : : 24 hou r(s) once 0.1% and 10 % solution s Con t rol grou p Observ ation period Resul t : : : less than 1% of applied d o s e is a b sorb e d into the ski n. Metho d : Year : 1993 GLP : Test s ubs ta nce : other TS: C-14 label ed Tromethami ne hydro c hlo r ide Metho d : In vitro percutaneo us a b sorption wa s stu d ied u s ing d e r matomi zed h u man ski n (obtai ne d from pla s tic surg ery bio p s ie s of the ab domen a nd froze n until use ) pla c ed in F r an z diffusion cell s to permit con t act of the dermis with a pre s e rvatio n liquid of so dium chl o ri de and bovine serum al bumin . The ski n wa s tha w ed, defatted , and cut to 0.3mm thickne ss. A cutan e ous biop sy is mai n tained h o ri zontally betwe en the dermis and epid e rmi s
. The system i s su rroun ded with a warm wate r jacket to maintain the temperatu r e at 37C. The air and liq uid in the syst em are ci rculate d contin uou sl y to insu re con s ist ent temperature. The test mate rial, C-14- la b e led trom etha mine hydroch l oride, was furnished by L'Oreal un der the same S1 (0.1%) and S 2 (10%). The test material (100 uL) was a ppli ed to 0.635 cm^2 ski n. At regul ar time i n tervals (2, 4, 6, 8, 10 hours), the totality ofthe liquid contain e d in the derm a l comp artme n t is take n by lateral adj ustm e n t and repl aced by new liq uid. At t=24 ho urs, the pre s e rvati on liquid i s re moved and th e surfa c e of t he biop sy is wa shed with 1 00 uL of differentsolvents: 1st wa sh - Cet avlon / doubl y-distilled wat er 2nd wash- do ubly-di stilled water 3rd wash- Ce tavlon / doubl y-distilled wat er 4th wa sh- d o ubly-di stilled water 5th wa sh- d o ubly-di stilled water The appli c ati on zo ne was wipe d with co tton rolls, the dermi s an d e pidermis were se pa rat ed mechani cally with a scalpel, and di g e sted in Solu ene TM for 24 hou rs at 37C. The dete c tion of remainin g radio a ctivity wa s determin ed via liquid sci ntillation counter. The pre s e rvation l i quid re move d, cotton roll s, and glass cylin der are also co u nted for ra dio a ctivity. Counting value s were corre c ted by the method of exte rnal sta n dard to obtai n the dpm. Resul t : Abso rption ofthe test material is lo w and variable from one skin sa mple to anothe r. At th e end of 24 h ours, 0. 506 +- 0.765 for th e 0.1% soluti on and 0.797 +- 0.69 1 for the 10% solution were determi ned. There is n o signifi cant differen c e b e twe en the per ce n t ages, sho w in g that the increase in test materi al con c e ntrai on doe s not a l ter cutan eou s permea bility under these te st co ndition s. The fluxes in the 2 ca se s reache s a ma ximum value after 4 hou rs and remai n co nst ant durin g the rest of the experim ent. After wa shi ng, the retention of tromethami ne hydro c hlo r id e in the dermi s and epid e rmi s is lo w (0.13

- 0.14% for the 2 solutions in t he e pi dermi s an d 0.69- 0.22% in the dermi s). The test material is not retain e d in the hornylayer. The washi ng waters contai ned mo re tha n 90% of the applie d dose. Source : Do w Ch emical, TERC Mi dland, MI 44 / 60

## 5. Toxicity Id 77-8 6 -1 Dat e 14.11.20 06

Conclu sion : Percutane ou s ab sorption throu gh hu ma n ski n in vitro is low, on ave r age less than 1% of the applied dose rema i n s at the end o f24 hours, an d the finding is in de pend ent of the con c e n trati on of trometh a mine hyd r o c hlorid e applie d. The test mate rial is alm o st totally eliminated by wa shin g the ski n after 24 hours. The test mate rial is not reta ined in the ho rny layer. Reliabilit y : (2) v alid with rest rictio ns Meets ge ne ra lly acce pted scientific stand ard s , well -do c ume nted an d accepta b le fo r asse ssment

. 09.11.20 06 (46 )
5.10 EXPOSURE EXPERIENCE

T y pe of experience : Dire ctob se rv at ion, clini c al ca se s Metho d : THAM (3 6g i n 5% dextrose water) was infuse d in 4 o fthe subje c ts over a perio d of 30 minutes to 24 hours. One patient re ceiv ed 9g THAM over 30 minutes. Resul t : In all cases, there wa s a substa ntial rise in the blood pH and a sig n ificant increa se in th e total CO2 content of the blood. Ho we ver, none of the patients sho w ed any clini c a l improveme n t. The increa se in blo od p H and total CO2 con t ent appea re d to persist fo r >48 hou rs i n most of the patients and continu e d to remain el evated for a p e riod of several days in 2 o fthe patients. Three of five patients showed 2 major complications following THAM admi n i s tration: hype rkal emia a nd oliguri a
. Source : Do w Ch emical, TERC Mi dland, MI Test c onditi on : Five patients manifestin g 3 types of rena l disea s e: po stpartum bilate ral renal co rtical necro sis, a c u t e and ch roni c glome r ul on ephriti s , and diabeti c intracapilla ry glome r ulon ep hro scl erosi s
. All subject s had severe impairm ent of renal fun c tio n s a s manife sted cli n ically by oliguria, e levated blood u r ea nit r oge n, and a bnormal uri n a ry findings. Tw o were a n u r ic and 2 others we re o liguri c
. Conclu sion : In 2 of the cases of com p lications , the subjects were acutely ill and manifeste d a severe re nal failure that co uld have re su lted in the hyperkale miaand oligu r ia. Howeve r, the third patient , who re ceive d the THAM admi n i s tration m o re rapidly than t he other 4 p a t ients, pre s en ted initially with normal u r ine o u tput and no rmal se rum po tassi um, and within 12 hou rs of T H AM admini s tration sh owe d marked oli g uria an d hyperkale miaand an eleva t ion of blood urea nit r og en level, although the mech ani sm o fsuch com p lication s was n o t kno w n at the time of publicatio n. THAM is a n e ffective agentin the prom ptcorre c tion of metaboli caci dosi s
. In compa r i s o n with so dium bicarbon ate or lactate, it may also offe r the advantag e of being a b le to buffer the intracell u lar hyd r ogen io n. THAM may therefore be a more suitable ag ent to corre c t aci dosi s when sodium intake i s a co nce r n. The a uthors su gge st a maximu m dose of 50 0mg/kg (0.3M T H AM in dextrose) g iven over24 hours with ca reful monito ri ng of urina ry output , serum p otassium, and bl o od ure a nitro gen for 48 h o urs following THAM administ r ation. Reliabilit y : (2) v alid with rest rictio ns Meets ge ne ra lly acce pted scientific stand ard s , well -do c ume nted an d accepta b le fo r asse ssment

. 23.03.20 04 (47 ) T y pe of experience : Dire ctob se rv at ion, clini c al ca se s Rem a r k : Re sults of thi s study (Sa m i y , et al., 1961) are typical o fmany publications evaluating clinical treatment of ill patients with THAM as a buffer for 45 / 60

## 5. Toxicity Id Dat e 77-8 6 -1 14.11.20 06

acid osi s
. The followin g re feren c e supp orts the g ene ral finding s ofthe authors of this arti cle.

Source Reliabilit y

23.03.20 04 : : Authors su pp ort THAM' s u s e a s treatme nt of metabolic aci d o s is, no ting gene rally po sitive outcome s and n o adv erse effect s a s soci ated wit h THAM. Do w Ch emical, TERC Mi dland, MI (2) v alid with rest rictio ns Meets ge ne ra lly acce pted scientific stand ard s , well -do c ume nted an d accepta b le fo r asse ssment

. (48 ) T y pe of experience : Dire ctob se rv at ion, clini c al ca se s Rem a r k : Re sults of thi s study (Sa m i y , et al., 1961) are typical o fmany publications evaluating clinical treatment of ill patients with THAM as a buffer for acid osi s
. The followin g re feren c e supp orts the g ene ral finding s ofthe authors of this arti cle.

Source Reliabilit y

23.03.20 04 : : The autho rs suppo rt THAM as treatme nt of respi r ato ry acid osi s , and addition ally noting a slig ht decrea s e in respi r ation rat e followin g treatment. Do w Ch emical, TERC Mi dland, MI (2) v alid with rest rictio ns Meets ge ne ra lly acce pted scientific stand ard s , well -do c ume nted an d accepta b le fo r asse ssment

. (49 ) T y pe of experience : Dire ctob se rv at ion, clini c al ca se s Rem a r k : Re sults of thi s study (Sa m i y , et al., 1961) are typical o fmany publications evaluating clinical treatment of ill patients with THAM as a buffer for acid osi s
. The followin g re feren c e supp orts the g ene ral finding s ofthe authors of this arti cle.

Source Reliabilit y

09.11.20 06 : : For tre atmentof acido s is d uring ca rdia c bypass proce dure s an d du ring total circulato ry arre st, these authors ad d itionally note that THAM is even more effe ctive whe n CO 2 is allo wed a m ean s to leave the blood (vi a respiration, etc), and can b e use d wh en sodi um bicarbonate can n o tbe (wh en a patie nt is on total cardiop u lmon ary bypass). THAM ha s no t been asso ciated wi th any toxic effects and i s r apidly excreted in the urin e. It has however, bee n noted to ha ve transie nt effects on blo o d sug a r, se ru m potassium, in cre a sed u r ina ry (wate r) lo ss, and de pre s sed respiratory rates. Do w Ch emical, TERC Mi dland, MI (2) v alid with rest rictio ns Meets ge ne ra lly acce pted scientific stand ard s , well -do c ume nted an d accepta b le fo r asse ssment

. (50 ) T y pe of experience : Dire ctob se rv at ion, clini c al ca se s Rem a r k : Re sults of thi s study (Sa m i y , et al., 1961) are typical o fmany publications evaluating clinical treatment of ill patients with THAM as a buffer for acid osi s
. The followin g re feren c e supp orts the g ene ral finding s ofthe authors of this arti cle. Authors note that Trometh a mine NF is an effective b l ood buffer fo r

Source Reliabilit y

23.03.20 04 : : metaboli can d respiratory acid osi s
. They sugg est a d ministe r in g sufficie n t amount s of water with tre a tment may pre v ent hypero s molarity and avoid tissu e dehyd r ation, and tha tcaution sh o u ld be u s ed t o avoid hyperkalemi a, hypoglycemia , and depression of the respirato ry cente r
. Do w Ch emical, TERC Mi dland, MI (2) v alid with rest rictio ns Meets ge ne ra lly acce pted scientific stand ard s , well -do c ume nted an d accepta b le fo r asse ssment

. (51 ) 46 / 60

## 5. Toxicity Id 77-8 6 -1 Dat e 14.11.20 06

T y pe of experience : Dire ctob se rv at ion, clini c al ca se s Rem a r k : Re sults of thi s study (Sa m i y , et al., 1961) are typical o fmany publications evaluating clinical treatment of ill patients with THAM as a buffer for acid osi s
. The followin g re feren c e supp orts the g ene ral finding s ofthe authors of this arti cle. Authors re co mmend T H A M for treatme nt of diabetic acid osi s , wh e n patients a r e u nable to tolerate sodi um bi carbon ate or l a ctate the r ap y due to severe ca rdi a c and/o rso di um-retainin g renal failu re. The autho rs sugge st that THAM may redu ce the insulin requi rements fo r m anag ement of diabeti ccom a s. The patie nts in this a rticle, whil e re spondi ng favorably to THAM treatm ent, eventuall ysuccumb e d to other com p licatio ns of their con d ition s , and upo n po st-mortem exam ination s there were n o findi ngs that sug g e s ted T H AM co ntribu ted to the cau s e s of death. Source : Do w Ch emical, TERC Mi dland, MI Reliabilit y : (2) v alid with rest rictio ns Meets ge ne ra lly acce pted scientific stand ard s , well -do c ume nted an d accepta b le fo r asse ssment

. 23.03.20 04 (52 ) T y pe of experience : Dire ctob se rv at ion, clini c al ca se s Metho d : The patient b r eathe s oxyg en in a non breathing syste m to remove nitroge n from the lung s, after whi c h a barbitu rate is use d to re nder the p a tie n t uncon sci ou s durin g the proce dure. THAM is adminis tered IV (0.33M solutio n in 0.2% NaCl ) to investigate th e buffering eff e ct of THAM. Succi nylch oli ne is inje cted and the bron chosco pe intro duced when t he patient is co mpletely para lyzed. O xygen is co ntinuo usly admini s t e red to the patients t o ensure com plete apn ei c oxygenation of the blood d u ring the 6-min u te pro c edure. Like wise, no carb o n dioxide is al lowe d to exit the lung s. Arterial blood sam p les were dra w n befo r e an d immediatel y followin g the 6-min u te apn eic pe riod, an d at 5 minutes and 2 h ours postpro c ed ure. All patients were maintaine d on ventilation until recove ry of norm al sp ont aneo us respiration. Blood wa s analy z ed for pH (o ne anae robi callyand two at known CO 2 t ensio ns) and p C O2. Actual HCO3- wa s cal c ul ate d according t o Hen der so n-Ha sselbal ch f ornul a, and p O2 wa s determi ned with a modified Clark ele c tro de. Brachi al arteri al blood pre s sure wa s measu r e d using an inflata b le cuff and mercury ma n o meter. Resul t : Duri ng the 6 minute proce dure, the Pa CO2 in crea se d from 38 to 66 mmHg, with a fall in pH from 7.41 t o 7.24 and a slight in cre a se in actual bica rbo nate from 24 to 28 mEq/L. Whe n THAM was admini s tered, the arteri al pH was kept nea rl y consta nt, with a rise of al most identi c al prop ortio ns in H2CO3 an d HCO3- in all ca se s. The PaCO 2 ro se from 37 to 42 mmHg an d the HCO3- from 24 to 28 mEq/L. In all subje c ts, the arterial blood wa s co mpletely satu rated with oxygen throu gh out the pro c e dure. The two subj ects in p oor condition p r ior to the pro c ed ure were cya notic whe n bre a thi ng air, ho wev e r breathin g pure O 2 for 5 minutes relie ved the cyano si s. Analysis of arte rial blood in b o th subj ect s showed an al most-fully comp en sated respi r ato ry acido s i s
. PaO2 durin g brea thing su gge st ed a shu n ting of n onoxygen ate d blood in the lungs. Six minutes of cont rolled ventilation pre-p r o c ed ure i n crea sed t he PaO2 slig htly and ca used a drop in PaCO2 to no rmal values wi th a marked a lkalin e shift in pH. Both faired simila rly to the subj ect s in better initial h ealth duri ng the apn eic p e riod. Both sho w ed improve d re spirato ry statu s upo n rem o val of secretio ns from the airways. In patients no t receiving T H AM, there was a con s iste nt rise in dia s tolic blood p r e s sure, not seen when the re spi r atory acid osi s was b uffered with THAM. No p a tient receiving THAM rep o rted any co mplaints o r 47 / 60

## 5. Toxicity Id 77-8 6 -1 Dat e 14.11.20 06
compli catio n s that could be attributed to the buffer o rthe apn ea. Source : Do w Ch emical, TERC Mi dland, MI Test c onditi on : Twelve ad ult subj ect s we re treated with THAM in the study. The chang es in acid -ba s e status we re compa r ed with those o c curri ng in 6 su bje c ts not receiving T H AM. Ten of the su bje c ts h ad lung tumo rs or small tub e rculou s cavities a nd u nderwe nt bro nch osco py as a routine p r e operative pro c ed ure. They we re all in good ge ne ral co ndition without re spi r atory distress. Two subj ect s , deal t with sepa rat e ly, were in p oor conditio n with ventilatory insuffici en cy prio r to the broncho scopy. Conclu sion : Becau s e all CO2 p r od uce d by the body remain ed in the body du rin g the apnei c pe riod , a nearly-p erfect stoic hom etric relation ship wa s reve aled betwe en CO2 produ ce d an d the THAM need ed to buffer it. The buffer wa s found to not o n ly redu ce th e increa se in arteri al blood pre s sure, but also redu ce the typical in crea se in cere brospi nal fluid pre s sure se en in respiratory acidosi s
. THA M was not found to ca use any hypoglycemia, nor were any toxic manife stations n o ted foll owin g THAM treatment. THAM is con s ide r ed by th e author to b e helpful in counteractin g respi r ato ry acid osi s e s pe cially in patie nts wh ere a b l ood pressu re rise o rCSF pre s sure rise i s not de sire d, duri ng pro c ed ur es where a deq uat e ventilation cannot be maintained at all times. Reliabilit y : (2) v alid with rest rictio ns Meets ge ne ra lly acce pted scientific stand ard s , well -do c ume nted an d accepta b le fo r asse ssment

. 23.03.20 04 (53 ) T y pe of experience : Dire ctob se rv at ion, clini c al ca se s Rem a r k : Re sults of thi s study (Holm dahl, 196 1) a r e typical of many publi c a t ions evaluating clinical treatment of ill patients with THAM as a buffer for acid osi s
. The followin g re feren c e s su p port the gen e r al finding s ofthe authors of this arti cle. For tre atmentof respi r atoryacido s i s ca u s ed by chro ni c alveola r hypoventilatio n , the followin g authors cau t ion use of T H AM, becau se, even though a n effective buffer, they noted it cau s e d in so me patients a n accentu a tion of anoxemia that became very seve re, a nd su gge sted that THAM should not be use d to treat su ch case s unl ess o t her mea ns of oxygenation are suppli ed (mech ani cal ai d). The auth o rs d o not su pport gene ral u s e o fTHAM as a treatme nt for respi r ato ry aci dosi s cau s ed by chroni c alveol ar hypoventil ation. Source : Do w Ch emical, TERC Mi dland, MI Reliabilit y : (2) v alid with rest rictio ns Meets ge ne ra lly acce pted scientific stand ard s , well -do c ume nted an d accepta b le fo r asse ssment

. 23.03.20 04 (54 ) T y pe of experience : Dire ctob se rv at ion, clini c al ca se s Resul t : Re sults of thi s study (Holm dahl, 196 1) a r e typical of many publi c a t ions evaluating clinical treatment of ill patients with THAM as a buffer for acid osi s
. The followin g re feren c e supp orts the g ene ral finding s ofthe authors of this arti cle. For tre atmentof respi r atoryacido s i s , the authors likewi s e caution th e use of THAM to buffer the bloo d unle ss sup p lemental oxygen is given due to a resulting de crease in minut e ventilati on rate. The authors advan ce the theory that the decrea s e in minute ventila tion rate is d ue to the increase in pH of the blo od and n o t du e to a redu ction of CO2 te nsio n per se. They repo rt sp urio us findin gs ofkidney (swell ing and hyd r o pic de gen erat ion of the lining of p r oximal tubul es) a nd/o rliver (hyd ropi c d egen eratio n o fthe cell s) chan ge s in 2 patient s re ceiv ing T H AM, however the autho rs are relu ctant to attribute the finding s dire ct ly to THAM ba sed on othe rs i n their 48 / 60

## 5. Toxicity Id Dat e 77-8 6 -1 14.11.20 06

Source Reliabilit y

23.03.20 04 : : sampli ng thatdo not pre s e nt such sig ns. Do w Ch emical, TERC Mi dland, MI (2) v alid with rest rictio ns Meets ge ne ra lly acce pted scientific stand ard s , well -do c ume nted an d accepta b le fo r asse ssment

. (55 ) T y pe of experience : Dire ctob se rv at ion, clini c al ca se s Rem a r k : Re sults of thi s study (Holm dahl, 196 1) a r e typical of many publi c a t ions evaluating clinical treatment of ill patients with THAM as a buffer for acid osi s
. The followin g re feren c e supp orts the g ene ral finding s ofthe authors of this arti cle.

Source Reliabilit y

23.03.20 04 : : For tre a tmentof respi r atoryacido s i s , the authors find T H AM an effective blood b u ffer, noting al so a decrea s e in v entilation rate , and addition ally a con s i s tent rise in arteri al bl ood p H and acon s ide r abl e incre a se in u r ina ry pH with a n asso ciated in crease in urin ary excretion ofbicarbon ate. They noted no a d verse effect s a ttributed to THAM treatme nt, and did no t note hypoglycemiaas othe r re se arche s have, likely due to l o we r do se le vels. Do w Ch emical, TERC Mi dland, MI (2) v alid with rest rictio ns Meets ge ne ra lly acce pted scientific stand ard s , well -do c ume nted an d accepta b le fo r asse ssment

. (56 ) T y pe of experience : Dire ctob se rv at ion, clini c al ca se s Rem a r k : Re sults of thi s study (Holm dahl, 196 1) a r e typical of many publi c a t ions evaluating clinical treatment of ill patients with THAM as a buffer for acid osi s
. The followin g re feren c e supp orts the g ene ral finding s ofthe authors of this arti cle.

Source Reliabilit y

23.03.20 04 : : For tre a tmentof respi r atoryacido s i s , the authors find T H AM an effective blood b u ffer, noting al so a decrea s e in v entilation rate , and addition ally a con s i s tent rise in arteri al bl ood p H and acon s ide r abl e incre a se in u r ina ry pH with a n asso ciated in crease in urin ary excretion ofbicarbon ate. They noted no a d verse effect s a ttributed to THAM treatme nt, and did no t note hypoglycemiaas othe r re se arche s have, likely due to l o we r do se le vels. Do w Ch emical, TERC Mi dland, MI (2) v alid with rest rictio ns Meets ge ne ra lly acce pted scientific stand ard s , well -do c ume nted an d accepta b le fo r asse ssment

. (57 ) T y pe of experience : Dire ctob se rv at ion, clini c al ca se s Rem a r k : Re sults of thi s study (Holm dahl, 196 1) a r e typical of many publi c a t ions evaluating clinical treatment of ill patients with THAM as a buffer for acid osi s
. The followin g re feren c e supp orts the g ene ral finding s ofthe authors of this arti cle.

Source Reliabilit y : : In the treatment of artificia lly-indu ced re spiratory a c id osi s (in patie nts recoveri ng fro m pulmon a ry tubercu lo si s with a normal l y sensitive respiratory ce nter), T H AM is found to be an effective b u ffer that pre v ents hyperventilati on whil e bre a t hing CO 2. No toxicity has been n o ted, a nd respiratory toxicity was su gge sted bythe authors to b e unlikely due to rapidly -reve rsible effects o n ventilation rates. Do w Ch emical, TERC Mi dland, MI (2) v alid with rest rictio ns Meets ge ne ra lly acce pted scientific stand ard s , well -do c ume nted an d accepta b le fo r asse ssment

. 49 / 60

## 5. Toxicity Id Dat e 77-8 6 -1 14.11.20 06

09.11.20 06 (58 ) T y pe of experience : Dire ctob se rv at ion, clini c al ca se s Rem a r k : Tromet hamin e occurs in K etorola c Tr o methamin e at approxim atel y 32% of the formulation to improve so lubility. While it is not possible to differentiate the side effe cts of the active dr ug from T r ometh ane, there we re no clini c ally-signi ficant finding s that w ould sugge st it is inhere n tly toxic.

Resul t : Re sults of thi s study a r e typical of manypublication s evaluating clinical treatment of post-ope rativ e patient s with Ketorola c T r ometh amine. Ketorola c Tro m ethamin e is a non-steroi d a l anti-inflam matory dru g (NSAID) indicated for the treatme nt of acut e pai n, rece ntly approved by the FDA (U.S. Food a nd Drug Admi nistratio n ) for IM (intramu scular) admini s t r ation. It is a pyrrolo -pyrrol e com p ound related t o tolmetin an d zome pirac. The tromethami n e salt in Ketorolac enhan ces its solubility; there are 6.8mg ket or ola c in 1 0mg ket o rol a c t r omet h ami ne. Clini c al trial s of Ketorola c Tromet hamin e have pro du c ed p ositive a nalge si c effects in po st-ope rative pa tients with qui ck o n set of re lief lasting lon ger than com p a r a b le do se s of morp hine. In addition, Keto rola c Tro m et hamine treatment p r o v ides a no n d epen den cy -fo r ming alte rnat ive to opiates. Authors note d that some p a tients ex pe ri enced a de creased platel e tcount and ble eding time that was statistically, but not clini c a lly, significant , althoug h it is not recomme nded for tr eat ment of patients with existi ng bleedi ng diso rde r s. It has, like mo st NS AIDS, the potential to cau s e gastri c muco sal inj u ry. Diarrhe a , dizzine s s, sw eating, and p a in at the inje ction site were noted in 1-3% of patients. There have bee n no drug inte ra ctions noted. Although anim a l st udie s have shown no evid ence of mutagene si s, carcin oge ne sis, or fertility impairm ent, it is not re com m ende d for p r egn ant or lactatin g women. Re spi r atory de pre s sion h a s n o t been n o ted in patients usin g the dru g
.

Source Conclu sion

Reliabilit y

23.03.20 04 : :

: While IM adm inistratio n ha s bee n app ro ved by the USFDA, intravenou s admini s tratio n wa s not ap proved whe n the article wa s publi s h ed. Do w Ch emical, TERC Mi dland, MI Ketorola c Tro m ethamin e h as be en eval uated by the USFDA to be a viable alternative to opiate s for pa in relief. Whil e side -effect s have been n o ted simila r to other NSAIDS, it can be safel yused a s a n on-h abit formi ng dru g to relieve pai n in most pati ents. (2) v alid with rest rictio ns Meets ge ne ra lly acce pted scientific stand ard s , well -do c ume nted an d accepta b le fo r asse ssment

. (59 ) T y pe of experience : Dire ctob se rv at ion, clini c al ca se s Rem a r k : Results of thi s study (Resman-T a rgoff, 1990) are typical of many publi c ation s e v aluating clini c al treatm ent of post-ope ra tive patients with Ketorola c Tro m ethamin e. The followi ng refere nce su pport s the ge neral finding s of the authors of this arti cle.

Source Reliabilit y

23.03.20 04 : : Noting a n increa se in liver function te st result s, the followin g autho rs have addition ally noted that the drug m a y not be ideal for u s e in patie nts with prio r liver da mage. Do w Ch emical, TERC Mi dland, MI (2) v alid with rest rictio ns Meets ge ne ra lly acce pted scientific stand ard s , well -do c ume nted an d accepta b le fo r asse ssment

. (60 ) T y pe of experience : Dire ctob se rv at ion, clini c al ca se s 50 / 60

## 5. Toxicity Id 77-8 6 -1 Dat e 14.11.20 06

Rem a r k : Results of thi s study (Resman-T a rgoff, 1990) are typical of many publi c ation s e v aluating clini c al treatm ent of post-ope ra tive patients with Ketorola c Tro m ethamin e. The followi ng refere nce su pport s the ge neral finding s of the authors of this arti cle. A transie nt re nal insufficien cy wa s not ed by the followi ng autho rs, di rectly attributed to Ketorola c Tro m ethamin e treatment. De cre a sed u r ine output and an in crea sed serum creatinine con c entration we re measured. There wa s no hype rkalemi a note d

. These o b servation s , ho wever, a r e in discord with ob servati ons of hype rkalemia an d in cre a sed u r ine output noted with THAM infu sio n (minu s Ketorola c) in oth e r patient s, sugge sting tha tthe observation s noted by these authors co u l d be prim aril y attributed to the Ketorolac

. Source : Do w Ch emical, TERC Mi dland, MI Reliabilit y : (2) v alid with rest rictio ns Meets ge ne ra lly acce pted scientific stand ard s , well -do c ume nted an d accepta b le fo r asse ssment

. 23.03.20 04 (61 ) T y pe of experience : Dire ctob se rv at ion, clini c al ca se s Metho d : Four h ealthy adults received THAM vi a IV over 30-60 minutes in 0. 03M Na CL / 0.005 M KCl durin g 2.3 or 3.4% CO2 bre a thing. Resul t : 64% of THA M admini s tered wa s elimin ated in the uri ne over 2 day s, and 77% over 3 d a ys. Major side eff ects n oted in the s ubj ect gi ven 8.8 mM/kg bodyweight inclu ded: hyp o tensi on, hyp o xia, hunge r, sweating, pe riodic b r eathi n g , wea k n e ss, so mnolen ce, di arrhea, inten s e wret chin g a nd vomiting, sen s atio ns ofheat, swelli n g and num bn ess of face. In all other su bject s , water di uresi s wa s note d with de cre ase blood seru m gluco s e con c e n tration s
. Urin e pH a nd CO 2 co ntent rose as th e THAM was eliminated. Source : Do w Ch emical, TERC Mi dland, MI Reliabilit y : (4) n ot assign able Do cume ntation insufficientfor asse ssm ent. 24.03.20 04 (62 ) T y pe of experience : Dire ctob se rv at ion, clini c al ca se s Metho d : Venou s bloo d sample s were taken from the other a r m 10, 20, and 3 0 minutes afte r starting the in fusion, an d 5, 10, 20, and 40 minute s at 1, 2, 4, 8, 12, 20, and 24 hours afte r it had ende d

. Urin e wa s col l ected fro m the begin ning o fthe infusion until 30 minut es after its end, from 30 min to 4 hours, from 4 to 8 hours, an d from 8 to 24 hours. Five patients requi re d hae modialysi s o rhaemofiltratio n as a result of acute anuri a
. In two patients, ga stri c juic e s were d r ain ed continuo usly via a plasti c tube a nd we re colle cted for 4 8 ho urs a nd an alyzed for T R IS. In one patient, a drai n had be en i m planted in t he com m on b ile duct diri ng chol edo cholit hotomy and the bile was collecte d over two 24 ho ur p e riod s. All sample s were an alyze d for TRIS co ncentration via gas chromato grap hy. Resul t : In three patie nts, unide ntified pea ks we re see n with the sam e rete ntion times a s TRI S due to addi tional medi ca tion. The hae mofiltrates of these patients wa s sub s e que ntly not anal yze d for TRIS con c entration s. The re sult s from the 6 heal thy subje c t were po oled. At the end of infusio n , the TRIS plasma con c e n tra t ion averag ed 565 ug/mL. There wa s a biexpon ential declin e of plasma T R IS le vels and after 24 hours, the level 51 / 60

## 5. Toxicity Id 77-8 6 -1 Dat e 14.11.20 06

wa s only 3.8 ug/mL. The half-life of t he terminal ph a s e was 5.6 h ours. T R IS c o nc en tr a t io ns in e ryth r oc yt es rose more sl owly, rea c hin g a maximum 20 minutes afte r the end of infu sio n s. After 2 hours, drug levels in erythro c yte s we re ab out 1.5 times gre a ter than tho s e in plasm a , and they remain e d well ab ove the co rrespon ding pla s ma l evels du ring t he re st of the obse rvation peri od. Pharma co kin etic paramete r s were cal c ul ated from the individual con c e n tration
- time cu rves usin g a two-compa rtment model with eli m ination from the c e ntral c o mpartment. TRIS is mainl yexcreted bythe kidney

. Already 30 mi nutes afte r the end of infusio n , 25% of the TRIS wa s found in the urine, an d after 24 hou rs, 82% of the TRIS had bee n elimi nated in that way. Infusion of th e stron g ly alkaline solution (pH 1 0 .9) wa s well tole rate d by all patients witho u t adverse re action s. The half-life o fTRIS in normuri c patient s (in c ludi ng some with po o rren a l function ) wa s longer tha n in healthy pati ents (1 6-45 h ours), an d the volumes of di stributio n we re much la rg er. Up to 72% of the TRIS was eliminated in the urine afte r 24 hou rs, and an ad ditio nal 2-5% ex creted durin g the ne xt 24 hours. The half-life o fTRIS in anuric patie nt s ra nged 1 5 -5 8 h ours. 25-6 6 % of the infuse d TRIS left the plasm a in the first 2 4 hours, and t he cle a ra nce averag ed 16. 7 mL/kg/h r
. Period s of ha emodialy s is o rhaem ofiltration did not affect pla s ma T R IS level. The amou nt of TRIS eliminated via these pro c ed ures could not be m easure d as th e fluid coul d not be coll ect ed for analysi s
. Less than 0.2 % of the infused do se ofT R IS wa s foun d durin g 24 h ours in gastri cjui c e o rbile. Source : Do w Ch emical, TERC Mi dland, MI Test c onditi on : Six healthy voluntee rs (5 males, 1 fem a le, age s 27-37, weig hing 50-9 0 kg ) and 20 p a tien ts in a su rgical intensive care unit (diag noses rang ed from traumati clesi ons, inte stinal bleedin g , perforated ap pe ndiciti s, and pan cre atitis to rectal a nd g astri c cancer, and aorti c an eurysm) were infused with 121 mg/ k g T R IS of 0.3 mol/L soluti on at a pH of 7.4 over 30 m i nutes in an anteb ra chi a l vein. Conclu sion : TRIS is prim a r ily eliminated via the kidne ys in the urin e. There were no advers e effects related to TRIS treatment in any s ubjec t. Accu mulatio n of TRIS may occur in the b ody if patients with impai re d renal funtion are re peatedly give n TRIS treat ment. Reliabilit y : (2) v alid with rest rictio ns Meets ge ne ra lly acce pted scientific stand ard s , well -do c ume nted an d accepta b le fo r asse ssment

. 09.11.20 06 (63 )
5.11 AD DITIO N A L REMA RKS

52 / 60

## 6. Analyt . Meth. for Detection and Identification Id 77-8 6 -1 Dat e 14.11.20 06

6.1 ANALYTI C AL METHO D S

6.2 DETECTIO N AN D IDEN TIFICATIO N

53 / 60

## 7. Eff. Against Target Org. and Intended Uses Id Dat e 77-8 6 -1 14.11.20 06

54 / 60 7.1 FUNCTIO N

7.2 EFFECTS O N OR GA NIS MS TO BE C O NT ROL LED

7.3 ORGANISMS TO BE PROTECTE D

7.4 USER

7.5 RESISTANCE

## 8. Meas. Nec. to Prot. Man, Animals, Environment Id Dat e 77-8 6 -1 14.11.20 06

55 / 60 8.1 METH OD S HA ND LING AN D STOR ING

8.2 FIRE GUID A NCE

8.3 EMERGENCY MEASURE S

8.4 POSSIB. OF RENDERI N G SUBST. HARMLESS

8.5 WASTE M A NAGEMENT

8.6 SIDE-EFFEC TS DETECTI O N

8.7 SUBSTANCE REGISTERED AS DA NGER OU S FOR GROUND W A TER

8.8 REA C TIVITY TOW A R D S CO NTAI NER MATERI AL

## 9. References Id 77-8 6 -1 Dat e 14.11.20 06

(1) Frau so n, L. (2004 ) Perso nal e-m a il to C. Houtma n dated Ma rch 19, 2004. (2) The Me rck In dex (Eleventh Edition). Merc k& Co., INC. Rahway, NJ , USA. 1989. (3) ANGUS Material Safety Data Sheet. (4) Eilerman, D. and Rudma n , R. (1980 ) Polymo rp hism of crystallin e poly(hyd r oxymethyl) compounds. III. The Struct ur es of cry s talline and plast ic tris(hydroxym ethyl)amin om ethane. J. Chem . Phys. 72(10 ): 565 6-5 666. 15 May 1980. (5) The Me rck In dex (Ninth Ed ition). Me rck & Co., Inc. Rahway, NJ, USA. 1976. (6) Nah a s, G. (1 962) Th e Pharma col ogy o f Tris(hydroxy methyl)amin o methane (THAM). Pharma col o g yReview, 14: 447-472. (7) Gabri e l, C.L. (1940 ) The Nitrop araffins: New Synthe tics for Synth e si s. Ind. Eng. Chem. 32:887 -89 2

. (8) U.S. EPA. (2000) EPIWI N softw are, version v3.11. Un ited States Environmental Protectio n Agency, Office o fPollution Prevent ion an d Toxics, Wa sh ington, D. C. Available at: http://www.epa.g o v/op pt/exposu r e/d o cs/epi suitedl .htm (9) Lhota k , H. (1 996) Partitio n Coeffici ent of Variou s Produ cts (AMP D APD). ANG U S Che m ie GmbH. Repo rt of The Do w Chemi c al Co mpany. (10 ) Lhota k , H. (1 997) Partitio n Coeffici ent of Variou s Produ cts (AMP D APD). ANG U S Che m ie GmbH. Determination of Partition Co effici ent Pow (n-octan ol/wate r ) of AMPD, APD and Corrg uard B-90 acco rdin g to OECD met hod 10 7. Rep o rt of The Do w Ch emical Comp any. (11 ) We st, R.J. (2 004) A re po rt of The Do w Chemical Co m pany, Usi ng ACD Lo g D S uite softwa r e, version 5.0 (200 1). Advance d Chem is try DevelopmentInc ., Toronto, Ontario. (12 ) Troe ste r , L. (2006 ) ANG U S Techni cal d a ta she e t. Perso nal comm unication. (13 ) Bates, R.J. a nd Pinchi ng, G. (194 9) Di ssoc i ation of wea kba se s from ele c tro m otive force measurement s of solutio n s of partially hydrolyz ed salts. J. Res. nat. Bur. Stand. 43:519
­ 526. (14 ) Troe ste r , L. (2006 ) Person al comm uni cation. (15 ) Gon s ior, S. (2006 ). Predi ction of the En vironm ental Distri bution a nd Tra nsport Between Environme n tal Comp artm ents for T r is(hydroxymeth y l)aminom eth ane Usin g Le vel I and Level III Fugacity Models. Draft Repo rt of the Dow Chemical Com pany. (16 ) We st, R.J. (2 004) A re po rt of The Do w Chem ical Co m pany usi ng L e vel I model versio n 2.11, Obtaine d from the Ca nadia n Enviro nm ental Mo d e ling Centre, Trent University, Peterbo r ou gh , Ontario, Ca nada. (17 ) IWL (1 990 ) Institut für gewerbli c he Wa sserwi rtschaft und Leftrei nh altung e. V. IWL - Auftragsnum mer: Lab 59 4. (18 ) McFa rlan d, W.N. and No rris, K.S. (1958) The C ontrol of pH by Buffers in Fi sh Tran sp ort. California Fi sh and Gam e
. 291-3 10. (19 ) Akute Toxizit ä t für Fische OECD Guidel ine 203, 96 h, statisch (1 99 7). AMPD für ANGUS Chemi e GMB H . Steinbeis-Tran sfe r ze ntrum Ange wan d te und Um welt-Ch e mie. (20 ) ANGUS MS DS # AT58, dated 9/13/20 01.

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## 9. References Id 77-8 6 -1 Dat e 14.11.20 06
(21 ) Young, B.E. and Tap p , J. F. (1983 ) Th e Toxici ty of AMP 95 to the bro w n shri mp (Cran gon cra ngo n) a nd plaice (Pl eurone ctes pl ate s sa). Repo rt K349/B for the Central Ele c tri c ity Gene rating B oard, Ma nche ster, UK by Imperi a l Ch e m ical Indu stri es, PLC. (22 ) Pare kh, C. (1 980) AMP-95 Acute Toxicit yE ffects on Dap hnia, 48 Hr L C 50. Rep o rt #255 61 for the Intern ational Mine rals & Che m ical Corpo r atio n, Pharma col ogy Labo rato ries, Animal Scien c e Research by Anal ytical Bio Che m istry Lab ora t ories, Colum bus, MO, US A. (23 ) Adams, N., G ouldin g , K.H., and Do bb s, A.J. (1985 ) Toxicity of Eight Wate r-Sol uble Orga nic Che m ical s to Selena strum cap r ico r n u tum: A Study of Method s for Cal c ulating ToxicValues Us ing Different Growth Paramete rs. Arch. Environ. Contam. Toxi col. 14:33 3­ 345. (24 ) Coe nen, T. M. M. (1990) A c ute Ba cteria Cell Mu ltipli cation Inhibitio n Test with T R IS AMINO 40%. Repo rt of the RC C N O TOX B.V. RCC NOT O X Proje c t 0383 28. The Neth erlan d s. (25 ) Bakteri ente s t (Zellvermeh r ung shem mte s t) v on TRIS AMINO 40% (1990) IWLIns titut für gewerbli c h e Wa sserwirt schaft und Leftreinhaltu ng e. V. IWL - Auftrag s nu mme r: IB/35/90. (26 ) Acute Toxiciti es of Amine s . (1955 ) Hill T op Re se arch Intitute Inc., Miamiville, OH, USA. (27 ) Giroux, J. an d Beaulaton, I. (1962) Recherch e s Pha r macol ogiq u e s Sur Le T H A M

. Societe Pharma cie M ontpellie r. 21:206-217. (28 ) Pare kh, C. (1 980) De rmal Toxicity of P-1826 (AMP ). Repo rt PLR-113 of Pharm a col ogy Labo rato rie s Animal Scien c e Research, Terre Haute, IN. (29 ) Rob e rts, M. a nd Linn, S. (1 961) Acut e a nd Subchroni c Toxicity of 2-Amino

- 2­ hydroxymeth y l-1,3-p r op an ediol. In Vitro and In Vivo Effects of Amine Buffers, A nnal s of the Ne w York Academy of Sci ences. V 92:724-734. (30 ) Thomp s o n , S.W., Allen, M.R., and McDo well, M. E. (19 65) Toxicity Studies with Tris (hydroxymethyl) aminomet hane. U.S. Army M edical Re sea r ch an d Nutrition L a borato ry, Fitzsim o n s G eneral Ho spit al, Denver, CO, USA. 1 March 196 5. (31 ) Baldwin, R. (1961 ) Tris(h ydroxymethyl)amin ometha ne Prima ry Skin Irritatio n Studies. Internal corre s po nde nce of The Do w Ch emical Com p any. May 19, 1961. (32 ) Machl e , W., Scott, E. W. a nd Treon, J. F. ( 1940) The Physiologi cal Response of Animals to Some Simple Mono nitro paraffin s and to Certain De rivatives of These Com p o und s. Jou r nal of Ind u strial Hygien e and Toxicol ogy V22, No.8 (315 -3 32). (33 ) Powe r, C. (1 975) Eye Irrit a tion, Bioassa y Repo rt #2 6775. Pharm a col ogy Lab o r atori e s Re sea r ch an d Develo pme n t, Comme rci a l Solvents Corpo r atio n. May 21, 1975

. (34 ) Toxicolo gy data summ ary on TRIS AMINO, 199 2. In DR-00 1 7
- 352 4-01 9 and T o xicologi cal Profile on Tri s Amino. In DR-0 017 -3 524 -019. The Do w Ch emical Comp any. (35 ) Pare kh, C. (1 982) Intra dermal Sensiti z a t ion Potential of AMP (P-18 26). Rep o rt No. PLR265 of Bio/dynamics Inc, E a st Millston e , NJ. Spon so red by the Internation a l Min e ral s & Chemi c al Co rporatio n, Terre Haute, IN. (36 ) Pare kh, C. (1 982) To pical Sensitization Po tential of AMP (P-182 6). Report No. PLR-266 of Bio/dynamics In c, East Millstone, NJ. Sponso r ed by the Interna t ional Mine ral s & Chemi c al Co rporatio n, Terre Haute, IN. (37 ) CIR: Co smeti cIngre d ient Revie wBoard (199 0)

. A Final Report on the Safety As s e ss ment of Aminomethylprop anol a nd Aminomet hylprop ane di ol. J. Am. Coll. of Toxicol. 9:203-228

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(38 ) Kociba, R. (2 003). Asse ssment of the Potentia l for Skin Sensitization of Structu r allyRelate d Anal ogs Rep r e s e n ted by 2-Am ino-2
- (Hyd roxy methyl)-1,3 -Propa nediol, 2-Amino
- 2
­ Methyl-1-P ro panol, an d 2-Amino-2
- Met h yl-1,3-P rop a nediol. Writte n comm uni ca tion. (39 ) Pare kh, C. (1 981) A Three Month In Die tToxi city Stu d y of 2-Amino-2

- methyl-1-prop anol in Rats. Study 80-2 503 of Bi o/dynamics Inc, East Millstone, NJ. Sp onsore d by the Internation a l Minerals & Chemical Co rp oration, Te rre Haute, IN. (40 ) Griffin, T. (19 90) A One

- YearO r al Toxi city Study of AMP in Dogs. Coulsto n Re sea r ch Incorpo r ated, White Sand s Research Ce nter, Alamog ordo, NM. Amende d by the origin al author o n April 20, 1993. (41 ) Livtak, K.M. a nd McEvoy, G.K. (1990) Keto rola c, an injectabl e no nnarcoti c ana lgesi c
. Clini c al Pharmacy. 9:921-935. (42 ) Ray, T. and Dyso n, P. Tris as a bifu ncti onal cl eaving agent of Stre ptomyce s lividan s DNA. (199 2) Muta g ene sis: 7(5):3 84. (43 ) Ketkar, M. , Gree n, U., Schneid e r, P., and Moh r , U. (1979 ) Investi gation on the carcinogenic burden by air pollution in m an. Intratrach eal instillation studies with ben zo[a]pyre ne in a mixture of Tris buffe r and saline i n Syrian gold en ham sters. Can c e r Letters, 6 2 7 9
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- Methyl-2 -Am i nopropa nol Hydro c hl orid e: A Repro d u c tive / Develo pmental Toxic i ty Sc reening Tes tin CD Rats

. Rep o rt DR-0 038-318 6-0 0 1 of the Dow Chemi c al Co mpany. (45 ) Carney, E.W. and Tho r sru d , B. (2006) AMP: Dermal Developm en tal Toxicity Study in CRL:CD(S D) Rats. The Dow Chemi c al Comp any Re port No: DR-0399 -39 40-0 01 (46 ) Wepi erre, J. and Noel-Hu d so n, M.S. Percutane ou s Abso rption ofTrometh amo ne Hydro c hl orid e: Study on Human Ski n in In Vitr o Diffusion Cell s (1 99 3) Unive r sity of ParisSud, Faculty of Pharma cy. August 9, 1993. (47 ) Samiy, A.H., Ram s ay, A.G., Rees, S.B., and Me rrill, J.P. (1961) Th e Use of 2-A mino-2­ hydroxymeth y l-1,3-p r op an ediol in the M anag ement ofRenal Aci d o s is. In Vitro and In Vivo Effects of Amine Buffers, A nnal s of the Ne w York Academy of Sci ences. V 92:802-812. (48 ) Con ant, J.S. and Hugh es, R.E. (1961 ) The Usefuln e ss of T H AM in Metaboli cAcido s i s
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- 2-hy d r oxymethyl-1,3-pro p a nediol (TRIS) Durin g Cardia c Bypa ss Procedu re s. In Vitro and In Vi vo Effects of Amine Buffers, Anna ls of the Ne w York Academy of Sci ences. V 92:765-782. (51 ) Hoove r , J.E. (1975 ) Remi n g ton's Ph arm a ce utic al Sci ences, 15th E d ition. Mack Publishi ng Comp any, Easton, Penn sylvania, USA. (52 ) Ree s , S.B., Young er, M.D., and Fre edle nder, A. E. (1961) Some I n Vivo and In Vitro Observation s on the Effects of Tris(hyd roxymet hyl)a minometh ane in Diabeti cAcido s i s
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(54 ) Luchsi nge r, P.C. (1961 ) T he Use of 2-A m i no-2-hyd r o xymethyl-1,3-prop ane diol i n the Manag eme n t of Respirato ry Acidosi s
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. (55 ) Brinkm an, G. L., Brunswick, F.W., and Whitehou se , F.W. (19 61) Th e Use of 2-A mino-2­ hydroxymeth y l-1,3-p r op an ediol in the Correctio n of Metaboli cand Re spiratory Acido s i s
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. (56 ) Brinkm an, G. L., Remp, D. G., Coates, E

. O., and McCall Priest, E. (1960 ) The Treatment of Re spiratory Acido s i s with T H AM. Am. J. Med. Sci. 239:341-346. (57 ) Brown, E.S., Bennett, T.E., Bunnell, I.L., Elam , J.O., Evers, J.L., Greene, D.G., Janney, C.D., Lowe, H.J., Nah a s, G.G., and Tarail, R. (1959 ) Effects of THAM Du ring CO2 Breathin g in Man: Ventilation and CO2 Exchang e. The Physiolo gi st. 2:18-19. (58 ) Brinkm an, G. L. (1960 ) Th e Use of THA M to Prevent Hyperve ntilation and Aci d o s is While Breathin g Ca rbon Dioxide. Am. J. Med. Sci. 239:728 -731. (59 ) Resman-Targoff, B.H. (1990) Keterolac: A Parent eral Nonsteroidal Antiinflammatory Drug. DICP, The A nnal s of Phar macoth erapy. 24:1098-11 04. (60 ) Dipalm a , J.R. (1991 ) Ketorola c: An Inject able NSAID. American F a mily Physici an. 4 3 ( 1) :2 07
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. (61 ) Corelli, R.L. and Geri cke, K.R. (1993 ) Renal Insufficie n cy Asso ciated with Intra m iscula r Administratio n of Ketorola c Tro m ethami ne. The Ann a ls of Pharm a coth era p y. 27:105 5­ 1057. (62 ) Tarail, R., Bennett, T.E., Brown, E.S., Bunnell, I.L., El am, J.O., Evers, J.L., Greene, D.G., Jan ney, C.D., Lowe, H. J., and Na ha s, G.G. (195 9) Effects of T H AM Durin g CO 2 Breathin g in Man: Meta bolic a nd Tox ic Effects. Th e Physiologi st, 2:114. (63 ) Brasch, H., T h ies, E., and Iven, H. (1982 ) Pharma co kinetics of TRI S (hydroxym ethyl­ )amin o metha ne in He althy Subject s and in Pa tients wi th Metabolic Acido s is. Europea n Jou r nal of Cli n ical Pha r ma colo gy 22:25 7-26 4.

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## 10. Summaryand Evaluation Id Dat e 77-8 6 -1 14.11.20 06

10.1 END POINT SUMMARY

10.2 HAZ A RD S U MMA RY

10.3 RISK ASSES SMENT

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