2.113 

r::--::--:----::---:-----:--------~----_..___________, 

15.00 20.00 25.00 30.00 35.00 40.. 00 

Solution of 3MBA (100 mg/L) 	 22-Feb-200 

14711b031 2: Diode Array 

Range: 1.443e+1 

1.08+1 

/ 	 Range: 1.966e+1 

1.oe+1 15.73 

10.00 15.00 	 30.00 

[1] Water + test Item (LC·MS sample -2) 

14711b024_' 	 2: DIode Array 

Range: 2.506e+1 

7. 

~ 5. 

0.:'0~1'h1""---

·~rlh~~~~~~~~~~~~~~~~~~~~~ThM 30.00 35.00 40.00 

IIf '11\ 

Solution of test item (100 mg/L) 22.Feb.200 

14711b029 2: Diode Array 

Range: 1.298e+1 

Test item 

I 

1.0e+1 

7. 

~ 5. 25.72 

2. 

O. 

"-r....,...,J,........."TT"1.."...,...,....,TTT'T"T"TTTT.,........,.....,.,..,.,......,......TTT".......-,-rrrrrrr-r"..,.,,..,..,...,l"""T'1rrr"'1'"T"M..,.,.,.,....,.,...,..,.T1 TIme 

6.00 10.00 

7. 

~ 5. 

O. 

20.00 25.00 30.00 

seA 2: Diode Array 

22·Feb-200 

10.00 15.00 20.00 25.00 

Fig. 15-1 	 UV chromatograms of LC-MS analysis for test solution (205 - 300nm) 

(LC·MS sample -2, qualitative analysis of water-soluble converted products), 

-82­

7. 

2. 

o. 

S. 0 

945 
j[2] Sludge + test Item (LC~MS sample ~~ i14711b025 

22-Feb·200 

2: Diode Array 

Range: 2.667e+1 

------------------~~--------

~~nT~~~~~~~~Tn~~~~~~nT~~~Tn~~nTi~ 

a.GO 10.00 15.00 20.00 

[3] Sludge + test Item (LC·MS sample -2) 

14711b026 

1.0e+1 

:it 5. 

2. 

a. . 

25.00 3MID 35.00 40.00 

22·Feb·200 

2: Diode Array 

Range: 2.664e+1 

~~~MT~~~~~~MT~~~~~~~~~~~~~~~nma 

.00 10.00 1 . '20.00 25.00 30.00 

[4) Sludge + test Item (LC·MS sample -2) 

14711b027 

1.0&+1 

7 2.57 

~ 5. 

2. 

0.. 

5.00. 10.00 15.00 

[5J Control blank (LC-MS sample -2) 

14711b028 

1.0e+1 

20.00 25.00 .00 

35.00 40.00 

22-Feb..20 

2: Diode Array 

Range: 2.7241+1 

2: Diode Array 

Range: 2.676e+1 

7. 

2. 

5;00 10.00 15.00 20.00 25.00 30.00 55.0.0 

l't'll \ 2a-elL 1, 12. 

Fig. 15-2 	 UV chromatograms of LC-MS analysis for test solution (205 - 300nm) 

(LC-MS sample -2, qualitative analysis of water-soluble converted products). 

-83­

946 
olution of test item (100 mg/L) 22·Feb·200 

14711b029 1: Scan ES+ 

TIC 

Test Item 2U2; 1.50e8 

5.00 10.00 15.00 

Solution of 3MBA (100 mg/L) 

20.00 

14711b031 1: Scan ES+ 

TIC 

1.5Oea 

3MBA 

I :13.16 

5.00 10.00 15.00 

olution of SBA (100 mglL) 

20.00 25.00 3000 

14111b030 1: ScanES+ 

TIC 

1.50e8 

S8A 

I 

16.78 

5. 10.00 15.00 20.00 

[1] Water + test item (Le-MS sample -2) 

25.00 30.00 

14711b024_1 

Test item 

Fig. 16-1 	 Total ion chromatograms of LC-MS analysis for test solution 

(LC-MS sample -2, qualitative analysis of water-soluble converted product~47 

-84­
[2) Sludge + test item (LC·MS sample -2) 

14711b025 

1.66 

A 

\ 

o Testitem 8 C \E / 

22-Feb-20071,' 

1: Scan ES+ 

TIC 

1.5086 

\ / af 

I ..,. T~~~~~8~.50~~~~~~~~~~~~~~~~~~~~:; I TJme ,........___-O50,;.;.0=-0 10.00 15.00 20.00 35.00 40;00 

[3] Sludge + test Item (LC-MS sample -2) 22·Feb·200 

14711b026 1: Scan ES+ 

A 

1.10 \ 

5.00 10.00 15.00 20.00 25,00 

[6] Control blank (LC.MS sample .2) 

as.1<1 

30.00 

TIC 

1.60ea 

1: Scan ES+ 

TIC 

i.50eS 

14711b02S 1: Scan ES+ 

nc 

1.50e8 

1.88 33.18 

20.00 25;00 ao.OO 35.00 

M, 'TaYlA.K,' 

Fig. 16·2 Total ion chromatograms of LC-MS analysis for test solution 

(LC-MS sample -2. qualitative analysis of water-soluble converted products). 

-85­

948 
[3] Sludge + test item (LC-MS sample ·2) 

14711b02e 

2.67 

6. 

.~ 

o. 

14711b026 1.68 

eo 

UV chromatogram 

(205 - 300 nm) 

5.00 10.00 

Total ion chromatogram 

A 

\ 

11.64 

15.00 20.00 25.00 30.00 

22-Feb-200 

2: Diode Array 

Range: 2.664e+1 

33.0$::-_--, 

35.00 40.00 

1: Scan ES+ 

TIC 

2.68e7 

-~~TM~~~nT~~~~~~~~MT~~~~~~~MT~~~nme 

5. 10.00 15.00 .00 

[3] Sludge + test item (LC·MS sample .2) 

14111 b026 462 (8.539) Cm (452:4n.(438:451 +480:492)x1.5OO) 

1 221 m 

14711 b02S 915 (18.912) em (901 :922-(885:893+931 :936)x1.500) 

1 m 

25.m 

ill 600 BOO 1 1200 

30.00 35.00 40.00 

22·Feb-20l> 

1:ScatI ES+ 

8.22e6 

Mass spectrum of peak A 

1::ScanES+ 

1;81e5 

~~!\Im of peak B 

1400 

Fig. 17-1 	 Mass spectra of water-soluble converted products 

(LC-MS sample -2, sludge + test item, vessel [3]). 

-86­

949 
Mass spectrum of peak E 

14711b0261207 (22.309) Cm (1202:1213-(1196:1198+1219:1224)xl.500) 1: Scan ES+ 

10 460 8.0785 

Fig. 17-2 Mass spectra of water-soluble converted products 

(LC-MS sample -2, sludge ... test item, vessel [3]). 

13] Sludge + test item (LC-MS sample -2) 

14711 b026 946 (17.465) Cm (942:954-(931 :938+959:968)Xl .500) 

1 ~1 

.~ 

1 458 

423 

14711b026 1168 (21.958) Om (1174: 1194-(1166:1171+1196:1201)x1.500) 1: Sean ES+ 

3.82e5 

Mass spectrum of peak D 

22.Feb_200~ 1; Scan ESta.56e5 

Mass spectrum of peak C . 

-87­

950 
[2] Sludge + test item (LC-MS sample -=2) 	 22-Feb~200 

14711bO,~~ 

8 

A 

\ 

9,50 

Total ion chromatogram 1: Scan ES+ 

TIC 

2.72e7 

.~ 

11 5.00 10.00 15.00 20.00 25.00 30.00 35.00 40.00 

14711b025 	 1: Scan ES+ 

8·50 	 m/z5~~~ 2.18 

- 0 "A... 0 roo 0., ,-,,0 t "1 0 0 " , '0 00'" ", .. ",. 0 0 0 , 0 0 " , 0 .... 0 .. ,' 0'''" 0 .., ...., 

5.00 10.00 15.00 20.00 25.00 30.00 35.00 40,00 

14711b025 1: Scan ES+ 

1j 3.<41 m/z = 221 

~J. 0 A, .. 0 .. 0 .. , ..!.,0 0 .. ,. .. 0 .. 0 :or 0 0 , 0 0 .., .. 0 ., 0 0 0 0 .. 0 .. , ' , , .... 0 0 ,. , , 0 .. 0 .. ,'.­

5.00 '10.00 15.00 20.00 25.00 30.00 35.00 4000 

14711b025 1: Scan ES+ 1J ~ ~=

J, 0 ... , 0 .. " ......1.,......'.. ,'0 0 0 10 , , ., , .. , , ' , " .. " 0 ,. 0 .. , .. 0 0 , .. 0 0 , 0 0 0 0 ,. " 0 ,'..... 

5.00 10.00 15.00 20.00 25;00 30;00 35.00 40.00 

14711b025 1: Scan ES+ 

100 Ut aa.14 mlz =281 

14711b025 

-J .""5;00 '1"" 0 ,1.""~,, 10.00 15.00 ",,,..,. 0"" 

20.00 o",! _0'2s.oo .",,., M.,...l,pi!f'¥~ 30.00 35.00 

1: Scan 

40,00 ~ ES-+ 

1.81 	 2.27.5 11 	 '1' mlz =321 

.. "1", 5.00 'I' " 0' , 00 10.00 0 "I\'~ ,0 ..15.0() ,~'-1"''''' 20.00 0 1~....25.oo30J)!) ,""''''~\i4!, If....oM, 35.00 ~..I~I ".~40.00 ;5.... 	 ,... '~,=E:;' 

:100.., .. 0' ,,,, 0" o"'~'~, 0'0" "t. 0 0"0""" ,"0 .,' 0 "1'" 0,0 0 ~ ,n"'" 0 0'" ,,:.47 5.00 10.00 1'5.00 20.00 25.00 30.00 35.00 40.00 , 1: Scan ES+ 

m/z =361 

1.16 

Fig. 17-3 	 Mass spectra of water-soluble converted products 

(LC-MS sample -2, sludge + test item, vessel [2]. mass chromatograms). 

-88­

951 
----- [2] Sludge + test item (LC-MS sample .2) 22·Feb·200 

14711b025 Total ion cnromatooram 1: Scan ES+ 

5.00 

11.50 

'# 

1.1& 

1.66 ­ TIC 

2.ne7 

\ 

A B 33.12 

8.50 

10.00 15.00 25.00 30.00 35.00 40.00 

1: Scan ES+ 

mlz =221 

1.48e6 

5.00 10.00 16.00 20,00 25.00 30.00 36.00 40.00 

147110025 1; Scan ES+ 

1 ~ mlz = 239 

1.5486 

.'# 

11.82 

1.79 

5.00 10.00 15.00 20.00 26;00 30.00 35.00 40.00 

14711b025 1: Scan ES+ 

. 1 17.<18 mlz =323 

7.26$5 

22,31

.,. 

6.0 10.00 15.00 20.00 25.00 30.00 35.00 40,00 

14111b025 	 1: Scan ES+ 

174 m/z =341 1 7.G485 

'if. 

5.00 15.00 25.00 30.00 35.00 40.00 

14711b025 1: Scan ES+ 

1 17.4& mlz::: S63 

4.46e 

5.00 10.00 15.00 20.00 25.00 30;00 36.00 40.00 

14711b025 1: Scan ES+ t7J~4 mlz =379 

1.41e5 

.,. us 

39.41) 

5.00 10.00 15.00 20.00 25.00 

!'t'll\ lao'? . 2.1.2. h .-rq~k,\ 

Fig. 17-4 	 Mass spectra of water-soluble converted products 

(LC-MS sample -2, sludge + test item, vessel [2}. mass chromatograms). 

-89­

1 

952 
7 

[2] Sludge + test Item (LC-MS sample -2) 22-Feb-200 14711b025 . 1.66 	Talal Ion chromatogram 1: Scan ES+ 

TIC 

2.72e7 A B 

\ 	 3$..12 

.~ 

6.50 

5.00 10.00 15.00 20.00 25.00 30.00 35.00 40.00 j14711bOZ5 1: Scan ES+ 

16.111 m/z '" 321 1.81 ! ' 2.27e5 220t 

~ 

5JlO 10.00 15.00 :mOO 15.00 30.00 35.00 40.00 

14711b025 1: Scan ES+ 

1 21.98 mlz '" 42$ 

4.02e5 

~ 

M.1. 

5.00 10.00 15.00 20.00 25.00 30:00 35.00 40.00 

14111bG25 1: Scan ES+ 

Fig. 17-5 	 Mass spectra afwater-soluble converted products 

(LC-MS sample -2. sludge + test item. vessel [21. mass chromatograms). 

953 

-90­

1 

5.00 35.00 

ILf11 \ 

2U18 m/z =441 

4.66e5 

if. 

84 

5.00 10.00 15.00 20.00 25.00 30.00 35.00 40.00 

14711b025 1: Scan ES+ 

21.81 	 m/z =456 

7.57e5 

'tit 

6.00 10.00 20.00 25.00 40.00 

14711.b025 1; Scan ES+ 

1 21.98 mtz=~ 2.46eS 

'tit 

aM 

5.00 10.00 15.00 . 20:00 30.00 35.00 40.00 

14711b025 1: ScanES+ 

21.$& 	 mtz ;::479 

1.8185 
[2] Sludge + test item (LC-MS sample -2) 

14711b025 . 1.66 	Total Ion chromatogram 


A 


\ 


8.50 

5.00 10.00 15.00 20.00 

14111b025 

1 

5.00 10.00 15.00 20.00 

14711b025 

1 

15.00 20.00 

14711b025 

1 

5..00 10.00 15.00 20.00 

14711b025 

1 

5.00 ie.oo 16.00 20.00 

14711b02& 

1 

5.00 10.00 15.00 20.00 

14711b02S 

1 

26.00 30.00 

25.00 30.00 

25.78 

25.00 30.00 

2UO 

22-Feb-200 

1: Scan ES+ 

TIC 

2.72e7 

35.00 40.00 

1: ScanES+ 

7.2565 

35.00 40.00 

1:ScanES+ 

mIz=42& 

9.7885 

35.00 40.00 

1: Scan ES+ 

mlz= 

1.3586 

17 •• ' mlz=323 

26.00 30.00 36;0040.00 

1: Scan ES+ 

m/z =480 

1.2Se6 

26.00 30.00 3ttOO 40.00 

1: Scan ES+ 

mlz =-465 

5.78e5 

25.00 SO.OO 35.00 40.00 

1: Scan ES+ 

m/z =481 

2.SSe5 

5.00 10.00 15.00 20.00 26.00 SO.OO 

I~f] 1 \ Mo7, )..., 12.. 

Fig. 17-6 	 Mass spectra of water-soluble converted products 

(LC-MS sample -2. sludge + test item, vessel [21. mass chromatograms). 

-91­

954 
1 

[3] Sludge +test Item (LC·MS sample -2) 22-Feb-200 

14711b026 

~S4 \7.{ \.1 2578 38,06 

--.......II.M.I.".,....,.. 

lu.....-~ 

Total ion chromatogram Test item 1: Scan E5+­

1M BeE TIC 

A D I 33,14 26897

a# 

1j 5.00 10,00 15.00 20.00 25.00 30.00 3500 40,00 

14111 b026 	 , 1: Scan ES+ 

8~'52 m/Z6~3!;~ 2.18 

# .. l,... OJ'" ",~" ""''''"" ", .. ,1" "'"' .• ''' 'j' .. .,,,,,,,, .. r.... ' .. ,,' 5.00 10.00 15.00 20.00 25.00 30.00 35.00 40.00 

14'l11 b026 1: Scan ES+ 1:1 	 8.54 mlz :: 221

"J., AI' "'t' .. ",t""".. ,', .. ,~r",,, "'1''' ", .. ,""""" ,,,,, , .... " , .. ,".'" 

5.00 10.00 15.00 20.00 25.00 SO.OO 35.00 40.00 

14711 b026 1: Scan ES+ 1:) 8.52 m/z::' 

j ,,'" ',', ,,.,,,,,!.j , , , , , , • , , , ' , , , t· ,,,,',,.,,,,,,,,,,,'.,.". ,.,.,,,,',,,,,,,•,"',.. 

5.00 10.00 15.00 20.00 25.00 30.00 35.00 40.00 

14711b026 1; Scan E8+ 

10J 	 T 33'10 mlZa~

#J fI J\.rz'~ \f'! tI, II'~"I ,','b, i • ••t" ,'I,. III f Ii, 1'1'; I., "ftj" 11'./; 1'.*"1'L1"~ ",'1./1'1'1

5.00 10.00 16.00 20.00 25.00 30.00 35.00 40.00 

14711b02e 1: Scan ES+ 

1~ ~ t ~ ~~ ~::m ~~ 

",J"...", "t'''"" '" ,., 'I' ,t,.,r,l..,II'" 10104• .,.,...,.., ",... " 

5.1)0 10.00 15.00 20.00 25.00 30.00 35.00 40.00 

14711b026 1: Scan E8+ 

':] ,.... 	 mlzo.,. 

"J,,.It, ,'''' ,.. ,,' •,'r' ,."",.,.. ,L,..,,,......."'..,,, "~,, ..,,
......... I" .".,.....,'­ .5,00 10.00 15.00 .20.00 25.00 30.00 35.00 40.00 

14711b026 . 1: Scan ES+ 

1j''. ""1$$ j1Ua 	 . ml29~

#.. 	 . . 2U2 31.0 . f 

,ifIJ\.,Wl't.· ,,~~tf'" jt:AI:!\-:-i;' ~~ 1"I~1t~"/'U1¥1:,.;Jt'111''''''';fi\·.f.I~~~~h~ TIme I 

5,00 10.00 .~. .--15.0020.:..00 .:.25.00_ 30.00 ~....!§.,~__ 40.00 . 

it{ 'll , 1"'0 '1 , '1, 2.1.. ~L To-rtA.k\ 

Fig. 17-7 	 Mass spectra of water-soluble converted products 

(LC-MS sample -2, sludge + test item, vessel [3], mass chromatograms). 

-92­

955 
[3] Sludge + teat Item (LC-MS sample -2) 

14711b026 Total ion chromatogram 1.118 

A 

E Test item 

BCD I 

14711b026 

1 ex:! 

Ul 

1-4711b026 

1 _10 

1.1' 

14711b026 

1 

14111b028 

14711b026 

1 

'" 

14711b028 

1 

5.00 

5.00 

5.00 

5.00 

5.00 

lit'll\ 

\ 

8,.5,4 

10.00 

10.00 

10.00 

'0.00 

10.00 

10.00 

\ / ~u( 26.78 

20.00 25.00 30.00 

15.00 20.00 25.00 30.00 

17.60 

.21.1' 

15:.00 20.00 ~5.00 30.00 

Wt 

1!1.oo 2tl00 26.00 30.00 

15.00 20.00 25.00 30.00 

17M 

tUO 20.00 26.130 30.00 

17$2 

22-Feb-200 

1:ScanES+ 

TIC 

2.8887 

35.00 40.00 

1: Scan ES+ 

mlz =221 

1.6186 

35.00 40.00 

1: Scan ES+ 

m1z= 

1;7 

35.00 ' 40.00 

1: Scan ES+ 

m/Z= 

6.17eS 

36.00 40.00 

35;00 

35.00 

1: Scan ES+ 

mlz =341 

6. 

40.00 

1: SCan ES+ 

mIz= 383 ... 

40.00 

1: SCan ES+ 

mJz .. &1$ 

1.6385 

Fig. 17-6 Mass spectra of water-soluble converted products 

(LC-MS sample -2. sludge + test item, vessel [3], mass chromatograms). 

-93­ 956 
[3] Sludge + test item (LC·MS sample -2) 

14711b()26 . tee ToIallQn chromatogram 

B C 

\1 33.14 

.l#! 

5.00 10.00 15.00 20.00 25.00 30.00 35.00 

14711b026 

10 1.86 

111.73 21.9<1 

5.00 10.00 15.00 20.00 30.00 35.00 

14711b026 

21M 1 

5.00 10.00 15.00 20.00 26.00 30.00 35.00 

14711b026 

lit 

25.77 3$.811 

21:88 1 

'# 

3&;17 

.,. 

5.00 10.00 15;00 20.00 2$.00 30.00 

14711b026 

21.88 1 

5.00 10.00 15.00 20.00 26.00 ·30.00 35.00 

14711b026 

21.98 1 

35.00 

35.00 

5.00 

(tt 111\ 

'# 

28.," 

5.00 10.00 15.00 20.00 25.00 3O~00 

14711b026 

21;" '1 

~ 

1.77 

22-Feb-200 

1: Scan ES+ 

TIC 

2.6687 

40.00 

1: Scan ES+ 

mlz =321 

21985 

40.00 

1: Scan ES+ 

m/z= 4 

4.00eS 

40.00 

1: Scan ES+ 

m/z:;; 441 

4.6485 

40.00 

1: Scan ES+ 

m/z =458 

6.5ge6 

40.00 

1: Scan E$+ 

mlz =483 

1.S1e6 

40.00 

1: Scan ES+ 

mlz =47t 

1.58e5 

Fig. 17·9 Mass spectra of water-soluble converted products 

(LC·MS sample -2. sludge + test item, vessel [3J. mass chromatograms). 

-94­

957 
[3] Sludge + test item (LC-MS sample ·2) 22-Feb-200 

14711b026 . 1: Scan ES+ 

A 

\ 

8.54 

1.68 Totailon chromatogram TIC 

2me1 

.~ 

5.00 10.00 15.00 20.00 25.00 30,00 35.00 40.00 

14711b026 1: Scan ES+ 

17.>48 m/z=323 

6.i7e5 

5.00 10.00 15.00 20;00 25.00 30.00 36.00 40.00 

141'flb026 1: Scan ES+ 

14711b026 1: Scan ES+ 

1 m/z= 

1.1 

26.77 

6.00 10.00 1S.00 20.00 25.00 30.00 35.00 40.00 

14711b026 	 1: Scan es+ 

22JJ1 mlz =4&0 10 1.4118 

6.00 10.00 15.00 20.00 25.00 30.00 35.00 40.00 

14711b026 1: Scan ES+ 

m/z :;:4$$ 1 ~U1e5 

15.00 20.00 25.00 30.00 35.00 40:00 

1: Scan ES+ 

(lC-MS sample -2. sludge + test item. vessel [3], mass chromatograms). 

958 

-95­

1 

5.00 10.00 

22.31 

15.00 20.00 25.00 

m/z=425 

1.02 

40.00 

mIz =481 

U8e5 

Fig.17-10 Mass spectra of water-soluble converted products 
[4] Sludge +test item (LC-MS sample -2) 22-Feb-200 

14711 b~~:o Total ion chromatogram 1: Scan ES+ 

1 A C TIC 

2.76e7 

3U4 

.;fl \ 

8.1>4 

5.00 10.00 15.00 20.00 25.00 30.00 35.00 40.00 

14711b027 1: Scan ES+ 

1J 8.54 mlz =137 

'}..l ...,'··",l,.,,,'1 .. " , ' ,,,I ' , , •I ' " ., " • ., ••', " ••• ., " " , ' ... , ' " , , ' .. , 5.00 10.00 15.00 20.00 25.00 30.00 35.00 40.00 

14711b027 1: Scan ES+ 

'j,,A,. •. 'I , , •• ,I,,..,",".,...:r.",..".. "., ".. ,........,,.........,':t 221 


5.00 10.00 15.00 20.00 2.5.00 30..00 35.00 
 40.00

14711b027 1: Scan ES+ 

8,6l1 mlz =239 

'j.....""'I ' , .. , ,t,,'",,,,, 'I' , "I" .,," ,.,'.. "" " ,•, ". ,,' " " ,,'. ," ' " ,.'8' 

5..00 10.00 15.00 20.00 25.00 30.00 35.00 40.00 

14711b027 1:ScanES+ 1j Sf M .1O mlz3~ 

'3" '".·,.I' ",jL,,,.,"~ ,",",,,,,r'" ,t, •to" ..'f' ....'/. ",.., 1....iI. "I, 5.00 10.00 15.00 20J)0 25.00 30.00 35.00 40.00 

14711b027 1: Soan ES+ 1j 1.85 m/z =321 t 11l.l1l1 :lUI 2.00e 

'" ..!1m ., ,.."'.,,.,,....... ".,.,1..,.1.,"'"" ,.,\ .~,. "",.....,
...1\ 

6.00 10.00 15.00 20.00 25.00 30.00 35.00 40.00 

14711b027 1: Soan ES+ 1:] 18,91 m/z = 339


"'1 ....""" ," """" ........\'.,.' t.".",,.., ",·,,·""".To"'" ,.• ",. ,..,'"....­ 5.00 10.00 16.00 20.00 25.00 30.00 35.00 40.00 

~4j711b0121':1 16.95 1:~~::~

\lUl1 

."., It'\''N t,l/IjAI';""'uf,'*4 r'1·tA~¥r'~ 4014 '''flI1.'I't·Jt~'''''jlIr\''''I'¥~+''''t~.;'''·f.~ TIme 

5,00 10.00 15.00 20.00 2s.o0 30.00 35.00 40.00 

IIf 11 l \ 19o7, 9... 1.."l.. h \\o..r-L\k,' 

Fig, 17-11 Mass spectra of water-soluble converted products 

(LC-MS sample -2, sludge + test item, vessel [4J. mass chromatograms). 

959 

-96­
[4] Sludge +test item (LC·MS sample -2) 22·Feb·200 

14711b027 1: Scan ES+ 1.70 Total ion chromatogram 

\1 30.69 \ 22.29 21i.77

D 

\/E 

I 	

TIC 

B c Test item 	 2.78e7 

33.14 

.at 

7 

6.00 10.00 15.00 20.00 25.00 30.00 35.00 40.00 

14711b027 1: Scan ES+ ~ 17.60 m/z =221 

1.40et1 

1.77 "#­

5.00 10.00 15.00 20.00 25.00 30.00 35.00 40.00 

1: Scan ES+ 

17.50 	 mlz =239 

1.51t6 

5.00 10.00 16.00 20.00 30.00 35.00 40.00 

14711b027 1: Scan ES+ 

17.110 mlz =$23 

5.21e5 

22.31 

il 

5.00 10.00 15.00 20.00 25.00 30.00 35.00 40.00 

14711b027 	 1: Scan ES+ 

1 17.48 m/z =341 

S.53e5 

MO 10.00 15.00 20.00 26.00 ~OO 85.00 40.00 


14111b027 1: Scan ES+ 


5.00 10.00 15.00 20.00 25;00 30.00 35.00 40.00 


141.11b027 1: Scan ES+ 
 17.62 	 m/z =379 10 1.48e6 

IP. U1 

15.00 

l(Jot'] ... 1, 1.l.. tv\, lcnr~k\ 

Fig. 17-12 Mass spectra of water-soluble converted products 

(LC-MS sample -2, sludge"" test item, vessel [4], mass chromatograms). 

960 

-97­

1 17.50 mlz =363 

a 

'* ,,1$1 

25;()OSG.QO 
1 

[4] Sludge + test item (LC~MS sample .2) 22-Feb.200 

14711b027 1.70 Total ion chromatogram 1: Scan ES+

TIC B C 278e7 A 33.14 

.# \ 

8.54 

T 

\! 

5.00 10.00 15.00 20.00 25.00 30.00 35.00 40.00 14711bO:27 1: Scan ES+ 1.85 	 mlz;;; 321 

21.98 2.0065 

16.89 

it 

5.00 10.00 15.00 20.00 25.00 30.00 35.00 40.00 

14711b027 1: Scan ES+ 

1 2U8 m/z =<423 

3, 

~ 

5.00 10.00 15.00 20.00 25.00 30.00 35.00 40.00 

14711b027 1: Scan ES+ 

1 218& m/z =441 

4.4985 

~ 

••1" 

5,00 10.00 15.00 20.00 25.00 30.00 35.00 40.00 

14711b027 1: Scan ES+ 

1 2U4 mlz =458 

S.06a 

'/I. 

5.00 10.00 16.00 20.00 26.00 30.00 35.00 40.00 

14711b027 1: Scan ES+ 

5.00 10.00 15;00 20.00 25.00 30.00 35.00 40.00 

14711b027 	 1: Scan ES+ 21 .... m/z =47. 1.~ 

.,. 

1.&7 

3O.U 

{ l.{- fj l\ lo07, L, ~2. 

10.00 15.00 20. 

1 2U8 m/z =<183 

2.18es 

.,. 

1.17 WIG 

Fig. 17-13 Mass spectra of water-soluble converted products 

(LC-MS sample -2, sludge + test item, vessel [4J. mass chromatograms). 

-98­

961 
7 

[4] Sludge + test item (LC-MS sample -2) 22-Feb~200 . 14711b027 . 1: Scan ES+ 1.70 Total Ion chromatogram TIC 

\ 

B 

\1 

C 2.78e7 A 3U4 

.<It 

8.5.4 

5.00 10.00 15.00 20.00 25.00 30.00 35.00 40.00 14711b027 1: Scan ES+ 

17.50 1 m/z =323 

5.21e5 

22.31 

'it 

5;00 11100 15.00 20.00 25.00 30.00 35.00 40.00 

14711b027 1: Scan ES+ 

1 22.31 mlz :;425 

9.11e5 

~ 

21$.77 

5.00 10.00 15.00 20.00 25.00 30.00 35.00 40.00 

14711bG27 1: Scan ES+ 

1 22.28 mlz=44S 

1.18e8 

<It 

5.00 1D.OO 15.00 20.00 2s.o0 30.00 as.oo 40.00 

14711b027 	 1: Scan ES+ mlz ::_ 1 22.28 

tileS 

~ 

6.00 10,00 15•.00 20.00 26.00 30.00 as.oo 40.00 

14711b027 1: Scan ES+ 22.21 m/z =465 

5.9385 

'at 

5.00 to.no 15.00 20.00 25.00 30.00 35.00 4D.CO 

14711b027 1: Scan ES+ 

ZUt 	 mlz =46.1 

3.2ge5 

~ 

Time 

10.00 16.00 20.00 25.00 	 40.00 

l.o-o~, 1. 12.. 111\ _TQ.. r1A KI' 

Fig. 17-14 Mass spectra of water-soluble converted products 

(LC-MS sample -2, sludge + test item, vessel [41. mass chromatograms). 

-99­

1 

962 
Study No.14711 

Peak A 

...... -.... -_ ..................... -_ ............ -............ --_ .................................................... ---. 


· · 


· 
 · · · · 

m/z Fragment ions 


261 (M + Na)+ 

239 (M + H)+ 

221 

137 

(M - H20 + H)+ 

( M - 3MBA + H20+ H t 

Fig.17 - 15 Mass spectra of water-soluble converted products (LC-MS sample -2, 

sludge +test item, presumed fragment ions of peak A). 

Date J..C907·S-J I Name 7. I<ff~~p.../ 

-100­

963 
6 

[5] Control blank (LC-MS sample -2) 	 22-Feb-200 

14711b028 	 1: Scan ES+ 

'~l TTol"'oooh,,,,,,log"'m 	 ,.i:.~ 

"~,, '1""1 ""1""' .•• ., n. ;;:" ii·;; ;,;;; ....2::::::: 5~OO 10.00 15,00 20.00 25.00 30.00 .35.00 40.00 

14111 b028 1: Selin ES+ 111.83 miL =13., 

.. \.L••~~~~,..~663" 5.00 1OJlO 15.00 2tlOO 25.00 30.00 35.00 40,00 

14711b026 1: Scan ES+ 1:11Ul 

$ 	 m1Z1~8: 

:1 ,....,...,..,.,.,.,'" 5.00 10.,00 ..

1.5.00 

... 

20.00 

1\...." ...25.00 """,.4\'10, 80.00 ~,L.""~''''M-II 35.00 40.00 

14711b028 1: Scan ES+ j T 	 mll,~l: 

"'ll.¥.."_,,, 5,00. 10.00 """"t .... 1.6.00 'I..W!I'No 

20.00 

/!~.1.,...,~1"" 28.00 ...

30.00 .,..!~~ 35.00 40,00 

14711bD28 1: Scan ES+ 10J ~3.16 m/z =261 

J " ~, ,I I I' ,.." '" I' " " ",,,"'rO II """ "I 't'"'' " •.t¥I • ., 'Io/'fL11."'11...... 5.00 . 10,00 15.00 20.00 25.00 30.00 35.00 40.00 

14711b028· 1: SCln ES+ 'I ~=~ '1,-,.,..,,-""'1",,11'."'......"'.,." .... /'I .... " ....., "..,14\ooj ....,./w..,..,.:­

5.00 10.00 16.00 20.00 25.00 30.00 35.00 40.00 

14711b028 	 1: Scan ES+ 

t ~=_ 

2.77e4 

['+t-t \\ loa!\ '1,-22.. M .1CAY"Vl.k,' 

Fig. 18-1 	 Mass chromatograms of water-soluble converted products 

(LC-MS sample -2, control blank). 

-101­

964 
Control blank (LC·MS sample -2) 

4711b028 

Total ion chromatogram 

5.00 10.00 15.00 20.00 30.00 

5.00 10.00 15.00 20.00 30.00 

Mass chromatograms of water-soluble converted products 

1: Scan ES+ 

mlz =221 

35.00 40.00 

1: Scan ES+ 

m/z= 

1 

35.00 40.00 

1: ScanES+ 

mlz= 

(LC-MS sample -2, control blank). 

-102­ 965 
[5} Control blank (LC·MS sample ·2) 

14711b028 1.88 Total ion chromatogram 

_ilt 

8 

5.00 1O,GO 15.00 20.00 25.00 30.00 

147f1bC28 

1 1.$6 

\\1: 

5.00 10.00 15.00 20.00 25.00 30.00 

14711b028 

~ 

5.00 10.00 16.'00 20.00 26.00 30.00 

14711b028 

1 UlS 

~ 

5.00 10.00 20.00 26.00 SO.OO 

14711b028 

1 

'tit 

5..00 10.00 16.00 20.00 25.00· 30.00 

leel \, 

Fig. 18-3 Mass chromatograms of water-soluble converted products 

(LC-MS sample -2, control blank). 

-103­

22·Feb·200 

1: Scan ES+ 

TIC 

2.66e7 

35.00 40.00 

1: Scan ES+ 

m/z '" 321 

1.86e5 

35.00 40.00 

1: Scan ES+ 

m/z =423 

3.00e4 

40.00 

1: Scan ES+ 

mlz = 441 

1.248 

35.00 40.00 

1: Scan ES+ 

m/z= 458 

1.1685 

35.00 40.00 

1: Scan ES+ 

mlZ= ...es e.611a4 

3$.00 40.00 

1: Scan ES+ 

m/z =47 

6.15e4 

966 
Control blank (LC·MS sample -2) 

14711bG28 . 1.68 Total ion chromatogram 

.'lIt 

14711b028 

5.00 

5.00 

10.00 15.00 

10.00 15.00 

20.00 25.00 3CI.OO 

20.00 25,00 3Q.00 

Mass chromatograms of water-soluble converted products 

(LC-MS sample -2, control blank). 

35.00 40.00 

1: Scan ES+ 

mil. = 

35.00 40.00 

1: Scan ES+ 

m/z = 

.00 

1: ScanES+ 

m/z= 

967 

14711b028 

10 1.118 

-104­
StudyNo. 14711 Wavelength 190.00 - 340.00 

Date 

Sample 

Solvent 

Reference 

Scp. IS, 2006 

Test item 

Acetonitrile 

Scale Limit 

SlitWUlth 

Scan Speed 

Sampling Pitch 

I.MOO O.lHIOO 

(UV)2.0nm 

200mnlmin 

0.500000 

Mooo- -0,\000 

:...0... ~.''I 

).. .. ~,. ',flill 

Cell 10mm x lOmm, quartz 

Instrument rASCO V-560 

Photometric Mode Abs 

Analyst 

Note H.~~ 

SO.OmgIL 

Chemicals Evaluation and Research Institute, Japan Kurume Laboratory 

1.5r-------------------------------------____________~ 

Abs 

A forHPLC 


(2IOnm)
 0.5 

o - - - --- ---- ----- - -- ••:.:"::_"!"-""-----------------1 

-0.1 L-L___....;L___-L___....L-___-'-___...I..-_--' 

190 200 250 300 340 

Wavelength[nm] 

PeakLlst 

1: t94.00 ( 0.9057) 

Fig.19 UV spectrum of test item. 

-105­

968 
Standard solution 25.0 mgll 	 10-Dec-200 

1471';005 	 4: ScanES~ 

1 TIC 

1.24&5 

Total ion chromatogram 

/ 

% 

tI,(11 

.2.00 400 8.00 

Standard solution 26.0 mgIL 

14711a005 35 (3.B39) em (33:38-(40:42+30:32)x1.500) 	 4:ScanES1 185.2 	 9.41!i3 

Mass spectrum of pentaerythritol 

(M + CH3COof 

I·_·_··_··· ___ ______ ··t ___ ... , ··~ 

, 


(M + HCOO)­

969 

Fig. 20 - 1 Mass spectrum of pentaerythritol, 

-106­
tandard solution 25.0 mg/L 10-Dec-200 

14711a005 4: Scan ES10 Total ion chromatogram TIC 

1.24eS 

3.01 

18~~~~~~~~~~~~~~~~~~~~~~~ 

14711a005 4: Scan ES3.114 m/z = 171 

9.2263 

1 

% 

o~~~~~~~~~~~~~~~~~~~~~~~~~ 

14711a005 4: Scan ES10 m/z =181 

2.3084 

% 

o~~~~~~~~~~~~~~~~~~~~~~~ 

14711a005 4: Scan ES1 3 m/z =195

3.8764 

Fig. 20 - 2 Mass spectrum of pentaerythritol. 

(mass chromatograms) 

-107­

970 
Solution of 3MSA (50.0mg/L) 

14711xoo2 

23-F.IJ..200 

1: Scan ESTIC 

4.0184 

1 

% 

% 

Total ion chromatogram 

/ 

Elli::1 ~~~~~~~~~~~~~~~~~~~~~~TIme 

. (M - Hf 

17.50 20.00 

23-Feb-200 

1: Scan E8­

9.63e3 

Mass spectrum of 3MBA 

.-............................................., . .. 

~~~.rnM~Mm.~~mm~~~~~~~~~~~~~~~~ 

100 120 140 160 180 200 220 240 2$0 280 300 320 340 

itt'1ll It>(1 , 2.~ 1.3 r"TaI"L.,.k~ 

Fig. 21 - 1 Mass spectrum of 3MBA. 

-108­

971 
Solution of 3MBA (50.0mg/L) 23..feb-2.00 

14711xOOZ 1: Scan ES10 4.85 Total ion chromatogram TIC 

4.0184 

16~~~~~~~~~~TN~~~~~~~~~~~~ 

14711x002 ,1: Scan ES1 m/z =11 

2.7 

% 

0.00 2.50 5.00 7;50 

2007, 1.1) 

Fig. 21 - 2 Mass spectrum of 3MBA 

(mass chromatogram) 

-109­

972 
10 

Solution of SBA (60.0mg/L) 23-Feb-200 

14711x001 1: Scan ES11.37 TIC 

268e4 

Total ion chromatogram 

% 

fB;;:1 

15.00 

Mass spectrum of SBA 

I ?J H0yYS,-~. & f S OH 

MaSs number (M);238 

I tr11 \ 2ao~.. 1.~).3 

Fig. 22 - 1 Mass spectrum of SSA. 

-110­

973 
Solution of SBA (50.0mg/L) 23-Feb-200'i! 

14711xOO1 1: Scan Es-l 

11.37 10 Total ion chromatogram TIC 

2.68&4 

• -.4 

1a~~~~~~~~~~~~~~~~~~~~~~~~ 

14711><001 1: Scan ES1 11.37 m/z:: 237

1.74e4 

0.00 2. 5.00 7.50 10;00 12.50. 15.00 

{tf'lll "loo?, L 1.l 

Fig. 22 - 2 Mass spectrum of SBA. 

(mass chromatogram) 

-111­

974 
IT 

60 

50 

C:-H 

~tr, 

-~-~--.------~--.--.--.- I .. I 

, 

,

, 

I 

__ ~ __ ~ ! __ ~ ___ ~ _____ l __ • __ ~wP_. ___ ~ __ 

0:0 

Str. 

, I 

-------------~-~- .. --~.--~ .-------.-~~-~----------~--.-------------~-- • 

4000 3000 2000 1500 1000 800 

l/em 

Instrument ~ Shimadzu IRPrestige-21 

Study No. : 14711 

Sample : Test item 

Method : Neat 

Date ! September 19, 2006 

Name :H\(::.~ 

Fig.23 ·1 IR spectrum oftest item measured before experimental start. 

-112­

975 
• • 

40 

100 

80 

, 
 8O~--··---·----i 

29' 


--.--------- -_._--­

.•• -._.-•. _••.•-------­

------- ----.---­

------------.. -----------­

, 

..! 

- ... ---- --- --"'1-- - ,.., - - - .. - - - ­ ..... - - - - - ......... - - ­ , 

... ---­ ---­' ... -- -:-­

I

, 

I 

. . -.. -..... --------... ---s- .. 

r... ---.... -... --.. -~ --- ---.... -------:- "!' 

" 

I 

• 

I 

I 


I 


OI-r~~~r_~4r_r~~.-r_+_~_r~~~r_r_,__r_r~--r_r_~or_r_; 

4000 3000 

Instrument 

Study No. 

Sample 

Method 

Date 

Name 

2000 1500 1000 500 

lIcm 

: Shimadzu IRPrestige-21 

: 14711 

.: Test item 

: Neat 

: February 26, 2007 

: fjzt ak 

Fig.23- 2 IR spectrum oftest item measured after experimental completion. 

-113­

976 
", 

Reference IR spectrum supplied by sponsor. 

-114­

977 
Mff:I yiiA1t~~W~ffiffi1iJf5E~~ 

~$* iIIS !fftg£ ~ 

{~~~WO)~~& v~~~O)m}fllJ h: MT 6 r~f=tn-r 

{~6 ~~Mff~~~MT 6 £~~-fr1ltiB~1itl£~f1~~~£ 

~, r~e~~1J&~~~~v'--Ct'j:, ~m{~¥:~ji~~f*6 

~~slfLr.J~~~-fl~m{~¥:~j(& V~=fI~tJ!{~* 

~:w,~t* 61f1ft1:O)wm~0)~ ~ ~~7EfJ) 6~'~m 4 

~~~mJET 6 ~~1itl£~ ~~ MT 6lttf§~cil~ L- --Cv' 6 

2:. C: :fr1it~ L *To 

t.ljO, 1lt~O)fr~WjrFl~t'j:, *1lt~.O)~ft s 7J~ G 

1m:' L- -C 3 iFrFl~ C: L-*To 

~JJt 1 6 &f: 1 2 J1 2 2 F.3 

MfB$A{~~~j{~fnIi."6jf~#l~ 

!Aii*.~m 

.~.!A.*$g/.-T§2.7% 

~~~~~,~~~~~&V~~~~. 

978 

,,' CERI 


Receipt No. 827-08-D-3387 

STUDY CODE: XlS-0908 

FINAL REPORT 

STABILITY ANALYSIS OF KarenzMT (Mixture of PEMB and PE3MB), 


HOMOGENEITY, STABILITY AND CONCENTRATION ANALYSES OF THE 


TEST SUBSTANCE FORMULATION 


March 2009 

Hita Laboratory 


Chemicals Evaluation and Research Institute, Japan 


-i 979 
X18-0908 

STATEMENT 


TITLE OF STADY 

Stability Analysis of KarenzMT (Mixture of PEMB and PE3MB), Homogeneity, 

Stability and Concentration Analyses of the Test Substance Fonnulation (Study Code: 

Xl 8-0908) 

I, the undersigned, hereby declare that this report provides a correct English translation of the 

final report (Study Code: X1S-0908, issued on March 16,2009). 

Yuji Kusune, M.S. Date 

Hita Laboratory 

Chemicals Evaluation and Research Institute, Japan 

-ii­ 980 
XlS-0908 

GLP STATEMENT 


Hita Laboratory 


Chemicals Evaluation and Research Jnstitute, Japan 


Sponsor: _____ Sho:wa Denko K.K. 

Title: Stability Analysis of KarenzMT (Mixture of PEMB and PE3MB)' Homogeneity. 

Stability and Concentration Analyses ofthe Test Substance Formulation 

Studv Code: Xl &:-_Q908 

I, the undersigned, hereby declare that this study was conducted in compliance with "Concerning 

Standard of the Testing Facilities Conducting the Test Relating to the New Chemical 

Substances" on Japanese GLP (Notification No. 1121003 of the PharmaceuticaJ and Food Safety 

Bureau, MHLW, No.3 (November 17,2003) of the Manufacturing Industries Bureau, METl & 

No. 031121004 oftile Environmental Health Department, MOE (November 21,2003»). 

I also confirmed that this report accurately reflected the raw data and the test data were valid. 

Study Director: Signed in original __ March 16, 2009 

Yuji Kusune, M.S_ 

-iH­ 981 
X18·0908 

QUALITY ASSURANCE STATEMENT 

Hita Laboratory 


Chemicals Evaluation and Research Institute, Japan 


Sponsor: Showa Dcnko K.K. 

Title: Stability Analvsis of K,!!:cnzMT (Mixture of PEMB and PE3MB). lIomogeneity, 

Stability and Concentration Analvses of the Test Substance Formulation 

Study Code: X 18-0908 

This study was inspected by Quality Assurance Unit of Hita Laboratory, Chemicals Evaluation 

and Research Institute, Japan. The dates inspected and the dates reported these res lilts to the 

study director and management are as follows 

Date Reported to Study Phase Date of Inspection Director and Management 

Protocol October 10, 2008 

lR Spectrum of Test S October 14, 2008 

Homogeneity and Stability 

Analyses of Test Substance October 14,2008 October 14, 2008 

Formulation 

Confinnation of the Answer from October 17, 2008 October 17, 2008 Study Director (Protocol) 

Amendment to Protocol October 27,2008 October 27, 2008 

Concentration Analysis cfTest November 6, 2008 November 6, 2008 Substance Formulation 

Amendment to Protocol (No.2) November 10, 2008 November 10,2008 

Raw Data and Draft Final Repmt February 24, 2009 February 24, 2009 

ReinspeClion of Raw Data and February 27,2009 February 27, 2009 Draft Final Report 


Amendment to Protocol (No.3) 
 March 12,2009 March 12, 2009 

Draft Final Report (Second Time) March 12,2009 March 12, 2009 

Final Report March 16, 2009 March 16, 2009 

I, the undersigned, hereby declare that this report provides an accurate description ofthe methods 

and procedures used in this study, and that the reported results accurately reflect obtained raw 

data. 

Head, Quality Assurance Unit: March 16, 2009 

Ryuichiro Mizuguchi, B.S. 

iv 982 
X18-0908 

TABLE OF CONTENTS 

Page 


TITLE ................................................................................................................................ 3 


SPONSOR ......................................................................................................................... 3 


'l'ESTING FACILITY ............................................................_ ........ _............................... 3 


PURPOSE OF STUDY ..................................................................................................... 3 


GLP COMPLIANCE ........................................................................................................ 3 


PERIOD OF STUDY ........................................................................................................ 3 


STORAGE AND RETENTION PERIOD OF DATA ........................................................ 4 


RETENTION OF ORIGfNAL DOCUMENTS ................................................................ 4 


STUDY DIRECTOR AND PERSONS CONCERNED WITH THE STUDY AND THE 


OPERATION ...............on.................................................................................................. 4 


APPROVAL BY AUTI·IOR .............................................................................................. 4 


SUMMARY ...................................................................................................................... 5 


MA TERlALS ............... ..... ................................................................................................ 6 


1. 	 TEST SUBSTANCE ................................................................................................... 6 


METHODS ........................................................................................................................ 7 


1. 	 SUBJECT STUDY ..................................................................................... ;............... 7 


2. 	 STADlLITY ANALYSIS OF THE TEST SUBSTANCE ......................................... 7 


3. 	 HOMOGENEITY, STABILITY Al\D CONCENTRATION ANALYSES OF THE 


TEST SUBSTANCE FORMULA TION .. ;.................................................................. 8 


THE ENVIRONMENTAL FACTORS THAT MIGHT IIA VE AFFECTED THE 


RELIABILITY OF THE STUDY RESULTS ................................................................. 12 


RESULTS AND DISCUSSION ....................................................................................... 12 


1. 	 RESULTS ............................................................................... ;................................. J2 


2. 	 DISCUSSION ............................................................................................................ 13 


-1­

983 
Xl 8-0908 

TABLES 


Homogeneity and stability analyses ofthe test substance formulation 14 


2 Concentration analysis ofthe dose formulation ........................................................ 14 


FIGURES 


IR spectrum measured prior to the administration period ......................................... 15 


2 IR spectrum measured after the end of the administration period ............................ 15 


REFERENCE 


IR spectrum provided by the sponsor ....... ...... ....................... ..... ........................ ...... 15 


-2­

984 
X18-0908 

Study Code: X 18-0908 

Test Substance Code: HR7645 

Sponsor Code: S-0170 

TITLE 

Stability Analysis of KarenzMT (Mixture of PEMB and PE3MB), Homogeneity, Stability and 

Concentration Analyses of the Test Substance Formulation 

SPONSOR 

Showa Denko K.K. 

13-9, Shiba Dalmon I-chome, Minato-ku, Tokyo 105-8518, Japan 

TESTING FACILITY 

Hita Laboratory 

Chemicals Evaluation and Research Institute, Japan 

822, 3-chome, Ishii-machi, Hita, Oha 877-0061, Japan 

PURPOSE OF STUDY 

The purpose of this study is to determine the stability of the test substance during the dosing 

period, and homogeneity, stability and concentration of the test substance in formulation in 

"Twenty-Eight-Day Repeated-Dose Oral Toxicity Study of KarenzMT (Mixture of PEMB and 

PE3MB) in Rats" (Study Code: B 11-0908). 

GLP COMPLIANCE 

This study was conducted in compliance with "Concerning Standard of the Testing Facilities 

Conducting the Test Relating to the New Chemical Substances" on Japanese GLP (Notification 

No. 1121003 of the Pharmaceutical and Food Safety Bureau, MHLW, No.3 (November 17, 

2003) of the Manufacturing Industries Bureau, MET! & No. 031121004 of the Environmental 

Health Department, MOE (November 21, 2003)). 

PERIOD OF STUDY 

Commencement of Study: October 9,2008 

Initiation of Examination (Initiation ofAnalysis): October 14, 2008 

Termination of Examination (Tennination ofAnalysis): December 26, 2008 

Completion of Study: March J6, 2009 

-3 985 
X1S-090& 

STORAGE AND RETENTION PI<:RIOD OF DATA 

The raw data, protocol, study contract documents, test substance information, final report and 

other record documents will be retained in the archive of the Hita Laboratory of our organization 

for the same period of B 11-0908 paper data. After termination of the retention period, any 

measures taken will be done so with the approval of the sponsor. 

RETENTION OF ORIGINAL DOCUMENTS 

An original protocol and an original final report will be retained at Hita Laboratory. The copies 

of their original that the study director will be recognized to be accurate copy will be sent to the 

sponsor. 

STUDY DIRECTOR AND PERSONS CONCER'l'iED WITH TH.E STUDY AND THE 

OP.ERATION 

Study director: Yuji Kusune, M.S. 

Study staff: 	 Masaru Suzuki, B.S. 

(Analysis ofthe test substance) 

Saori Takakura, 1.L.A.T., I.A.C.L.C.T. 

(Preparation of the test substance formulation) 

APPROVAL BY AUTHOR 

Study director: Signed in original March 16, 2009 

Yuji Kusune, M.S. 

Analytical Chemistry Section 

Hita Laboratory 

-4­

986 
Xl 8-0908 

SUMMARY 

The test substance (KarenzMT (Mix.ture of PEMB and PE3MB)) was stable during the dosing 


period of subject study (Study Code: B 11-0908). 


The test substance in 10.0 and 0.25 w/v% formulations was stable for 8 days after preparation at 


cold and dark place and showed good homogeneity. The concentration oftest substance in 10.0, 


1.5 and 0.25 w/v% dose fomlUJations for subject study was acceptable level. 

-5 

987 
XI 8-0908 

MATF.RIALS 

..... ~:..... !~~T~.~r~.~T.~~~~.(l.~.:t:!?~~I!Q~.f.~.9.Y.!IJ.~~lJ.#X.lli~~r.9.N~9.~)' ......__ ...__ ._ .._.._.... ; . , 

-6­

988 
X1S-0908 

1.8 	 Storage Conditions 

Stored at room temperature (cabinet No. 1 in test substance storage room, tolerance 

temperature: IO-30°C). Actual temperature between received of test substance to 

termination of examination was 19.0-22.4°C. It satisfied tolerance. Avoid oxidizing 

agent. 

1.9 	 Handling Precaution 

Glove, mask, cap, goggles and lab coat were worn. The test substance was treated in the 

draft since the test substance has characteristic odor. 

METHODS 

1. 	 SUBJECT STUDY 

Twenty-Eight-Day Repeated-Dose Oral Toxicity Study of KarcnzMT (Mixture of PEMB 

and PE3MB) in Rats (Study Code: B11-0908) 

2. 	 STABIUTY ANALYSIS OF THE TEST SUBSTANCE 

The infrared (JR) spectrum was measured by IR spectrophotometer before and after the 

dosing period of subject study. 

2.1 	 Measurement of IR 

I) rnstrument 

JR spectrophotometer: FT-nO (HORlBA, Ltd.) 

2) Condition 

Wave number: 4600 cm-I - 400 cm-' 

3) 	 Pre-Treatment 


Potassium bromide neat 


2.2 	 Criteria for Judgment 

IR spectrum orthc tcst substance that measured prior to dosing in our laboratory should be 

identical with provided from the sponsor. The test substance was judged to be stable 

when there are no differences in the IR spectrums at before and after dosing period. 

-7­

989 
XIS-0908 


3. 	 HOMOGENEITY, STABILITY AND CONCENTRATION ANALYSES OF THE 

TEST SUBSTANCE }<'ORMULATION 

In the homogeneity analysis, the samples were taken (n=l) from the upper, middle and 

lower layers of formulations immediately after preparation, respectively. These samples 

were pretreated and measured (11=1) with high performance liquid chromatography 

(HPLC). 

[n the stability analysis, the formulations were stored at cold and dark place for 8 days, and 

the sample was taken (n= I) from the middle layer of the formulations at point of 

measurement (3 days and 9days after preparation). These samples were pretreated and 

measured (n=l) with HPLC. 

III the concentration analysis, the samples were taken (n=l) from the middle layer of dose 

formulations immediately after preparation for subject study. These samples were 

pretreated and measured (n=l) with HPLC. 

3.1 	 The Test Substance Formulation 

I) Homogeneity and Stability Analyses 

(1) Concentration 

10.0 and 0.25 w/v% 

(2) Preparation Method 

Weighed 5.00 g of tile test substance in a beaker, and mixed with olive oil to make 

50 mL, and used as 10.0 w/v% fOimulation. Accurate 2.5 mL of 10.0 w/v% 

formulation was diluted with olive oil to make toO mL, and used as 0.25 w/v% 

formulation. 

Vehicle: olive oil (Lot No. 0460YS, Fujimi Pharmaceutical) 

2) 	 Concentration Analysis 

The 10.0, 1.5 and 0.25 w/v% dose formulations at first preparation for subject study 

were used. 

3.2 	 Outline of Analytical Method 

The analytical method was decided, according to results of validation of the analytical 

method 011 non-GLP at the test facility. 

I) Validation of the Analytical Method 

(!) Preparation for Measurement Sample 

a) 	 Standard Stock Solution for Validation of the Analytical Method 

Weighed 0.10187 g of the test substance, dissolved in 2-propanol to make 

100 mL, and used this solution as 1018.7 llg!mL standard stock solution for 

validation of the analytical method. 

b) 	 Sample for Specificity 

The standard stock solution for validation of the analytical method was diluted 

with 2-propanol to make 50.9 /lg/mL standard solution and 50.9 /lg!mL 

8 

990 
X18-0908 

vehicle-containing standard solut ion (containing 2 v/v% vehicle (olive oil)). 

2-Propanol was lIsed as solvent blank, and vehicle blank (containing 2 v/v% 

vehicle) was prepared. 

c) Sample for Linearity 

The standard stock solution for validation of the analytical method was diluted 

with 2-propanol to make 25.5, 50.9'1 and 1 02 ~lglmL standard solutions. 

*I: It was used 50.9 ~lglmL standard solution of sample for specificity. 

d) Sample for Accuracy and Repeatability 

Standard solutions (concentration: 25.5, 50.9 and 102 ~glmL, eaeh 

concentration: n=3) were prepared in the same way of sample for linearity. 

(2) Specificity 

Samples for specificity were measured with HPLC. The variation of detection 

value (total peak area) of the test substance between standard solution with and 

without vehicle was -2.0%. Therefore, it was confirmed that the result of 

variation satisfied criteria for judgment (within ±5%). In the results of HPLC 

analysis of solvent blank and vehicle blank, it was confirmed there were no 

background and interfering peaks at the elution peak position of test substance. 

(3) Linearity 

Samples for linearity were measured with HPLC. The calibration curve was 

made by the concentration of the test substance in the horizontal line and the 

detection value of test substance in the vertical line. The coefficient of 

correlation of calibration curve which was obtained from least square wa~ R=0.999. 

Therefore, it was confirmed that the result of linearity satisfied criteria for 

judgment (more than 0.999). 

(4) Accuracy and Repeatability 

Samples for accuracy and repeatability were measured with HPLC. The 

concentrations of the test substance were calculated with regression formula that 

was obtained at linearity. Accuracy and repeatability were calculated with these 

values. 

Accuracy of25.5 llg/mL standard solution was 0.4, -3.1 and -0.4%. 

Accuracy of 50.9 IlglmL standard solution was -0.4, -1.2 and -1.0%. 

Accuracy of 102 IlglmL standard solution was 0.0, 0.0 and 1.0%. 

Repeatability of25.5, 50.9 and I021lglmL standard solution was 1.9, 0.4 and 0.6%, 

respective ly. 

It was confirmed that the result of accuracy and rcpeatability satisfied criteria for 

judgment (accuracy: within ±IO%, repeatability: less than 5%). 

-9­

991 
Xl 8-0908 

2) 	 Preparation for Standard Solution 

Weighed 0.10 113 g ofthe test substance dissolved in 2-propanoJ to make 100 mL, and 

used this solution as 1011.3 J.LglmL standard stock solution. Accurate 1 mL of 

standard stock solution diluted with 2-propanol to make 20 mL, and 50.6 ~lglmL 

standard solution was prepared. 

3 ) 	 Pre-Treatment 


Formulations were mixed well using a magnetic stirrer. 


(I) 10.0 w/v% Formulation (Homogeneity, Stabil ity and Concentration Analyses) 

Accurate 0.5 mL of formulation was dissolved in 2-propanol to make 50 mL. 

Accurate 1 mL of this solution was diluted with 2-propanol to make 20 mL, and 

served as an analytical sample (dilution rate: 2000). 

(2) 	1.5 w/v% Formulation (Concentration Analysis) 

Accurate 0.5 mL of formulation was dissolved in 2-propanol to make 30 mL. 

Accurate 2 mL of this solution was diluted with 2-propanol to make 10 mL, and 

served as an analytical sample (dilution rate: 300). 

(3) 0.25 w/v% Formulation (Homogeneity, Stability and Concentration Analyses) 

Accurate 0.5 mL of formulation was dissolved in 2-propanol to make 25 mL, and 

served as an analytical sample (dilulion rate: 50). 

4) 	 Conditions of HPLC Analysis 

(l) Instruments (HPLC7) 

Interface: D-7000IF (Ilitachi, Ltd.) 

Pump: L-7100 (Hitachi, Ltd.) 

Auto sampler: L-7200 (Hitachi, Ltd.) 

UV-VIS detector: L-7400 (Hitachi, Ltd.) 

Column oven: L-7300 (Hitachi, Ltd.) 

Data processor: D-7000 HPLC System manager (Hitachi, Ltd.) 

(2) Conditions 

Column: L-column ODS (4.6 mm I.D. x 50 mm, CERI) 

Column temperature: 40°C 

Mobile phase: 0.5 v/v% phosphoric acid solution/acetonitrile 45/55 (v/v) 

Flow rate: 1 mUm in 

Wavelength: 210 nm 

Injection volume: 

3.3 	 J)ata Processing 

1) Detection Value 

A total peak area was used as the detection value. 

2) 	 Quantitative Analytical Method 

In validation of the analytical method, the result of linearity was a straight line range 

-10­

992 
XI 8-0908 

of25.5, 50.9 and 102 !!glmL standard solutions, and it passed through the origin of the 

coordinates. Therefore, the concentrations of analytical samples were determined by 

single level calibration. 

3) 	 Calculation ofthe Test Substance Concentration in Formulation 

Concentration of test substance in each sample (C: w/v%) was calculated with the 

equation shown below and rounded off to three significant figures. 

C =0 	CsxAxD 


Asx 10000 


Cs: Test substance concentration in standard solution (/lglmL) 


As: Detection value of test substance in standard solution 


A: Detection value of test substance in each analytical sample 

D: Dilution rate in each analytical sample 

3.4 Criteria ror Judgment 

1) 	 Homogeneity Analysis 

The test substance was judged as homogeneous dispersion in vehicle if a coefficient of 

variation (CV) was within 5%. The CV was calculated using the following equation: 

CV(%) =Standard deviation for concentration of test substance in each x 100 

Mean concentration of test substance in each layer 

2) 	 Stability Analysis 

The test substance was judged as stable state in vehicle if a rate to the nominal 

concentration for the actual concentration (R.N.) and a rate to the mean concentration 

immediately after preparation for the actual concentration (R.P.) were within the range 

of 1OO± I0%. The R.N. and R.P. were calculated using the following equation: 

Actual concentration x] 00 R.N.{%) Nominal concentration 

R.P.(%) '" Actual concentration x I 00 

Mean concentration immediately after preparation 

3) 	 Concentration Analysis 

It was confirmed that R.N. was within the range of 100±10%. The R.N. was 

calculated using the following equation: 

11 ­ 993 
XIS-0908 

R.N .(%) "". Actual concentration xl 00 


Nominal concentration 


ENVIRONMENTAL FACTORS THAT MIGHT HAVE AFFECTED RELIABILITY OF 

STUDY RESULTS 

There were no factors that might have affected the reliability of the study data. 

RESULTS AND DISCUSSION 

1. 	 RESULTS 

1.1 	 Stability Analysis of the Tellt Substance 

IR spectrum of test substance provided by the sponsor (Reference I) was identical with 

that measured before dosing period for subject study (Figure 1). 

There were no differences in the IR spectra between before and after dosing period 

(Figures 1, 2). 

1.2 	 Homogeneity, Stability and Concentration Analyses of tbe Test Substance 

Formulation 

J) Homogeneity and Stability Analyses 

The results of homogeneity and stability analyses of the test substance formulation are 

shown in Table I. 

(1) Homogeneity Analysis 

CY of 10.0 and 0.25 w/v% formulations were 0.6 and 1.0%, respectively. The 

results satisfied criteria for judgment. 

(2) Stability Analysis 

a) 	 10.0 w/v% Formulation 

At immed iately after preparation, R.N. were 101 to 102%. 

At 3 days after preparation, R.N. was 100%, and R.P. was 98.0%. 

At 9 days after preparation, R.N. was 101%, and R.P. was 99.0%. 

All the results of R.N. and R.P. satisfied criteria for judgment. 

b) 	 0.25 w/v% Formulation 

At immediately after preparation, R.N. were 102 to 104%. 

At 3 days after preparation, R.N. was 102%, and R.P. was 99.6%. 

At 9 days alter preparation, R.N. was 102%, and R.P. was 100%. 

All the results of R.N. and R.P. satisfied criteria for judgment. 

-12­

994 
X18-0908 

2) 	 Concentration Analysis 

The results of concentration analysis of the test substance formulation are shown in 

Table 2. 

R.N. of 10.0, 1.5 and 0.25 w/v% dose fomlUlations were 99.1 to 101%. All the 

results satisfied criteria fot" judgment. 

2. 	 DISCUSSION 

The test substance was stable during the dosing period of subject study. 

The test substance ill 10.0 and 0.25 w/v% formulations was stable for 8 days after 

preparation at cold and dark place and showed good homogeneity. The concentration of 

test substance in 10.0, 1.5 and 0.25 w/v% dose formulations for subject study was 

acceptable level. 

13­

995 
XIS-090S 


, I I . Tablel Homoenelty and stabT1 ltv ana vses 01 t le test substance ormu atlon 

Nominal 

cone. 

(w/v%) 

Time point 

of 

measurement 

Layer of 

measurement 

Actual 

cone. 

(w/v%) 

R.N. 

(%) 

Mean cone. 

(w/v%) 

R.P. 

(%) 

CV 

(%) 

10.0 

Immediately 

after 

prcparation 

Upper 10.2 102 

Middle 10.2 102 10.2 - 0.6

Lower 10.1 101 

3 days 

after 

preparation 

Middle 10.0 100 - 98.0 -

9 days 

after 

preparation 

Middle 10.1 101 . 99.0 -

0.25 

Immediately 

aftcr 

preparation 

Upper 0.259 104 

Middle 0.254 102 0.256 . 1.0

Lower 0.256 102 

3 days 

after 

preparation 

Middle 0.255 102 - 99.6 -

9 days 

after 

preparation 

Middle 0.256 102 - 100 -

R.N.: Rate to the nominal concentration 

R.P.: Rate to the concentration measured immediately after preparation 

CV; Coefficient of variation 

T ]'fhd f; I . able 2 ConcentratlOn analYSIS 0 t e ose ormu atlOn 

Date of analysis 

Nominal cone. 

(w/v%) 

Actual cone. 

(w/v%) 

R.N. 

(%) 

10.0 9.91 99.1 

November 6, 2008 1.5 1.51 101 

0.25 0.250 100 

R.N.: Rate to the nom inal concentration 

14 

996 
Xl 8-0908 

~~,,~,.~" 

lOOO 400.0 

Figul'e I IR spectrum measured prior to the administration period 

Figure 2 IR spectrum measured after the end of the administration period 

_L'__-' 

1000 

Reference I lR spectrum provided by the sponsor 

- J5­

997 
PAGE 1 OF 25 PAGES 

Mixture of PEMB and PE3MB: 

MICRONUCLEUS TEST IN olliE MOUSE 

PRO..IECT NUMBER: 0145/0219 

AUTHOR: L Flanders 

STUDY SPONSOR: TEST FACILITY: 

Showa Denko K.K. Harlan Laboratories Ltd 

13-9, Shiba Daimon 1-Chome Shardlow Business Park 

Minato-ku Shardlow 

TOKYO 105-8518 Derbyshire 

JAPAN DE722GD 

UK 

Telephone: +44 (0) 1332 792896 

Facsimile: +44 (0) 1332 799018 

0145-02191LF 

998 
PROJECT NUMBER: 014510219 PAGE 2 

QUALITY ASSURANCE REPORT 


This study type is classed as short-term. The standard test method for this study type 

("General Study Plan" in OECD terminology) was reviewed for compliance once only on 

initial production. Inspection of the routine and repetitive procedures that constitute the 

study is carried out as a continuous process designed to encompass the major phases at 

or about the time this study was in progress. In addition, inspection of general facilities 

not specifically related to this study are done monthly or annually in accordance with QA 

Standard Procedure. 

This report has been audited by the Quality Assurance Unit, and is considered to be an 

accurate account of the data generated and of the procedures followed. 

In each case, the outcome of QA evaluation is reported to the Study Director and 

Management on the day of evaluation. Audits of study documentation, and process 

inspections appropriate to the type and schedule of this study were as follows: 

17 November 2008 Standard Test Method Compliance Audit 

03 June 2009 Test Material Preparation 

03 June 2009 Animal Preparation 

03 June 2009 Dosing 

05 June 2009 Assessment of Response 

05 June 2009 Cell Harvest 

05 June 2009 Staining/Slide Preparation 

§ 29 July 2009 Draft Report Audit 

§ Date of QA Signature Final Report Audit 

§ Evaluation specific to this study 

04 AUG 2009 ...M~..S0..~..............
DATE: .............................................. .. 

For the Quality Assurance Unit* 

*Authorlsed QA Signatures: 

Manager, Quality Assurance: J G Riley SSe (Hons) MRQA 

Deputy Head of Department JM Crowther MIScT MRQA 

Senior Audit Staff: G Wren ONC MRQA 

999 

PROJECT NUMBER: 014510219 PAGE 3 

GLP COMPLIANCE STATEMENT 


The work. described was performed in compliance with UK GLP standards (Schedule 1. 

Good Laboratory Practice Regulations 1999 (SI 1999/3106 as amended by SI 

2004/0994)}. These Regulations are in accordance with GLP standards published as 

OECD Principles on Good Laboratory Practice (revised 1997, ENVIMC/CHEM{98)17): 

and are in accordance with. and implement, the requirements of Directives 2004/9/EC 

and 2004/10/EC. 

This report fully and accurately reflects the procedures used and data generated. 

. 03 AUG 2009 DATE .................................................. .. 

L Flanders 

Study Director 

This report may be presented In final form as a digital (pdf) document. Such documents are prepared by scanning the paper original, 

and are considered ofequivalent integrity and authenticity to versions produced by optical photocopy. However, In all cases the 

hand-slgned paper Original, held in secure archives. is the definitive document 

1000 
PROJECT NUMBER: 0145/0219 PAGE 4 

CONTENTS 

QUALITY ASSURANCE REPORT 2 


GLP COMPLIANCE STATEMENT 3 


CONTENTS 4 


SUMMARY 5 


1. INTRODUCTION 7 


2. PRINCIPLES OF INVESTIGATION 7 


3. TEST AND CONTROL MATERIALS AND EXPERIMENTAL PREPARATION 8 


3. 1 Test Material 8 


3.2 Positive Control Material 8 


3.3 Vehicle Control 9 


4. METHODS 9 


4.1 Animals and Animal Husbandry 9 


4.2 Procedure 10 


5. ARCHIVES 13 


6. RESULTS 14 


6.1 Range-finding Toxicity Test 14 


6.2 Micronucleus Test 15 


7. CONCLUSION 15 


Table 1 
 Micronucleus Test - Summary of Group Mean Data 16 


Table 2 


. Table 3 


Table 4 


Table 5 


Table 6 


Table 7 


Table 8 


Appendix 1 
 Historical Control Data from 45 Studies (2007 and 2008) 24 


Appendix 2 
 Statement of GLP Compliance in Accordance with Directive 2004/9IEC 25


Micronucleus Test - Individual and Group Means and Standard 


Deviations: Vehicle Control (10 mllkg) 48-Hour Sampling Time 17 


Micronucleus Test -Individual and Group Means and Standard 


Micronucleus Test -Individual and Group Means and Standard 


Micronucleus Test -Individual and Group Means and Standard 


Micronucleus Test - Individual and Group Means and Standard 


Micronucleus Test - Individual and Group Means and Standard 


Micronucleus Test -Individual and Group Means and Standard 


Deviations: Vehicle Control (10 mllkg) 24-Hour Sampling Time 18 


Deviations: Cyclophosphamide (50 mglkg) 24-Hour Sampling Time 19 


Deviations: Test Material (600 mglkg) 48-Hour Sampling Time 20 


Deviations: Test Material (600 mglkg) 24-Hour Sampling Time 21 


Deviations: Test Material (300 mglkg) 24-Hour Sampling Time 22 


Deviations: Test Material (150 mg/kg) 24-Hour Sampling Time 23 


1001 

PROJECT NUMBER: 0145/0219 PAGE 5 

Mixture of PEMB and PE3MB: 


MICRONUCLEUS TEST IN THE MOUSE 


SUMMARY 

Introduction. The study was performed to assess the potential of the test material to 

produce damage to chromosomes or aneuploidy when administered to mice. The 

method was designed to comply with the DECO Guidelines for Testing of Chemicals 

No.474 "Micronucleus Test", Method 812 of Commission Regulation (EC) No. 44012008 

of 30 May 2008, the USA EPA, TSCA and FIFRA guidelines and the Japanese 

METI/MHLW guidelines for testing of new chemical substances. 

Methods. A range-finding test was performed to find suitable dose levels of the test 

material, route of administration and to investigate to see if there was a marked 

difference in toxic response between the sexes. There was no marked difference in 

toxicity of the test material between the sexes; therefore the main test was performed 

using only male mice. The micronucleus test was conducted using the oral route in 

groups of seven mice (males) at the maximum tolerated dose (MTD) 600 mglkg and with 

300 and 150 mglkg as the two lower dose levels. Animals were killed 24 or 48 hours 

later, the bone marrow extracted, and smear preparations made and stained. 

Polychromatic (PCE) and normochromatic (NCE) erythrocytes were scored for the 

presence of micronuclei. 

Further groups, each of 7 mice, were given a single oral dose of arachis oil or dosed 

orally with cyclophosphamide (5 mice). to serve as vehicle and positive controls 

respectively. Vehicle control animals were killed 24 or 48 hours later, and positive 

control animals were killed after 24 hours. 

Results. There were no premature deaths seen in any of the dose groups. Clinical 

signs were observed in animals dosed with the test material at 600 mg/kg in both the 24 

and 48-hour groups, these were as follows: Hunched posture, lethargy. ataxia and 

ptosis. 

No statistically significant decreases in the PCE/NCE ratio were observed in the 24 or 

48-hour test material dose groups when compared to their concurrent control groups. 

1002 
PROJECT NUMBER: 014510219 PAGES 

However, the observation of clinical signs was taken to indicate that systemic absorption 

had occurred and exposure to the target tissue had been achieved. 

There was no evidence of a significant increase in the incidence of micronucleated 

polychromatic erythrocytes in animals dosed with the test material when compared to the 

concurrent vehicle control groups. 

The positive control group showed a marked increase in the incidence of micronucleated 

polychromatic erythrocytes hence confirming the sensitivity of the system to the known 

mutagenic activity of cyclophosphamide under the conditions of the test. 

Conclusion. The test material was considered to be non-genotoxic under the conditions 

of the test. 

1003 

PROJECT NUMBER: 0145/0219 PAGE 7 

Mixture of PEMB and PE3MB: 


MICRONUCLEUS TEST IN THE MOUSE 


1. INTRODUCTION 

The micronucleus test is a mammalian in vivo test that detects damage to the 

chromosomes induced by chemicals. In addition, numerical changes due to 

chromosome loss during cell division can be detected in this test. 

The study was performed according to a method that was designed to comply with the 

1997 OECD Guidelines for Testing of Chemicals No.474 "Micronucleus Tesf', Method 

812 of Commission Regulation (EC) No. 440/2008 of 30 May 2008, the USA EPA, TSCA 

and FIFRA guidelines and the Japanese METIIMHLW guidelines for testing of new 

chemical substances. The results of the test are believed to be of value in predicting the 

mutagenic potential of the test material to man. The test system was chosen because 

the mouse has been shown to be a suitable model for this type of study and is 

recommended in the test method. 

The experimental phases of the study were performed between 13 May 2009 and 

03 July 2009. 

2. PRINCIPLES OF INVESTIGATION 

In mitotic cells in which chromosome damage has been caused by the test material or its 

metabolites, fragments (centric or acentric) or whole chromosomes tend to lag behind in 

the anaphase stage of cell division. After telophase a large proportion of the fragments 

are not included in the nuclei of the daughter cells and hence form a single or multiple 

micronuclei (Howell-Jolly bodies) in the cytoplasm of these cells. These micronuclei are 

seen in a wide variety of cell types, but erythrocytes are chosen since micronuclei are 

easily detected in these cells. 

A few hours after the last mitosis is completed, erythrocytes expel their nuclei. Immature 

erythrocytes, less than 24 hours old, stain blue with May-GrOnwald/Giemsa due to the 

presence of minute fragments of nuclear material in the cytoplasm. This material is 

mainly ribonucleic acid (RNA), which gradually disappears so that more mature 

erythrocytes (normochromatic erythrocytes) stain pink with May-GrOnwald/Giemsa. The 

1004 
PROJECT NUMBER: 014510219 PAGES 

immature blue staining cells are known as polychromatic erythrocytes and mauve 

stained micronuclei are easily detected in this cell type. If scoring is restricted to 

polychromatic erythrocytes, all the chromosomal damage detected will have been 

caused during the final cell cycle of the nucleated precursor cells. Thus by examining 

polychromatic cells at various periods after administration, the effect of the test material 

over the previous 30 hours can be monitored. 

Any toxic effects of the test material on the immature nucleated cells may lead to a 

reduction in cell division and cell death. This in turn leads to a reduction in cell volume 

and to compensate for this, peripheral blood is shunted into the bone marrow. If the ratio 

of polychromatic to normochromatic erythrocytes is scored and found to be significantly 

lower than the control value, this is taken as being indicative of cytotoxicity. 

3. TEST AND CONTROL MATERIALS AND EXPERIMENTAL PREPARATION 

3.1 Test Material 

Sponsor's identification w ••Mixture _~ __ •• _ ••• of ______ PEMB •• ~~ __ and • PE3MB , , , . Description , , , . , . . Batch number . . .. -~ ..-....-... --.-.---~ 

Date received 15 December 2008 

Storage conditions Room temperature in the dark 

The integrity of supplied data relating to the identity, purity and stability of the test 

material is the responsibility of the Sponsor. 

For the purpose of this study the test material was freshly prepared as required as a 

solution at the appropriate concentration in arachis oil. 

Detennination by analysis of the concentration, homogeneity and stability of the test 

material preparations was not appropriate because it was not specified in the Standard 

Test Method. 

3.2 Positive Control Material 

The positive control material was supplied by Acros Organics, as follows: 

1005 

PROJECT NUMBER: 014510219 PAGES 

Supplier's identification Cyclophosphamide 

Supplier's lot number A0164185 

Date received 10 March 2009 

Storage conditions Approximately 4°C in the dark 

For the purpose of this study the positive control material was freshly prepared as 

required as a solution at the appropriate concentration in distilled water (supplied by 

PAM Medical Supplies Ltd. batch no. 300522801). 

The concentration, homogeneity and stability of the positive control material and its 

preparation were not determined by analysis. 

3.3 Vehicle Control 

The vehicle was supplied by Lab 3 Ltd, as follows: 

Supplier's identification Arachis Oil BP 

Supplier's lot number W72 

Date received 05 February 2008 

Description Pale straw coloured slightly viscous liquid 

Storage conditions Room temperature 

4. METHODS 

4.1 Animals and Animal Husbandry 

Sufficient albino Hsd: leR (CD-1~ strain mice were obtained from Harlan UK. At the 

start of the main test the male mice weighed 23 to 30g and were approximately five to 

eight weeks old. After a minimum acclimatisation period of five days the animals were 

selected at random and given a number unique within the study by tail marking and a 

number written on a colour coded cage card. 

The animals were housed in groups of up to seven, by sex, in solid-floor polypropylene 

cages with wood-flake bedding. Free access to mains drinking water and food (Harlan 

Teklad 2014 Rodent Pelleted Diet) was allowed throughout the study. 

1006 

PROJECT NUMBER: 014510219 PAGE 10 

The temperature and relative humidity were set to achieve limits of 19 to 25°C and 30 to 

70% respectively. Any occasional deviations from these targets were considered not to 

have affected the purpose or integrity of the study. The rate of air exchange was 

approximately fifteen changes per hour and the lighting was controlled by a time switch 

to give twelve hours light and twelve hours darkness. 

4.2 Procedure 

4.2.1 Range-finding Toxicity Test 

A range-finding toxiCity test was performed to determine a suitable dose level and route 

of administration for the micronucleus test. The dose level selected should ideally be the 

maximum tolerated dose level or that which produces some evidence of toxicity up to a 

maximum recommended dose of 2000 mg/kg. The range-finding toxicity test was also 

used to determine if the main test was to be performed using both sexes or males only. 

Groups of mice were dosed orally as follows: 

Dose Level Concentration DoseVofume Number of Mice 

(mg/kg) {mg/mO (mllkg) Male Female 

2000 

1000 

800 

600 

600 

200 

100 

80 

60 

60 

10 

10 

10 

10 

10 

1 

1 

1 

1 

2 

1 

1 

1 

1 

0 

All animals were dosed once only at the appropriate dose level by gavage using a metal 

cannula attached to a graduated syringe. The volume administered to each animal was 

calculated according to its bodyweight at the time of dosing. 

Animals were observed within the time periods of '%, 1 and 2 hours after dosing and 

subsequently once daily for up to two days. Any deaths and evidence of overt toxicity 

were recorded at each observation. No necropsies were performed. 

4.2.2 Micronucleus Test 

Groups, each of seven mice, were dosed once only via the oral route with the test 

material at 600, 300 or 150 mg/kg. One group of mice from each dose level was killed 

1007 
PROJECT NUMBER: 0145/0219 PAGE 11 

by cervical dislocation 24 hours following treatment and a second group dosed with test 

material at 600 mg/kg was killed after 48 hours. In addition, three further groups of mice 

were included in the test; two groups (each of seven mice) were dosed via the oral route 

with the vehicle alone (arachis oil) and a third group (five mice) was dosed orally with 

cyclophosphamide. Cyclophosphamide is a positive control material known to produce 

micronuclei under the conditions of the test. The vehicle controls were killed 24 or 48 

hours fo.llowing dosing and positive control group animals were killed 24 hours following 

dosing. The experimental design is summarised as follows: 

Dose Group 

Dose 

Level 

(mglkg) 

Concentration 

(mg/ml) 

Dose 

Volume 

(mVkg) 

Kill Time 

(Hours 

After 

Dosing) 

Animal 

Numbers 

1. Vehicle Control (Arachis oil) 0 0 10 48 1-7 

2. Vehicle Control (Arachis oil) 0 0 10 24 8 -14 

3. Positive Control 

(Cyclophosphamide) 50 5 10 24 15 -19 

4. Mixture of PEMB and PE3MB 600 60 10 48 20 -26 

5. Mixture of PEMB and PE3MB 600 60 10 24 27 -33 

6. Mixture of PEMB and PE3MB 300 30 10 24 34-40 

7. Mixture of PEMB and PE3MB 150 15 10 24 41-47 

All animals were observed for signs of overt toxicity and death one hour after dosing and 

then once daily as applicable and immediately prior to termination. 

4.2.3 Slide Preparation 

Immediately following termination (i.e. 24 or 48 hours following dosing), both femurs 

were dissected from each animal, aspirated with foetal calf serum and bone marrow 

smears prepared following centrifugation and re-suspension. The smears were air-dried, 

fIXed in absolute methanol, stained in May-GrOnwald/Giemsa, allowed to air-dry and a 

cover slip applied using mounting medium. 

1008 

PROJECT NUMBER: 014510219 PAGE 12 

4.2.4 Slide Evaluation 

Stained bone marrow smears were coded and examined blind using light microscopy at 

x1000 magnification. The incidence of micronucleated cells per 2000 polychromatic 

erythrocytes (PCE-blue stained immature cells) per animal was scored. Micronuclei are 

normally circular in shape. although occasionally they may be oval or half-moon shaped, 

and have a sharp contour with even staining. In addition, the number of normochromatic 

erythrocytes (NCE-pink stained mature cells) associated with 1000 erythrocytes was 

counted; these cells were also scored for incidence of micronuclei. 

The ratio of polychromatic to normochromatic erythrocytes was calculated together with 

appropriate group mean values and standard deviations. 

4.2.5 Interpretation of Results 

A comparison was made between the number of micronucleated polychromatic 

erythrocytes occurring in each of the test material groups and the number occurring in 

the corresponding vehicle control group. 

A positive mutageniC response was demonstrated when a statistically Significant, doseresponsive, toxicologically relevant increase in the number of micronucleated 

polychromatic erythrocytes was observed for either the 24 or 48-hour kill times when 

compared to their corresponding control group. 

If these criteria were not fulfilled, then the test material was considered to be nongenotoxic under the conditions of the test. 

A positive response for bone marrow toxiCity was demonstrated when the dose group 

mean polychromatic to normochromatic ratio was shown to be statistically significantly 

lower than the concurrent vehicle control group. 

All data were statistically analysed using appropriate statistical methods as 

recommended by the UKEMS Sub-committee on Guidelines for Mutagenicity Testing 

Report. Part III (1989). The data was analysed following a .j(x+1) transformation using 

Student's t-test (two tailed) and any significant results were confirmed using the one way 

analysis of variance. 

1009 
PROJECT NUMBER: 0145/0219 PAGE 13 

5. ARCHIVES 


Unless instructed otherwise by the Sponsor, all original data and the final report will be 

retained in the Harlan Laboratories Ltd, Shard low, UK archives for five years, after which 

instructions will be soUght as to further retention or disposal. 

1010 

6.1 

PROJECT NUMBER: 014510219 PAGE 14 

6. RESULTS 

Range-finding Toxicity Test 

The mortality data are summarised as follows:­

Dose Level 

(mg/kg) Sex 

Number of 

Animals 

Treated 

Route 

Deaths on Day Total Deaths 

0 1 2 

2000 

1000 

Male 1 

oral 

oral 

1e - - 212 

012 

Female 

Male 

1 

1 

1 

0 

-

0 

-

0 

Female 1 0 a 0 

800 

Male 1 

oral 

0 0 0 

1/2 

Female 1 1 i - -

600 

Male 1 

oral 

0 0 0 

0/2 

Female 1 a 0 a 

600 

Male 2 

oral 

a 0 0 

0/2 

Female 0 - - -

In animals dosed with the test material via the oral route premature deaths occurred at 

800 and 2000 mg/kg. and clinical signs were observed at and above 600 mg/kg as 

follows: Hunched posture. ptosis. ataxia, lethargy. pilo-erection, splayed gait, decreased 

respiratory rate. laboured respiration. elevated tail and tonic convulsions. 

The test material showed no marked difference in its toxicity to male or female mice, it 

was therefore considered to be acceptable to use males only for the main test. 

Adequate evidence of test material toxicity was demonstrated via the oral route of 

administration, therefore, this was selected for use in the main test. The maximum 

tolerated dose (MTD) of the test material, 600 mg/kg, was selected for use in the main 

test, with 300 and 150 mg/kg as the lower dose levels. 

- =No data 

e =Killed in extremis 

1011 
PROJECT NUMBER: 014510219 PAGE 15 

6.2 Micronucleus Test 

6.2.1 Mortality Data and Clinical Observations 

There were no premature deaths seen in any of the dose groups. Clinical signs were 

observed in animals dosed with the test material at 600 mglkg in both the 24 and 

48-hour groups, these were as follows: Hunched posture, lethargy. ataxia and ptOSiS. 

6.2.2 Evaluation of Bone Marrow Slides 

A summary of the results of the micronucleus test is given in Table 1. Individual and 

group mean data are presented in Tables 2 to 8. 

There were no statistically significant decreases in the peE/NCE ratio in the 24 or 

48-hour test material groups when compared to their concurrent vehicle control groups. 

However, the observation of clinical signs was taken to indicate that systemic absorption 

had occurred and exposure to the target tissue had been achieved. 

There were no statistically significant increases in the frequency of micronucleated PCE 

in any of the test material dose groups when compared to their concurrent vehicle control 

groups. 

The positive control group showed a marked increase in the incidence of micronucleated 

polychromatic erythrocytes hence confirming the sensitivity of the system to the known 

mutagenic activity of cyclophosphamide under the conditions of the test. 

The test material was found not to produce a Significant increase in the frequency of 

micronuclei in polychromatic erythrocytes of mice under the conditions of the test. 

7. CONCLUSION 

The test material was considered to be non-genotoxic under the conditions of the test. 

1012 

PROJECT NUMBER: 014510219 PAGE 16 

Mixture of PEMB and PE3MB: MICRONUCLEUS TEST IN THE MOUSE 

Table 1 Micronucleus Test - Summary of Group Mean Data 

Treatment Group 

Number of PCE with 

Micronuclei per 2000 PCE PCE/NCE Ratio i 

Group Mean SO Group Mean SO 

1. Vehicle Control 

10 mllkg 

48-hour Sampling Time 

0.4 0.5 1.02 0.18 

2. Vehicle Control 

10 mVkg 

24-hour Sampling Time 

0.7 0.8 0.78 0.16 

3. Positive Control 

50 mgJkg 

24-hour Sampling Time 

30.0-* 4.7 0.64 0.14 

4. Mixture of PEMB and PE3MB 

600 mglkg 

48-hour Sampling Time 

0.7 0.8 0.99 0.12 

5. Mixture of PEMB and PE3MB 

600 mg/kg 

24-hour Sampling Time 

0.6 0.8 1.00 0.32 

6. Mixture of PEMB and PE3MB 

300 mg/kg 

24-hour Sampling Time 

1.6 1.5 0.81 0.15 

7. Mixture of PEMB and PE3MB 

150 mglkg 

24-hour Sampling Time 

1.0 1.0 0.79 0.30 

PCE = Polychromatic erythrocytes 

NCE = Nonnochromatic erythrocytes 

SO =standard deviation *.. =P < 0.001 

1013 

Mixture of PEMB and PE3MB: MICRONUCLEUS TEST IN THE MOUSE 

Table 2 	 Micronucleus Test - Individual and Group Means and Standard Deviations: Vehicle Control (10 ml/kg) 48~Hour 

Sampling Time 

BODYWEIGHT 

POLYCHROMATIC ERYTHROCYTES NORMOCHROMATIC 

TREATMENT GROUP ANIMAL (g) WHEN (PCE) ERYTHROCYTES (NCE) PCE/NCE 

NUMBER NUMBER NUMBER RATIO DOSED PCE+MN %PCE+MN NCE+MN SCORED SCORED 

1 26 2000 a 0.00 445 a 1.25 ---­ -----­

2 28 2000 0 0.00 446 0 1.24 -_. - .. .-,-~ 

- . ­ '­ ........ -" -~.-- , .­ ___ H • 

3 27 2000 1 0.05 482 0 1.07 

1. 4 27 2000 1 0.05 498 0 1.01 

VEHICLE CONTROL 5 23 2000 1 0.05 544 0 0.84 10 mllkg 48-hour 

sampling time 6 27 2000 0 0.00 530 1 0 0.89 

7 26 2000 0 0.00 552 1 0.81 

Group Mean 26.3 2000 0.4 lu­ 0.02 500 0.1 1.02 !----­ ----­

SO 1.6 a 0.5 0.03 44 0.4 0.18 --­ j --­ , --­

o 

...... 

.1/10. 

PROJECT NUMBER: 014510219 	 PAGE 17 
Mixture of PEMB and PE3MB: MICRONUCLEUS TEST IN THE MOUSE 

Table 3 	 Micronucleus Test - Individual and Group Means and Standard Deviations: Vehicle Control (10 ml/kg) 24-Hour 

Sampling Time 

BODVWEIGHT 

POLYCHROMATIC ERYTHROCYTES NORMOCHROMATIC 

TREATMENT GROUP ANIMAL (g) WHEN (PCE) ERYTHROCYTES (NCE) PCE/NCE 

NUMBER NUMBER NUMBER RATIO DOSED PCE+ MN %PCE+ MN NCE+ MN SCORED SCORED 

8 27 2000 0 0.00 594 2 0.68 

9 29 2000 1 0.05 486 0 1.06 , -<. __ ..._­ - ----.~.-- ..__. - ---­ - - .__., -_..­ ~.~ ,-_..._­ - . ... - ._-, , -------,. 

10 25 2000 1 0.05 580 0 0.72 .. 

2. 11 26 2000 0 0.00 577 0 0.73 

VEHICLE CONTROL -------­

10 mllkg 24-hour 12 30 2000 0 0.00 619 0 0.62 

sampling time 13 29 2000 2 0.10 588 0 0.70 ~--

14 29 2000 1 0.05 521 1 0.92 

Group Mean 27.9 2000 0.7 0.04 566 0.4 0.78 ----­

SO 1.9 0 0.8 0.04 46 0.8 0.16 

o.... 

c.n 

PROJECT NUMBER: 0145/0219 	 PAGE 18 
Mixture of PEMB and PE3MB: MICRONUCLEUS TEST IN THE MOUSE 

Table 4 	 Micronucleus Test -Individual and Group Means and Standard Deviations: Cyclophosphamide (60 mg/kg) 

24-Hour Sampling Time 

POLYCHROMATIC ERYTHROCYTES NORMOCHROMATIC 

ANIMAL BODYWEIGHT (PCE) ERYTHROCYTES (NCE) PCE/NCE TREATMENT GROUP (g) WHEN NUMBER DOSED NUMBER NUMBER RATIO 

SCORED PCE+MN °Ji,PCE + MN SCORED NCE+MN 

15 23 2000 32 1.60 703 0 0.42 

15 23 2000 25 1.25 577 0 0.73 , ..,._--.. - "­ '. --_._._-­ ~- .-. ~-" -­1---,..... -, ----- .. ,­ .. .. -~ 

3. 17 24 2000 33 1.55 607 0 0.65 

CYCLOPHOSPHAMIDE -

50 mglkg 18 28 2000 35 1.75 563 0 0.78 

24-hour sampling time 19 24 2000 25 1.25 625 0 0.50 

Group Mean 24.4 2000 30.0 1.50 615 0.0 0.64 ..... 

I SD 2.1 0 4.7 0.23 55 0.0 0.14 

o 

en 

PROJECT NUMBER: 014510219 	 PAGE 19 
Mixture of PEMB and PE3MB: MICRONUCLEUS TEST IN THE MOUSE 

Table 5 	 Micronucleus Test -Individual and Group Means and Standard Deviations: Test Material (600 mg/kg) 48-Hour 

Sampling Time 

BODYWEIGHT POLYCHROMATIC ERYTHROCYTES (PCE) NORMOCHROMATIC 

TREATMENT GROUP ANIMAL (g) WHEN 

ERYTHROCYTES (NCE) PCE/NCE 

NUMBER DOSED NUMBER NUMBER RATIO 

SCORED PCE+ MN %PCE+MN SCORED NCE+MN 

20 28 2000 1 0.05 514 0 0.95 --­ -­ 21 26 2000 0 0.00 485 0 1.06 ---­

22 29 2000 1 0.05 506 0 0.98 

4. 

Mixture of PEMB and 23 24 2000 1 0.05 454 0 1.20 --~ 

PE3MB 24 23 2000 0 0.00 558 0 0.79 

600 mg/kg ---­ --­ -­ --­ -

48-hour sampling time 25 27 2000 0 0.00 504 0 0.98 

26 25 2000 2 0.10 509 1 0.96 

Group Mean 26.0 2000 0.7 0.04 504 0.1 0.99 

SD 2.2 0 0.8 0.04 31 0.4 0.12 -­

o 

...... 

....., 

PROJECT NUMBER: 0145/0219 	 PAGE 20 
Mixture of PEMB and PE3MB: MICRONUCLEUS TEST IN THE MOUSE 

Table 6 	 Micronucleus Test -Individual and Group Means and Standard Deviations: Test Material (600 mg/kg) 24-Hour 

Sampling Time 

BODYWEIGHT POLYCHROMATIC ERYTHROCYTES (PCE) NORMOCHROMATIC 

TREATMENT GROUP ANIMAL (g) WHEN ERYTHROCYTES (NCE) PCEINCE 

NUMBER DOSED NUMBER NUMBER RATIO 

SCORED PCE+MN %PCE+MN SCORED NCE+ MN 

27 28 2000 1 0.05 508 0 0.97 

28 28 2000 2 0.10 533 0 0.88 ---­

29 26 2000 0 0.00 377 0 1.65 

5. --­

Mixture of PEMB and 30 27 2000 0 0.00 604 0 0.66 

PE3MB 31 25 2000 1 0.05 519 0 0.93 

600 mg/kg 32 24 2000 0 0.00 24-hour sampling time 482 a 1.07 -~-

33 29 2000 0 0.00 548 0 0.82 

Group Mean 26.7 2000 0.6 0.03 510 0.0 1.00 --­

SO 1.8 0 0.8 0.04 70 0.0 0.32 --­ -

o.... 

CIO 

PROJECT NUMBER: 0145/0219 	 PAGE 21 
Mixture of PEMB and PE3MB: MICRONUCLEUS TEST IN THE MOUSE 

Table 7 	 Micronucleus Test - Individual and Group Means and Standard Deviations: Test Material (300 mg/kg) 24-Hour 

Sampling Time 

BODYWEIGHT POLYCHROMATIC ERYTHROCYTES (PCE) NORMOCHROMATIC I 

TREATMENT GROUP ANIMAL (g) WHEN 

ERYTHROCYTES (NCE) PCE/NCE 

NUMBER DOSED NUMBER NUMBER RATIO 

SCORED PCE+MN O/OPCE + MN SCORED NCE+ MN 

34 25 2000 1 0.05 598 0 0.67 --­ -­ -­

35 24 I 2000 0 0.00 520 0 0.92 

36 25 2000 4 0.20 604 0 0.66 

6. 

Mixture of PEMB and -­ 37 26 2000 1 0.05 525 a 0.90 

PE3MB 38 24 2000 0 0.00 501 0 1.00 

300mglkg 

24-hour sampling time 

40 

39 

25 

25 2000 

2000 2 

3 --­

0.15 

0.10 

--­

604 

530 0 

0 o~0.89 

Group Mean 24.9 2000 1.6 0.08 555 0.0 0.81 

SD 0.7 0 1.5 0.08 45 0.0 0.15 -­ -~ --­ I 

o 

co 

PROJECT NUMBER: 0145/0219 	 PAGE 22 
Mixture of PEMB and PE3MB: MICRONUCLEUS TEST IN THE MOUSE 

Table 8 	 Micronucleus Test -Individual and Group Means and Standard Deviations: Test Material (150 mg/kg) 24~our 

Sampling Time 

BODYWEIGHT POLYCHROMATIC ERYTHROCYTES (PCE) NORMOCHROMATIC 

TREATMENT GROUP ANIMAL (g) WHEN 

ERYTHROCYTES (NCE) PCE/NeE 

NUMBER DOSED NUMBER NUMBER RATIO 

SCORED PCE+MN %PCE+MN SCORED NCE+MN 

41 25 2000 0 0.00 705 0 0.42 -­

42 24 2000 2 0.10 584 0 0.71 ----. 

43 24 2000 2 0.10 603 0 0.66 

7. -1------­ --­

Mixture of PEMB and 44 26 2000 0 0.00 542 0 0.85 -­

PE3MB 45 26 2000 1 0.05 603 0 0.66 

150 mglkg --­

24-hour sampling time 46 26 2000 0 0.00 533 0 0.88 

47 29 2000 2 0.10 420 a 1.38 

Group Mean 25.7 2000 1.0 0.05 570 0.0 0.79 

SO 1.7 0 I 1.0 0.05 87 0.0 0.30 

o 

N 

o 

PROJECT NUMBER: 0145/0219 	 PAGE 23 
PROJECT NUMBER: PAGE 24 

Mixture of PEMB and PE3MB : MICRONUCLEUS TEST IN THE MOUSE 

Appendix 1 Historical Control Data from 45 Studies (2007 and 2008) 

48-HourVehicle Control Group Mean Data 

! 

PCE + MN Per 2000 PCE PCEINCE 1000 Erythrocytes 

Mean 0.90 1.09 

SO 0.54 0.37 

Minimum 0.00 0.62 

Maximum 2.60 2.54 

24-Hour Vehicle Control Group Mean Data 

PCE + MN Per 2000 PCE PCElNCE 1000 Erythrocytes 

I 

Mean 0.99 1.05 

SO 0.60 0.32 

Minimum 0.10 0.60 

Maximum 3.20 2.32 

24·Hour Positive Control Group Mean Data 

PCE + MN Per 2000 PCE PCEfNCE 1000 Erythrocytes 

Mean 42.76 1.17 

SO 13.55 0.31 

Minimum 17.60 0.72 

Maximum 66.40 2.03 

PCE =Polychromatic erythrocytes 

NCE =Normochromatic erythrocytes 

SO =Standard deviation 

1021 

PROJECT NUMBER: 014510219 PAGE 25 

Appendix 2 Statement of GLP Compliance in Accordance with Directive 

2004/9IEC 

,DH') De.'partment , of Health 

THE DEPARTMENT OF REALTH OF THE GOVERNMENT 


OF TIm UNITED KlNGDOM 


GOODLABOllATORYPRACTICE 

STATEMENT OFCOMPLIA.."iCE 


IN ACCORDANCE WlTflDJR.ECm1B 2004J91EC 


'T)';ST FACILrrv TES'fTYPE 

Harlan Laboratories Ltd. Analytical/Clinkal ChelJ!isby 

Shardlaw BllSiness Park Environmental Fate 

.L&ndon Rood Environmental Toxicology 

Sliardlow Mutag'nicity 

Derby Pby&lCbem

DE72 2GD Toxicology 

19th August: 2008 

Agen~ral inspeco<m for compliance with the Principles of Good LabQcatory Pracrice 

was c~ out a! the ftbove1:est facility as part oftbe UK. G.LPCompliance PrOgrnmme. 

At the ~rile of inspection DO deviations were fQuod of sufficient magnitude to affect 

the validity olnon-clinical slUdlesperfonned attitesefacilities. 

Dr. Andrew J. Gray 

Head, UK GLPMonitoringAuthorllJ 

1022 

PAGE 1 OF 68 PAGES harlannl 

.... 

Mixture of PEMB and PE3MB: 

DETERMINATION OF ABIOTIC DEGRADATION, 

HYDROLYSIS AS A FUNCTION OF pH 

PROJECT NUMBER: 014510218 

AUTHORS: 	 R E Butler 

D FWhite 

STUDY SPONSOR: 	 TEST FACILITY: 

Showa Denko K.K. Harlan Laboratories Ltd 

13-9, Shiba Daimon 1-Chome Shardlow Business Park 

Minato-ku Shard low 

TOKYO 105-8518 Derbyshire 

JAPAN DE722GD 

UK 

Telephone: +44 (O) 1332792896 

Facsimile: +44 (0) 1332 799018 

1023 

PROJECT NUMBER: 014510218 PAGE 2 

QUALITY ASSURANCE REPORT 


This study type is classed as short-tenn. Inspection of the routine and repetitive 

procedures that constitute the study is carried out as a continuous process designed to 

encompass the major phases at or about the time this study was in progress. In 

addition, inspection of general facilities not specifically related to this study are done 

monthly or annually in accordance with QA Standard Procedure. 

This report has been audited by the Quality Assurance Unit, and is considered to be an 

accurate account of the data generated and of the procedures followed. 

In each case, the outcome of QA evaluation is reported to the Study Director and 

Management on the day of evaluation. Audits of study documentation, and process 

inspections appropriate to the type and schedule of this study were as follows: 

§ 30 April 2009 Protocol Compliance Audit 

03,06,09,10 July 2009 Abiotic Degradation 

§ 07 September 2009 Draft Report Audit 

§ Date of QA Signature Final Report Audit 

§ Evaluation specific to this study 

\ 


'"Authorised QA Signatures: 

Manager, Quality Assurance: J G Riley SSe (Hons) MRQA 

Deputy Head of Department: JM Crowther MIScT MRQA 

Senior Audit Staff: G Wren ONC MRQA 

1024 
PROJECT NUMBER: 014610218 PAGE 3 

GLP COMPLIANCE STATEMENT 


The work described was performed in compliance with UK GLP standards (Schedule 1, 

Good Laboratory Practice Regulations 1999 (SI 1999/3106 as amended by 

SI 2004/0994»). These Regulations are in accordance with GLP standards published as 

OECD Principles on Good Laboratory Practice (revised 1997, ENVIMC/CHEM(98}17): 

and are in accordance with, and implement, the requirements of Directives 2004/9/EC 

and 2004/10/EC. 

This report fully and accurately reflects the procedures used and data generated . 

. 2 It SEP 2009 ................. ......................... DATE ................................................ . 

R E Butler GRSC 

STUDY DIRECTOR 

This report may be presented in final form as a digital (Pdf) document. Such documents .ue prepared by scanning the paper Original, 

and are considered ofequivalent integrity and authenticity to versions produced by optical photocopy. However, in all cases the 

hand-signed paper original, held in secure archives. Is the definitive document 

1025 

PROJECT NUMBER: 0145/0218 PAGE 4 

CONTENTS 


QUALITY ASSURANCE REPORT 2 


GLP COMPLIANCE STATEMENT 3 


CONTENTS 4 


SUMMARY 5 


1. INTRODUCTION 6 


2. TEST MATERIAL 6 


2.1 Description, Identification and Storage Conditions 6 


2.2 Chemical Structure 7 


3. ARCHIVES 7 


4. HYDROLYSIS AS A FUNCTION OF pH 8 


4.1 Method 8 


4.2 Procedure 8 


4.3 Calculation 15 


4.4 Results 17 


4.5 Validation 66 


4.6 Discussion 66 


4.7 Conclusion 67 


Appendix 1 Statement of GLP Compliance in accordance with Directive 


2004l9/EC 68 


1026 

PROJECT NUMBER: 0145/0218 PAGE 5 

Mixture of PEMB and PE3MB: 


DETERMINATION OF ABIOTIC DEGRADATION, 


HYDROLYSIS AS A FUNCTION OF pH 


SUMMARY 


Assessment of hydrolytic stability was carried out using Method C7 Abiotic Degradation, 

Hydrolysis as a Function of pH of Commission Regulation (EC) No 44012008 of 30 May 

2008 and Method 111 of the OECD Guidelines for Testing of Chemicals, 13 April 2004. 

The results are as follows: 

pH 

Rate constant (S·1) Estimated half-life at 25"C 

PE3MB PEMB PE3MB PEMB 

4 not applicable not applicable >1 year >1 year 

7 not applicable not applicable >1 year >1 year 

9 2.07 x 10-5 2.91 x 10.5 9.33 hours 6.61 hours 

No definitive signs of hydrolysis at pH 4 or pH 7. Testing performed at various 

temperatures indicated that losses observed were not pseudo-first order and therefore 

most likely to be due other mechanisms or effects rather than hydrolysis. For further 

information refer to Discussion (Section 4.6). 

1027 

2.1 

PROJECT NUMBER: 014510218 PAGE 6 

Mixture of PEMB and PE3MB: 


DETERMINATION OF ABIOTIC DEGRADATION, 


HYDROLYSIS AS A FUNCTION OF pH 


1. INTRODUCTION 

The abiotic degradation. hydrolysis as a function of pH of the test material has been 

determined. 

The method employed complies with those specified in Commission Regulation (EC) No 

440/2008 of 30 May 2008. Part C: Methods for the determination of ecotoxicity and the 

current OECD Guidelines for Testing of Chemicals. 

Testing was conducted between 01 May 2009 and 29 July 2009. 

2. TEST MATERIAL 

Description, Identification and Storage Conditions 

Sponsor's identification Mixture of PEMB and PE3MB .-----------.-.-..------------.. ----~ 

Description 

Purity 

Batch number . Date received ··-10·0ecEiiiioer-200S·········, 

Storage conditions room temperature in the dark 

The integrity of supplied data relating to the identity, purity and stability of the test 

material is the responsibility of the Sponsor. 

1028 

2.2 

PROJECT NUMBER: 014510218 PAGE 7 

Chemical Structure 

The Sponsor provided the foll?~i.~~.c?~~.,,!!i.C?~I.~~~I:I~I:I!~.~9rJ~.~J~~trJ:I~~rj~L ........... 
 , .. ----.......................................... ---...... --............ f


· .
 · .
 · .
 · .
 · .
 · .
 · .
 · 

. :................... __ ....................................... _ .............................. __ ........ _._ .. _ ............................... _._ ............................. _ ................ _ ...... _ .. _ .. _t 


3. ARCHIVES 

Unless instructed otherwise by the Sponsor, all original data and the final report will be 

retained in the Harlan Laboratories Ltd, Shardlow, UK archives for five years, after which 

instructions will be sought as to further retention or disposal. 

1029 

PROJECT NUMBER: 014510218 	 PAGE 8 

4. HYDROLYSIS AS A FUNCTION OF pH 

4.1 Method 

The determination was carried out using Method C7 Abiotic Degradation, Hydrolysis as a 

Function of pH of Commission Regulation (EC) No 44012008 of 30 May 2008 and 

Method 111 of the OECD Guidelines for Testing of Chemicals, 13 April 2004. 

4.2 Procedure 

4.2.1 	 Specification of buffer solutions 

Table 4.1 

Buffer solution 

(pH) Components Concentration 

(moldm-3) 

4 

Citric acid 

Sodium chloride 

0.061 

0.044 

7 

Disodium hydrogen orthophosphate 

(anhydrous) 

Potassium dihydrogen orthophosphate 

Sodium chloride 

0.03 

0.02 

0.02 

9 

Disodium tetraborate 

Sodium chloride 

0.01 

0.02 

The buffer solutions were filtered through a 0.2 IJm membrane filter to ensure they were 

sterile before commencement of the test. Also these solutions were subjected to 

ultrasonication and degassing with nitrogen to minimise dissolved oxygen content. 

4.2.2 Preparation of samples 

Sample solutions were prepared in capped glass bottles at a nominal concentration of 

3.7 x 10-4 gil in the three buffer solutions. A 1% co-solvent of acetonitrile was used to aid 

solubility. 

The test solutions were split into individual vessels for each data point. 

The solutions were shielded from light whilst maintained at the test temperature. 

1030 

PROJECT NUMBER: 0145/0218 PAGE 9 

4.2.3 Preliminary testJTier 1 

Sample solutions at pH 4, 7 and 9 were maintained at 50.0 ±O.soC. 

4.2.4 Tier 2 

Results from the Preliminary testITier 1 showed it was necessary to undertake further 

testing at pH 4, pH 7 and pH 9. Testing at the three pHs was performed as follows; pH 4 

at 40, 50 and 60°C, pH 7 at 20, 30, 40, 50 and 60°C, and pH 9 at 20, 30 and 40°C. The 

time intervals at which hydrolysis were monitored are given in Tables 4.20 to 4.36. 

4.2.5 Analysis of sample solutions 

The sample solutions were taken from the water baths at various times and the pH of 

each solution recorded. 

The concentration of the sample solution was determined by high performance liquid 

chromatography (HPLC). 

Samples 

An aliquot (100 ml) of each sample was extracted with three portions (3 x 10 ml) 

of methyl tertiary butyl ether (MTBE). The combined extracts were evaporated to 

dryness and the residue redissolved in 1 ml of acetonitrile: water (1:1 v/v). 

Standards 

Duplicate standard solutions of test material were prepared in acetonitrile water 

(1:1 v/v) at a nominal concentration of 37 mg/l. 

Matrix blanks 

An aliquot (100 ml) of the relevant buffer solution was extracted with three portions (3 x 

10 ml) of MTBE. The combined extracts were evaporated to dryness and the residue 

redissolved in 1 ml of acetonitrile: water (1:1 v/v). 

1031 

PROJECT NUMBER: 0145/0218 	 PAGE 10 

Analysis 

The standard 

conditions*: 

and sample solutions were analysed by HPLC using the following 

HPLC System Agilent Technologies 1100, incorporating autosampler 

and workstation 

Column Phenomenex 00S3 5lJm (250 x 4.6 mm id) 

Column tempe

Mobile phase 

rature 40°C 

Solvent A: acetonitrile (0.1% orthophosphoric acid) 

Solvent B: water (0.1 % orthophosphoric acid) 

, Time (mins) 

omins 

Solvent A (%) 

50 

Solvent B (%) 

50 

25 mins 80 20 

I 26 mins 100 0 

31 mins 100 0 

Flow-rate 1.0 mllmin 

UV detector wavelength 200 nm 

Injection volume 100 IJI 

Retention time 	 Component A: -12 mins 

Component B: - 22 mins 

• Analysis of pH 4. 40°C samples at time pOints, initial, 48. 70.5 and 144 hours were analysed with the 

following amendments to the conditions: 

1. Column: Phenomenex ODS3 5IJm (150 x 4.6 mm id) 

2. Mobile Phase: gradient modified to 75% Solvent A. 25 % Solvent B at 25 minutes. 

3. Retention times: Component A -9 mlns, Component B -19 mins 

1032 

PROJECT NUMBER: 014510218 PAGE 11 

Typical Chromatography 


Standard: 37.7 mgn 


VWD 1 A. Wavelength=200 nm (PGT3221001-0101.0) 

mAU 

175 

150 

125 

100 

75 

.... '" SO N

<II 

2S 

0 A ~Jl J\ 

n ~ 10 15 

Sample: pH 4, 50°C, 24 Hour 

---_.._-­ VW01 A. Wavelengtll=200 nm (PGT5S41007-0701.0) 

mAU ~ 

200 


175 


150 

~ 125 ~ 

100 

75 

... 50 ... 

~ 25 

10 15 2 I I 20 

1033 

PROJECT NUMBER: 0145/0218 PAGE 12 

Typical Chromatography 


Matrix Blan k: pH 4 


mllU 

VW01 A. Wavelenglh=200 nm (PGT5941021-2102.0) 

200 

175 

15D 

125 

100 

75 

50 

I 

25 

0 

;; 

\.. 

1Q 15 20 2S 

--./ 

Sample: pH 7, 60°C, 43 Hour 

VW01 A. Wavelenglh-200 lim (PGT48BIO07.o701.0) 

mAU 

200 

175 

150 

125 

100 

75 

so 

25 

o 

1034 

PROJECT NUMBER: 014510218 PAGE 13 

Typical Chromatography 


Matrix Blank: pH 7 


mAU 

VWOl A, Wavelengtlr-2OO om (PGT486\D()4..0401.0) 

200 

175 

1!ill 

125 

100 

75 

50 

26 

o 

.M .A ~ \ v-. lLJ-! 

I) Ii HI :IS 2!l 211 '" 

Sample: pH 9, 30GC, 2 Hour 

VW01 A. Wavelength=200 nm (PGT32211l11-1101.0) 

mAU 


175 


150 


125 


100 


75 


50 
 ~ 

25 

~~~A~A~~ '" ________~J~~---~ __~QL-________~5L-________~1~Q__________'~5________~2wO__________.~L-________~m~!a 

1035 

150 

I 

PROJECT NUMBER: 014510218 PAGE 14 

Typical Chromatography 


Matrix Blank: pH 9 


VW01 A. Wawlanglh"'200 nm (PGT3221OD4-0401.D} 


mAU 
1 


175 


125 


100 


75 


25 


~~~~l,~.~.~__~----------------~ 

10 


1036 

PROJECT NUMBER: 014510218 PAGE 15 

Treatlnentofnesults 

Graphs of the common logarithm of the concentration (gil) versus time (hours) were 

plotted for each pH at each temperature (see Figures 4.1 to 4.22) and where applicable, 

the rate constants and half-lives calculated using Equation 4.2 and Equation 4.3 

respectively. 

By plotting the natural logarithm of the rate constants against the reciprocal of the 

temperature (K), the rate constants and half-lives at 25°C were obtained by interpolation 

or extrapolation (see Figures 4.23 and 4.24). 

4.3 Calculation 

4.3.1 Sample solution concentration 

The mean peak height or area* of each standard was corrected to a nominal 

concentration of 37 mgll and the mean value taken. 

The concentration of the sample solutions (gil) was calculated using Equation 4.1. 

Equation 4.1 

PSpI 1 C xDx-­ SPI =-xCstd Psld 1000

where: 

Cspl = sample concentration (g/l) 

Pspl = mean peak height or area of sample solution 

Pstd = mean peak height or area of standard solution, corrected to nominal 

standard concentration 

Cstd = nominal standard concentration (37 mg/l) 

D = sample dilution factor (0.01) 

.. Peak area was used for the pH 4 and 9 calculations and peak height was used for the pH 7 calculations 

(see Discussion, Section 4.6). 

1037 
PROJECT NUMBER: 0145/0218 	 PAGE 16 

4.3.2 Estimation of half-life 

From the graph of log10 concentration (gIl) versus time, the rate constant was calculated 

using Equation 4.2. 

Equation 4.2 

kobs =-slopex2.303 

and the half-life was obtained from the rate constant using Equation 4.3. 

Equation 4.3 

where: 

kObs = 	 rate constant (hours-') 

half-life (hours) 

1038 

PROJECT NUMBER: 014510218 PAGE 17 

4.4 Results 

4.4.1 Preliminary testITier 1 


The mean peak areas relating to the pH 4 and pH 9 standard and sample solutions are 


shown in the following table: 

Solution 

Standard 46.2 mgll 


Standard 46.7 mgll 


Initial Sample A. pH 4 


Initial Sample B. pH 4 


2.4 hour Sample A. pH 4 


2.4 hour Sample B. pH 4 


4 hour Sample A. pH 4 


4 hour Sample B, pH 4 


Standard 46.2 mgll 


Standard 46.7 mgll 


24 hour Sample A. pH 4 


24 hour Sample B. pH 4 


Initial Sample A, pH 9 


Initial Sample B. pH 9 


2.4 hour Sample A. pH 9 


2.4 hour Sample B, pH 9 


24 hour Sample A, pH 9 


24 hour Sample 8, pH 9 


Standard 46.2 mglJ 


Standard 46.7 mgll 


120 hour Sample A, pH 4 


120 hour Sample B, pH 4 _I 


ND - none detected 

Table 4.2 

Component A 


Mean Peak Area 


376.809 

387.843 

349.774 

351.318 

364.178 

240.039 

353.094 

330.860 

384.043 

397.867 

146.740 

278.857 

256.161 

272.743 

NO 

44.970 

NO 

NO 

360.395 

354.935 

65.561 

92.852 

Component B 


Mean Peak Area 


2.570 x 103 


2.587 x 103 


2.672 x 103 


2.882 x 103 


2.852 x 103 


1.607 x 103 


3.062 x 103 


2.725 x 103 


2.511x103 


2.616 x 103 


1.412 x 103 


2.930 x 103 


1.330 x 103 


1.463 x 103 


NO 


76.761 


NO 


NO 


2.490 x 103 


2.496 x 10' 


479.563 


860.987 


1039 

PROJECT NUMBER: 014510218 	 PAGE 18 

The mean peak heights relating to the pH 7 standard and sample solutions are shown in 

the following table: 

Tabla 4.3 

Solution Component B 

Mean Peak Height 

Standard 46.2 mg/l 234.921 

Standard 46.7 mgll 248.334 

Initial Sample A, pH 7 156.610 

Initial Sample S, pH 7 145.797 

2.4 hour Sample A, pH 7 18.970 

2.4 hour Sample S, pH 7 41.456 

50 hour Sample A, pH 7 NO 

50 hour Sample B, pH 7 NO 

96 hour Sample A, pH 7 NO 

96 hour Sample S, pH 7 NO 

NO - none detected 

The test material concentrations at the given time points are shown in the following 

tables: 

Table 4.4 pH 4 at 50.0 ±O.SoC M Component A 

Time (Hours) 

Concentration (gIl) Logio [concentration (gll)1 I % of mean initial 

concentration 

A B A B A S 

0 4.25 X 10"" 4.27 X 10"" -3.37 -3.37 - - 2.4 4.43 x 1.r 2.92 x 10"'" -3.35 -3.54 104 68.5 

4 4.29 x 10"'" 4.02 X 10"" -3.37 -3.40 101 94.4 

24 1.74x10"" 3.31 x 10"" -3.76 -3.48 40.9 77.8 

120 8.51 x 10-5 1.12x 10"'" -4.07 -3.92 20.0 28.3 

Result: 	 Slope = -4.99 x 10-3 (see Figure 4.1) 

r2 = 0.850 

kobs = 1.15 x 10-2 hou(1, 3.19 X 10-6 second-1 

t% = 6004 hours, 2.52 days 

1040 
PROJECT NUMBER: 0145/0218 PAGE 19 

Table 4.5 pH 4 at 50.0 ±O.5°C - Component B 

Time (Hours) Concentration (g/l) L0910 [concentration (gil)] % of mean initial 

concentration 

A 8 A 8 A 8 

0 4.81 X 10"" 5.19x10·4 -3.32 -3.29 - 2.4 5.14x10"" 2.90 x 10"" -3.29 -3.54 103 57.9 

4 5.52 x 10"" 4.91 X 10"" -3.26 -3.31 110 98.1 

24 2.56 x 10"" 5.31 X 10"" -3.59 -3.28 51.1 106 

120 8.94 x 1005 1.60 X 10"" -4.05 -3.80 17.9 32.1 

Result: Slope = -5.00 x 10-3 (see Figure 4.2) 

~ = 0.794 

1.15 x 10-2 hou(1, 3.20 x 10-6 second-1 

60.2 hours, 2.51 days 

Table 4.6 pH 7 at 50.0 ±O.SOC • Component B 

Time (Hours) Concentration (gil) Log10 [concentration (gIl)] % of mean initial 

concentration 

A B A B A B 

Initial 3.01 2.80 -3.52 -3.55 - - 2.4 3.65 7.97 -4.44 -4.10 12.5 27.4 

50 - - - - - - 96 - - - - - -

Result: Slope = -0.305 (see Figure 4.3) 

~ = 0.903 

kobs = 0.702 hour-1, 1.95 x 10-4 second-1 

t~ = 0.988 hours, 4.12 x 10-2 days 

1041 

PROJECT NUMBER: 014510218 	 PAGE 20 

Table 4.7 pH 9 at 50.0 ±O.soC ~ Component A 

Time (Hours) 

Concentration (gil) Log10 [concentration (gil)] % of mean initial 

concentration 

A B A B A B 

Initial 3.04 x 10-4 3.24 X 10"" -3.52 -3.49 - -

2.4 - 5.34 x 10-5 - -4.27 - 17.0 

24 - - - - . -

Result: 	 Slope = -0.321 (see Figure 4.4) 

r2 = 1.00 

0.738 hOUf1

, 2.05 x 10-4 second-1 

0.939 hours, 3.91 x 10.2 days 

Table 4.8 	 pH 9 at 50.0 ±O.soC - Component B 

Time (Hours) 

Concentration (gil) Log10 [concentration (gil)) % of mean initial 

concentration 

A B A B A B 

Initial 2.41 X 10-4 2.65 x 10"" -3.62 -3.58 - 2.4 - 1.39 x 10-5 . -4.86 - 5.50 

24 - - - - . -

Result: 	 Slope = -0.525 (see Figure 4.5) 

r2 = 1.00 

kObl = 	 1.21 hOU(l, 3.36 X 10-4 second-1 

0.574 hours, 2.39 x10-2 days 

For each pH, the extent of hydrolysis indicated that further tests (Tier 2). conducted at 

three temperatures' were required to estimate the rate constants and half-lives. 

* Testing at pH 7 was initially performed at three temperatures (20, 30 and 40°C). Due to inconclusive 

data. further testing was pertormed at elevated temperatures of 50 and 60°C plus some testing at 25°C. 

1042 

PROJECT NUMBER: 0145/0218 PAGE 21 

Figure 4.1 pH 4 at 50.0 ± O.soC - Component A 


Graph of log10 Concentration (gil) versus Time (Hours) 


pH4\PreUmlnary Test at so·e ~-3. 

S 

c 

,8

i ·3, 

§ 

£ 

Q 

~-3.5 

-3.S 

-3.7 

• 

• 

.3. 

-4.0 

-.0 4.)72 _ ~'/O 

l.4OO .uN 2.A4IO 4 ... 4.00II ..... ,.DaD ..... 

- .....
 2'- 4AIG 120JIIIG ....,., 

• -4.1~------------------------""'-------------""'----- o 20 40 60 80 100 120 

Ttme (hours) 

1043 

• 

• • 

• 

PROJECT NUMBER: 014510218 PAGE 22 

Figure 4.2 pH 4 at 50.0 ±0.5OC - Component B 


Graph of IOg10 Concentration (gIl) versus TIme (Hours) 


pH4\Prelimlnary Test at 50'e 

~-3.2 

S 

:;:s

a 

g

c: 

8

1-3.4 

0 

..... 

~ 

-3.6 

-3. 

• 

-4. 

• 

-4.2~------------------------------------------- o 20 40 60 80 100 120 

Time (hours) 

1044 
PROJECT NUMBER: 0145/0218 PAGE 23 

Figure 4.3 pH 7 at SO.O ±O.soC - Component B 


Graph of 10g1o Concentration (gil) versus Time (Hours) 


-3.8 

-4.0 

-4.2 

-4.4 

pHnPrelimlnary Test at SOOC 

• 

• 

-4.6'------------------...-..----. 

0.0 0.5 1.0 1.5 2.0 2.5 

TIme (hours) 

1045 

-3.8 

PROJECT NUMBER: 014510218 PAGE 24 

Figure 4.4 pH 9 at 50.0 :t: 0.5°C - Component A 


Graph of 10910 Concentration (gil) versus Time (Hours) 


pH9\Preliminary Test at 50°C 

......uo:s ..--.

.. 

-4. 

-4.2 

-4.4k-------------------------------~--------

0.0 0.5 1.0 1.5 2.0 2.5 

Time (hours) 

1046 

PROJECT NUMBER: 014510218 PAGE 25 

Figure 4.5 pH 9 at SO.O ::I: 0.5°C - Component B 


Graph of IOg10 Concentration (gil) versus Time (Hours) 


pH9\Preliminary Test at so·e 

::::::-3.2 

~ c 

i 

~ 

~.3. 

J 


-4.0 

-4.4 

-4.8 

-5.2'----------------------­

0.0 0.5 1.0 1.5 2.0 2.5 

Time (hours) 

1047 

PROJECT NUMBER: 014510218 PAGE 26 

4.4.2 Tier 2 

The mean peak areas relating to the standard and sample solutions are shown in the 

following table: 

Table 4.9 - pH 4, 40.0°C 

Solution Component A 

Mean Peak Area 

Component B 

Mean Peak Area 

Standard 36.5 mgll 363.676 2.425 x 103 

Standard 38.4 mgll 384.687 2.571 x 103 

Initial Sample A, pH 4, 40°C 326.388 2.012 x 103 

Initial Sample B, pH 4, 40°C 306.091 1.896 x 103 

Standard 35.5 mgll 349.483 2.340 x 103 

Standard 35.4 mgll 347.767 2.329 x 103 

48 hour Sample A, pH 4, 40°C 305.432 2.073 x 103 

48 hour Sample B, pH 4, 40°C 319.634 1.965 x 103 

Standard 39.0 mgll 396.670 2.680 x 103 

Standard 37.8 mgll 369.230 2.474 x 103 

70.5 hour Sample A, pH 4, 400C 305.548 1.975 x 103 

70.5 hour Sample B, pH 4, 40°C 271.469 1.673 x 103 

Standard 36.1 mgll 359.705 2.330 x 103 

Standard 35.4 mglJ 356.530 2.313 x 103 

144 hour Sample A, pH 4. 400C 307.226 1.775 x 103 

144 hour Sample B. pH 4. 400C 306.714 1.753 x 103 

Standard 36.4 mgll 283.989 2.026 x 103 

Standard 35.1 mg/l 293.127 2.017 x 103 

168 hour Sample A. pH 4, 40°C 148.711 1.338 x 103 

168 hour Sample B, pH 4, 40°C 163.450 1.178 x 103 

Standard 38.9 mgll 329.985 2.495 x 16' 

Standard 36.6 mg/l 309.613 2.138 x 103 

192 hour Sample A. pH 4, 40°C 234.394 1.968 x 103 

192 hour Sample B. pH 4, 40°C 199.665 1.675 x 103 

216 hour Sample A. pH 4. 400c 272.301 1.832 x 103 

216 hour Sample B. pH 4, 400C 276.868 1.999 x 103 

240 hour Sample A. pH 4. 400C 251.460 1.920 x 103. 

240 hour Sample B, pH 4, 400C 256.364 1.803 x 103 

1048 

PROJECT NUMBER: 014510218 PAGE 27 

Table 4.10 - pH 4, 50.0°C 

Solution eomponentA 

Mean Peak Area 

Component B 

Mean Peak Area 

Standard 36.1 mgll 

Standard 36.9 mg/l 

284.882 

304.104 

2.001 x 103 

2.096 x 103 

Initial Sample A, pH 4, 50Ge 260.503 1.250 x 103 

Initial Sample B, pH 4. 50Ge 257.228 1.227 x 103 

17.5 hour Sample A, pH 4, 50Ge 

17.5 hour Sample B, pH 4, 50Ge 

257.282 

205.585 

1.004 x 103 

994.525 

24 hour Sample A, pH 4, 500e 212.452 1.238 x 103 

24 hour Sample B, pH 4, 50Ge 237.786 1.122 x 103 

42 hour Sample A, pH 4, 500C 198.590 890.156 

42 hour Sample B, pH 4, 50Ge 176.134 934.834 

72 hour Sample A, pH 4, 50Ge 

72 hour Sample B, pH 4, sOGe 

164.029 

220.916 

665.490 

922.371 

Standard 36.1 mgtl 280.907 1.948 x 103 

Standard 36.9 mg/l 297.772 2.021 x 103 

144 hour Sample A, pH 4, 50Ge 201.211 800.869 

144 hour Sample B, pH 4, 50Ge 199.580 905.924 

162 hour Sample A, pH 4, 50Ge 208.545 813.875 

162 hour Sample B, pH 4, 50Ge 

209 hour Sample A, pH 4, 50Ge 

209 hour Sample B, pH 4, 5QGe 

221.548 

223.379 

236.429 

i 

I 

994.144 

909.384 

983.451 

1049 

PROJECT NUMBER: 014510218 PAGE 28 

Table 4.11 - pH 4, 60.DoC 

Solution Component A 

Mean Peak Area 

ComponentB 

Mean Peak Area 

Standard 38.9 mgll 329.985 2.495 x 103 

Standard 36.6 mgll 309.613 2.138 x 103 

Initial Sample A. pH 4, 60°C 252.557 2.203 x 103 

Initial Sample B, pH 4, 60°C 256.376 2.612 x 103 

Standard 36.6 mgll 314.073 2.180 x 103 

Standard 36.7 mgll 321.039 2.149 x 103 

5.5 hour Sample A, pH 4, 60°C 217.485 1.455 x 103 

5.5 hour Sample B, pH 4, 60°C 218.362 1.44S x 103 

27 hour Sample A. pH 4, 600C 22S.215 1.407 x 103 

27 hour Sample Bt pH 4, SooC 81.543 349.624 

Standard 36.S mgll 307.341 2.029 x 103 

Standard 36.7 mg/l 314.859 2.062 x 103 

45.5 hour Sample A, pH 4, SooC 219.999 1.368 x 103 

45.5 hour Sample B, pH 4, 60°C 207.938 1.388 x 103 

52.5 hour Sample A, pH 4, eooe 218.841 1.210 x 103 

52.5 hour Sample B. pH 4, 60°C 221.278 1.205 x 103 

72 hour Sample A. pH 4, 60°C 200.475 926.899 

72 hour Sample B, pH 4. 60°C 204.481 1.367 x 103 

96 hour Sample A, pH 4, 600C 224.059 1.559 x 103 

9S hour Sample B, pH 4, 60°C 240.450 2.S89 x 103 

Standard 40.6 mg/l 330.393 2.222 x 103 

Standard 43.0 mg/l 357.839 2.402 x 103 

168 hour Sample A, pH 4, sooe 211.106 1.401 x 103 

168 hour Sample B, pH 4, eo°c 254.353 1.184x 103 

1050 

PROJECT NUMBER: 014510218 PAGE 29 

The mean peak heights relating to the standard and sample solutions are shown in the 

following table: 

Table 4.12 - pH 7, 20.0°C 

Solution Mean Peak Height 

Standard 36.7 mgll 205.480 

Standard 35.3 mgn 185.033 

Initial Sample A. pH 7. 20°C 98.889 

Initial Sample S, pH 7, 20°C 146.413 

2 Hour Sample A. pH 7, 20°C 54.478 

2 Hour Sample S, pH 7, 20°C 71.160 

4 Hour Sample A, pH 7, 20°C 78.544 

4 Hour Sample B. pH 7, 20°C 123.681 

24 Hour Sample A. pH 7, 20°C 105.604 

24 Hour Sample B. pH 7, 20°C 51.597 

Standard 34.0 mg/l 200.386 

Standard 35.0 mg/l 193.892 

96 Hour Sample A, pH 7, 20°C 154.810 

96 Hour Sample B. pH 7, 20°C 207.335 

Standard 36.5 mgJI 198.865 

Standard 36.0 mgll 205.550 

120 Hour Sample A, pH 7, 20°C .53.262 

120 Hour Sample S, pH 7. 20°C 168.125 

144 Hour Sample A, pH 7, 20°C 157.104 

144 Hour Sample B. pH 7, 20°C 42.115 

1051 

PROJECT NUMBER: 014510218 PAGE 30 

Table 4.13 - pH 7, 30.0°C 

Solution Mean Peak Height 

Standard 39.2 mgll 224.389 

Standard 39.3 mg/l 201.859 

Initial Sample A, pH 7, 300 e 230.982 

Initial Sample B, pH 7, 300 e 241.923 

0.5 Hour Sample A, pH 7, 300e 177.151 

0.5 Hour Sample B, pH 7, 300e 178.177 

1 Hour Sample A, pH 7, 300G 217.271 

1 Hour Sample B, pH 7I 300 e ND 

2 Hour Sample A, pH 7, 300 e 174.125 

2 Hour Sample B, pH 7. 300 e 173.772 

3 Hour Sample A, pH 7. 300 e 239.311 

3 Hour Sample B, pH 7, 300e 204.052 

4 Hour Sample A, pH 7, 300 e 187.011 

4 Hour Sample B. pH 7, 300 e 214.148 

6 Hour Sample A, pH 7. 300 e 83.174 

6 Hour Sample B. pH 7, 30ee 229.365 

Standard 39.2 mgll 237.978 

Standard 39.3 mgil 206.333 

24 Hour Sample A, pH 7, 30ee 111.432 

24 Hour Sample B. pH 7, 300 e 239.600 

30 Hour Sample A, pH 7. 300G 221.399 

30 Hour Sample B, pH 7, 30ee 184.277 

Standard 36.7 mgil 208.555 

Standard 35.3 mg/l 184.920 

48 Hour Sample A. pH 7, 300G 42.437 

48 Hour Sample B, pH 7. 30e

e 79.174 

Standard 34.0 mgll 200.386 

Standard 35.0 mg/l 193.892 

120 Hour Sample A, pH 7, 300e 187.426 

120 Hour Sample B, pH 7, 300 e 80.119 

NO - none detected 

1052 

PROJECT NUMBER: 014510218 PAGE 31 

Table 4.14 - pH 7, 40.0°C 

Solution Mean Peak Height 

Standard 37.1 mgll 209.086 

Standard 36.5 mgll 190.380 

Initial Sample A, pH 7, 40°C 173.506 

Initial Sample B, pH 7I 40°C 208.041 

Standard 37.6 mgll 208.951 

Standard 37.5 mg/t 205.367 

72 Hour Sample A, pH 7, 40°C 74.405 

72 Hour Sample B, pH 7, 400C 166.715 

Standard 40.7 mgll 219.627 

Standard 36.5 mgll 182.629 

96 Hour Sample A, pH 7, 40°C 109.314 

96 Hour Sample B, pH 7, 40°C 115.875 

Standard 36.5 mg/! 206.654 

Standard 38.4 mgl1 211.145 

120 Hour Sample A, pH 7, 40°C 86.306 

120 Hour Sample B. pH 7, 40°C 66.804 

Standard 35.5 mg/I 198.470 

Standard 35.4 mg/l 195.271 

120 Hour Sample A, pH 7, 40°C 139.362 

120 Hour Sample B, pH 7, 40°C NO 

Standard 39.0 mg/l 239.926 

Standard 37.S mg/l 226.913 

168 Hour Sample A, pH 7, 400C 90.951 

168 Hour Sample B, pH 7. 40°C 73.880 

Standard 39.0 mg/l 259.826 

Standard 37.8 mg/I 233.015 

240 Hour Sample A. pH 7. 40°C 45.363 

240 Hour Sample B, pH 7. 40°C 76.631 

Standard 36.4 mgll 189.465 

Standard 35.1 mg/l 191.769 

264 Hour Sample A, pH 7, 40°C 31.819 

264 Hour Sample B. pH 7, 40°C 73.540 

Standard 40.0 mg/l 226.175 

Standard 39.3 mgll 217.131 

288 Hour Sample A, pH 7, 40°C 34.121 

288 Hour Sample B. pH 7. 40°C 118.684 

312 Hour Sample A. pH 7, 40°C 84.052 

312 Hour Sample B, pH 7, 40°C 41.059 

Standard 37.1 mg/l 218.160 

Standard 39.1 mg/I 214.240 

336 Hour Sample A, pH 7, 40°C 7.965 

336 Hour Sample B, pH 7, 400C 14.076 

NO - none detected 

1053 
PROJECT NUMBER: 014510218 PAGE 32 

Table 4.15 - pH 7, 50.0oe 

Solution Mean Peak Height 

Standard 34.0 mgll 200.386 

Standard 35.0 mg/I 193.892 

Initial Sample A. pH 7. 500 e 175.853 

Initial Sample B, pH 7. 50Ge 140.206 

2 Hour Sample A. pH 7. 50Ge 62.632 

2 Hour Sample B, pH 7. 500 e 33.762 

Standard 34.0 mg/l 213.735 

Standard 35.0 mg/l 202.226 

4 Hour Sample A, pH 7, 500 e 139.173 

4 Hour Sample B, pH 7, 50Ge 97.991 

6 Hour Sample A, pH 7. 50·e 65.654 

6 Hour Sample B. pH 7, 500e 80.473 

Standard 36.5 mgtl 188.231 

Standard 36.0 mg/l 200.644 

22.75 Hour Sample A. pH 7. 50·e 47.585 

22.75 Hour Sample B, pH 7, 50Ge 50.283 

28 Hour Sample A, pH 7, 50Ge 52.376 

28 Hour Sample B, pH 7. 50·e 50.177 

46 Hour Sample A. pH 7, 50Ge 58.953 

46 Hour Sample B. pH 7, 500

e 47.833 

52 Hour Sample A, pH 7. 50Ge 43.774 

52 Hour Sample B. pH 7. 500 e 41.472 

Standard 36.5 mgtl 198.865 

Standard 36.0 mgll 205.550 

70.5 Hour Sample A. pH 7, 500e 44.187 

70.5 Hour Sample B. pH 7, 500e 46.049 

Standard 37.1 mgtl 209.086 

Standard 36.5 mgt! 190.380 

96 Hour Sample A, pH 7. 50Ge 49.737 

96 Hour Sample B. pH 7, 500 e 53.893 

Standard 37.6 mg/l 208.951 

Standard 37.5 mg/I 205.367 

168 Hour Sample A. pH 7, 500 e 47.802 

168 Hour Sample B, pH 7, 500 e 126.163 

1054 

PROJECT NUMBER: 014510218 PAGE 33 

Table 4.16 - pH 7, 60.0oe 

Solution Mean Peak Height 

Standard 37.6 mgll 208.951 

Standard 37.5 mg/I 205.367 

Initial Sample A, pH 7, aooc 214.411 

Initial Sample B, pH 7, BOoC 228.369 

Standard 40.7 mgJl 219.627 

Standard 36.5 mgll 182.629 

18.75 Hour Sample A, pH 7, 60°C 73.Sn 

18.75 Hour Sample B, pH 7, 60°C 58.289 

Standard 36.5 mgJI 206.654 

Standard 38.4 mgll 211.145 

24 Hour Sample A, pH 7. 60°C 140.374 

24 Hour Sample B. pH 7, 60°C 135.720 

43 Hour Sample A. pH 7. 60°C 169.007 

43 Hour Sample B, pH 7. 60°C 157.198 

48 Hour Sample A. pH 7. 600c 9S.770 

48 Hour Sample B, pH 7, BOoC 41.722 

Standard 35.5 mg/l 198.470 

Standard 35.4 mgJI 195.271 

67 Hour Sample A, pH 7, 600C 30.025 

67 Hour Sample B, pH 7, 60°C 34.356 

Standard 39.0 mgll 239.926 

Standard 37.S mgll 226.913 

72 Hour Sample A, pH 7. 600c 120.256 

72 Hour Sample B, pH 7, 60°C 7.533 

91 Hour Sample A, pH 7, 60°C 50.857 

91 Hour Sample B, pH 7, 60°C 86.965 

96 Hour Sample A, pH 7. 600c 21.370 

96 Hour Sample B. pH 7, 60°C 14.264 

1055 

PROJECT NUMBER: 0145/0218 PAGE 34 

The mean peak areas relating to the standard and sample solutions are shown in the 

following table: 

Table 4.17 - pH 9, 20.O°C 

Component A ComponentB Solution Mean Peak Area Mean Peak Area 

306.851 2.028 x103 Standard 36.0 mgll 

Standard 37.2 mgll 276.116 1.825 x103 

Initial Sample A, pH 9, 20ClC 230.835 1.357 x103 

Initial Sample B, pH 9, 20°C 217.788 1.390 x103 

1 hour Sample A, pH 9, 20°C 51.504 605.362 

1 hour Sample B, pH 9, 20°C 70.532 1.028x103 

2 hour Sample A, pH 9, 20°C 249.740 1.386 x10a 

2 hour Sample B, pH 9, 20°C 224.057 1.287 x103 

4 hour Sample A, pH 9, 20°C 227.952 1.192 x103 

4 hour Sample B, pH 9, 20ClC 191.379 1.025 x103 

Standard 36.0 mgll 327.155 2.170 x103 

Standard 37.2 mg/l 294.037 1.937 x103 

22.5 hour Sample A, pH 9, 20°C 89.477 346.558 

22.5 hour Sample B, pH 9, 20ClC 79.429 318.134 

25 hour Sample A, pH 9, 20°C 61.524 254.378 

25 hour Sample B, pH 9, 20°C 61.814 256.067 

28 hour Sample A, pH 9, 20°C 60.499 317.926 

28 hour Sample B, pH 9, 200C NO NO 

48 hour Sample A, pH 9, 20°C NO 74.124 

48 hour Sample B, pH 9, 200C NO 31.385 

50 hour Sample A, pH 9, 20°C 17.319 112.637 

50 hour Sample B, pH 9, 200C NO 147.497 

ND - none detected 

1056 

PROJECT NUMBER: 014510218 PAGE 35 

Table 4.18 - pH 9, 30.0°C 

Solution Component A 

Mean Peak Area 

eomponentB 

Mean Peak Area 

Standard 37.7 mgll 296.928 2.040 x 103 

Standard 35.3 mg/l 298.854 2.026 x 103 

Initial Sample A, pH 9, 30De 216.380 1.676 x 103 

Initial Sample B, pH 9, 30De 239.002 1.607 x 103 

0.5 hour Sample A. pH 9, 30De 213.732 1.267 x 103 

0.5 hour Sample B. pH 9, 30°C 218.453 1.308 x 103 

1 hour Sample A, pH 9, 30°C 230.713 1.250 x 103 

1 hour Sample B, pH 9, 30°C 220.130 1.429 x 103 

2 hour Sample A, pH 9, 30De 220.375 1.006 x 103 

2 hour Sample S, pH 9, 30De 210.096 983.161 

3 hour Sample A, pH 9, 30°C 105.259 493.0n 

3 hour Sample B, pH 9, 30°C 221.040 933.354 

4 hour Sample A, pH 9, 30°C 175.047 649.668 

4 hour Sample S, pH 9, 30°C 145.732 569.211 

6 hour Sample A, pH 9, 30De 135.529 496.333 

6 hour Sample B, pH 9, 30°C 169.313 574.470 

24 hour Sample A, pH 9, 30°C NO NO 

24 hour Sample B, pH 9, 30De NO NO 

27 hour Sample A, pH 9, 300e NO NO 

27 hour Sample B, pH 9, 30°C NO NO 

30 hour Sample A, pH 9, 30De NO NO 

30 hour Sample B, pH 9, 300e NO NO 

NO - none detected 

1057 
PROJECT NUMBER: 014510218 PAGE 36 

Table 4.19 - pH 9, 40.0°C 

Solution Component A 

Mean Peak Area 

ComponentB 

Mean Peak Area 

Standard 36.8 mg/l 305.932 2.020 x 103 

Standard 34.8 mgll 285.237 1.837 x 103 

Initial Sample A, pH 9, 40°C 230.879 1.343 x 103 

Initial Sample B, pH 9, 40°C 206.530 1.237 x 103 

0.5 hour Sample A, pH 9, 40°C 232.431 1.175 x 103 

0.5 hour Sample B, pH 9, 40°C 254.164 1.339 x 103 

1 hour Sample A, pH 9, 40°C 177.409 787.260 

1 hour Sample B, pH 9, 400C 205.872 904.013 

2 hour Sample A, pH 9, 40°C 173.851 598.211 

2 hour Sample B, pH 9, 40°C 180.180 641.124 

3 hour Sample A, pH 9, 40°C 12.819 124.355 

3 hour Sample B, pH 9, 40°C 95.648 358.425 

4 hour Sample A, pH 9, 40°C 28.042 153.363 

4 hour Sample B, pH 9, 40°C 43.125 187.116 

5 hour Sample A, pH 9. 40°C NO 15.913 

5 hour Sample B. pH 9. 40°C 12.487 85.739 

6 hour Sample A, pH 9. 40°C 

6 hour Sample B, pH 9, 40°C I 

I 

NO 

NO 

NO 

NO 

NO - none detected 

1058 

PROJECT NUMBER: 0145/0218 	 PAGE 37 

The test material concentrations at the given time points are shown in the following 

tables: 

Table 4.20 pH 4 at 40.0 ± 0.5°C • Component A 

Time (Hours) 

Concentration (gIl) L0910 [concentration (gll)J % of mean initial 

concentration 

A B A B A B 

Initial 3.27 x 10"'" 3.06 x 10"'" -3.49 -3.51 - - 48 3.11 x 10"'" 3.25 x 10"" -3.51 -3.49 98.1 103 

70.5 3.07 x 10"" 2.72 x 10-4 -3.51 -3.57 96.8 86.0 

144 3.07 x 10-4 3.06 x 10"" -3.51 -3.51 96.9 96.7 

168 1.84 x 10"'" 2.02 x 10"" I -3.74 -3.69 58.2 63.9 

192 2.77 x 10-4 2.36 x 10"" -3.56 -3.63 87.4 74.5 

216 3.21 x 10"'" 3.27 X 10"" -3.49 -3.49 102 103 

240 2.97 x 10"'" 3.03 X 10"'" -3.53 -3.52 93.8 95.6 

Result: 	 No definitive signs of hydrolysis. Half-life expected to be greater than 

1 year at 25°C. 

Table 4.21 pH 4 at 40.0 t 0.5°C - Component B 

Time (Hours) Concentration (g/l) L0910 [concentration (g/l)1 % of mean initial 

concentration 

A B A B A B 

Initial 3.01 x 10"" 2.84 X 10"" -3.52 -3.55 - - 48 3.15 x 10"" 2.98 X 10"'" -3.50 -3.53 107 102 

70.5 2.94 x 10"" 2.49 X 10"" -3.53 -3.60 100 85.1 

144 2.73 x 10"" 2.70 X 10'" -3.56 -3.57 93.3 92.1 

168 2.37 x 10"'" 2.08 X 10-4 -3.63 -3.68 80.8 71.1 

192 3.21 x 10"'" 2.73 X 10-4 -3.49 -3.56 110 93.3 

216 2.99 x 10-4 3.26 x 10"" -3.52 -3.49 102 111 

240 3.13x10"" 2.94 x 10"" -3.50 -3.53 107 100 

Result: No definitive signs of hydrolysis. Half-life expected to be greater than 

1 year at 25°C. 

1059 

PROJECT NUMBER: 0145/0218 	 PAGE 38 

Table 4.22 pH 4 at 50.0 ±0.5OC • Component A 

Time (Hours) Concentration (gil) LOQ10 [concentration (gil)} % of mean initial 

concentration 

A B A B A B 

Initial 3.23 X 10-4 3.19 X 10-4 -3.49 -3.50 - - 17.5 3.19 X 10-4 2.55 X 10-4 -3.50 -3.59 99.4 79.4 

24 2.63 x 10-4 2.95 X 10-4 -3.58 -3.53 82.1 91.9 

42 2.46 x 10-4 2.18 X 10-4 -3.61 -3.66 76.7 68.0 

72 2.03 x 10"" 2.74 X 10-4 -3.69 -3.56 63.4 85.3 

144 2.54 x 10-4 2.52 X 10-4 -3.60 -3.60 79.1 78.5 

162 2.63 x 10-4 2.80 X 10-4 -3.58 -3.55 82.0 87.1 

209 2.82 x 10-4 2.98 X 10-4 -3.55 -3.53 87.8 93.0 

Result: 	 No definitive signs of hydrolysis. Half-life expected to be greater than 

1 year at 25"C. 

Table 4.23 pH 4 at 50.0 ± O.SoC • Component B 

Time (Hours) 

Concentration (gIl) LOQ10 (concentration (gil)] % of mean initial 

concentration 

A B A B A B 

Initial 2.23 x 10"" 2.19x 10-4 -3.65 -3.66 - -

17.5 1.79x10-4 1.77 x 10-4 -3.75 -3.75 81.1 80.3 

24 2.21 x 10-4 2.00 X 10-4 -3.66 -3.70 99.9 90.6 

42 1.59 x 10-4 1.67x10-4 -3.80 -3.78 71.9 75.5 

72 1.19x10-4 1.64 x 10-4 -3.93 -3.78 53.7 74.5 

144 1.47 X 10-4 1.67 x 10-4 -3.83 -3.78 66.7 75.5 

162 1.50 x 10-4 1.83 X 10-4 -3.83 -3.74 67.8 82.8 

209 1.67 x 10-4 1.81 X 10-4 -3.78 -3.74 75.8 81.9 

Result: No definitive signs of hydrolysis. HalfRlife expected to be greater than 

1 year at 25"C. 

1060 

PROJECT NUMBER: 014510218 	 PAGE 39 

Table 4.24 pH 4 at 60.0 ± 0.5°C - Component A 

Time (Hours) 

Concentration (g/l) Logl0 [concentration (gil)] % of mean initial 

concentration 

A B A B A B 

Initial 2.98 x10-4 3.03 X 10-4 -3.53 -3.52 - - 5.5 2.54 x 10-4 2.55 X 10-4 -3.59 -3.59 84.7 85.0 

27 2.65 x 10-4 9.54 X 10.5 -3.58 -4.02 88.1 31.8 

45.5 2.63 x10-4 2.48 X 10-4 -3.58 -3.61 87.5 82.7 

52.5 2.61 x10-4 2.64 X 10-4 -3.58 -3.58 87.0 88.0 

72 2.39 x10-4 2.44 X 10.4 -3.62 -3.61 79.7 81.3 

96 2.68 x 10-4 2.87 X 10-4 -3.57 -3.54 89.1 95.6 

168 2.57 x10-4 3.09 X 10-4 -3.59 -3.51 85.4 103 

Result: 	 No definitive signs of hydrolysis. Half-life expected to be greater than 

1 year at 25 QC. 

Table 4.25 pH 4 at 60.0 ±O.SoC - Component B 

No definitive signs of hydrolysis. Half-life expected to be greater than 

1 year at 25Q C. 

Time (Hours) 

: 

Initial 

5.5 

27 

45.5 

52.5 

72 

96 

168 

Result: 

Concentration (gil) Log10 [concentration (glOJ % of mean initial 

concentration 

A B A B A B 

3.60 x10-4 4.26 X 10-4 -3.44 -3.37 - - 2.50 x10-4 2.48 X 10-4 -3.60 -3.61 63.5 63.1 

2.41 x10-4 6.00 X 10-0 -3.62 -4.22 61.4 15.3 

2.48 x 10-4 2.52 X 10-4 -3.61 -3.60 63.2 64.2 

2.20 x10-4 2.19 X 10-4 -3.66 -3.66 55.9 55.7 

1.68 x 10-4 2.48 X 10-4 -3.77 -3.61 42.8 63.2 

2.83 x10-4 4.88 X 10-4 -3.55 -3.31 72.0 124 

2.53 x 10-4 2.14 X 10.4 -3.60 -3.67 64.5 54.5 

1061 

PROJECT NUMBER: 014510218 	 PAGE 40 

Table 4.26 pH 7 at 20.0 ± 0.5°C M Component B 

I 

Time (Hours) • 

Concentration (gil) Log10 {concentration (gIl)] % of mean initial 

concentration 

A B A B A B 

Initial 1.82 x 10'" 2.70 X 10'" -3.74 -3.57 - - 2 1.01 X 10'" 1.31 X 10'" -4.00 -3.88 44.4 58.0 

4 1.45 x 10'" 2.28 X 10'" -3.84 -3.64 64.0 101 

24 1.95 x 10'" 9.52 X 10.5 -3.71 -4.02 86.1 42.1 

96 2.71 x 10'" 3.63 X 10'" -3.57 -3.44 120 160 

120 9.55 x 10-5 3.01 X 10'" -4.02 -3.52 42.2 133 

144 2.82 x 10'" 7.55 X 10.5 -3.55 -4.12 124 33.4 

Result: 	 No definitive signs of hydrolysis. Half-life expected to be greater than 

1 year at 25°C. 

Table 4.27 pH 7 at 30.0 ± O.soC - Component B 

Time (Hours) 

Initial 

0.5 

1 

2 

3 

4 

6 

24 

30 

48 

120 

Result: 

Concentration (gil) L0910 [concentration (g/l)] % of mean initial 

concentration 

A B A B A B 

4.25 x 10'" 4.46 X 10'" -3.37 -3.35 - - 3.26 x 10'" 3.28 X 10'" -3.49 -3.48 74.9 75.4 

4.00 x 10'" - -3.40 - 91.9 - 3.21 X 10'" 3.20 X 10'" -3.49 -3.50 73.6 73.5 

4.41 x 10'" 3.76 X 10'" -3.36 -3.43 101 86.3 

3.44 x 10'" 3.94 X 10'" -3.46 -3.40 79.1 90.6 

1.53 x 10'" 4.22 X 10'" -3.82 -3.37 35.2 97.0 

1.97 x 10'" 4.23 X 10'" -3.71 -3.37 45.2 97.2 

3.91 x 10'" 3.26 X 10'" -3.41 -3.49 89.8 74.8 

7.77 x 10-4 1.45 X 10'" -4.11 -3.84 17.8 33.3 

3.28 x 10'" 1.40 X 10'" -3.48 -3.85 75.3 32.2 

No definitive signs of hydrolysis. Half-life expected to be greater than 

1 year at 25°C. 

1062 

72 

PROJECT NUMBER: 014510218 PAGE 41 

Table 4.28 pH 7 at 40.0 ± O.SOC - Component B 

Result: 

Time (Hours) 

Initial 

96 

120 

144 

168 

240 

264 

288 

312 

336 

Concentration (gIl) L0910 [concentration (gIl)] I % of mean initial 

concentration 

A B A B A B 

3.20 x 10-4 3.83 X 10-4 -3.50 -3.42 - - 1.35 x 10-4 3.02 X 10-4 -3.87 -3.52 38.4 85.9 

2.10 x10-4 2.23 x 10-4 -3.68 -3.65 59.8 63.4 

1.55 x 10-4 1.20 x 10-4 -3.81 -3.92 44.0 34.0 

2.51 x 10-4 - -3.60 - 71.7 - 1.50 x 10-4 1.22 x 10-4 -3.83 -3.92 42.6 34.6 

7.07 x 10-6 1.20 X 10-4 -4.15 -3.92 20.1 34.0 

5.97 x 10-5 1.38 x 10...( -4.22 -3.86 17.0 39.2 

6.10 X 10-6 2.12 x 10...( -4.21 -3.67 17.4 60.4 

1.50 x 10-4 7.35 X 10-5 -3.82 -4.13 42.8 20.9 

1.40 x 10-5 2.48 X 10.5 -4.85 -4.61 3.99 7.05 

No definitive signs of hydrolysis. Half-life expected to be greater than 

1 year at 25°C. 

Table 4.29 pH 7 at 50.0 ±O.soC - Component B 

I 

Time (Hours) Concentration (gil) LOQ10 [concentration (gIl)) % of mean initial 

concentration 

A B A B A B

Initial 3.08 x 10.4 2.45 X 10-4 -3.51 -3.61 - - 2 1.10 X 10-4 5.91 X 10-5 -3.96 -4.23 39.6 21.4 

4 2.31 x 10...( 1.62 X 10-4 -3.64 -3.79 83.5 58.8 

6 1.09 x 10-4 1.33 X 10-4 -3.96 -3.88 39.4 48.3 

22.75 8.87 x 10-6 9.37 x 10-6 -4.05 -4.03 32.1 33.9 

28 9.76 x 10-5 9.35 x 10-6 -4.01 -4.03 35.3 33.8 

46 1.10 x 10-4 8.92 x 10-6 -3.96 -4.05 39.7 32.3 

52 8.16 x 10-5 7.73 X 10.5 -4.09 -4.11 29.5 28.0 

70.5 7.92 x 10-6 8.25 x 10-6 -4.10 -4.08 28.7 29.9 

96 9.17 x 10.5 9.93 x 10.5 -4.04 -4.00 33.2 35.9 

168 8.67 x 10-5 2.29 X 10-4 -4.06 -3.64 31.3 82.7 

Result: No definitive signs of hydrolysis. Half-life expected to be greater than 

1 year at 25°C. 

1063 

PROJECT NUMBER: 0145/0218 PAGE 42 

Table 4.30 pH 7 at 60.0 ± 0.5°C - Component B 

Time (Hours) Concentration (g/l) L0910 [concentration (gil)} % of mean initial 

concentration 

A 8 A 8 A 8 

Initial 3.89 x 10-4 4.14 X 10.4 -3.41 -3.38 - -

18.75 1.42 x 10-4 1.12 X 10-4 -3.85 -3.95 35.4 27.9 

24 2.52 x 10-4 2.43 X 10"" -3.60 -3.61 62.7 60.6 

43 3.03 x 10-4 2.82 X 10-4 -3.52 -3.55 75.5 70.2 

48 1.77 x 10-4 7.48 X 10.5 -3.75 -4.12 44.1 18.6 

67 5.41 x 10-5 6.19x10·s -4.26 -4.21 13.5 15.4 

72 1.98 x 10"" 1.24 x 10-5 -3.70 -4.91 49.3 3.09 

91 8.37 x 10.5 1.43 X 10"" -4.08 -3.84 20.9 35.7 

96 3.52 x 10-5 2.35 X 10-5 -4.45 -4.63 8.76 5.85 

Result: No definitive signs of hydrolysis. Half-life expected to be greater than 

1 year at 25°C. 

1064 

PROJECT NUMBER: 0145/0218 	 PAGE 43 

Table 4.31 pH 9 at 20.0 :t O.SOC • Component A 

Time (Hours) 

Concentration (gil) Log10 [concentration (911)] % of mean initial 

concentration 

A B A B A B 

Initial 2.90 x 10-4 2.73 X 10-4 ·3.54 -3.56 - - 1* 6.46 X 10-5 8.85 X 10-6 4.19 -4.05 23.0 31.4 

2 3.13x10-4 2.81 X 10-4 -3.50 -3.55 111 99.9 

4 2.86 x 10-4 2.40 X 10-4 -3.54 -3.62 102 85.3 

22.5 1.05 x 10-4 9.35 X 10-6 -3.98 -4.03 37.4 33.2 

25 7.24 x 10.5 7.28 x 10-6 4.14 -4.14 25.7 25.9 

28 7.12 x 10.5 - 4.14 - 25.3 . 

48 - - - - - - 50 2.04 X 10.5 - -4.69 - 7.25 -

Result: 	 Slope = -2.36 X 10-2 (see Figure 4.17) 

f = 0.988 

kobs = 5.43 x 10-2 hOU(1, 1.51 x 10-5 second-1 

t% = 12.8 hours, 0.532 days 

Table 4.32 	 pH 9 at 20.0 ±O.SoC - Component B 

TIme (Hours) 

Initial 

1 

2 

4 

22.5 

25 

28 

48 

50 

Concentration (gil) L0910 [concentration (gil)] 

A 

2.58 x 10-4 

1.15 X 10"" 

2.63 x 10-4 

2.26 x 10-4 

6.17 X 10-fi 

4.53 x 10-6 

5.66 x 10-5 

1.32 x 10.5 

2.01 x 10.5 

B A B 

2.63 X 10"" 3.59 -3.58 

1.95 X 10-4 -3.94 -3.71 

2.44 X 10-4 -3.58 -3.61 

1.95 X 10-4 -3.65 -3.71 

5.66 x 10-6 4.21 4.25 

4.56 X 10-6 -4.34 -4.34 

- 4.25 - 5.59 X 10-6 4.88 -5.25 

2.63 x 10.5 4.70 4.58 

-2.49 X 10.2 (see Figure 4.18) 

0.897 

5.73 x 10-2 hOUf1. 1.59 X 10-5 second-1 

12.1 hours, 0.504 days 

% of mean initial 

concentration 

A B 

- - 44.1 74.9 

101 93.7 

86.8 74.6 

23.7 21.7 

17.4 17.5 

21.7 - 5.06 2.14 

7.69 10.1 

Result: Slope = 

f = 

kobs = 

t% = 

.. 1 hour time pOint considered to be an outlier and not used in the calculations. 

1065 

PROJECT NUMBER: 0145/0218 	 PAGE 44 

Table 4.33 pH 9 at 30.0 ± O.SoC • Component A 

Result: 	 Slope = 

r2 = 

kobs = 

Concentration (gil) Log10 [concentration (g/l)1 % of mean initial 

Time (Hours) concentration 

A B A B A B 

Initial 2.65 x 10-4 2.93 X 10-4 -3.58 -3.53 - - J 

0.5 

1 

2 

3 

4 

6 

2.62 x 10-4 

2.82 X 10-4 

2.70 X 10-4 

1.29 x 10-4 

2.14 x 10-4 

1.66 X 10-4 

2.67 X 10-4 -3.58 -3.57 

2.69 X 10-4 -3.55 -3.57 

2.57 X 10-4 -3.57 -3.59 

2.71 X 10-4 -3.89* -3.57 

1.78 X 10-4 -3.67 -3.75 

2.07 X 10'" -3.78 -3.68 

-3.18 X 10-2 (see Figure 4.19) 

0.731 

7.32 x 10-2 hour1 

, 2.034 x 10-5 second-1 

9.47 hours, 0.394 days 

93.9 95.9 

101 96.7 

96.8 92.3 

46.2 97.1 

76.9 64.0 

59.5 74.4 

I 

I 

Table 4.34 	 pH 9 at 30.0 ± O.SOC - Component B 

Time (Hours) 

I 

! 

Concentration (g/l)* 

A B 

Log10 [concentration (gIl)] 

A· I B 

% of mean initial 

concentration 

A B 

Initial 3.01 x 10-4 2.88 X 10'" -3.52 -3.54 - - 0.5 2.27 x 10-4 2.34 X 10-4 -3.64 -3.63 77.2 79.7 

1 

2 

3 

2.24 x 10'" 

1.81 x 10'" 

8.85 x 10.5 

2.56 X 10-4 

1.76 X 10-4 

1.67 X 10'" 

-3.65 

-3.74 

-4.05* 

-3.59 

-3.75 

-3.78 

76.2 

61.3 

30.0 

87.0 

59.9 

56.9 

4 

6 

1.17 x 10-4 

8.90 x 10.5 

1.02x10'" 

1.03 X 10'" 

-3.93 

-4.05 

-3.99 

-3.99 

39.6 

30.2 

34.7 

35.0 

Result: 	 Slope = -8.26 X 10-2 (see Figure 4.20) 

r2 = 0.935 

kobs = 0.190 hour\ 5.28 x 10.5 second-1 

t~ = 3.65 hours, 0.152 days 

* Components A and B. 3 hour time points, replicate A, considered to be outliers and not used in the 

calculations. 

1066 
PROJECT NUMBER: 0145/0218 PAGE 45 

Table 4.35 pH 9 at 40.0 ±0.5°C • Component A 

Time (Hours) 

Concentration (gil) Log10 [concentration (g/l)1 % of mean initial 

concentration 

A B A B A B 

Initial 2.80 x 10-4 2.50 X 10-4 -3.55 -3.60 - - 0.5 2.82 x 10-4 3.08 X10-4 -3.55 -3.51 106 116 

1 2.15 x 10-4 2.49 x 10'" -3.67 -3.60 81.1 94.1 

2 2.11 x 10-4 2.18 X 10-4 -3.68 -3.66 79.5 82.4 

3 1.55 x 10-5 1.16x10-5 -4.81* -3.94 5.86 43.7 

4 3.24 x 10-5 4.99 X 10.5 -4.49 -4.30 12.2 18.8 

5 - 1.44 X 10-5 - -4.84 - 5.45 

Result: Slope = -0.238 (see Figure 4.21) 

f = 0.872 

0.549 hou(1, 1.53 x 10-4 second-1 !cobs = 


t% = 1.26 hours, 5.26 x 10-2 days 


Table 4.36 pH 9 at 40.0 ±O.5°C - Component B 

Time (Hours) 

Concentration (g/l)* Log10 [concentration (gIl)] % of mean initial 

concentration 

A B A B A B 

Initial 2.50 x 10-4 2.30 X10-4 -3.60 -3.64 - - 0.5 2.18x10-4 2.49 x 10-4 -3.66 -3.60 91.1 104 

1 1.46 x 10-4 1.68 X 10-4 -3.84 -3.78 61.0 70.1 

2 1.11 x 10-4 1.19x10'" -3.95 -3.92 46.4 49.7 

3 2.31 x 10.5 6.66 X 10-5 -4.64t -4.18 9.64 27.8 

4 2.69 x 10-5 3.28 X 10-5 -4.57 -4.48 11.2 13.7 

5 2.79 x 10-6 1.51 X 10.5 -5.55t -4.82 1.17 6.28 

Result: Slope = -0.240 (see Figure 4.22) 

f = 0.972 

kobs = 0.551 houf1, 1.53 x 10-4 seconcf1 

1.26 hours, 5.24 x 10-2 days 

.. Component A. 3 hour time point, replicate A. considered to be an outlier and not used in the calculations. 

t Component B, 3 and 5 hour time points, replicate A. considered to be outliers and not used In the 

calculations. 

1067 
PROJECT NUMBER: 014510218 	 PAGE 46 

Figure 4.6 pH 4 at 40.0 ±O.soC - Component A 


Graph of 10g1o Concentration (gil) versus Time (Hours) 


o.CIOIE40

~ 

-
-
-
'IUGII 

-

144.C1111 

,_
 -
-
-
 21U11O 

21U11O

-


• -3.7·5'--~----------------------

o 	 50 100 150 200 250 

TIme (hours) 

1068 
-- • • 

PROJECT NUMBER: 0145/0218 PAGE 47 

Figure 4.7 pH 4 at 40.0 ::t O.soC - Component B 

Graph of log10 Concentration (gil) versus Time (Hours) 

• 

• 

• 


• 


!:=::-3.48 

0:::: 

S 

Co • 0 

:.1:1 • ~ 1-3 53

• • 0 • -

ClII 

.S! 

_to 4.121 

4,I4C 

.ua2 

Q,OQIII!" 

-

4.. oIl1111 -

7O.IIDI oUIt 144_ 01_

~ 

t.._ 

,.­

4_ t._ 1.._ ••1 

1­ .3._ 

tlUGO ... 21"_ .uIl

ItuaO .3.4111 

: ....4,I4C 1-..-.. 

.f~ -3.56 

• 

-3.60 • 

• 

-3.64 

-3.68 • 

-3.72 

0 50 100 150 200 250 

Time (hours) 

1069 

• • 

PROJECT NUMBER: 0145/0218 PAGE 48 

Figure 4.8 pH 4 at 50.0 ±O.5°C- Component A 


Graph of log10 Concentration (gIl) versus Time (Hours) 


~-3.48 a_


! 

i 

• 

g-3.62 

~ • ..... 

d •

CD 

..!2 

• • 

-3.58 • 

•

-3.60 

•

• 

• 

-3.84 

• 

-3.68 

• 

-3.72 

a-.o 17_ 11_

----

T.tOIIO 

11_ 

,

I....

..­

-

1__ 

= 

4AM 

.;tMI 

4AtII 

.UN 

4Jl7I 

4.QO

.s.tCIII 

-1.111

--

.:a_

-

4.­

A• .:aJIM = - ':1: 

0 40 80 120 160 200 240 

Time (hours) 

1070 

--

PROJECT NUMBER: 0145/0218 	 PAGE 49 

Figure 4.9 pH 4 at SO.O :t O.soC- Component B 


Graph of 10910 Concentration (gIl) versus Time (Hours) 


• 

-3.915'----------------------­

o 	 40 80 120 160 200 240 

Time (hours) 

~-3.6 

i • 

L7 • .... 

~ 

r 

• • 

~ .,. 0.-..0 17_ 17_ 4'­

4:1" 

- 4.11' 

-- 4 .• 

of••

4_ 

.ani 1'lD -3.121 1'lD -UN

.... ..... ;~ ­ 4.11'1

182.GOO 4._ l_ ...m - A-4.m

.....-

oll_ 

1071 
Figure 4.10 pH 4 at 60.0 1: O.soC- Component A 


Graph of 10g1o Concentration (gn) versus Time (Hours) 


PROJECT NUMBER: 014510218 	 PAGE 50 

~..a. ..
 • -C 

1 • 	

• 

• 

g -3. • 

I .... 

~ 

I 

-3.7 

-3.8 

_ 

0.-.. 

IJ,.$GO 

UCiO 21_ 

%1I/l10

... 

.u. 

IUGII 

IaJII1II 

~ 

...

... 

'''0IIII 

<UIII ".It, 

"J84 ".u71 ...

""-ClIO 

4M1

,,­ 4._ 

.u71 

4 ..' .un 

..un 

.ua 

4MI 

A.~

... 

• 

~.1~-------------------------------------------- o 	 80 120 160 200 

TIme (hours) 

1072 

--3.7 

PROJECT NUMBER: 014510218 PAGE 51 

Figure 4.11 pH 4 at 60.0 ± O.S°C- Component B 


Graph of log10 Concentration (gil) versus Time (Hours) 


2;:-3. 

S •

i 

8 

I::: J,w3.4 

..... 

~ 

D_ 

~ 

-II,"" 

-2.:1'10 

-.... 

..­ .­ ..­ .... 11'_ "flY 

-.uos 

.uN 

IUCII 4.l1li.

.­ 4MO 

-

~ 4.774

n­ .... IUD 4_4:at1

-".;&JIIt 

.uta 

.......... 


• • 

• 

·3.8'-----------------------..... 
 o 40 80 120 160 200 

nme(hours} 

1073 

PROJECT NUMBER: 014510218 PAGE 52 

Figure 4.12 pH 7 at 20.0 :t 0.5°C- Component B 


Graph of IOg10 Concentration (gIl) versus Time (Hours) 


~-3.4 

i - 8

i -3.5 

C· 

.§. 

C

... ..S'-3.6 

• 

• 

• 

• 

• 

-3.7 • 

- -3.8 

• 

-3.9 • 

-4.0 • 

• • 

-4.1 

• 

-4.2 

0 40 80 .120 160 

Time (hours) 

1IJIIIIII.a 

~ 

2.GOO 

2.GOO 

.a.ooo 

4.Il00 

<iNJIIIt 

:wIlD

120.Il00 -­

-­

:: -

4..". 

4MI 

4_ 

4_ 

4.m 

4.1Nl 

4.7tII 

-4J1i11 

4.H7 

4...cI 

"'-"20 

4_ 

~ ...4.7111 •• 3.......

•-li.1D4 

1074 

--

PROJECT NUMBER: 014510218 	 PAGE 53 

4.m 

4:18' 

4MI 

4MI 

..u. 

4MI 

<IMt!I 

4.:l1li 

4Aa 

.:JAG 

4_ 

':'.l1S 

4.114 

.... ".37:1 ­

~ 

-',417 "".
 4MI

""­

Figure 4.13 pH 7 at 30.0 ± O.S°C-Component B 


Graph of log1D Concentration (gil) versus Time (Hours) 


0._
 !I.IIOIIEtG 

o.faO 

o.faO

,.­

UGO 2._

3.0IIII 

~ 

4JIIIG 

1.000 

-

uoo 

24.1100 

-.... 


....


--
A-4A11

..,. ~ 

• 
 • 

• 


-42.~----------------------------------------- o 	 20 40 60 80 100 120 

Time (hours) 

~-3.2 

-­i 

.c 

8 •

1-3.4 • .... 

I 

83. 

~.8 • 

-4. 

• 

• 

• 

• 

1075 

--

-4.4 

PROJECT NUMBER: 0145/0218 	 PAGE 54 

Figure 4.14 pH 7 at 40.0 :t O.SOC- Component B 


Graph of log10 Concentration (gil) versus Time (Hours) 


~-3.2 

c: 

L~ • 0 •

I 

..­

-4. 


0_ 4.... 

~tO ~AIt 

n.ooo 4.1111 n.ooo <IJI2II

oUTI 

,_­ 4M2

4.1\1 

1_1_­ 4_ 

4AOO 

1M_ 4.111!1 1_ 4.115 

-4.tI!II _OlIO 4.123 

-

2M_ -4.ZM 

-1M! 

- ..uu 

.1m 

-

JIZ..GIJII ...­ :n:I.GIIO -4.IM 

-4.153 

-A ......«II

..
.~ 

• 

• 

-5.2'-------------------------' 


o 	 100 200 300 400 

Time (hours) 

1076 

• • 

• • • 

--

-4.1 

PROJECT NUMBER: 014510218 PAGE 55 

Figure 4.15 pH 7 at 50.0 ±O.soC- Component B 


Graph of 10g1o Concentration (g/l) versus Time (Hours) 


~ 4.11l 

o.!ICIIIIM - ­4.l1G

2.lIIIO 4JIIII

Z.QlII .... f. 4.IS7 4JIIIO 4.711 UIII 4.1113

.... 4.m 

.. 

-4

- lUOII • lUOII • 3... ..­ ..unG 2&lIIIO 

.... 4_

.... .... - .... 

-

- -4.1t:/ 

".101 

",au -..­ ...

".GO! 

=: 1:: "'-4.121 

a....?'77II.f 

o4.f!1l! 

• 

• 

-4.2 

-4.3.~-------------------------------------------

o 40 80 120 160 200 

TIme (hours) 

• 


1077 

6 

PROJECT NUMBER: 0145/0218 PAGE 56 

Figure 4.16 pH 7 at 60.0:t 0.6OC- Component B 


Graph of log10 Concentration (gil) versus Time (Hours) 


~-3.2 

~ 

J 

E: 

Br: 

~-3. ..... 

F 

• 

• 

-4.4 

-4.8 

• 

-5.2~l-------,----------------

..-.0 G_'" ,a.TiIQ 

lilJ110 

IN.IIGO 

U.llGO 

~lGCIG 

GJIGO 

8.11CO

...., .,­ .,,­

,,_
 ?2.DIIO

1l.OOO 

fI.IIGO

.... 


4.41& 

.ut:! 

.....7 ...... 

4_ 

4.1tl

4.$11 .:1_

....,., ..... 


.......,. 


"-*

..... -4._1111 

.un' 

4.144 "'..... ••"IMI) 

.....a1... . 


o 20 40 60 80 100 

Time (hours) 

1078 

PROJECT NUMBER: 014510218 PAGE 57 

Figure 4.17 pH 9 at 20.0 :t O.SOC- Component A 


Graph of log10 Concentration (gIl) versus Time (Hours) 


::;::-3.4

c:::: 

.9 

~ • 

i

i-3

.

a 

0

.... 

B' 

-3.8 

0_0.-.. -UIII 

-- ."....

uoo -

-

-

~ 

ZJIlI) -UII' ~ .".... UOII 

-uN 


22.5CD 


-

111,000 ....­

"',1. 

- .....Q 

-..."--IJIIII' 

4.1. 

.

~ 

-4.0 

-4.2 

-4.4 

-4.6 

4.8:~----------------------------------------- o 10 20 30 40 50 

Time (hours) 

1079 

---

PROJECT NUMBER: 014510218 PAGE 58 

Figure 4.18 pH 9 at 20.0 ± O.S°C- Component B 


Graph of 10910 Concentration (g/l) versus Time (Hours) 


:;::-3.2 

i 

S 

! 

t:: 

§

8 --3. 

a'

.... 

j 

-4. 

-4. 

-5.2 

• 

-5..6'----------------------­

. 

_-

1_.. -....... 


4.II1II 1._ "L7tG 

2JIOII ..... 2JIOII _I

~.oao 4.... 

4.(11l0 41f' 11_ 2UOII -4.no 

-urr 

~~ 

- ~M1 

-

&ClIO ..c.a41 

...... ".I11III .a.2D 

--A-4AMO

..,. ~ 

7 

o 10 20 30 40 50 

TIme (hours) 

1080 

PROJECT NUMBER: 014510218 PAGE 59 

Figure 4.19 pH 9 at 30.0 t 0.6°C- Component A 


Graph of 10910 Concentration (gil) versus Time (Hours) 


~-3.5 

1I.CCCE<o 4.177 :§ 1I.CCCE<o 4.Q1 

~ 

! 

c; 4.112 4IMI) ~ 1.oao 4Mt ,.oao ~ 

c; 2.lDI -aMI 2.lIIIO ''''Il10 

~ -3.55 • -

UCG ..AlII 

UCG -

4Mt .§. 

• ~.oao 4.748 0 .1.710

~ 

.... • -A·4.511/

.'·UIGM . 

-3.7 

, • 

-3.8 

• 

• 


0 2 3 4 5 6 

Time (hours) 

1081 

• • 

PROJECT NUMBER: 014510218 PAGE 60 

Figure 4.20 pH 9 at 30.0 :t 0.5°C- Component B 


Graph of 10910 Concentration (gn) versus Time (Hours) 


lI.IlOIIIEtO 0_


o.soo 

o.soo 

\.000 

UIIIO 

2.000 

2.000

-

1.000 

"-000 

_.IICO 

UDIl 

oU4C 4A2'Z

.".dO 

.a.­

."... 

03.111 

4.744 

.1.110&

-

4.771 

-uu 

03,1It\ 

........ 

-3.7 

• 


-3.8 

-3.9 

-4. 

-4.1'----------------------­

o 1 2 3 4 5 6 

Time (hours) 

1082 

PROJECT NUMBER: 014510218 PAGE 61 

Figure 4.21 pH 9 at 40.0 ±O.S°C- Component A 


Graph of log'lo Concentration (gIl) versus Time (Hours) 


~-3.2 

~ 

I 

8 

1-3. 

0

-

J 

-4. 

-4.8 

CI.OOGI;+O _.. IUIIQ 

IUIIQ 

1.@ 

1.000 

2.000 

2.000

-

UGe 

..coo 

..coo

• ..-.uot .--G.13I . 

• 

• 

.I&.AA 4._ 

4Ma 

4.612 

.~ 

4.ICII 

4.f11 

4,1111

-

03'­

... o4JI«I ­

... 

-5.2:1--------------~-------...... o 1 2 3 4 5 

Time (hours) 

1083 

-4.2 

PROJECT NUMBER: 014510218 	 PAGE 62 

Figure 4.22 pH 9 at 40.0 ± 0.5°C- Component B 


Graph of log10 Concentration (gIl) versus Time (Hours) 


II.IIDaEtO 4.11i11 

II.DCOI<O ~ 

Q.$III -1./111 

0JlCIII 4.101 

1.0011 4.as 

1_ 4175 

.LIIIIO 4.114 2.CIIIII 4.n4

-

3.tIIlU ­"",177 

tJIIO ....170 .­ ...­ -A-4M8 ­ ".0.240

. 

• 

• 

• -4. 

• 

•.~----------------------------------------- o 	 1 2 3 4 5 

Time (hours) 

1084 
PROJECT NUMBER: 014510218 PAGE 63 

The Arrhenius plots were constructed using the data shown in the following tables: 

Table 4.37 pH 9 Arrhenius Data - Component A 

T (0C) T (K) 

1 

T(K) kobS (h(l) In kob$ 

20 293 3.41 x 10-3 5.43 X 10-2 ~2.91 

30 303 3.30 x 10-3 7.32 X 10-2 -2.61 

40 313 3.19 x 10-3 0.549 -0.599 

From the graph (Figure 4.23) of the above data, the rate constant and half-life at 25'-'C 

have been estimated to be as follows: 

kObS = 7.43 X 10.2 hou(l, 2.07 x 10-5 second-1 

t~ = 9.33 hours, 0.389 days 

Table 4.38 pH 9 Arrhenius Data - Component B 

T (OC) T (K) 1 

T(K} kobs (hf1) In kobs 

20 293 3.41 x 10-3 5.73 X 10-2 -2.86 

30 303 3.30 x 10-3 0.190 -1.66 

40 313 3.19 x 10-3 0.551 -0.595 

From the graph (Figure 4.24) of the above data, the rate constant and half-life at 25°C 

have been estimated to be as follows: 

0.105 hou(1, 2.91 x 10-5 second-1 

6.61 hours, 0.275 days 

1085 

PROJECT NUMBER: 014510218 PAGE 64 

Figure 4.23 pH 9 - Component A 


Graph of In.kobs versus 11T (K) 


• 

-3 

0.00316 0.00320 0.00324 0.00328 0.00332 0.00336 0.00340 0.00344 

1I[Temperature{K}} 

1086 

PROJECT NUMBER: 014510218 PAGE 65 

Figure 4.24 pH 9 - Component B 


Graph of In.kobs versus 11T (K) 


--1 

I 

I 

-2 

0.00316 0.00820 0.00324 0.00328 0.00332 0.00336 0.00340 0.00344 

1/[Temperature(K)} 

1087 

PROJECT NUMBER: 0145f0218 PAGE 66 

4.5 Validation 

The linearity of the detector response with respect to concentration was assessed over 

the nominal concentration range of 0 to 50 mgt!. This was satisfactory with a correlation 

coefficient of 1.00 being obtained for Components A and B. 

Recovery of analysis of the sample procedure was assessed and proved adequate for 

the test, with the exception of Component A in pH 7 buffer. 

Recovery data is shown in the following table: 

Table 4.39 

Buffer 

Nominal 

concentration 

(mgll) 

! 

I 

Recovery range (%) 

Component A Component B 

Mean recovery (%) 

Component A Component B 

pH4 0.37 87.3 to 92.7 75.8 to 82.3 90.8 78.4 

pH 7 0.37 0 62.0 to 74.1 0 66.2 

pH9 0.37 62.9 to 80.3 78.2 to 84.7 72.7 82.0 

Concentrations have not been corrected for recovery of analysis. 

4.6 Discussion 

Initial experiments demonstrated poor recoveries of analyte in buffer solutions, but 

acceptable recoveries in pH adjusted water. Therefore, preliminary testing was 

performed in reverse osmosis water adjusted to pH 4. 7 and 9 using 0.1 M hydrochloric 

acid or 0.1 M sodium hydroxide. However, due to the absence of buffering, the pH of 

these solutions was unstable. Further investigation revealed that acceptable recoveries 

could be achieved in the buffers specified in Section 4.2.1, with the exception of pH 7 

phosphate buffer in which no recovery of Component A could be achieved. This 

anomaly could not be readily explained since the pH 7 matrix blank did not demonstrate 

interfering peaks and the analytical procedure was shown to be successful for both 

analytes in pH 4 and pH 9 buffers. Therefore, no data for Component A at pH 7 could be 

generated. 

Tier 2 testing at pH 7 demonstrated a significant degree of variation in the analytical 

results for duplicate samples and across the time points over the period of the test. 

EVen though this test was performed at five different temperatures plus some limited 

testing at 25°C, consistent data could not be obtained. The variation in analytical data 

was not considered to be due to solubility issues. It was concluded that there was no 

evidence of hydrolysis at pH 7, since losses observed were not of a pseudo-first order 

kinetic nature and were considered to be due to other mechanisms or effects. 

1088 
PROJECT NUMBER: 014510218 PAGE 67 

For additional information, an estimation of the hydrolytic stability of the test material was 

performed using the specialised predictive software, HYDROWIN v 2.00 (September 

2008), © 2000 US Environmental Protection Agency. The estimated half-lives at 25°C 

are shown in the following table: 

Component pH7 pHS 

A (PE3MB) 13.1 years 1.31 years 

B (PEMB) 9.85 years 360 days 

For calculation of the test material concentration, peak area was used throughout the 

pH 4 and pH 9 tests, whereas peak height was the pH 7 tests. The use of peak height in 

the pH 7 calculations gave initial concentrations consistent with the fortified 

concentrations. 

The kinetics of the study for pH 9 has been detennined to be consistent with that of a 

pseudo-first order reaction as the graphs of 10910 concentration versus time are straight 

lines. 

As the test material was determined to be hydrolytically stable under acidic conditions 

(t~ > 1 year at 25°C). no additional testing was performed at pH 1.2, 37.0 ± O.5°C. 

4.7 Conclusion 

The rate constants and estimated half-lives at 25°C of the test material are shown in the 

following table: 

pH 

I 

I 

Rate constant (S·1) 

PE3MB PEMB 

Estimated half-life at 25°C 

PE3MB PEMB 

4 not applicable not applicable >1 year >1 year 

7 

9 

not applicable 

2.07 x 10.5 

not applicable 

2.91 X 10-6 

>1 year 

9.33 hours 

>1 year 

6.61 hours 

No definitive signs of hydrolysis at pH 4 or pH 7. Testing performed at various 

temperatures indicated that losses observed were not pseudo-first order and therefore 

most likely to be due other mechanisms or effects rather than hydrolysis. 

1089 

PROJECT NUMBER: 0145/0218 PAGE 68 

Appendix 1 Statement of GLP Compliance in accordance with Directive 2004/9/EC 

,DH) Department 

~ of Health 

THE DEPARTMENT OF HEALTH OF THE GOVERNMENT 


OF THE UNITED KINGDOM 


GOOD LABORATORY PRACTICE 

STATEMENT OF COMPLIANCE 


IN ACCORDANCE WlTIf DJRECTI\'E 2004191EC 


TESTFACIL1TV TE!>'TTYI'E 

Karlun Lnboratories Ltd. Analytical/CUnical Cbemlstry 

Shal'dJow Business Purl{ Environmontal Fate 

London Road EDl'iroDmcnfal Toxicology 

Shardlol\' Mutagenicity 

Derby PbyslCbem 

DE7l2GD Toxicology 

DATE OF INSPECTION 

19th August 2008 

A general inspection for compliance with the Principles of Good Laboratory Practice 

was curried out at the above test facility as part of the UK GLP Compliance Programme. 

At the time of inspection no deyiations were found of sufficient magnitude to affect 

the validity of non-clinical studies perfunned at these facilities . 

Dr. Andrew J.Gray 

Head, UK GLP MOllitoring Authority • 

1090 

[KarenzMT PEl] [SHOWA DENKO K.K.J [SCA-1279] [02/10/2009] [1/6] 

SHOWA DENKO KK 

13-9, Shiba Daimon l-Chome unA 

OEln Minato-Ku, Tokyo 105-8518, Japan

Date ofpreparation : March 31, 2006 

Date ofrevision October 2, 2009 

MATERIAL SAFETY DATA SHEET 

1. PRODUCT AND COMPANY IDENTIFICATION 

Product name KarenzMTTM PE 1 

Company name SHOWA DENKO K.K. 

Address 5-1, Ogimachi, Kawasaki-Ku, Kawasaki-shi, Kanagawa 210-0867 Japan 

Section Chemicals Division, Chemicals Sector 

Phone number +81-44-329-0726 

Facsimile number +81-44-329-0782 

Product code PEl 

Emergency phone number : +81-90-7239-0335( cell phone) 

Recommended use ofthe chemical and restrictions on use 

Industrial Purpose Only 

MSDS number : SCA-1279 

2. HAZARDS IDENTIFICATION 

GHS classification 

Physical hazards Any of"classification not possible" and "Not classified" 

Health hazards Sensitization -Skin : Hazard category 1 

Other Hazard Class : Any of "classification not possible" and "Not 

classified" 

Environmental hazards Hazard to the Aquatic Environment-acute hazard : Hazard category 1 

Hazard to the Aquatic Environment-chronic hazard: Not classified 

GHS label elements 

Pictograms (Symbols) 

Signal words 

Hazard statements 

Precautionary statements 

Main symotoms 

Emergency overview 

Warning 

May causes an allergic skin reaction 

Very toxic to aquatic life 

[Prevention] 

Avoid breathing dust/fumelgaslmist/vaporslspmy. 

Contaminated work clothing should not be allowed out ofthe workplace. 

Wear protective gloves. 

[Response] 

IF ON SKIN: Wash with plenty ofsoap and water. 

IF Skin irritation or rash occurs: Get medical advice/attention. 

Wash contaminated clothing before reuse. 

[Disposal] 

Dispose of contents/container to licensed waste disposal site in 

accordance with 10cal/regional/national/intemational regulation. 

No information available 

No information available 

1091 
[KarenzMT PEl] [SHOWA DENKO K.K.J [SCA-1279J [02/10/2009J [2/6J 

3. 	COMPOSITION/INFORMATION ON INGREDIENTS 

Single substance or mixture : Mixture 

---..-.-.-.----------....-..-.--...-.....-... -~- ....-.-----.-..-...--.. ..-..... . Common chemical name 

(generic name) 

Synonym(s) 

Chemical formula " 

CAS number 

Content 

Publication number in Chemical Substances Control Law: Low production volume chemical 

Japanese official gazette 	 substances 


Industrial Safety and Health Law: 2-(6)-1770 


4. FIRST-AID MEASURES 

Inhalation Move to a well-ventilated space to get fresh air and immediately obtain 

medical aid regardless ofpatient's sYmptoms. 

Skin contact 	 Remove contaminated clothing, and wash affected area with plenty of 

water or lukewarm water using soap. If there is the change in the 

appearance or irritation persists, obtain medical aid. 

Eye contact Immediately flush with plenty of fresh water for at least 15 minutes, and 

immediately obtain medical aid. 

Ingestion Immediately obtain medical aid. 

Briefdescription ofthe most Not specified. 

important symptoms 

Protection offlTst-aiders Remove polluted clothes. 

Wear gloves and be careful not to touch the toxic substance. 

Special notes to a physician If the victim has a medical wound, after having removed a pollutant, treat 

as a normal burn allowance. 

5. 	FIRE-FIGHTING MEASURES 

Extinguishing media Powder, carbon dioxide, halides, foam 

Inappropriate extinguishing Not specified. 

media 

Specific hazards with regard Besides the gases such as carbon monoxide etc, sulfur compounds may be 

to fire-fighting measures included in the combustion gas ofthe product. 

Specific methods offire­ Stop supply of source of fire to the burning spot and put out the fire using 

fighting and special an extinguishing medium. The fire-fighting operation should be carried out 

equipment from the upwind. 

Protection of firefighters Wear the protective cloth, goggles for chemicals and gas mask, to prevent 

from contact and inhalation ofthe vapor and harmful gases. 

6. ACCIDENTAL RELEASE MEASURES 

1092 
[KarenzMT PElJ [SHOWA DENKO K.K.J [SCA-1279J [02110/2009J [3/6J 

Personal precautions, 

protective equipment and 

emergency procedures 

Environmental precautions 

Methods for cleaning up 

Prevention of secondary 

hazards 

7. HANDLING AND STORAGE 

Handling 

Technical measures 

Local exhaust ventilation! 

general ventilation 

Precautions 

Storage 

Appropriate storage 

conditions 

Safe packaging materials 

Wear appropriate protective equipment during operations to prevent from 


contact of splashed product and the like with the skin. 


Do not allow concentrated effluent to flow into public water conduits or 


rivers. 


For small spills, recover the spills with saw dust or the like and incinerate 


them. 


In case of large spills, recover as much of the spills as possible and be 


careful NOT to flow into public water or rivers. 


No infonnation available 


Take measures against static electricity and wear conductive clothes and 


shoes. 


Handle in the local exhaust ventilation or at a place with total ventilation 


facilities. 


Handle the product where adequate local exhaust ventilation is provided. 


Store the product in a dry, cool, and dark place. Do not allow high 


temperature objects, sparks or flame in the vicinity ofthe product. 


Not specified. 


8. EXPOSURE CONTROLS/PERSONAL PROTECTION 

Engineering measures to reduce exposure 

Provide local exhaust ventilation equipment. 

Administrative control level : Not established. 

Exposure guidelines 

Japan Society for Occupational Health (2008) 

Not established. 

ACGIH (2009) Not established. 

OSHA (2004) Not established. 

Personal protective equipment 

Respiratory protection Gas mask for organic gas. Air respirator 

Hand protection Rubber gloves 

Eye protection Protective goggles 

Skin and body protection Apron and boots to avoid contact to the skin. 

Specific hygiene measures Do not eat or smoke while working. 

9. PHYSICALAND CHEMICAL PROPERTIES 

Physical state 

Appearance 

Color " 

-----------------------, 

Odor (odor threshold) : Slightly sulfur odor 

pH : No data available .-------------------------­ Boiling point .. : ·~------------------- ______I 

Initial boiling point and No data available 

boiling range 

Melting point/freezing point 

Decomposition temperature 

FJashpoint 

Auto-ignition temperature 

1093 
[KarenzMT PElJ [SHOWA DENKO K.K.J [SCA-l279J [02110/2009J [4/6J 

Upperllower flammability or No data available 


explosion limits 


Vapor pressure 


Vapor density 


Evaporation rate 


Relative density 


Solubility 


Water .'

.,
,

,

,

,

,
 Other solvents ., 


n-octanollwater partition 


coefficient , , 
 ~ .......... ~ -..... _........ _.......... _.. -...... _................................................ _...... .. _' 


10. STABILITY AND REACTIVITY 

Stability Stable in usual handling. 

Possibility ofhazardous Reacts with oxidizing compounds, and forms disulfide compounds .. 

reactions Causes addition reactions to ethylene-type double bonds 


Conditions to avoid Contact with oxidizing agents. 


Incompatible materials No information available 


Hazardous decomposition Oxidizing agents 


products 


11. TOXICOWGICAL INFORMATION 

Acute toxicity Oral Rat LD50 >2000mg!kg 3) 

Dermal Rat LD50 >2000mg!kg 4) 

Skin corrosion/irritation Non-Irritant to rabbit skin (Draize classification scheme) 


No corrosive effects were noted. 


P.l.L 0.0 5

) 

Serious eye damage/irritation 	 Minimal irritant to eyes of rabbits. (Kay and Calandra Classification 

scheme). 

Mean scores were 8.0. 6) 

Respiratory or skin Positive in the local lymph node assay (LLNA) in mice. (OECD T0429) 7) 

sensitization 

Oerm cell mutagenicity 'Negative in the Ames test for Salmonella typhimurium TA98, TAIoo, TA 

1535, TA 1537 and Escherichia coli WP2uvrA with and without metabolic 

activation. 8) 

•Positive 	in the Chromosome aberration test for Chinese hamster lung 

fibroblasts with and without metabolic activation. 14) 

•Negative in the Micronucleus test for mice. (OECD T0474) 15) 

Carcinogenicity No data available 

Reproductive toxicity No data available 

Specific target organ toxicity No data available 

-single exposure 


Specific target organ toxicity No data available 


-repeated exposure 


Aspiration hazard No data available 


12. ECOLOGICAL INFORMATION 

Eco-toxicity 


Fish toxicity Medaka LC50 (96hr) 0.16mgIL (Semi-static system) 9) 


Others Daphinia magna EC50 (48hr) 9.0mgIL (Semi-static system) 10) 


Algae EC50 (12hr) OAOmgIL III 

Persistence/degradability Readily biodegradation under the condition of OECD T0301B, CO2 

evolution test (70% degradation after 28 days) 12) 

1094 
[KarenzMT PEIJ [SHOWA DENKO K.K.J [SCA-1279J [02/10/2009J [5/6J 

Bioaccumulative potential 	 Bioconcentration potential (after 60 days) using Carp was low. 13) 

High exposure level {2.51lg/L):BCF 70 

High exposure level (I Ilg/L):BCF 170 

Mobility in soil 	 No data available 

13. DISPOSAL CONSIDERATIONS 

Because this product contains a sulfur compound, please bum up in an appropriate incinerator or dispose of 

contents/container to licensed waste disposal site in accordance with locallregionallnationallintemational 

regulation. 

Note: When disposing of the product outside Japan, conform to applied laws and regulations in that country 

or territory. 

14. TRANSPORT INFORMATION 

International regulations for transport 


UN hazard class Not applicable 


UN number Not applicable 


UN Proper Shipping Name Not applicable 


Packing group Not applicable 


Marine pollutant Not applicable 


Additional regulations for Not applicable 


transport 


Specific precautionary transport measures and conditions 

When transporting the product, inspect the containers for leakage and load 

then securely to prevent from topping, failing, or breakage, and take 

preventive measures against collapsing ofcontainers. 

Emergency Response 	 Not applicable 

Guidebook number 

Note: When transporting the product outside Japan, conform to applied laws and regulations in that country 

or territory. 

15. REGULATORY INFORMATION 

Laws and regulations applied in Japan: 


The Fire Service Law: Designated inflarnmables{Flammable liquids) 


Note: When using the product outside Japan, it must be handled in accordance with applied laws and 

regulations in that country or territory. 

16. OTHER INFORMATION 

Content ofPRTR Law chemicals : Not contained 

TSCA Inventory Not included 


EINECS number Not included 


References 	 1) Showa Denko K.K. test report 014510193 (2009) 


2) Showa Denko K.K. test report 1000855956 (2009) 


3) Showa Denko K.K. test report 014510194 (2009) 


4) Showa Denko K.K. test report 014510195 (2009) 


5) Showa Denko K.K. test report 014510198 (2009) 


6) Showa Denko K.K. test report 014510199 (2009) 


7) Showa Denko K.K. test report 014510200 (2009) 


8) Showa Denko K.K. test report KO 1-4229 (2009) 


9) Showa Denko K.K. test report 94746 (2009) 


10) Showa Denko K.K. test report 94745 (2009) 


11) Showa Denko K.K. test report 94744 (2009) 


12) ShowaDenko K.K. test report 014510215 (2009) 


13) Showa Denko K.K. test report 44929 (2008) 


14) Showa Denko K.K. test report K06-1279 (2009) 


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[KarenzMT PEl] [SHOWA DENKO K.KJ [SCA-1279] [02/10/2009J [6/6J 

15) Showa Denko K.K. test report 014510219 (2009) 

For further information. please contact 

Chemicals Division. Chemicals Sector. Showa Denko K.K. 

Telephone number +81-44-329-0726 

Facsimile number +81-44-329-0782 

The information in this MSDS was obtained from sources which we believe are reliable, but no warranty or 

representation as to its accuracy or completeness is hereby given. Users should consider the information 

herein only as a supplement to other information gathered by them and must make independent determinations 

of suitability and completeness of information from all sources to assure proper use and disposal, the safety 

and health ofemployees and customers and the protection ofthe environment. 

This is a translation oforiginal Material Safety Data Sheet prepared in Japanese. 

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