Document O16o7qpz3b7KxmOj483mZ0g8Q
`CENTRE FOR XENOBIOTIC RESEARCH
University of Dundee, Biomedical Research Centre, Level 5, Ninewells Hospital and Medical School, Dundee, DD1 9SY
Study Report
Determinationofcyanide insensitive Palmitoyl-CoA oxidation in samples from 3M Environmental Laboratory -- Covance Studies 6329-183 and 6329-212
Study Number XR0108
w
pater: So fore lion .~"BMElcombe-
Approved: CC.R ElFcoambse--nie
rots
Pace 10f3
owe J5/2/14
002401
1 Introduction
`liTvheerpsuarmppolseesofptrhoivsidsetdudbyyw3aMs EtonviinrveosntmiegnattealthLeableovrealtsoorfyPfCroomACooxviadnatcieonstiunditehse6f3r2oz9e-n183 and 6329-212.
2 Test Substance
6329-183: 6329-212:
PFOS, 20ppm, administered by EFOSE-OH, 100 and 300ppm,
Covance until week 4. administered by Covance
until
week
4.
3 Testing Facility
Centre for Xenobiotic Research, UniversityofDundee, Biomedical Research Centre, Level 5, Ninewells Hospital and Medical School, Dundee, DDI 9SY, UK.
4 Homogenate Preparation
41 Reagents
SET buffer: 0.25M Sucrose, SmM EDTAand20mM Tris-HCI, pH 7.4
42 Instruments
Potter-Elvehjem homogeniser
43 Method
Frozen liver samples were thawed on ice, blotted and weighed. A 25% homogenate was prepared in ice cold SET using a Potter-Elvehjem homogeniser (6 passes)
5 Determination of Cyanide-insensitive Palmitoyl-Co Oxidation
`The 25% homogenate liversampleswere diluted 1 part in 10 parts with ice cold SET. `These diluted samples were used to determine the PCoA activity,followingCXR SOPoir
6 Statistical Analysis
A Student's T-Test (2-sided) control p<0.05; ** =p<0.01;
w*as=*pepr*<f0o.r0m0e1d
on
the
results;
*
=
statistically
different
from
XRO108
Pace 20f3
002402
7 Results
E
Study /Group Dose (ppm)
WeaenekS4D.
%of Control
.
[e20170 TTss:r0ass| i000:i2s |
[e227
0
[632021274 177 100
| 513085 | 1000x166 | |990140800 | 19214155 |
PCoA Oxidation 6329-183 (xro108)
i 0
-
i.
i] ||3:2
|
0
20
|
Dose PFOS (ppm)
I i SPeCcoAaoOxaidnationt6a3z29e-a2t12s (xR0108)
|
[w |[f& +
PE. |i E
h HB | 3 -
.
100
200
Dose EtFOSE-OH (ppm)
XRO108
:
Pace 3of 3
|
|
|
|
{
|
|
| |
002403
sEEeEsas.] wn ITE wie TRIS
3FS 2S3s
Rm xe ww mam
wry 25% homogenates 6329-183, diluted 1/10
JRRELATION COEFF TERCEPT OPE RANGE YRANGE
0 0.031 0 0.033 0.025 0.081 0.025 0.085 0.05 0.113 0.05 0.119 0.075 0.166 0.075 0.161 0.1 0.205 0.1 0.200 0.15 0.283 0.15 0.283
099857408 0.0326 1686
Y CALC 0033 0033 0075 0075 0.117 0.117 0159 0159 0201 0201 0.286 0.286
No C2508M C92624M Ce2526M C92529M C92546M C2752M C92759M C2779M C92793M C92798M
GIVENY1 0.231 0.336 0.283 0.319 0.261 0.231 0.245 0.243 0.256 0.244
Standard curve
030,
1s1din. GIVENY2
0.237 0.250 0.261 0251 0256 0231 0233 0270 0.249 0.246
XR0108 protein mg/ml 6.09 7.88 7.24 7.63 6.83 6.00 6.24 6.77 665 6.42
_020
E
goss!
a
010 J
00s |
=
000 0 0.025
rss
.
ns
005 0075
01
0125 ots |
Protein mg/ml
|
------------
002405
faE orts,w omn on
@Se
Tf rn to et
na ww xe ew erlei
23
El TE
owry 25% homogenates 6329-212, diluted 1/10
SORRELATION COEFF NTERCEPT SLOPE XRANGE ~ YRANGE
0 0.031 0 0.033 0.025 0.081 0.025 0.065 0.05 0113 0.05 0.119 0.075 0.166 0.075 0.161 01 0.205 01 0200 015 0.283 015 0283
0.99857408 0.0326 1.686
~~ YCALC 0033 0033 0075 0075 0117 0117 0159 0159 0201 0201 0286 0286
No CS071SM C90725M C90732M C90753M C90775M C9093M (C90942M C90961M C909%6M C90984M C91005M C91024M Co103M CO1037TM
GIVENY1 0.220 0255 0.252 0.263 0.266 0235 0253 0.256 0261 0.244 0253 0.291 0261 0.251
2)030
Standard curve
_fo020 1
c: 010
| eos |
|| 000
|
Protein mgimi
stdin. GIVENY2
0235 0.249 0.255 0.262 0.261 0.235 0.256 0.253 0.263 0.232 0.260 0.256 0.251 0.256
XR0108 protein mgiml 6.03 6.64 6.68 6.95 6.98 6.12 6.71 671 6.94 621 677 729 6.76 668
| | ||
002407
: