Document by27Zbrg5O16NnnMG42bqvby6
.
SPONSOR Elf Atochem S.A. Cours Michelet
La Dfense 10 92091 Paris-la-Dfense CEDEX
France
AR226-3068
v-^iNriuciN i IAL
STUDY TITLE ACUTE DERMAL IRRITATION
IN RABBITS TEST SUBSTANCE
STUDY DIRECTOR Stphane de Jouffrey
STUDY COMPLETION DATE 3 February 1997
PERFORMING LABORATORY Centre International de Toxicologie (C.I.T.)
Miserey - 27005 Evreux - France
LABORATORY STUDY NUMBER 14887 TAL
-errp an y Sanitized. Does not contain TS'CA CB
% CONTENTS
STATEMENT OF THE STUDY DIRECTOR OTHER SCIENTISTS INVOLVED IN THIS STUDY. STATEMENT OF QUALITY ASSURANCE UNIT SUMMARY RESUME
4 5 6
1. INTRODUCTION
2. MATERIALS AND METHODS
2.1. TEST SUBSTANCE
2.1.1 Identification 2.1.2 Preparation
2.2. TEST SYSTEM
2.2.1 Animals 2.2.2 Environmental conditions
2.2.3 Food and water
2.3. TREATMENT 2.3.1 Preparation and selection of the animals
2.3.2 Study design 2.3.3 Application of the test substance
.
2.3.4 Date of treatment
2.4. CUTANEOUS EXAMINATIONS
2.5. DESCRIPTION AND EVALUATION OF CUTANEOUS REACTIONS
2.6. INTERPRETATION OF RESULTS AND CLASSIFICATION OF SUBSTANCES
2.6.1 Interpretation of the results
2.6.1.1 Criteria for irritation
2.6.1.2 Criteria for corrosion
*
.
2.6.2 Classification of the test substance
2.6.2.1 Irritant substances
2.6.2.2 Corrosive substances
'
2.7. ARCHIVES
3. RESULTS (table 1)
8
8
8
8
9 9 9 9
10 10 10 10 10 11 11 11
11
11 .
12 12
12 12 12
13
4. CONCLUSION
13
Table 1: Mean values of the scores recorded at each reading (24, 48 and 72 hours) for all three
animals (4-hour exposure)
14
5 n y San,feed. Does not con,a|n t s c a ^
APPENDICES
1. Test article description 2. Diet formula
15 18 and 19
ComPany sanitized. Does n t contain TSCA CB|
4
STATEMENT OF THE STUDY DIRECTOR
The study was performed in compliance with the following Principles of Good Laboratory Practice Regulations: . O.E.C.D. Principles of Good Laboratory Practice, Decision Concerning Mutual Acceptance of
Data in the Assessment of Chemicals, C(81)30(final) Annex 2. May 12, 1981. . Dcret N 90-206 du 7 mars 1990 concernant les Bonnes Pratiques de Laboratoire (Journal
Officiel du 9 mars 1990), Ministre de l'Industrie et de l'Amnagement du Territoire.
I declare that this report constitutes a true and faithful record of the procedures undertaken and the results obtained during the performance of the study.
This study was performed at the Centre International de Toxicologie (C.I.T.), Miserey, 27005 Evreux, France.
Toxicology
S. de Muflfrey
Date: 3 February 1997
Study"Director .
Doctor of Veterinary Medicine
Head of Short-term and Environmental
Toxicology
OTHER SCIENTISTS INVOLVED IN THIS STUDY
For Pharmacy: J. Richard t Doctor of Pharmacy
For Toxicology: C. Pelcot Study Supervisor
Company Sanitized. Does not contain TSCA CBS
5
STATEMENT OF QUALITY ASSURANCE TIMT
1. Specific study inspections
Type of inspections
Inspections
Protocol Report
21 Oct. 96 20 Jan. 97
Dates '
Report to Study Director (*)
29 Oct. 96 21 Jan. 97
Report to Management (*)
29 Oct. 96 21 Jan. 97
2. Routine inspections performed on other studies of the same type according to a frequency defined in Q.A.U. procedures
Inspected phase
....... Inspections
Treatment/test substance 13 Aug. 96 Preparation/test substance 25 Sept. 96
Dates
Report toReport to
Study Director (*)
Management (*)
22 Aug. 96 25 Sept. 96
22 Aug. 96 25 Sept. 96
The inspections were performed in compliance with C.I.T. Quality Assurance Unit procedures and the Good Laboratory Practice Regulations.
(*) The dates mentioned correspond to the dates of signature of audit reports by Study Director and Management.
L. Valette-Talbi Date: 3 February 1997 Doctor of Biochemistry Head of Quality Assurance Unit and Scientific Archives
Company Sanitized. Does not contain TSCA CBt
6
SUMMARY
At the request of Elf Atochem S.A., Paris-la-Dfense, France, the potential of the test substance
to induce dermal irritation was evaluated in rabbits according to O.E.C.D. (No. 404, 17th July 1992) and E.C. (92/69/E.E.G, B4, 31st July 1992) guidelines. The study was conducted in compliance with the Principles of Good Laboratory Practice Regulations.
Methods
The study design was established according to available information on the test substance and the above guidelines.
The test substance was applied for 4 hours to three male New Zealand White rabbits.
A single dose of 0.5 ml of the test substance in its original form was applied to the closelyclipped skin of the flank.
The test substance was held in contact with the skin by means of a semi-occlusive dressing. Cutaneous reactions were observed approximately one hour, 24, 48 and 72 hours after removal of the dressing.
The mean values of the scores for erythema and oedema were calculated for each animal.
Results
Very slight or well-defined erythema was noted in all animals at the 1-hour reading. It persisted for 24 hours in one animal, and for 48 hours in the two other animals. No oedema was observed.
Mean scores over 24, 48 and 72 hours for individual animal were 0.3, 1.0 and 0.7 for erythema and 0.0 for oedema.
Conclusion
Under our experimental conditions, the test s u b s t a n c e ! ^ _____ __ considered non-irritant when administered by cutaneous route in rabbits.
was
Company Sanitized. Does not contain TSCA CBi
RESUME
A la demande de Elf Atochem_S.A., Paris-la-Dfense, France, l'irritation cutane pouvant tre
induite par le produit
a t value chez le Lapin conformment
aux lignes directrices de l'O.C.D.E. (No. 404, 17th July 1992) et de la C.E.E. (92/69/E.E.C.,
B4, 31st July 1992). L'tude a t ralise conformment aux rgles de Bonnes Pratiques de
Laboratoire.
n
Mthodes
L'tude a t ralise selon les informations disponibles sur le produit et les lignes directrices mentionnes ci-dessus.
Le produit a t appliqu pendant 4 heures sur 3 lapins mles New Zealand White. Une dose unique de 0,5 ml de produit tel quel a t applique sur une surface de peau tondue sur le flanc.
Le produit a t maintenu en contact avec la peau au moyen d'un pansement semi-occlusiL Les
iactions cutanes ont t observes environ 1 heure, 24, 48 et 72 heures aprs l'enlvement du
pansement.
La moyenne des scores a t calcule pour chaque animal pour l'rythme et pour l'oedme.
Rsultats
Un i ythme trs lger ou bien dfini est observ chez tous les animaux la lecture 1 heure. Il persiste pendant 24 heures chez 1 animal, et pendant 48 heures chez les 2 autres. Aucun oedme n'est not.
La moyenne des scores sur les temps 24, 48 et 72 heures pour chaque animal est de 0,3 ; 1,0 et
0,7 pour l'rythme et de 0,0 pour l'oedme.
'''
Conclusion
Dans nos conditions exprimentales, le produit, non irritant par voie cutane chez le Lapin.
est considr
Company Sanitized. Does not contain TSCACBg
1. INTRODUCTION
The objective of this study was to evaluate the potential of the test substance to induce dermal irritation following a single administration in rabbits.
In the assessment of the toxic characteristics of a test substance, determination of the irritant and/or corrosive effects on the skin of mammals is an important initial step.
Information derived from this test serves to indicate the possible existence of hazards to Man likely to arise from cutaneous exposure to the test substance.
This study was conducted in compliance with: . O.E.C.D. guideline No. 4 0 4 ,17th July 1992. . E.C. Directive No. 92/69/E.E.C., B4, 31st July 1992.
2. MATERIALS AND METHODS
2.1. TEST SUBSTANCE
2.1.1 Identification The test substance '
jsed in the study was supplied by Elf Atochem S.A.
Documentation supplied by the Sponsor identified the test substance as follows: . name:
- protocol and labelling: J . batch number:
- protocol and labelling: . Elf Atochem filing number!] . description: dark brown liquid . container: one plastic flask . date of receipt: 11 October 1996
. storage conditions: at room temperature and protected from light.
Data relating to the characterization of the test substance are documented in a test article
description (presented in appendix 1) provided by the Sponsor. At the beginning of the study.,
the analytical certificate was not available.
*
The pH of the test substance as mentioned in the safety data sheet was 8.5.
2.1.2 Preparation
'
The test substance was applied in its original form.
C om pan y Sanitized. B oss not contain T S C A c b i
9
2.2. TEST SYSTEM
2.2.1 Animals
Sex, species, strain: male New Zealand White rabbits.
Reason for this choice: species commonly requested by the international regulations for this type of study. Breeder: Elevage Cunicole de Val de Selle, 80160 Prouzel, France.
Number of animals and identification: three animals were used, as recommended by the international regulations and taking into account that a good correlation of results can be obtained with either three or six animals (i). The animals were identified individually with a metal tag in the ear.
Weight: on the day of treatment, the animals had a mean body weight standard deviation of 2.2 0.2 kg.
Acclimatization: at least five days before the beginning of the study.
2.2.2 Environmental conditions
During the acclimatization period and during the main test, the environmental conditions in the animal room were set as follows:
. temperature: 183C
. relative humidity: 30 to 70%
. light/dark cycle: 12 h/12 h
'
. ventilation: approximately 12 cycles/hour of filtered, non-recycled air.
The temperature and relative humidity were recorded continuously and records retained.
The housing conditions (temperature, relative humidity and ventilation) were checked monthly.
The animals were housed individually in polystyrene cages (35 cm x 55 cm x 32 cm or
48.2 cm x 58 cm x 36.5 cm). Each cage was equipped with a food container and a water bottle.
2.2.3 Food and water
All the animals had free access to 112 C pelleted diet (U.A.R., 91360 Villemoisson-sur-Orge,
Ef!
France). Each batch of food was analysed (composition and contaminants) by the supplier. The diet formula is presented in appendix 2.
Drinking water filtered by a F.G. Millipore membrane (0.22 micron) was provided ad libitum. : Bacteriological and chemical analysis of the water and diet and detection of possible
contaminants (pesticides, heavy metals and nitrosamines) are performed periodically. Results are archived at C.I.T.
r
[t It was verified that no contaminants in the diet'or water at levels likely to influence the outcome of the study were present.
(I) Hatouin, N.S.; Leach, C.L.; Talsma, D.M.; Gibbons, R.D.; Garvin, P.J.: A statistical basis for using fewer Rabbits in dermal irritation testing. Journal of the American College of Toxicology. 9: 49-60 (1990).
Company Sanitized. Does not contain TSCA CBlj
10
2.3. TREATMENT
2.3.1 Preparation and selection of the animals
The day before treatment, the flanks of each animal were clipped using electric clippers The
skin of each animal was examined in order to use only animals without any signs of cutaneous
irritation. Animals showing signs of cutaneous irritation, cutaneous defects or pre-existin
dermal injury were not used.
.
2.3.2 Study design
The study design was established according to available information on the test substance and following the O.E.C.D. and E.C. guidelines.
As no irritant effects were anticipated, the test substance was applied for 4 hours to three animals.
2.3.3 Application of the test substance The test substance was used in its original form.
A single dose of 0.5 ml of the test substance was prepared on a 6 cm2 dry gauze pad
(Cooprative Pharmaceutique Franaise, 77000 Melun, France) which was then applied to the
right flank of the animals for 4 hours.
'
The test substance and the gauze pad were held in contact with the skin by means of an adhesive
hypoallergenic aerated semi-occlusive dressing (Laboratoires de Pansements et d'Hygine,
21300 Chenove, France) and a restraining bandage (Laboratoires 3M Sant, 92245 Malakoff,'
France).
'
The untreated skin served as control.
No residual test substance was noted at removal of the dressing. 2.3.4 Date of treatment
Animal number 01 02 03
Date of treatment (day 1) 5 November 1996 5 November 1996
5 November 1996
End of the observation period 8 November 1996 8 November 1996 8 November 1996
2.4. CUTANEOUS EXAMINATIONS
lessin" " ' Xami"ed w roxim a,e' " c < 24, 48 and 72 hours after removal of ,he
Following the O.E.C.D. and E.C. guidelines:
. when there is no evidence of dermal irritation after 72 hours, the studv is ended
. when there is persistent cutaneous irritation after 72 hnnrc rhn nke y ,
to a maximum of 14 days (until dav 15) in order tn der ' bservatl0n Penod 1S extended
their reversibility.
y J rder t0 deternune the Progress of the lesions and
when severe irritant effects are observed, the animals are killed on humane ground.
Any change in the animals' behaviour was noted.
2.5. DESCRIPTION AND EVALUATION OF CUTANEOUS REACTIONS
Dermal irritation was evaluated for each animal according to the following scoring scale:
Erythema and eschar formation: . no erythema.............................
very slight erythema (barely perceptible) . . . Z Z Z Z ^ Z .............. ..................
. well-defined erythema .......................
..............................................
. moderate to severe erythema
...........................................
. severe erythema (beet redness) s i i h U s c t o ' f t , ^ ^ ^ " in diptii)
? 1 ^
4
Oedema formation . no oedema .........................
very slight oedema (barely perceptible)
....................................................
slight oedema (edges of area well-defined i n f i n i t e raisin*)............................
. moderate oedema (raised approximately 1 millimetre)
.........................
9 %
' olexpoTureT * * *
* * 1 miUimetre and ^tending be'y'onda're'a.......
..................................................................................................
Any other lesions were noted.
4
2.6. INTERPRETATION OF RESULTS AND CLASSIFICATION OF SUBSTANCES
S ' " , ' S S r re eVa'Uated ,aki" S im " " N a t i o n the nature and the reversibility of
2.6.1 Interpretation of the results 2.6. 1.1 Criteria for irritation
,An " a ^ 24 hours or more after the end of theexp'osure period
if. 'hen it is applied to heahhy n " " ' M * >"*" for
Company Sanitized. Does not contain TSCAc
12
2.6.1.2 Criteria for corrosion
MimaTskin, h p T c t e full the test for skin irritation, or if alkaline reactions.
^ ^ * iS
t0 hea,thy intact
f '" ^ le3St ne animal durinS P edlcted>fr example: from strongly acid or
2.6.2 Classification of the test substance 2.6.2.1 Irritant substances - symbol Xi: indication of danger: "irritant", - phrase indicating the nature of special risks:
R 38: "Irritating to skin" Inflammation is significant if:
' d e l a i c T ei`!" r e,y,heraa a"d " Char
or
animals if the scorn
the.IBt has
"*'' i-g Itae
two or more animals is equi valent to the value ofTwo m m ore^
f rmatl n observed in
' hyperplasia" d l ^ u a n r t ^ ^ s ^ l 'm t i o n '1fisu ^ bserhvation Period- SPe^ific effects such as
account.
discolouration, fisures, eschar and alopecia should be taken into
All scoies obtained at each readin time C24 48 3nH n l , c calculating the respective mean valueSs.tIme ' 48 ^ 72 h UrS) f r 30 effect are used b7
2.6.2.2 Corrosive substances - symbol C: indication of danger: "corrosive"
.
- phrases indicating the nature of special risks:
R 34: "Causes burns"
R 35: Causes severe burns"
2.7. ARCHIVES
The study documentation and materials, namely:
. protocol and possible amendments,
raw data,
'
. correspondence,
. final report and possible amendments,
are stored in the archives of C I T MiseTM ?7nns c
r- ,
of the in vivo phase of the study. At the end" o f f h k f r five >'ears after the end
returned to the Sponsor.
b s Penh. t"hKe- s.-t>uiuduyy duocumentation will be
Company Sanitized. Does not contain TSCA
fjg
3. RESULTS (table 1) Very slight or well-defined erythema (grade 1 or 2) was noted in all animals at the 1-hour reading. It persisted for 24 hours in one animal, and for 48 hours in the two other animals. No oedema was observed. Mean scores over 24, 48 and 72 hours for individual animal were 0.3, 1.0 and 0.7 for erythema and 0.0 for oedema. 4. CONCLUSION Under our experimental conditions, the test substance FORAFAC 1203 (batch No. 25-28) was considered non-irritant when administered by cutaneous route in rabbits.
Company Sanitized. Does not contain TSCACS1
Table
1: Individual cutaneous examinations and mean values of the scores reading (24, 48 and 72 hours) for each animal (4-hour exposure)
recorded
at
each
Rabbit number
01
Dermal Irritation
lh D1
Erythema Oedema Other
1 0 *
Scores
24h 48h D2 D3
Mean Interpretation
irritation
(+)
72h score (1)
(-)
D4
100 000 ***
0,3 0,0
(-) (-)
02
Erythema
22
10
1,0
(-)
Oedema Other
00 0 0 ** *
0,0 . (-)
03
Erythema 2 1 ! 0 0,7
(-)
Oedema
0000
0,0
(-)
Other
* **
( I) mean of scores on days 2. 3 and 4 li = hour D = day
(+) --irritant according to E.E.C. criteria (-) = non-irritant according to E.E.C. criteria * = None
^ " r s m u e d .,,^
contain tsc a c m
APPENDICES
not contain gfflpany anU,zed' 0 eS
Test article description SW &tiy Sanitized. Does not contain TSCA CBS
TOXICOLOGY DEPARTMENT
CONFIDENTIAL 10 October 1996
eiT a t o c h e m s . a La dfense 10, cedex 42 92091 Paris-la-Dfense, France
TESTARTICLE DESCRIPTION
CAS number EINECS number 3urity Origin and batch Batch ElfAtochem filing number
physical a n d o m c Xl ^
Appearance Melting point Boiling point Flash point Solubility
t o x ic o l o g ic a l 1n f o r m a t io n s =a n d =use=s a f e t y See safety data sheet
[STORAGE AND DISPOSAL
'Storage
Expiry.date Disposal
in dark and at room temperature December 1997
incineration
Company Sanitized. Does no, contain TSCACS
# 18
2. Diet formula Company Sanitized. Does noi coniato TSCA CS
19
Ref: 112 COMPLETE DIET RABBIT MAINTENANCE DIET Appearance: 4.5 mm diameter granules Conditioning: bags of 25 kgs
Daily portion: in accordance with race and body weight, Rabbits 100-150 g, water ad libitum.
FORMULA %
C e r e a ls ....................................
Grain biproducts and le g u m e s ...................................
43 g 49
MINERALS (calculated in mg/kg) Nat. CMV
val- val. Total
Vegetable proteins (soya bean meal, yeast) ............................
Vitamin and mineral mixture ..
42 3
p ..... ....... Ca .......... K .... .......
AVERAGE ANALYSIS %
Na ... ...... Mg .. ......
Mn .. ......
Calorific value (KCal/kg)....... 2200
Moisture ...................................... jq
Proteins........................................ 13
Lipids ...........................................
27
F e .... ......
Cu .... ...... Zn .... ...... Co .... .....
Carbohydrates (N.F.E.) ......... Fibre ............................................
Minerals (ash) .............................
49.3 17
g
1 ...... ..... C l .... .....
3500 4500 11600 400 2100
40 160 12 30
0.1 0 500
3500 4500
0 1600 100
40 140 15
,, 45 1.5 0
3000
7000 9000 11600 2000 2200
80 300 27 75
1.6 0 3500
AMINO ACID VALUES (calculated in mg/kg)
Arginine ................................. Cystine ...................................
Lysine .................................... Methionine ............................. Tryptophan ............................ Glycine ..................................
6800 2100
4600 1600 1400 5200
FATTY ACID.VALUES (calculated in mg/kg)
Palmitic acid............................ 6400 Palmitoleic acid.......................... 0 Stearic acid............................... 600 Oleic acid................................. 6400 Linolcic acid............................ 12100 Linolenic acid.......................... 2400
VITAMINS (calculated per kg)
Nat. CMV
val. val:
Total
Vitamin A 2850IU
Vitamin D3 30 IU
Vitamin B 1 4.3 mg
Vitamin B2 3.8 mg
Vitamin B3 16 mg
Vitamin B6
1 mg
Vitamin B 12 0 mg
Vitamin E
16 mg
Vitamin K3
6 mg
Vitamin PP 55 mg
Folic acid ' 0 mg
Biotin
0 mg
Choline
850 mg
VIeso-Inositol 0 mg
6500 IU 9350IU
1000 IU 1030 IU
0 mg 4.3 mg
0 mg 3.8 mg
0 mg 16 mg
1 mg 2 mg
0 mg
0 mg
10 mg 26 mg
1 mg 7 mg
5 mg 60 mg
0 mg
0 mg
0 mg
0 mg
200 mg 1050 mg
0 mg
0 mg
Available under quality "Control Ref.: 112 C"
U.A.R., 7 rue Gailini, 91360 Villemoisson -Tel: 69.04.03.57 - Fax : 69.04.81.97 (Ref. Doc. UAR: 1992)
i
Company Sanitized. Does not contain TSCACBI
