Toxicity Test

ACUTE ORAL TOXICITY STUDY OF PENETRON IN SPRAGUE-DAWLEY RATS  
 
Submitted to:    

ICS/Penetron International Ltd.
45 Research Way
Suite 203
East Setauket, NY 11733
FDRL Study No. 8375A

FDRL Study No. 8375A

Acute Oral Toxicity Study in Rats (FHSA) FDRL Test Article ID: 84-0909 Sponsor Test Article ID: Penetron    
SUMMARY  

The above referenced test article was evaluated for acute oral toxicity in male and female Sprague-Dawley rats. The test article was administered by gauge to each of ten rats at a level of 5.0 g/kg body weight.

One animal died on day 7 following administration of the test article. According to 16 CFR 1500, the above referenced test article is not considered to be toxic and does not require cautionary labeling.

FDRL Study No. 8375A

Acute Oral Toxicity Study

Purpose:
To determine the acute oral toxicity of the test article following a single dose by gauge to male and female Sprague-Dawley rats.

Compliance with Government Regulations:
This study was conducted in accordance with the regulations for Good Laboratory Practice as described by the FDA (21 CFR Part 58), 16 CFR 1500.3 and FDRL Standard Operating Procedures. All data and pertinent records have been retained by FDRL and are available upon request. Experimental Design:

Animals and Animal Care
Ten young adult Sprague-Dawley rats were obtained from Charles River Breeding Laboratories, Inc., Wilmington, MA.

All housing and care conformed to the standards established in the "Guide for the Care and Use of Laboratory Animals" DHEW Publication No. (NIH) 78-23. Animals were individually housed in wire mesh bottom cages in environment-controlled rooms and provided food and water ad libitum.

Procedure
After an acclimation period of at least 5 days, animals were assigned to the test. Food was withheld the night prior to dosing on the first day of the study, animals received a single oral dose by gauge of the test article at a level of 5.0 g/kg body weight.

Observations

Animals were weighed prior to dosing, at termination of the study or at necropsy. They were observed frequently on the day of dosing and daily thereafter for a total of 15 days. All external signs of toxicity or pharmacological effects were noted.

Necropsy

All animals that died during the study were subjected to a gross necropsy and abnormalities were recorded.

 

FDRL Study No. 8375A

 

Table 1 Mortality   

Dosage 

Level 

G/kg

 No. 

Rats 

Dosed

No. of Animal Deaths on Day

1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

Cumulative Mortality

5.0

10

0

0

0

0

0

0

1

0

0

0

0

0

0

0

0

1/10

Percent Mortality 10%

95% Confidence Intervala 0 to 40%

a E. L. Crow Biometrika 43, 423-435 (1956).

Table 2 Mean Body Weight Dataa  

Dosage 

Level 

G/kg

Body Weights (g)

Initialb

Day 15

Males 

5.0

 225.4 

+/-10.8(5)

 322.4 

+/-15.2(5)
Females 

5.0

 211.0 

+/-9.7(5)

 254.8 

+/-18.5(5)

 

a Values are means +/- S.D. for the nuinber of observations in ( ).

b Fasted body weight.

FDRL Study No. 8375A

Table 3 Incidence of Daily Observations

Dosage 

Level  

g/kg

Observations

No. Affected/No. Examined

Males

Females

5.0 
    Ataxia 
    Decreased activity 
    Diarrhea 
    Gasping 
    Rales 
    Salivation 
    Wet - abdomen 
    Deaths
    0/5
    0/5
    0/5
    0/5
    0/5
    0/5
    0/5
    0/5
    1/5
    2/5
    2/5
    1/5
    1/5
    2/5
    3/5
    1/5

Table 4 Incidence of Gross Necropsy Findings

Dosage 

Level  

g/kg

Findings

No. Affected/No. Examined

Males

Females

5.0  Intestines: contain blood like viscous liquid

N/A

1/1  

DOSING SCHEDULE

Study Title: Acute Oral FDRL Study No.: 8735A

Instructions: FHSA Design FDRL Test Article

Species: S.D. Rats I.D. No.: 84-0909

C.R. 11/7/84

Test Article

Diluent:Dist./Deionized H20 Density: N/A

Concentration: 95% Form: Suspension  

Animal Number & Sex

Time Dosed

Body Weight (g) At Day

Level Dosage G/KG

Dose ML (total)

Date of Death

Comments

1

15

WAD

D0750031M 1:26 227 303 5.0 1.19 DAW- Debbie Webster
0032 208 322 5.0 1.09 NT - Nancy Trout
0033 224 313 5.0 1.18 SB - Shirley Bennit
0034 236 341 5.0 1.24
0035 232 333 5.0 1.22
0036F 202 245 5.0 1.06
0037 216 -- -- 147 5.0 1.14 11/25 DAW
0038 223 274 5.0 1.17
0039 

 

 200 234 5.0 1.05
0040 214 266 5.0 1.13
Initials 

& Date

 NT DAW 

11/19/84

 DAW 

11/19/84

 SB 

12/3/84

Reviewed by: BR Date: 12/3/84 TEST ARTICLE PREPARATION DATA  

Study Title: Acute Oral                          FDRL Study No.: 8735A

Species: Rat                                          FDRL Test Article I.D.: 84-0909
                                                             Concentration Prepared: 95%
Test Article Density: N/A gm/ml              Diluent: Destilled/Deionized H20

Total Amount Dispensed: 25.0              Prepared Physical State: Susspension

PREPARATION PROCEDURES

(   ) Test article dispensed undiluted.

( X ) Weighed 23 75 gm test article, Q.S. to 25.0 ml with diluent

(   ) Test article ground with rnortar and pestle.

(   ) Test article blended at ______ speed for ______

DOSING INFORMATION
 

Level          Coefficient a           Level              Coefficienta

5.0g/kg          5.26
 

Special Instructions:

Shake prior to dosing

 Comments
   

Calculations:     LW 11/19/84         Preparation:      DS 11/19/84
                        Initials/Date                                      Initials/Date


Review         SB 11/19/84 BS 12/3/84
                    Initials/Date

a Coefficient x kg body weight = dose              N/A = Not Applicable
Study Title: Acute Oral FDRL Study No.: 8375A

FDRL Identification No.: 84-0909 Balance Calibration And Work Log  

Date

Weight

Difference

Scale No.

Technician Inititals

Applied

Read

11/19/84

200/500

200/500

0/0

1

DAW 8:30

11/25/84

200/500

200/500

0/0

1

DAW 8:20

12/3/84

200/500

200/500

0/0

1

SB 11:00

BR 12/3/84

Study Title: Acute Oral FDRL Study No.: 8375A

Dose Level: 5.0 g/kg

Dosed: 11-19-84 Species: S.D. Rats FDRL Test Article I.D. No.: 84-0909
DAILY OBSERVATIONS  

Animal No. 0031M

Animal No. 0032M

Animal No. 0033M

Animal No. 0034M

Animal No. 0035M

Initials/ Date

Day 1 - Initial

/

/

/

/

/

NT DAW 11/19

Day 1

Dosed late

No

2nd

obs

Day 1 3:12

/

/

/

/

/

DAW 11/19

Day 2 8:36

/

/

/

/

/

NT 11/20

Day 2 3:14

/

/

/

/

/

DAW 11/20

Day 3

/

/

/

/

/

DAW 11/21

Day 4

/

/

/

/

/

DAW 11/22

Day 5

/

/

/

/

/

NT 11/23

Day 6

/

/

/

/

/

DAW 11/24

Day 7

/

/

/

/

/

DAW 11/25

Day 8

/

/

/

/

/

SB 11/26

Day 9

/

/

/

/

/

SB 11/27

Day 10

/

/

/

/

/

SB 11/28

Day 11

/

/

/

/

/

SB 11/29

Day 12

/

/

/

/

/

SB 11/30

Day 13

/

/

/

/

/

NT 12/1

Day 14

/

/

/

/

/

NT 12/2

Day 15

/

/

/

/

/

SB 12/3

Reviewed By: BR Date: 12/3/84

Study Title: Acute Oral FDRL Study No.: 8375A

Dose Level: 5.0 g/kg

Dosed: 11-19-84 Species: S.D. Rats FDRL Test Article I.D. No.: 84-0909
DAILY OBSERVATIONS  

Animal No. 0036F

Animal No. 0037F

Animal No. 0038F

Animal No. 0039F

Animal No. 0040F

Initials/ Date

Day 1 - Initial

/ 

Sal.

 

Sal.

/

/

NT DAW 11/19

Day 1

Dosed late

No

2nd

obs

Day 1 3:12

/

/

/

/

/

DAW 11/19

Day 2 8:36

/

Rales, Ad

/

Ad

/

NT 11/20

Day 2 3:14

/

ClearRales, G, Ad Wetm-6

/

Ad Wetm-6

Di

DAW 11/20

Day 3

/

NC

/

NC

Clear Di Wetm-6

DAW 11/21

Day 4

/

G, Ad, Wetm-6,/Di

/

Clear Wetm-6 & Ad, V

Clear Wetm-6

DAW 11/22

Day 5

/

Clear Wetm-6,/Ad, G, Di

/

/

/

NT 11/23

Day 6

/

Ad, G, Di

/

/

/

DAW 11/24

Day 7

/

DEAD

/

/

/

DAW 11/25

Day 8

/

/

/

/

SB 11/26

Day 9

/

/

/

/

SB 11/27

Day 10

/

/

/

/

SB 11/28

Day 11

/

/

/

/

SB 11/29

Day 12

/

/

/

/

SB 11/30

Day 13

/

/

/

/

NT 12/1

Day 14

/

/

/

/

NT 12/2

Day 15

/

/

/

/

SB 12/3

Reviewed By: BR Date: 12/3/84

Study Title: Acute Oral FDRL Study No.: 8375A

Dose Level: 5.0 g/kg

Dosed: 11-19-84 Species: S.D. Rats FDRL Test Article I.D. No.: 84-0909
DAILY OBSERVATIONS  

Animal No. 0036F

Animal No. 0037F

Animal No. 0038F

Animal No. 0039F

Animal No. 0040F

Initials/ Date

Day 1 - Initial

/ 

Sal.

Sal.

/

/

NT DAW 11/19

Day 1

Dosed late

No

2nd

obs

Day 1 3:12

/

/

/

/

/

DAW 11/19

Day 2 8:36

/

Rales, Ad

/

Ad

/

NT 11/20

Day 2 3:14

/

ClearRales, G, Ad Wetm-6

/

Ad Wetm-6

Di

DAW 11/20

Day 3

/

NC

/

NC

Clear Di Wetm-6

DAW 11/21

Day 4

/

G, Ad, Wetm-6,/Di

/

Clear Wetm-6 & Ad, V

Clear Wetm-6

DAW 11/22

Day 5

/

Clear Wetm-6,/Ad, G, Di

/

/

/

NT 11/23

Day 6

/

Ad, G, Di

/

/

/

DAW 11/24

Day 7

/

DEAD

/

/

/

DAW 11/25

Day 8

/

/

/

/

SB 11/26

Day 9

/

/

/

/

SB 11/27

Day 10

/

/

/

/

SB 11/28

Day 11

/

/

/

/

SB 11/29

Day 12

/

/

/

/

SB 11/30

Day 13

/

/

/

/

NT 12/1

Day 14

/

/

/

/

NT 12/2

Day 15

/

/

/

/

SB 12/3

Reviewed By: BR Date: 12/3/84 Abbreviations and Symbols used in Recording Observations   

AD – accidental death 

Ad – activity decreased 

Ai – activity increased 

Al – hair loss 

An – anorexia (loss of appetite) 

At – ataxia, wobbly gait 

B – blood 

C – clinicals 

COP – corneal opacity 

Cy – cyanosis 

D – died 

DB – dried blood (give location) 

DME – dark material around eyes 

DMN – dark material around nose 

Di – diarrhea 

Em – emaciated 

FD – found dead 

G – gasping 

Lac – lacrimation greater than normal (tearing) - 

NC – no change 

 / – normal 

Nd – nasal discharge 

M – respiratory rate decreased 

Ri – respiratory rate increased 

Ppp – pin point pupils 

S – sacrificed 

Sal – salivation 

Sb – appears to have blood in stools 

Sh – shallow breathing 

SM – sacrificed moribund 

SS – soft stools 

T – tremors 

Ub – appears to have blood in urine 

Us – urine stain (state location) 

V – vomiting 

X – escaped, lost, missing 

NNf – no noteworthy findings. 

Use L – left, R – right, M – middle plus numbers 1-12 for locations of nodules or other conditions where appropriate. Use +1, +2, +3 after appropriate abbreviations to indicate degree of severity of that condition (+1 – slight, +2 – moderate, +3 – .extreme), i.e. moderately increased activity would be Ai +2.

Appendix II Labeling Requirements Labeling Requirements* Acute Oral Toxicity 

If 50% or more of the animals die when given a single dose of 50 mg/kg or less, then the test article is considered "highly toxic" and appropriate cautionary labeling is required.

If 50% or more of the animals die when given a single dose greater than 50 mg/kg up to and including 5.0 gms/kg, then the test article is considered "toxic" and appropriate cautionary labeling is required.

If less than 50% of the animals die when given a single dose of 5 grams/kg or greater, then the test article is not considered to be toxic and does not require cautionary labeling.

As defined in 16 CFR 1500.3

    Appendix III

  • Quality Assurance Unit Statement

This final report was reviewed as required by Good Laboratory Practice Regulations for non-clinical laboratory studies, 21 CFR Part 58 and the study protocol. Inspections were accomplished as noted, and reported to the study director and inanagement immediately following their completion. Based on these inspections and the review of the report, this study was conducted and reported in conformance with the Good Laboratory Practice regulations.