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REACH

Acetic acid, chloro-, sodium salt, reaction products with 4,5-dihydro-2-undecyl-1H-imidazole-1-ethanol and sodium hydroxide

EC number: 271-794-6 | CAS number: 68608-66-2

Life Cycle description
Uses advised against

Toxicological information

Endpoint summary

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Administrative data

Description of key information

The data for repeated dose toxicity are read across from a substance analogue.

A reliable 90-day repeated dose oral toxicity study (performed according to OECD test guidelines and GLP principles) with rats was performed with a substance analogue, Amphoacetates C12-C14. The NOAEL was determined to be at least 1000 mg/kg bw/day in absence of any adverse effects.

The rationale to read across the data is attached in Section 13.

Key value for chemical safety assessment

Toxic effect type:: dose-dependent

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Endpoint:

sub-chronic toxicity: oral

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Justification for type of information:

The rationale to read across the data is attached in Section 13.

Reason / purpose for cross-reference:

read-across source

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

Abnormal breathing sounds were noted for animals dosed at all dose levels, with a dose dependent increase in incidence with increasing dose. Abnormal breathing sounds were also noted for one control female. For a few individual animals, additional deep, laboured or shallow breathing was incidentally noted across the test material-treated groups. Moreover, retching was noted for males at 100 and 1000 mg/kg bw/day and females at 100, 300 and 1000 mg/kg bw/day. Finally, hunched posture was noted for one male each at 300 and 1000 mg/kg bw/day and one female at 1000 mg/kg bw/day, and erected fur was noted for two males at 300 mg/kg bw/day, four males at 1000 mg/kg bw/day and one female each at 300 and 1000 mg/kg bw/day.
Salivation, on some occasions accompanied by ploughing, was noted in animals dosed at 300 and 1000 mg/kg bw/day. Taking into account the nature and minor severity of the effect and its time of occurrence (i.e., after dosing), salivation was considered to be a physiological response rather than a sign of systemic toxicity. Ploughing was considered to be an adaptive response to salivation, and not directly related to treatment with the test material.
During the Recovery Period, abnormal breathing sounds and/or laboured breathing were noted on few occasions for one female at 1000 mg/kg bw/day. At the incidence observed, and as this concerned the treatment-free period, this was considered to be unrelated to treatment with the test material. No test material-related clinical signs were noted during weekly arena observations for the Main and Recovery animals.
Any other clinical signs noted during the Treatment or Recovery Period occurred within the range of background findings to be expected for rats of this age and strain which are housed and treated under the conditions in this study and/or did not show any apparent dose-related trend. At the incidence observed, these were considered to be unrelated to treatment with the test material.
(see Table 29-30 under "Any other information on results incl. tables")

Mortality:

no mortality observed

Description (incidence):

No mortality occurred during the study period.

Body weight and weight changes:

effects observed, non-treatment-related

Description (incidence and severity):

Body weights and body weight gain were considered to have been unaffected by treatment with the test material.
At 1000 mg/kg bw/day, body weight gain was low for males over Days 78-85, and high for females over Days 1-8. For one male at 1000 mg/kg bw/day, a body weight loss of 9% was noted over Days 57-64. In addition, mean body weight gain over the complete Recovery Period was slightly higher for males at 1000 mg/kg bw/day, when compared to control. As they did not result in significant differences in mean body weights and showed no trend regarding duration of dosing, these slight and incidental changes were considered to be unrelated to treatment with the test material.
(see Table 5-12 under "Any other information on results incl. tables")

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

Food consumption was considered to have been unaffected by treatment with the test material.
At 300 mg/kg bw/day, food intake was slightly low for females during the Treatment Period. As this did not result in significant differences in mean food consumption, showed no trend regarding duration of dosing and did not result in a dose response, these slight and incidental changes were considered to be unrelated to treatment with the test material.
For females at 1000 mg/kg bw/day, food intake was slightly low during the Recovery Period (7.5% lower when compared to control). As this concerned the Recovery Period only, this was considered to be unrelated to treatment with the test material.

Ophthalmological findings:

no effects observed

Description (incidence and severity):

No ophthalmology findings were noted that were considered to be related to the test material.

Haematological findings:

effects observed, treatment-related

Description (incidence and severity):

Hematology parameters were considered not to have been affected by the test material in females up to 1000 mg/kg bw/day and in males up to 300 mg/kg bw/day.
In males at 1000 mg/kg bw/day, red blood cell (RBC) count and red blood cell distribution width (RDWG) were decreased (0.92x and 0.95x of control, respectively), and mean corpuscular volume (MCV) and mean corpuscular hemoglobin (MCH) were increased (1.03x and 1.05x of control, respectively) at the end of the Treatment Period. All reported changes reached statistical significance but were within historical control for this rat strain at the site.
At the end of the Recovery Period, RBC counts were comparable to control, while RDWG was still decreased, and MCV and MCH were still increased (not statistically significant) in males at 1000 mg/kg bw/day.
Any other differences in hematology parameters, regardless of statistical significance, were considered to be unrelated to the test material as these occurred in the absence of a dose-related trend or only at the end of the Recovery Period.

Coagulation parameters were considered not to have been affected by treatment with the test material.

Actual mean values:
Red Blood Cell Count males: control group mean = 9.302; high dose group mean = 8.558
Red Blood Cell Distribution Width males: control group mean =12.67 ; high dose group mean = 12.04
Mean Corpuscular Volume males: control group mean = 51.75 ; high dose group mean = 53.47
Mean Corpuscular Hemoglobin males: control group mean = 17.30; high dose group mean = 18.19

HCD
HCD Red Blood Cell Count males: mean = 8.802; P5-P1: 7.9–9.55
HCD Red Blood Cell Distribution Width males: mean = 427; P5-P1: 11.3–13.6
HCD Mean Corpuscular Volume males: mean = 51.61; P5-P1: 48.9–54.6
HCD Mean Corpuscular Hemoglobin males: mean = 17.21; P5-P1: 16.3–18.9

(see Table 13-16 under "Any other information on results incl. tables")

Clinical biochemistry findings:

effects observed, treatment-related

Description (incidence and severity):

Clinical chemistry parameters were considered not to have been affected by the test material in females up to 1000 mg/kg bw/day and in males up to 300 mg/kg bw/day.
In males at 1000 mg/kg bw/day, triglyceride concentration was increased (1.40x of control, not statistically significant and within historical control data) at the end of the Treatment Period.
At the end of the Recovery Period, triglyceride concentration in males at 1000 mg/kg bw/day was partly recovered.
Remaining differences in clinical chemistry parameters, regardless of statistical significance, were considered not test material-related based on the absence of a dose response, general overlap of individual values with the range of control values, variation in direction of change, were considered to have arisen as a result of slightly high control values, were seen at end of recovery only and/or were of a magnitude of change commonly observed in rats under similar study conditions.

Actual mean values:
Triglyceride males: control group mean = 0.956; high dose group mean = 1.338

HCD
HCD Triglyceride males: mean = 399; P5-P1: 0.36–1.52

(see Table 17-20 under "Any other information on results incl. tables")

Endocrine findings:

effects observed, treatment-related

Description (incidence and severity):

No effect was noted on T3 and TSH levels in males and females, and on T4 levels in males up to 1000 mg/kg bw/day.
In females at 100 and 1000 mg/kg bw/day, T4 levels were increased at the end of the Treatment Period (1.22x and 1.37x of control, not statistically significant at 100 mg/kg bw/day). Mean values remained within the historical control range .
At the end of the Recovery Period, T4 levels in females at 1000 mg/kg bw/day were comparable to control.

Since no histopathological changes were observed in the thyroid, the observed increase in the study was considered not toxicologically relevant.

Actual mean values:
T4 (ng/mL) – females: control group mean = 24.72; high dose group mean = 33.98

HCD
HCD T4 (ng/mL) – females: mean = 29.87; P2.5-P-97.5: 13.92 - 51.38

(see Table 17-20 under "Any other information on results incl. tables")

Behaviour (functional findings):

effects observed, treatment-related

Description (incidence and severity):

Hearing ability, pupillary reflex and static righting reflex were normal in all examined animals. Grip strength was considered unaffected by treatment with the test material.
Motor activity was considered unaffected by treatment with the test material in males at all dose levels and in females up to 300 mg/kg bw/day.
For females at 1000 mg/kg bw/day, the mean total movements and ambulations were both lower compared to control (0.59x and 0.61x of control, respectively; not statistically significant for ambulations). Mean values remained within the historical control range, albeit close to the lower limit.
At the end of the Recovery Period, mean total movements and ambulations were within the same range for control animals and animals treated at 1000 mg/kg bw/day.
All groups showed a similar motor activity habituation profile with a decreasing trend in activity over the duration of the test period.

Actual mean values:
Total movements – females: control group mean = 4615; high dose group mean = 2739
Total ambulations – females: control group mean = 1095; high dose group mean = 667

HCD
HCD Total movements – females: mean = 4740; P5-P95 = 2393-8076
HCD Total ambulations – females: mean = 1335; P5-P95 = 638-2271

(see Table 21-22 under "Any other information on results incl. tables")

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

Test material-related higher kidney and liver weights (absolute and relative to body weight) were noted in females at 1000 mg/kg bw/day (see Table 2 under "Any other information on results incl. tables"). These differences were within the historical control data of the test site.
Higher kidney weight (absolute and relative to body weight) was present in females at 1000 mg/kg bw/day at the end of the Treatment Period. Following the Recovery Period, there was no significant difference in the kidney weight of test material-treated females compared to the control females, suggesting full recovery.
Higher liver weight (absolute and relative to body weight) was present in females at 1000 mg/kg bw/day at the end of the Treatment Period. Following the Recovery Period, there was no significant difference in the liver weight of test material-treated females compared to the control females, suggesting full recovery.
Other statistically significant changes at the end of the Treatment Period (lower relative pituitary gland weight, females at 100 mg/kg bw/day) or Recovery Period (higher relative brain weight, females at 1000 mg/kg bw/day) were considered to be not test material-related due to the lack of a dose-related pattern (pituitary gland) or was regarded related to the lower final body weight (brain).

Since the higher kidney and liver weights in females at 1000 mg/kg bw/day remained within the historical control data, the changes were no longer observed after recovery and in the absence of any concomitant histopathological changes, these effects were considered non-adverse.

Actual mean values:
Liver weight: control group mean = 5.3753; high dose group mean = 6.4360
Liver relative to body weight: control group mean = 2.43901; high dose group mean = 2.79153
Kidney weight: control group mean = 1.4672; high dose group mean = 1.7272
Kidney relative to body weight: control group mean = 0.66594; high dose group mean = 0.74781

HCD
HCD Liver weight: Mean = 5.56; P5-P95 = 4.634-6.472
HCD Liver relative to body weight: Mean = 2.50; P5-P95 = 2.226-2.876
HCD Kidney weight: Mean = 1.53; P5-P95 = 0.526-1.80
HCD Kidney relative to body weight: Mean = 0.69; P5-P95 = 0.588-0.807

(see also Table 23-26 under "Any other information on results incl. tables")

Gross pathological findings:

effects observed, non-treatment-related

Description (incidence and severity):

There were no test material-related gross observations.
All the recorded macroscopic findings were within the range of background gross observations encountered in rats of this age and strain.

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

Test material-related microscopic findings were noted in the rectum of males at 1000 mg/kg bw/day and the stomach of females at 300 and 1000 mg/kg bw/day (see Table 3 under "Any other information on results incl. tables").
In the rectum, goblet cell hyperplasia was recorded in 3/10 males at 1000 mg/kg bw/day (minimal) at the end of the Treatment Period. Following the Recovery Period this finding was not present, suggesting full recovery.
In the stomach, in the non-glandular stomach, squamous cell hyperplasia often accompanied by hyperkeratosis was recorded in 1/10 females at 300 mg/kg bw/day (minimal) and in 3/10 females at 1000 mg/kg bw/day (up to mild degree) at the end of the Treatment Period. Following the Recovery Period this finding was not present, suggesting full recovery (see Table 4 under "Any other information on results incl. tables").
Focal erosion of the non-glandular and glandular portion of the stomach (minimal) was noted in 1/10 females at 1000 mg/kg/day (Main Group females) and 1/5 females at 1000 mg/kg bw/day (Recovery Group females). At this low incidence and severity, these findings were considered not to represent a test material-related effect but rather represent a procedure-related effect or an incidental finding.
There were no other test material-related histologic changes. The remainder of the recorded microscopic findings were considered to be incidental findings or were within the range of background pathology encountered in rats of this age and strain. There was no test material related alteration in the prevalence, severity, or histologic character of those incidental tissue alterations.

(see also Table 27-28 under "Any other information on results incl. tables")

Key result

Dose descriptor:

NOAEL

Effect level:

>= 1 000 mg/kg bw/day (actual dose received)

Based on:

test mat.

Remarks:

corrected for water content

Sex:

male/female

Remarks on result:

not determinable due to absence of adverse toxic effects

Key result

Critical effects observed:

The accuracy and homogeneity of the formulations of Group 4 were not determined because it was pure test material.
Accuracy
The concentrations analyzed in the formulations of Groups 2 and 3 were in agreement with target concentrations (i.e., mean sample concentration results were within or equal to 90-110% of target concentration).
No test material was detected in the Group 1 formulations.
Homogeneity
The formulations of Groups 2 and 3 were homogeneous (i.e., coefficient of variation ≤ 10%).

Table 2: Mean Percent kidney and liver Weight Differences from Control Groups

	Main Females			Recovery Females
Dose level (mg/kg bw/day):	100	300	1000	1000
KIDNEYS
Absolute	8	0	18**	-3
Relative to body weight	5	4	12**	2
LIVER
Absolute	1	-5	20**	-7
Relative to body weight	-2	-2	14**	-2

**: P≤0.01

Table 3: Summary Test Material-Related Microscopic Findings - Males

	Main Males				Recovery Males
Dose level (mg/kg bw/day):	0	100	300	1000	0	1000
RECTUM ^a	10	10	10	10	5	5
Hyperplasia, goblet cell
Minimal	-	-	-	3	-	-

^a = Number of tissues examined from each group.

Table 4: Summary Test Material-Related Microscopic Findings - Females

	Main Females				Recovery Females
Dose level (mg/kg bw/day):	0	100	300	1000	0	1000
STOMACH ^a	10	10	10	10	5	5
Hyperplasia, squamous cell, non-glandular
Minimal	-	-	1	2	-	-
Mild	-	-	-	1	-	-

^a = Number of tissues examined from each group.

Table 5: Body weights (gram) summary (males) -Main

Sex: Male		Day(s) Relative to Start Date
Sex: Male		1	8	15	22	29	36	43	50	57	64	71	78	85	91
0	Mean	193.5	235.9	270.2	296.3	317.1	338.5	350.3	366.9	377.4	387.4	395.7	401.7	411.6	415.7
mg/kg/day	SD	10.6	14.1	18.5	20.5	23.0	23.3	25.8	25.4	29.4	28.9	28.9	30.9	31.8	32.1
Group 1	N	15	15	15	15	15	15	15	15	15	15	15	15	15	15
100	Mean	199.5	241.6	275.0	299.5	321.2	342.0	356.8	374.6	383.8	393.9	404.5	412.1	420.6	423.6
mg/kg/day	SD	9.3	14.7	17.5	19.0	20.1	22.6	25.0	27.2	27.6	29.7	33.4	34.0	34.4	35.8
Group 2	N	10	10	10	10	10	10	10	10	10	10	10	10	10	10
	%Diff	3.1	2.4	1.8	1.1	1.3	1.0	1.8	2.1	1.7	1.7	2.2	2.6	2.2	1.9
300	Mean	197.8	237.0	268.0	292.0	312.3	334.5	350.6	365.2	377.2	386.4	397.7	406.6	414.0	418.6
mg/kg/day	SD	14.1	17.1	18.4	24.4	26.1	29.2	28.2	31.8	32.3	34.4	33.3	34.4	35.6	37.6
Group 3	N	10	10	10	10	10	10	10	10	10	10	10	10	10	10
	%Diff	2.2	0.5	-0.8	-1.4	-1.5	-1.2	0.1	-0.5	-0.1	-0.3	0.5	1.2	0.6	0.7
1000	Mean	196.5	236.8	269.8	298.0	318.3	337.9	350.3	365.3	373.1	381.3	390.4	398.5	401.1	408.1
mg/kg/day	SD	12.1	16.0	19.1	20.8	23.9	26.0	28.3	30.4	31.5	38.2	36.4	34.8	35.1	37.0
Group 4	N	15	15	15	15	15	15	15	15	15	15	15	15	15	15
	%Diff	1.6	0.4	-0.1	0.6	0.4	-0.2	0.0	-0.5	-1.1	-1.6	-1.3	-0.8	-2.5	-1.8

Table 6: Body weigths (gram) summary (Females) -Main

Sex: Female		Day(s) Relative to Start Date
Sex: Female		1 [G]	8 [G]	15 [G1]	22 [G]	29 [G]	36 [G]	43 [G1]	50 [G]	57 [G]	64 [G]	71 [G]	78 [G]	85	91
0	Mean	131.6	147.7	169.2	183.3	195.1	206.5	212.5	219.9	225.8	228.9	233.2	236.0	240.5	241.3
mg/kg/day	SD	9.6	9.4	11.2	14.5	13.8	13.5	12.2	15.3	14.7	13.7	14.5	16.4	16.0	15.1
Group 1	N	15	15	15	15	15	15	15	15	15	15	15	15	15	15
100	Mean	130.9	150.5	166.5	184.1	196.0	208.2	213.7	223.1	226.8	231.7	234.0	238.0	240.2	241.1
mg/kg/day	SD	7.9	10.7	11.8	10.7	12.3	14.4	15.3	13.9	15.2	16.5	17.9	16.7	15.6	17.0
Group 2	N	10	10	10	10	10	10	10	10	10	10	10	10	10	10
	%Diff	-0.5	1.9	-1.6	0.5	0.4	0.8	0.5	1.5	0.4	1.2	0.3	0.8	-0.1	-0.1
300	Mean	123.8	139.5	158.3	174.6	185.8	195.9	202.6	210.5	213.0	217.4	220.8	224.9	229.0	229.6
mg/kg/day	SD	11.0	11.1	15.5	15.7	17.4	18.4	17.3	18.4	18.2	18.6	20.3	18.3	17.1	17.0
Group 3	N	10	10	10	10	10	10	10	10	10	10	10	10	10	10
	%Diff	-5.9	-5.6	-6.4	-4.7	-4.8	-5.1	-4.7	-4.3	-5.7	-5.0	-5.3	-4.7	-4.8	-4.8
1000	Mean	135.7	156.1	175.5	192.5	205.7	214.1	219.3	231.4	234.9	236.6	239.5	243.7	245.0	244.8
mg/kg/day	SD	8.7	9.7	8.4	9.7	10.3	10.7	10.3	12.6	11.6	11.8	14.4	12.4	11.9	13.9
Group 4	N	15	15	15	15	15	15	15	15	15	15	15	15	15	15
	%Diff	3.1	5.7	3.7	5.0	5.4	3.7	3.2	5.2	4.0	3.4	2.7	3.3	1.9	1.5

[G] - Anova & Dunnett

[G1] - Kruskal-Wallis & Dunn

Table 7: Body weigths (gram) summary (Males) -Recovery

Sex: Male		Day(s) Relative to Start Date
Sex: Male		92	99	106	113	120
0	Mean	419.0	425.6	432.4	437.0	440.8
mg/kg/day	SD	31.8	30.6	30.8	33.4	30.9
Group 1	N	5	5	5	5	5
1000	Mean	414.4	424.2	433.6	441.2	443.2
mg/kg/day	SD	33.6	33.7	32.6	34.3	35.5
Group 4	N	5	5	5	5	5
	%Diff	-1.1	-0.3	0.3	1.0	0.5

Table 8: Body weigths (gram) summary (Females) -Recovery

Sex: Female		Day(s) Relative to Start Date
Sex: Female		92	99	106	113	120
0	Mean	254.0	254.2	260.0	259.8	259.0
mg/kg/day	SD	11.0	9.0	10.8	10.6	13.3
Group 1	N	5	5	5	5	5
1000	Mean	242.0	244.2	249.0	244.6	244.4
mg/kg/day	SD	12.4	16.7	17.0	15.4	15.9
Group 4	N	5	5	5	5	5
	%Diff	-4.7	-3.9	-4.2	-5.9	-5.6

Table 9: Body weight gain (%) summary (Males) -Main

Sex: Male		Day(s) Relative to Start Date
Sex: Male		1 → 8 [G]	8 → 15 [G]	15 → 22 [G]	22 → 29 [G]	29 → 36 [G]	36 → 43 [G1]	43 → 50 [G]	50 → 57	57 → 64	64 → 71	71 → 78	78 → 85	85 → 91	1 → 91
0 mg/kg/day Group 1	Mean	42.4	34.3	26.1	20.9	21.4	11.8	16.6	10.5	10.0	8.3	6.1	9.9	4.1	222.2
	SD	4.0	5.9	4.8	4.2	4.4	4.9	3.9	5.8	3.8	4.4	3.7	3.3	2.4	23.9
	N	15	15	15	15	15	15	15	15	15	15	15	15	15	15
100 mg/kg/day Group 2	Mean	42.1	33.4	24.5	21.7	20.8	14.8	17.8	9.2	10.1	10.6	7.6	8.5	3.0	224.1
	SD	7.5	4.9	3.6	3.0	3.0	3.1	5.4	3.6	3.7	5.5	4.1	4.1	3.1	29.0
	N	10	10	10	10	10	10	10	10	10	10	10	10	10	10
300 mg/kg/day Group 3	Mean	39.2	31.0	24.0	20.3	22.2	16.1	14.6	12.0	9.2	11.3	8.9	7.4	4.6	220.8
	SD	4.6	6.0	9.7	4.6	3.4	2.9	6.2	3.2	3.2	2.2	2.8	3.8	4.4	32.0
	N	10	10	10	10	10	10	10	10	10	10	10	10	10	10
1000 mg/kg/day Group 4	Mean	40.3	33.0	28.2	20.3	19.6	12.3	15.0	7.9	8.1	9.1	8.1	2.6**	7.0	211.6
	SD	5.0	5.8	5.6	7.7	4.5	5.5	4.6	4.5	11.3	5.8	5.4	3.2	4.8	30.7
	N	15	15	15	15	15	15	15	15	15	15	15	15	15	15

[G] - Anova & Dunnett

[G1] - Kruskal-Wallis & Dunn

Table 10: Body weight gain (%) summary (females) -Main

Sex: Female		Day(s) Relative to Start Date
Sex: Female		1 → 8 [G]	8 → 15 [G]	15 → 22 [G1]	22 → 29 [G]	29 → 36 [G]	36 → 43 [G]	43 → 50 [G]	50 → 57	57 → 64	64 → 71	71 → 78	78 → 85	85 → 91	1 → 91
0 mg/kg/day Group 1	Mean	16.1	21.5	14.1	11.9	11.3	6.1	7.3	5.9	3.1	4.3	2.8	4.5	0.7	109.7
	SD	4.3	4.9	4.6	5.3	4.5	4.4	4.6	5.4	4.3	5.8	4.2	4.3	4.0	8.5
	N	15	15	15	15	15	15	15	15	15	15	15	15	15	15
100 mg/kg/day Group 2	Mean	19.6	16.0	17.6	11.9	12.2	5.5	9.4	3.7	4.9	2.3	4.0	2.2	0.9	110.2
	SD	3.9	3.1	4.4	4.1	5.0	3.7	5.2	4.6	3.1	3.7	4.1	4.3	5.1	11.8
	N	10	10	10	10	10	10	10	10	10	10	10	10	10	10
300 mg/kg/day Group 3	Mean	15.7	18.8	16.3	11.2	10.1	6.7	7.9	2.5	4.4	3.4	4.1	4.1	0.6	105.8
	SD	4.6	5.4	1.5	3.5	5.0	3.3	3.0	3.9	4.3	5.1	3.7	3.7	7.0	12.9
	N	10	10	10	10	10	10	10	10	10	10	10	10	10	10
1000 mg/kg/day Group 4	Mean	20.5*	19.4	16.9	13.3	8.3	5.2	12.1	3.5	1.7	2.9	4.3	1.3	-0.2	109.1
	SD	5.1	5.7	4.5	4.0	6.5	5.3	6.9	6.4	4.8	9.4	5.4	4.1	5.8	11.6
	N	15	15	15	15	15	15	15	15	15	15	15	15	15	15

[G] - Anova & Dunnett: * = p ≤ 0.05
[G1] - Kruskal-Wallis & Dunn

Table 11: Body weight gain (%) summary (males) -Recovery

Sex: Male		Day(s) Relative to Start Date
Sex: Male		92 → 99 [G]	99 → 106 [G]	106 → 113 [G1]	113 → 120 [G]	92 → 120 [G]
0 mg/kg/day Group 1	Mean	6.6	6.8	4.6	3.8	21.8
	SD	1.9	1.3	4.8	2.9	5.0
	N	5	5	5	5	5
1000 mg/kg/day Group 4	Mean	9.8	9.4	7.6	2.0	28.8*
	SD	2.6	2.3	2.5	1.6	4.0
	N	5	5	5	5	5

[G] - Anova & Dunnett: * = p ≤ 0.05

Table 12: Body weight gain (%) summary (females) -Recovery

Sex: Female		Day(s) Relative to Start Date
Sex: Female		92 → 99	99 → 106	106 → 113	113 → 120	92 → 120
0 mg/kg/day Group 1	Mean	0.2	5.8	-0.2	-0.8	5.0
	SD	3.1	3.5	4.3	4.5	3.7
	N	5	5	5	5	5
1000 mg/kg/day Group 4	Mean	2.2	4.8	-4.4	-0.2	2.4
	SD	6.2	3.0	2.9	4.8	3.6
	N	5	5	5	5	5

Table 13: Haematology summary (males) - Day 92 (end of treatment)

Sex: Male		Reporting Hematology
Sex: Male		WBC (10^9/L) [G]	NEUT (10^9/L) [G]	LYMPH (10^9/L) [G]	MONO (10^9/L) [G]	EOS (10^9/L) [G]	BASO (10^9/L) [G]	LUC (10^9/L) [G]	RBC (10^12/L) [G]	RETIC (10^9/L) [G]	RDWG (%) [G]	HGB (g/L) [G1]	HCT (L/L) [G]	MCV (fL) [G]	MCH (pg) [G]	MCHC (g/L) [G]	PLT (10^9/L) [G]
0 mg/kg/day Group 1	Mean	6.008	1.074	4.658	0.096	0.116	0.008	0.053	9.302	206.11	12.67	160.9	0.4814	51.75	17.30	334.2	723.0
	SD	1.263	0.331	1.013	0.031	0.023	0.004	0.013	0.224	27.72	0.54	4.7	0.0186	1.07	0.42	6.7	35.2
	N	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10
100 mg/kg/day Group 2	Mean	5.138	1.106	3.785	0.079	0.128	0.005	0.033	9.142	188.16	12.23	159.9	0.4724	51.70	17.53	339.0	643.8
	SD	1.661	0.366	1.683	0.021	0.031	0.007	0.015	0.549	25.76	0.35	7.2	0.0267	0.94	0.44	7.3	80.9
	N	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10
	tCtrl	0.86	1.03	0.81	0.82	1.10	0.63	0.62	0.98	0.91	0.97	0.99	0.98	1.00	1.01	1.01	0.89
300 mg/kg/day Group 3	Mean	6.502	1.235	4.931	0.107	0.160	0.007	0.062	9.086	202.13	12.44	161.7	0.4768	52.52	17.83	339.6	674.3
	SD	1.688	0.259	1.532	0.042	0.040	0.005	0.022	0.645	38.11	0.44	7.5	0.0263	1.43	0.61	6.3	97.1
	N	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10
	tCtrl	1.08	1.15	1.06	1.11	1.38	0.88	1.17	0.98	0.98	0.98	1.00	0.99	1.01	1.03	1.02	0.93
1000 mg/kg/day Group 4	Mean	5.481	0.992	4.216	0.088	0.130	0.006	0.049	8.558 **	216.19	12.04 **	155.7	0.4581	53.47 *	18.19 **	340.2	726.4
	SD	2.253	0.199	2.146	0.028	0.043	0.005	0.038	0.563	26.23	0.38	12.1	0.0398	2.05	0.76	7.5	69.1
	N	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10
	tCtrl	0.91	0.92	0.91	0.92	1.12	0.75	0.92	0.92	1.05	0.95	0.97	0.95	1.03	1.05	1.02	1.00

[G] - Anova & Dunnett: ** = p ≤ 0.01
[G1] - Kruskal-Wallis & Dunn

Table 14: Haematology summary (Females) - Day 93 (end of treatment)

Sex: Female		Reporting Hematology
Sex: Female		WBC (10^9/L) [G]	NEUT (10^9/L) [G]	LYMPH (10^9/L) [G]	MONO (10^9/L) [G]	EOS (10^9/L) [G]	BASO (10^9/L) [G1]	LUC (10^9/L) [G]	RBC (10^12/L) [G]	RETIC (10^9/L) [G]	RDWG (%) [G]	HGB (g/L) [G]	HCT (L/L) [G]	MCV (fL) [G]	MCH (pg) [G]	MCHC (g/L) [G]	PLT (10^9/L) [G]
0 mg/kg/day Group 1	Mean	3.863	0.430	3.239	0.061	0.088	0.005	0.039	7.942	222.87	10.97	149.6	0.4435	55.86	18.86	338.0	723.9
	SD	0.933	0.165	0.733	0.031	0.034	0.005	0.021	0.363	24.72	0.29	5.9	0.0183	1.25	0.65	7.0	110.8
	N	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10
100 mg/kg/day Group 2	Mean	3.189	0.591	2.431	0.052	0.089	0.000 *	0.027	8.269	218.39	11.01	149.4	0.4477	54.17 **	18.08 **	333.7	755.7
	SD	0.724	0.239	0.546	0.023	0.034	0.000	0.008	0.290	25.11	0.35	5.1	0.0123	1.09	0.56	4.6	84.2
	N	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10
	tCtrl	0.83	1.37	0.75	0.85	1.01	0.00	0.69	1.04	0.98	1.00	1.00	1.01	0.97	0.96	0.99	1.04
300 mg/kg/day Group 3	Mean	3.715	0.815 *	2.664	0.077	0.117	0.000 *	0.039	8.222	212.56	11.18	152.7	0.4513	54.93	18.59	338.5	735.2
	SD	1.399	0.505	0.822	0.046	0.076	0.000	0.022	0.269	25.28	0.35	4.9	0.0150	1.27	0.59	6.2	57.6
	N	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	9
	tCtrl	0.96	1.90	0.82	1.26	1.33	0.00	1.00	1.04	0.95	1.02	1.02	1.02	0.98	0.99	1.00	1.02
1000 mg/kg/day Group 4	Mean	4.430	0.745 *	3.462	0.077	0.097	0.005	0.043	8.106	215.62	10.83	149.5	0.4425	54.60 *	18.45	338.1	801.0
	SD	1.372	0.369	1.159	0.029	0.025	0.007	0.026	0.347	45.04	0.29	5.3	0.0190	0.74	0.37	4.0	92.3
	N	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10
	tCtrl	1.15	1.73	1.07	1.26	1.10	1.00	1.10	1.02	0.97	0.99	1.00	1.00	0.98	0.98	1.00	1.11

[G] - Anova & Dunnett

Table 15: Haematology summary (males) - Day 121 (end of recovery)

Sex: Male		Reporting Hematology
Sex: Male		WBC (10^9/L) [G]	NEUT (10^9/L) [G]	LYMPH (10^9/L) [G]	MONO (10^9/L) [G]	EOS (10^9/L) [G]	BASO (10^9/L) [G]	LUC (10^9/L) [G]	RBC (10^12/L) [G]	RETIC (10^9/L) [G]	RDWG (%) [G]	HGB (g/L) [G]	HCT (L/L) [G]	MCV (fL) [G]	MCH (pg) [G]	MCHC (g/L) [G]	PLT (10^9/L) [G]
0 mg/kg/day Group 1	Mean	4.578	0.928	3.414	0.090	0.114	0.008	0.024	8.780	186.72	12.38	158.4	0.4568	52.14	18.14	347.6	751.0
	SD	1.457	0.335	1.192	0.033	0.040	0.004	0.009	0.892	14.08	0.59	11.6	0.0420	1.20	1.01	16.2	83.3
	N	5	5	5	5	5	5	5	5	5	5	5	5	5	5	5	4
1000 mg/kg/day Group 4	Mean	5.108	0.894	3.978	0.096	0.112	0.008	0.022	8.662	172.64	11.68 *	160.8	0.4676	54.02	18.60	344.4	730.0
	SD	2.131	0.347	1.732	0.063	0.023	0.004	0.013	0.557	31.43	0.26	4.9	0.0229	1.73	0.97	9.9	132.8
	N	5	5	5	5	5	5	5	5	5	5	5	5	5	5	5	5
	tCtrl	1.12	0.96	1.17	1.07	0.98	1.00	0.92	0.99	0.92	0.94	1.02	1.02	1.04	1.03	0.99	0.97

[G] - Anova & Dunnett: * = p ≤ 0.05

Table 16: Haematology summary (Females) - Day 121 (end of recovery)

Sex: Female		Reporting Hematology
Sex: Female		WBC (10^9/L) [G]	NEUT (10^9/L) [G1]	LYMPH (10^9/L) [G]	MONO (10^9/L) [G1]	EOS (10^9/L) [G]	BASO (10^9/L) [G1]	LUC (10^9/L) [G1]	RBC (10^12/L) [G]	RETIC (10^9/L) [G]	RDWG (%) [G1]	HGB (g/L) [G]	HCT (L/L) [G]	MCV (fL) [G]	MCH (pg) [G]	MCHC (g/L) [G]	PLT (10^9/L) [G]
0 mg/kg/day Group 1	Mean	2.426	0.602	1.670	0.050	0.086	0.004	0.008	7.516	181.82	10.54	140.0	0.4116	54.78	18.64	340.2	710.2
	SD	0.783	0.466	0.754	0.016	0.015	0.005	0.004	0.726	12.28	0.27	12.5	0.0396	1.42	0.43	5.2	80.4
	N	5	5	5	5	5	5	5	5	5	5	5	5	5	5	5	5
1000 mg/kg/day Group 4	Mean	1.698	0.310	1.270	0.030	0.076	0.004	0.008	8.260	153.40	10.40	149.6	0.4410	53.42	18.12	339.2	723.6
	SD	0.294	0.067	0.274	0.012	0.038	0.005	0.004	0.250	44.49	0.32	3.0	0.0124	0.90	0.29	4.1	40.8
	N	5	5	5	5	5	5	5	5	5	5	5	5	5	5	5	5
	tCtrl	0.70	0.51	0.76	0.60	0.88	1.00	1.00	1.10	0.84	0.99	1.07	1.07	0.98	0.97	1.00	1.02

[G] - Anova & Dunnett
[G1] - Anova & Dunnett

Table 17: Clinical biochemistry summary (males) - Day 92 (end of treatment)

Sex: Male		Reporting Biochemistry
Sex: Male		ALT (U/L) [G]	AST (U/L) [G]	ALP (U/L) [G1]	TPROT (g/L) [G]	ALB (g/L) [G]	TBIL (umol/L) [G]	UREA (mmol/L) [G1]	CREAT (umol/L) [G]	GLUC (mmol/L) [G]	CHOL (mmol/L) [G]	TRIG (mmol/L) [G]	HDL (mmol/L) [G]	LDL (mmol/L) [G]	NA (mmol/L) [G]	K (mmol/L) [G]	CL (mmol/L) [G]	CA (mmol/L) [G]	PHOS (mmol/L) [G]	T3 (ng/mL) [G]	T4 (ng/mL) [G]	TSH (mU/L) [G1]
0 mg/kg/day Group 1	Mean	51.7	71.7	102.8	65.23	40.96	1.59	5.38	28.2	9.830	1.875	0.956	1.266	0.340	146.3	4.15	107.4	2.562	1.617	0.324	37.52	0.1327
	SD	8.2	7.9	18.5	2.46	0.97	0.22	0.74	3.3	1.323	0.436	0.371	0.243	0.073	0.9	0.50	1.0	0.140	0.109	0.059	5.28	0.0737
	N	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10
100 mg/kg/day Group 2	Mean	49.2	75.0	85.4	63.59	40.15	1.94	5.37	29.6	10.179	1.892	0.959	1.274	0.333	146.5	4.07	107.3	2.597	1.494	0.316	36.09	0.1350
	SD	12.3	25.7	13.3	1.73	1.26	0.26	0.44	2.0	1.700	0.192	0.309	0.116	0.042	1.4	0.30	1.6	0.168	0.183	0.071	7.27	0.1308
	N	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10
	tCtrl	0.95	1.05	0.83	0.97	0.98	1.22	1.00	1.05	1.04	1.01	1.00	1.01	0.98	1.0	0.98	1.0	1.01	0.92	0.98	0.96	1.02
300 mg/kg/day Group 3	Mean	51.3	69.7	95.0	65.35	41.17	1.59	5.46	29.0	9.169	1.853	1.025	1.258	0.321	146.6	3.88	107.3	2.642	1.600	0.331	41.68	0.1354
	SD	9.8	5.8	31.1	2.41	1.41	0.32	0.50	3.0	1.359	0.202	0.336	0.165	0.040	1.2	0.29	1.6	0.105	0.182	0.070	5.78	0.0650
	N	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10
	tCtrl	0.99	0.97	0.92	1.00	1.01	1.00	1.01	1.03	0.93	0.99	1.07	0.99	0.94	1.0	0.93	1.0	1.03	0.99	1.02	1.11	1.02
1000 mg/kg/day Group 4	Mean	53.4	87.5	92.5	64.27	41.05	1.82	5.86	29.6	10.106	2.010	1.338	1.362	145.8	145.8	4.17	106.9	2.613	1.791	0.349	36.79	0.0863
	SD	10.0	55.3	22.8	3.24	2.15	0.49	1.34	4.6	1.409	0.190	0.438	0.131	0.046	1.1	0.37	1.5	0.135	0.238	0.082	5.52	0.0568
	N	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10
	tCtrl	1.03	1.22	0.90	0.99	1.00	1.14	1.09	1.05	1.03	1.07	1.40	1.08	1.06	1.0	1.0	1.0	1.02	1.11	1.08	0.98	0.65

[G] - Anova & Dunnett
[G1] - Kruskal-Wallis & Dunn

Table 18: Clinical biochemistry summary (females) - Day 93 (end of treatment)

Sex: Female		Reporting Biochemistry
Sex: Female		ALT (U/L) [G]	AST (U/L) [G1]	ALP (U/L) [G]	TPROT (g/L) [G]	ALB (g/L) [G]	TBIL (umol/L) [G]	UREA (mmol/L) [G1]	CREAT (umol/L) [G]	GLUC (mmol/L) [G]	CHOL (mmol/L) [G]	TRIG (mmol/L) [G]	HDL (mmol/L) [G]	LDL (mmol/L) [G]	NA (mmol/L) [G]	K (mmol/L) [G]	CL (mmol/L) [G]	CA (mmol/L) [G1]	PHOS (mmol/L) [G]	T3 (ng/mL) [G]	T4 (ng/mL) [G]	TSH (mU/L) [G1]
0 mg/kg/day Group 1	Mean	56.4	102.6	38.1	71.08	45.71	2.36	6.85	35.3	8.201	1.628	0.840	1.111	0.328	150.8	3.76	111.6	2.629	1.533	0.479	24.72	0.1075
	SD	17.9	58.1	11.8	2.19	1.64	0.34	0.71	3.2	0.851	0.300	0.317	0.192	0.050	1.0	0.20	1.4	0.082	0.200	0.107	5.03	0.0759
	N	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10
100 mg/kg/day Group 2	Mean	50.7	74.8	36.1	71.40	45.63	2.01	6.56	37.5	8.172	1.477	0.903	0.995	0.306	150.3	3.90	112.0	2.604	1.104**	0.495	30.04	0.0829
	SD	8.6	6.8	6.9	2.28	1.77	0.36	0.67	3.4	0.652	0.279	0.360	0.189	0.060	1.2	0.36	1.5	0.078	0.192	0.106	8.18	0.0525
	N	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10
	tCtrl	0.90	0.73	0.95	1.00	1.00	0.85	0.96	1.06	1.00	0.91	1.08	0.90	0.93	1.00	1.04	1.00	0.99	0.72	1.03	1.22	0.77
300 mg/kg/day Group 3	Mean	46.1	73.4	38.6	67.05 *	42.96 *	1.93	6.60	34.2	7.832	1.408	1.013	0.950	0.287	147.2**	3.85	109.5*	2.671	1.299	0.414	24.36	0.0687
	SD	9.1	11.3	18.4	4.85	2.56	0.36	1.02	3.3	1.440	0.358	0.501	0.250	0.075	2.6	0.21	2.3	0.192	0.296	0.051	6.66	0.0504
	N	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10
	tCtrl	0.82	0.72	1.01	0.94	0.94	0.82	0.96	0.97	0.96	0.86	1.21	0.86	0.88	0.98	1.02	0.98	1.02	0.85	0.86	0.99	0.64
1000 mg/kg/day Group 4	Mean	54.3	67.3 *	41.3	69.92	44.71	2.25	6.65	32.8	8.691	1.796	0.924	1.254	0.387	146.8**	3.81	108.5**	2.705	1.406	0.544	33.98*	0.0941
	SD	13.0	5.4	11.7	2.83	2.07	0.53	0.82	2.9	1.021	0.335	0.388	0.230	0.084	0.8	0.27	1.4	0.122	0.203	0.115	7.40	0.0400
	N	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10	10
	tCtrl	0.96	0.66	1.08	0.98	0.98	0.95	0.97	0.93	1.06	1.10	1.10	1.13	1.18	0.97	1.01	0.97	1.03	0.92	1.14	1.37	0.88

[G] - Anova & Dunnett: * = p ≤ 0.05; ** = p ≤ 0.01
[G1] - Kruskal-Wallis & Dunn

Table 19: Clinical biochemistry summary (males) - Day 121 (end of recovery)

Sex: Male		Reporting Biochemistry
Sex: Male		ALT (U/L) [G]	AST (U/L) [G]	ALP (U/L) [G]	TPROT (g/L) [G]	ALB (g/L) [G]	TBIL (umol/L) [G]	UREA (mmol/L) [G1]	CREAT (umol/L) [G]	GLUC (mmol/L) [G]	CHOL (mmol/L) [G]	TRIG (mmol/L) [G]	HDL (mmol/L) [G]	LDL (mmol/L) [G]	NA (mmol/L) [G1]	K (mmol/L) [G]	CL (mmol/L) [G]	CA (mmol/L) [G]	PHOS (mmol/L) [G]	T3 (ng/mL) [G]	T4 (ng/mL) [G1]	TSH (mU/L) [G1]
0 mg/kg/day Group 1	Mean	46.5	69.3	87.8	62.74	39.14	1.94	4.93	28.7	8.630	1.844	1.044	1.278	0.348	144.6	3.76	106.8	2.590	1.468	0.328	37.74	0.2830
	SD	9.6	9.2	20.5	2.36	1.34	0.33	0.50	1.4	0.482	0.338	0.541	0.208	0.047	0.5	0.25	1.1	0.082	0.104	0.070	3.13	0.2264
	N	5	5	5	5	5	5	5	5	5	5	5	5	5	5	5	5	5	5	5	5	5
1000 mg/kg/day Group 4	Mean	42.0	62.4	86.9	63.42	39.10	1.92	5.41	29.3	9.218	1.790	1.176	1.246	0.338	145.0	3.86	107.6	2.542	1.532	0.286	40.78	0.2070
	SD	4.5	8.7	16.4	3.40	1.15	0.40	0.35	0.8	0.686	0.170	0.786	0.130	0.059	1.4	0.22	1.3	0.072	0.102	0.067	10.51	0.0911
	N	5	5	5	5	5	5	5	5	5	5	5	5	5	5	5	5	5	5	5	5	5
	tCtrl	0.90	0.90	0.99	1.01	1.00	0.99	1.10	1.02	1.07	0.97	1.13	0.97	0.97	1.00	1.03	1.01	0.98	1.04	0.87	1.08	0.73

[G] - Anova & Dunnett
[G1] - Kruskal-Wallis & Dunn

Table 20: Clinical biochemistry summary (females) - Day 121 (end of recovery)

Sex: Female		Reporting Biochemistry
Sex: Female		ALT (U/L) [G]	AST (U/L) [G]	ALP (U/L) [G1]	TPROT (g/L) [G]	ALB (g/L) [G]	TBIL (umol/L) [G]	UREA (mmol/L) [G1]	CREAT (umol/L) [G]	GLUC (mmol/L) [G]	CHOL (mmol/L) [G]	TRIG (mmol/L) [G]	HDL (mmol/L) [G]	LDL (mmol/L) [G]	NA (mmol/L) [G]	K (mmol/L) [G]	CL (mmol/L) [G1]	CA (mmol/L) [G]	PHOS (mmol/L) [G]	T3 (ng/mL) [G]	T4 (ng/mL) [G]	TSH (mU/L) [G]
0 mg/kg/day Group 1	Mean	56.4	142.6	46.0	66.08	42.30	3.18	6.46	33.4	7.566	1.630	0.710	1.178	0.314	143.8	3.63	106.8	2.622	1.478	0.402	26.26	0.0754
	SD	30.7	155.3	12.3	2.79	2.06	0.66	0.69	3.5	0.732	0.374	0.247	0.261	0.103	1.5	0.41	1.3	0.098	0.172	0.038	3.65	0.0422
	N	5	5	5	5	5	5	5	5	5	5	5	5	5	5	5	5	5	5	5	5	5
1000 mg/kg/day Group 4	Mean	40.2	77.5	30.0 *	61.66 *	39.62	1.94 **	6.62	35.8	8.112	1.310	0.576	0.906	0.248	143.8	3.64	108.2	2.592	1.494	0.448	25.18	0.0398
	SD	8.6	15.4	5.8	2.05	1.90	0.23	1.02	5.3	0.340	0.311	0.227	0.242	0.062	0.4	0.23	0.4	0.146	0.311	0.060	5.00	0.0295
	N	5	5	5	5	5	5	5	5	5	5	5	5	5	5	5	5	5	5	5	5	5
	tCtrl	0.71	0.54	0.65	0.93	0.94	0.61	1.03	1.07	1.07	0.80	0.81	0.77	0.79	1.00	1.00	1.01	0.99	1.01	1.11	0.96	0.53

[G] - Anova & Dunnett: * = p ≤ 0.05; ** = p ≤ 0.01
[G1] - Kruskal-Wallis & Dunn

Table 21: Functional observations (males)

		GROUP 1 CONTROL	GROUP 2 100 MG/KG	GROUP 3 300 MG/KG	GROUP 4 1000 MG/KG
AT WEEK 13 HEARING	MEDIAN	0	0	0	0
SCORE 0/1	N	5	5	5	5
PUPIL L	MEDIAN	0	0	0	0
SCORE 0/1	N	5	5	5	5
PUPIL R	MEDIAN	0	0	0	0
SCORE 0/1	N	5	5	5	5
STATIC R	MEDIAN	0	0	0	0
SCORE 0/1	N	5	5	5	5
GRIP FORE	MEAN	1836	1753	1587	1620
GRAM	ST.DEV	144	126	272	180
	N	5	5	5	5
GRIP HIND	MEAN	1236	1025	1083	1061
GRAM	ST.DEV	215	104	193	131
	N	5	5	5	5
Total movements	MEAN	3194	2383	2374	2863
	ST.DEV	919	521	634	400
	N	5	5	5	5
Ambulations	MEAN	517	389	371	306
	ST.DEV	155	123	123	108
	N	5	5	5	5
END OF RECOVERY HEARING	MEDIAN	0			0
SCORE 0/1	N	5			5
PUPIL L	MEDIAN	0			0
SCORE 0/1	N	5			5
PUPIL R	MEDIAN	0			0
SCORE 0/1	N	5			5
STATIC R	MEDIAN	0			0
SCORE 0/1	N	5			5
GRIP FORE	MEAN	1385			1318
GRAM	ST.DEV	101			108
	N	5			5
GRIP HIND	MEAN	621			610
GRAM	ST.DEV	90			46
	N	5			5

Table 22: Functional observations (females)

		GROUP 1 CONTROL	GROUP 2 100 MG/KG	GROUP 3 300 MG/KG	GROUP 4 1000 MG/KG
AT WEEK 13 HEARING	MEDIAN	0	0	0	0
SCORE 0/1	N	5	5	5	5
PUPIL L	MEDIAN	0	0	0	0
SCORE 0/1	N	5	5	5	5
PUPIL R	MEDIAN	0	0	0	0
SCORE 0/1	N	5	5	5	5
STATIC R	MEDIAN	0	0	0	0
SCORE 0/1	N	5	5	5	5
GRIP FORE	MEAN	1318	1379	1385	1309
GRAM	ST.DEV	100	122	260	298
	N	5	5	5	5
GRIP HIND	MEAN	753	757	758	731
GRAM	ST.DEV	112	124	108	55
	N	5	5	5	5
Total movements	MEAN	4615	4448	3870	2739*
	ST.DEV	1238	1689	954	754
	N	5	5	5	5
Ambulations	MEAN	1095	968	1018	66
	ST.DEV	250	382	298	298
	N	5	5	5	5
END OF RECOVERY HEARING	MEDIAN	0			0
SCORE 0/1	N	5			5
PUPIL L	MEDIAN	0			0
SCORE 0/1	N	5			5
PUPIL R	MEDIAN	0			0
SCORE 0/1	N	5			5
STATIC R	MEDIAN	0			0
SCORE 0/1	N	5			5
GRIP FORE	MEAN	1141			1118
GRAM	ST.DEV	51			143
	N	5			5
GRIP HIND	MEAN	498			420
GRAM	ST.DEV	68			48
	N	5			5

* indicates a p-value <0.05

Table 23: organ weights (males) - end of treatment

Sex: Male Day(s) Relative to Start Date		0 mg/kg/day Group 1	100 mg/kg/day Group 2	300 mg/kg/day Group 3	1000 mg/kg/day Group 4
Terminal Body Weight (g) - [G]	Mean	390.2	397.5	392.7	379.9
	SD	34.3	31.1	35.0	38.2
	N	10	10	10	10
	%Diff	-	1.9	0.6	-2.6
Brain Weight (g) - [G]	Mean	2.0935	2.1146	2.0818	2.0894
	SD	0.0440	0.0863	0.0783	0.0921
	N	10	10	10	10
	%Diff	-	1.0079	-0.5589	-0.1958
Brain (%bw) - [G]	Mean	0.54047	0.53454	0.53342	0.55451
	SD	0.05097	0.04181	0.04581	0.05655
	N	10	10	10	10
	%Diff	-	-1.09685	-1.30435	2.59825
Epididymis Weight (g) - [G]	Mean	1.1058	1.2115	1.1425	1.1034
	SD	0.2424	0.1529	0.0569	0.0814
	N	10	10	10	10
	%Diff	-	9.5587	3.3189	-0.2170
Epididymis (%bw) - [G]	Mean	0.28625	0.30616	0.29311	0.29259
	SD	0.06911	0.04314	0.03155	0.03184
	N	10	10	10	10
	%Diff	-	6.95598	2.39837	2.21606
Gland, Adrenal Weight (g) - [G]	Mean	0.05769	0.05761	0.05602	0.04911
	SD	0.00851	0.00549	0.00635	0.01015
	N	10	10	10	10
	%Diff	-	-0.13867	-2.89478	-14.87259
Gland, Adrenal (%bw) - [G]	Mean	0.01484	0.01454	0.01438	0.01305
	SD	0.00216	0.00149	0.00222	0.00291
	N	10	10	10	10
	%Diff	-	-2.04209	-3.15592	-12.06055
Gland, Pituitary Weight (g) [G]	Mean	0.00824	0.00858	0.00785	0.00821
	SD	0.00105	0.00140	0.00140	0.00071
	N	10	10	10	10
	%Diff	-	4.12621	-4.73301	-0.36408
Gland, Pituitary (%bw) [G1]	Mean	0.00211	0.00215	0.00200	0.00217
	SD	0.00021	0.00027	0.00035	0.00014
	N	10	10	10	10
	%Diff	-	1.94015	-5.12563	2.58658
Gland, Prostate Weight (g) [G]	Mean	0.9458	0.9291	0.8763	0.8380
	SD	0.1549	0.1875	0.1589	0.1562
	N	10	10	10	10
	%Diff	-	-1.7657	-7.3483	-11.3978
Gland, Prostate (%bw) [G]	Mean	0.24348	0.23503	0.22478	0.22299
	SD	0.04065	0.05206	0.04463	0.04970
	N	10	10	10	10
	%Diff	-	-3.47163	-7.68092	-8.41815
Seminal Vesicle Weight (g) - [G]	Mean	1.2683	1.3261	1.2883	1.2644
	SD	0.1735	0.2229	0.1849	0.2585
	N	10	10	10	10
	%Diff	-	4.5573	1.5769	-0.3075
Seminal Vesicle (%bw) - [G]	Mean	0.32660	0.33502	0.32943	0.33592
	SD	0.04811	0.05917	0.04807	0.07580
	N	10	10	10	10
	%Diff	-	2.57825	0.86792	2.85589
Thyroid/Parathyroid Weight (g) - [G1]	Mean	0.01561	0.01824	0.01915	0.01891
	SD	0.00394	0.00529	0.00305	0.00751
	N	10	10	10	10
	%Diff	-	16.84817	22.67777	21.14029
Thyroid/Parathyroid (%bw) - [G]	Mean	0.00399	0.00460	0.00492	0.00491
	SD	0.00094	0.00133	0.00090	0.00171
	N	10	10	10	10
	%Diff	-	15.30492	23.42286	23.16273
Heart Weight (g) - [G]	Mean	1.0040	1.0086	1.0097	0.9653
	SD	0.0791	0.1207	0.0761	0.0657
	N	10	10	10	10
	%Diff	-	0.4582	0.5677	-3.8546
Heart (%bw) - [G]	Mean	0.25866	0.25349	0.25822	0.25538
	SD	0.02636	0.01924	0.02185	0.01998
	N	10	10	10	10
	%Diff	-	-1.99783	-0.17065	-1.26674
Kidney Weight (g) - [G]	Mean	2.3623	2.4359	2.4542	2.2696
	SD	0.2188	0.3319	0.1431	0.2500
	N	10	10	10	10
	%Diff	-	3.1156	3.8903	-3.9241
Kidney (%bw) - [G]	Mean	0.60731	0.61216	0.62858	0.59839
	SD	0.05371	0.05901	0.05800	0.04842
	N	10	10	10	10
	%Diff	-	0.79814	3.50222	-1.46983
Liver Weight (g) - [G]	Mean	8.7411	8.7143	8.7788	8.9585
	SD	0.9853	0.9765	1.0136	0.8868
	N	10	10	10	10
	%Diff	-	-0.3066	0.4313	2.4871
Liver (%bw) - [G]	Mean	2.23748	2.18929	2.23487	2.36098
	SD	0.10434	0.11829	0.15663	0.11509
	N	10	10	10	10
	%Diff	-	-2.15404	-0.11678	5.51955
Spleen Weight (g) - [G1]	Mean	0.5104	0.5840	0.5480	0.5271
	SD	0.0500	0.1114	0.0702	0.0474
	N	10	10	10	10
	%Diff	-	14.4201	7.3668	3.2719
Spleen (%bw) - [G]	Mean	0.13119	0.14625	0.14010	0.13916
	SD	0.01222	0.02020	0.01873	0.00983
	N	10	10	10	10
	%Diff	-	11.47773	6.78977	6.07030
Testis Weight (g) - [G]	Mean	3.2215	3.7755	3.6595	3.5242
	SD	0.9363	0.3880	0.3237	0.1842
	N	10	10	10	10
	%Diff	-	17.1970	13.5962	9.3962
Testis (%bw) - [G]	Mean	0.83486	0.95425	0.93987	0.93357
	SD	0.25480	0.11391	0.12907	0.07621
	N	10	10	10	10
	%Diff	-	14.30168	12.57932	11.82419
Thymus Weight (g) - [G]	Mean	0.2759	0.2790	0.3239	0.3061
	SD	0.0834	0.0786	0.0605	0.0959
	N	10	10	10	10
	%Diff	-	1.1236	17.3976	10.9460
Thymus (%bw) - [G]	Mean	0.06975	0.06991	0.08280	0.07997
	SD	0.01722	0.01779	0.01446	0.02148
	N	10	10	10	10
	%Diff	-	0.23340	18.70625	14.65698

[G] - Anova & Dunnett
[G1] - Kruskal-Wallis & Dunn: * = p ≤ 0.05

Table 24: organ weights (females) - end of treatment

Sex: Female Day(s) Relative to Start Date		0 mg/kg/day Group 1	100 mg/kg/day Group 2	300 mg/kg/day Group 3	1000 mg/kg/day Group 4
Terminal Body Weight (g) - [G]	Mean	220.6	226.2	213.3	231.0
	SD	13.7	15.3	15.3	13.0
	N	10	10	10	10
	%Diff	-	2.5	-3.3	4.7
Brain Weight (g) - [G1]	Mean	1.8958	1.9140	1.8589	1.8968
	SD	0.0612	0.0492	0.0271	0.0563
	N	10	10	10	10
	%Diff	-	0.9600	-1.9464	0.0527
Brain (%bw) - [G]	Mean	0.86149	0.84923	0.87619	0.82264
	SD	0.04436	0.05478	0.07360	0.03431
	N	10	10	10	10
	%Diff	-	-1.42301	1.70715	-4.50976
Gland, Adrenal Weight (g) - [G]	Mean	0.05907	0.06330	0.06361	0.06358
	SD	0.00971	0.01033	0.00735	0.00694
	N	10	10	10	10
	%Diff	-	7.16100	7.68580	7.63501
Gland, Adrenal (%bw) - [G]	Mean	0.02672	0.02794	0.02982	0.02754
	SD	0.00369	0.00378	0.00273	0.00268
	N	10	10	10	10
	%Diff	-	4.59109	11.62243	3.06385
Gland, Pituitary Weight (g) [G1]	Mean	0.01478	0.01014	0.01116	0.01126
	SD	0.00532	0.00255	0.00296	0.00167
	N	10	10	10	10
	%Diff	-	-31.39378	-24.49256	-23.81597
Gland, Pituitary (%bw) [G1]	Mean	0.00666	0.00448 *	0.00523	0.00488
	SD	0.00223	0.00103	0.00130	0.00073
	N	10	10	10	10
	%Diff	-	-32.71386	-21.48404	-26.68346
Thyroid/Parathyroid Weight (g) - [G]	Mean	0.01239	0.01440	0.01277	0.01443
	SD	0.00220	0.00284	0.00367	0.00354
	N	10	10	10	10
	%Diff	-	16.22276	3.06699	16.46489
Thyroid/Parathyroid (%bw) - [G]	Mean	0.00564	0.00638	0.00600	0.00630
	SD	0.00111	0.00132	0.00174	0.00182
	N	10	10	10	10
	%Diff	-	13.19007	6.42890	11.71956
Heart Weight (g) - [G]	Mean	0.6792	0.6821	0.6207	0.7181
	SD	0.0507	0.0757	0.0536	0.0448
	N	10	10	10	10
	%Diff	-	0.4270	-8.6131	5.7273
Heart (%bw) - [G]	Mean	0.30853	0.30129	0.29126	0.31131
	SD	0.02475	0.02313	0.01832	0.01892
	N	10	10	10	10
	%Diff	-	-2.34775	-5.59609	0.90247
Kidney Weight (g) - [G]	Mean	1.4672	1.5831	1.4736	1.7272 **
	SD	0.0838	0.1193	0.1311	0.1965
	N	10	10	10	10
	%Diff	-	7.8994	0.4362	17.7208
Kidney (%bw) - [G1]	Mean	0.66594	0.70065	0.69076	0.74781 **
	SD	0.03215	0.04251	0.03323	0.07497
	N	10	10	10	10
	%Diff	-	5.21116	3.72597	12.29292
Liver Weight (g) - [G]	Mean	5.3753	5.4101	5.0887	6.4360 **
	SD	0.3872	0.5482	0.4642	0.5899
	N	10	10	10	10
	%Diff	-	0.6474	-5.3318	19.7329
Liver (%bw) - [G]	Mean	2.43901	2.39304	2.38706	2.79153 **
	SD	0.14303	0.20287	0.15493	0.27667
	N	10	10	10	10
	%Diff	-	-1.88477	-2.12999	14.45347
Ovary Paired (g) - [G]	Mean	0.1361	0.1468	0.1421	0.1377
	SD	0.0210	0.0210	0.0243	0.0172
	N	10	10	10	10
	%Diff	-	7.8619	4.4085	1.1756
Ovary Paired (%bw) - [G]	Mean	0.06164	0.06522	0.06703	0.05966
	SD	0.00831	0.01101	0.01308	0.00699
	N	10	10	10	10
	%Diff	-	5.80168	8.74297	-3.22236
Spleen Weight (g) - [G1]	Mean	0.4167	0.4165	0.3773	0.4617
	SD	0.0763	0.0388	0.0371	0.0357
	N	10	10	10	10
	%Diff	-	-0.0480	-9.4552	10.7991
Spleen (%bw) - [G]	Mean	0.18857	0.18411	0.17756	0.20006
	SD	0.03104	0.01172	0.02108	0.01443
	N	10	10	10	10
	%Diff	-	-2.36212	-5.83761	6.09656
Thymus Weight (g) - [G1]	Mean	0.2929	0.2894	0.2506	0.2886
	SD	0.0820	0.0706	0.0479	0.0574
	N	10	10	10	10
	%Diff	-	-1.1949	-14.4418	-1.4681
Thymus (%bw) - [G1]	Mean	0.13204	0.12701	0.11827	0.12521
	SD	0.03288	0.02467	0.02659	0.02594
	N	10	10	10	10
	%Diff	-	-3.81140	-10.42849	-5.17288
Uterus/Cervix (g) - [G]	Mean	0.6704	0.6153	0.4746	0.5312
	SD	0.2816	0.2422	0.0977	0.1136
	N	10	10	10	10
	%Diff	-	-8.2190	-29.2064	-20.7637
Uterus/Cervix (%bw) - [G]	Mean	0.30787	0.27105	0.22254	0.23051
	SD	0.13656	0.09880	0.04277	0.05042
	N	10	10	10	10
	%Diff	-	-11.96024	-27.71662	-25.12699

[G] - Anova & Dunnett: ** = p ≤ 0.01
[G] - Anova & Dunnett

Table 25: organ weights (males) - end of recovery

Sex: Male Day(s) Relative to Start Date		0 mg/kg/day Group 1	1000 mg/kg/day Group 4
Terminal Body Weight (g) - [G]	Mean	416.2	417.8
	SD	30.4	33.6
	N	5	5
	%Diff	-	0.4
Brain Weight (g) - [G]	Mean	2.1002	2.0828
	SD	0.1227	0.1062
	N	5	5
	%Diff	-	-0.8285
Brain (%bw) - [G]	Mean	0.50524	0.50009
	SD	0.01655	0.03204
	N	5	5
	%Diff	-	-1.01955
Epididymis Weight (g) - [G]	Mean	1.2254	1.2330
	SD	0.1105	0.1300
	N	5	5
	%Diff	-	0.6202
Epididymis (%bw) - [G]	Mean	0.29438	0.29543
	SD	0.01556	0.02716
	N	5	5
	%Diff	-	0.35575
Gland, Adrenal Weight (g) - [G]	Mean	0.05264	0.05538
	SD	0.00576	0.00582
	N	5	5
	%Diff	-	5.20517
Gland, Adrenal (%bw) - [G]	Mean	0.01266	0.01333
	SD	0.00117	0.00178
	N	5	5
	%Diff	-	5.26824
Gland, Pituitary Weight (g) [G1]	Mean	0.00922	0.01498
	SD	0.00154	0.00829
	N	5	5
	%Diff	-	62.47289
Gland, Pituitary (%bw) [G1]	Mean	0.00221	0.00353
	SD	0.00027	0.00185
	N	5	5
	%Diff	-	59.90894
Gland, Prostate Weight (g) [G]	Mean	1.0258	0.8896
	SD	0.1418	0.1802
	N	5	5
	%Diff	-	-13.2774
Gland, Prostate (%bw) [G]	Mean	0.24663	0.21396
	SD	0.03056	0.04640
	N	5	5
	%Diff	-	-13.24820
Seminal Vesicle Weight (g) - [G]	Mean	1.3982	1.5228
	SD	0.2755	0.3414
	N	5	5
	%Diff	-	8.9115
Seminal Vesicle (%bw) - [G]	Mean	0.33727	0.36373
	SD	0.06901	0.06780
	N	5	5
	%Diff	-	7.84566
Thyroid/Parathyroid Weight (g) - [G1]	Mean	0.01610	0.01556
	SD	0.00533	0.00319
	N	5	5
	%Diff	-	-3.35404
Thyroid/Parathyroid (%bw) - [G1]	Mean	0.00388	0.00370
	SD	0.00129	0.00054
	N	5	5
	%Diff	-	-4.55710
Heart Weight (g) - [G]	Mean	1.0272	1.0704
	SD	0.0534	0.0746
	N	5	5
	%Diff	-	4.2056
Heart (%bw) - [G]	Mean	0.24756	0.25665
	SD	0.01706	0.01272
	N	5	5
	%Diff	-	3.66822
Kidney Weight (g) - [G]	Mean	2.3472	2.5980
	SD	0.2748	0.3611
	N	5	5
	%Diff	-	10.6851
Kidney (%bw) - [G]	Mean	0.56367	0.62134
	SD	0.04690	0.06600
	N	5	5
	%Diff	-	10.23148
Liver Weight (g) - [G]	Mean	8.8570	9.6144
	SD	1.0204	1.5403
	N	5	5
	%Diff	-	8.5514
Liver (%bw) - [G]	Mean	2.12282	2.29077
	SD	0.09992	0.21010
	N	5	5
	%Diff	-	7.91134
Spleen Weight (g) - [G]	Mean	0.6252	0.6066
	SD	0.0440	0.1345
	N	5	5
	%Diff	-	-2.9750
Spleen (%bw) - [G1]	Mean	0.15038	0.14470
	SD	0.00762	0.02485
	N	5	5
	%Diff	-	-3.77737
Testis Weight (g) - [G]	Mean	3.6980	3.7970
	SD	0.1267	0.1574
	N	5	5
	%Diff	-	2.6771
Testis (%bw) - [G]	Mean	0.89239	0.91195
	SD	0.07222	0.05657
	N	5	5
	%Diff	-	2.19240
Thymus Weight (g) - [G]	Mean	0.2932	0.2612
	SD	0.0699	0.0988
	N	5	5
	%Diff	-	-10.9141
Thymus (%bw) - [G]	Mean	0.07044	0.06192
	SD	0.01514	0.02120
	N	5	5
	%Diff	-	-12.09428

[G] - Anova & Dunnett
[G1] - Kruskal-Wallis & Dunn

Table 26: organ weights (females) - end of recovery

Sex: Female Day(s) Relative to Start Date		0 mg/kg/day Group 1	1000 mg/kg/day Group 4
Terminal Body Weight (g) - [G]	Mean	239.8	227.6
	SD	9.6	15.5
	N	5	5
	%Diff	-	-5.1
Brain Weight (g) - [G1]	Mean	1.9122	1.9748
	SD	0.0511	0.1385
	N	5	5
	%Diff	-	3.2737
Brain (%bw) - [G]	Mean	0.79809	0.86811 **
	SD	0.02740	0.03459
	N	5	5
	%Diff	-	8.77419
Gland, Adrenal Weight (g) - [G]	Mean	0.06100	0.07034
	SD	0.00904	0.01447
	N	5	5
	%Diff	-	15.31148
Gland, Adrenal (%bw) - [G]	Mean	0.02552	0.03068
	SD	0.00446	0.00444
	N	5	5
	%Diff	-	20.21525
Gland, Pituitary Weight (g) [G1]	Mean	0.01114	0.01256
	SD	0.00414	0.00060
	N	5	5
	%Diff	-	12.74686
Gland, Pituitary (%bw) [G]	Mean	0.00460	0.00554
	SD	0.00161	0.00042
	N	5	5
	%Diff	-	20.31781
Thyroid/Parathyroid Weight (g) - [G]	Mean	0.01410	0.01654
	SD	0.00103	0.00277
	N	5	5
	%Diff	-	17.30496
Thyroid/Parathyroid (%bw) - [G]	Mean	0.00589	0.00732
	SD	0.00057	0.00157
	N	5	5
	%Diff	-	24.25956
Heart Weight (g) - [G]	Mean	0.7156	0.6652
	SD	0.0564	0.0945
	N	5	5
	%Diff	-	-7.0430
Heart (%bw) - [G]	Mean	0.29813	0.29128
	SD	0.01336	0.02578
	N	5	5
	%Diff	-	-2.29654
Kidney Weight (g) - [G]	Mean	1.5562	1.5080
	SD	0.1430	0.1578
	N	5	5
	%Diff	-	-3.0973
Kidney (%bw) - [G]	Mean	0.64965	0.66137
	SD	0.06221	0.02983
	N	5	5
	%Diff	-	1.80335
Liver Weight (g) - [G]	Mean	5.8176	5.3962
	SD	0.5446	0.6612
	N	5	5
	%Diff	-	-7.2435
Liver (%bw) - [G]	Mean	2.42291	2.36682
	SD	0.14767	0.20047
	N	5	5
	%Diff	-	-2.31522
Ovary Paired (g) - [G]	Mean	0.1388	0.1400
	SD	0.0169	0.0182
	N	5	5
	%Diff	-	0.8646
Ovary Paired (%bw) - [G]	Mean	0.05802	0.06134
	SD	0.00809	0.00477
	N	5	5
	%Diff	-	5.72474
Spleen Weight (g) - [G1]	Mean	0.4208	0.3914
	SD	0.0223	0.0478
	N	5	5
	%Diff	-	-6.9867
Spleen (%bw) - [G]	Mean	0.17580	0.17162
	SD	0.01300	0.01342
	N	5	5
	%Diff	-	-2.37705
Thymus Weight (g) - [G]	Mean	0.2656	0.2180
	SD	0.0429	0.0323
	N	5	5
	%Diff	-	-17.9217
Thymus (%bw) - [G]	Mean	0.11060	0.09586
	SD	0.01639	0.01350
	N	5	5
	%Diff	-	-13.32804
Uterus/Cervix (g) - [G]	Mean	0.6696	0.5598
	SD	0.2254	0.1107
	N	5	5
	%Diff	-	-16.3978
Uterus/Cervix (%bw) - [G1]	Mean	0.27920	0.24616
	SD	0.09299	0.04749
	N	5	5
	%Diff	-	-11.83234

[G] - Anova & Dunnett: ** = p ≤ 0.01
[G1] - Kruskal-Wallis & Dunn

Table 27: microscopic pathology (males & females) - end of treatment

Removal Reason(s): TERMINAL EUTHANASIA Summary: Incidence Number of Animals:	0 mg/kg/ day Group 1	Male 100 300 mg/kg/ mg/kg/ day day Group Group 2 3		1000 mg/kg/ day Group 4	0 mg/kg/ day Group 1	Female 100 300 mg/kg/ mg/kg/ day day Group Group 2 3		1000 mg/kg/ day Group 4
	10	10	10	10	10	10	10	10
ARTERY, AORTA
Examined	10	0	0	10	10	0	0	10
No Visible Lesions	10	.	.	10	10	.	.	10
BONE MARROW, STERNUM
Examined	10	0	0	10	10	0	0	10
No Visible Lesions	10	.	.	10	10	.	.	10
BONE, STERNUM
Examined	10	0	0	10	10	0	0	10
No Visible Lesions	10	.	.	10	10	.	.	10
BRAIN
Examined	10	0	0	10	10	0	0	10
No Visible Lesions	10	.	.	10	10	.	.	10
CERVIX
Examined	.	.	.	.	10	0	0	10
No Visible Lesions	.	.	.	.	10	.	.	10
EPIDIDYMIS
Examined	10	0	0	10	.	.	.	.
No Visible Lesions	9	.	.	9	.	.	.	.
Sperm, decreased; lumen	1	.	.	0	.	.	.	.
.... marked	1	.	.	0	.	.	.	.
Infiltration, mononuclear cell;	0	.	.	1	.	.	.	.
interstitial
.... minimal	0	.	.	1	.	.	.	.
ESOPHAGUS
Examined	10	0	0	10	10	0	0	10
No Visible Lesions	10	.	.	10	10	.	.	10
EYE
Examined	10	0	0	10	10	0	0	10
No Visible Lesions	10	.	.	9	10	.	.	9
Hemorrhage; acute	0	.	.	1	0	.	.	1
.... minimal	0	.	.	1	0	.	.	1
GALT
Examined	10	0	0	10	10	0	0	10
No Visible Lesions	10	.	.	10	10	.	.	10
GLAND, ADRENAL
Examined	10	0	0	10	10	0	0	10
No Visible Lesions	10	.	.	9	10	.	.	10
Vacuolation; zona fasciculata	0	.	.	1	0	.	.	0
.... minimal	0	.	.	1	0	.	.	0
GLAND, CLITORAL
Examined	.	.	.	.	0	1	1	0
Dilatation; ductular	.	.	.	.	.	1	1	.
.... minimal	.	.	.	.	.	1	1	.
GLAND, COAGULATING
Examined	10	0	0	10	.	.	.	.
No Visible Lesions	10	.	.	10	.	.	.	.
GLAND, MAMMARY
Examined	10	0	0	10	10	0	0	10
No Visible Lesions	10	.	.	10	10	.	.	10
GLAND, PARATHYROID
Examined	10	0	0	10	10	0	0	10
No Visible Lesions	10	.	.	10	10	.	.	10
GLAND, PITUITARY
Examined	10	0	0	10	10	0	0	10
No Visible Lesions	10	.	.	10	10	.	.	10
GLAND, PREPUTIAL
Examined	0	0	1	0	.	.	.	.
Dilatation; acinar	.	.	1	.	.	.	.	.
.... minimal	.	.	1	.	.	.	.	.
GLAND, PROSTATE
Examined	10	0	0	10	.	.	.	.
No Visible Lesions	8	.	.	10	.	.	.	.
Infiltration, mononuclear cell;	2	.	.	0	.	.	.	.
interstitial
.... minimal	2	.	.	0	.	.	.	.
GLAND, SALIVARY,
MANDIBULAR
Examined	10	0	0	10	10	0	0	10
No Visible Lesions	10	.	.	10	10	.	.	10
GLAND, SALIVARY,
SUBLINGUAL
Examined	10	0	0	10	10	0	0	10
No Visible Lesions	10	.	.	10	10	.	.	10
GLAND, SEMINAL VESICLE
Examined	10	0	0	10	.	.	.	.
No Visible Lesions	10	.	.	10	.	.	.	.
GLAND, THYROID
Examined	10	0	0	10	10	0	0	10
No Visible Lesions	8	.	.	7	7	.	.	7
Cyst; ultimobranchial	0	.	.	0	2	.	.	2
.... minimal	0	.	.	0	2	.	.	2
Hypertrophy; follicular cell	1	.	.	2	0	.	.	0
.... minimal	1	.	.	2	0	.	.	0
Ectopia; thymic	1	.	.	1	1	.	.	1
.... minimal	1	.	.	1	1	.	.	1
HEART
Examined	10	0	0	10	10	0	0	10
No Visible Lesions	10	.	.	9	10	.	.	10
Infiltration, mononuclear cell;	0	.	.	1	0	.	.	0
myocardial
.... minimal	0	.	.	1	0	.	.	0
KIDNEY
Examined	10	0	3	10	10	0	0	10
No Visible Lesions	1	.	2	5	8	.	.	7
Basophilia; tubular	6	.	0	2	1	.	.	1
.... minimal	6	.	0	2	1	.	.	1
Accumulation; hyaline droplets	7	.	0	2	0	.	.	0
.... minimal	5	.	0	2	0	.	.	0
.... mild	2	.	0	0	0	.	.	0
Infiltration, mononuclear cell;	3	.	0	1	1	.	.	3
interstitial
.... minimal	3	.	0	1	1	.	.	3
Dilatation; cortical, tubular	0	.	0	1	0	.	.	0
.... minimal	0	.	0	1	0	.	.	0
Dilatation; pelvis	0	.	1	0	0	.	.	0
.... minimal	0	.	1	0	0	.	.	0
LARGE INTESTINE, CECUM
Examined	10	0	0	10	10	0	0	10
No Visible Lesions	10	.	.	10	10	.	.	10
LARGE INTESTINE, COLON
Examined	10	0	0	10	10	0	0	10
No Visible Lesions	10	.	.	10	10	.	.	10
LARGE INTESTINE, RECTUM
Examined	10	10	10	10	10	0	0	10
No Visible Lesions	10	10	10	7	10	.	.	10
Hyperplasia; goblet cell	0	0	0	3	0	.	.	0
.... minimal	0	0	0	3	0	.	.	0
LIVER
Examined	10	0	0	10	10	0	0	10
No Visible Lesions	5	.	.	5	8	.	.	10
Infiltration, mononuclear cell	5	.	.	5	2	.	.	0
.... minimal	5	.	.	5	2	.	.	0
Atrophy	2	.	.	0	0	.	.	0
.... minimal	1	.	.	0	0	.	.	0
.... mild	1	.	.	0	0	.	.	0
LUNG
Examined	10	0	1	10	10	0	0	10
No Visible Lesions	9	.	0	7	9	.	.	8
Aggregate; alveolar, macrophage	0	.	1	1	0	.	.	0
.... minimal	0	.	0	1	0	.	.	0
.... moderate	0	.	1	0	0	.	.	0
Infiltration, mixed cell;	1	.	0	1	1	.	.	2
perivascular
.... minimal	1	.	0	1	1	.	.	2
Metaplasia; osseous	0	.	0	1	0	.	.	0
.... minimal	0	.	0	1	0	.	.	0
LYMPH NODE,
MANDIBULAR
Examined	10	2	0	10	10	1	0	10
No Visible Lesions	8	0	.	9	7	0	.	10
Hemorrhage; acute	2	2	.	1	3	1	.	0
.... minimal	2	2	.	1	2	1	.	0
.... mild	0	0	.	0	1	0	.	0
LYMPH NODE, MESENTERIC
Examined	10	0	0	10	10	0	0	10
No Visible Lesions	10	.	.	10	10	.	.	10
MUSCLE, SKELETAL
Examined	10	0	0	10	10	0	0	10
No Visible Lesions	5	.	.	9	9	.	.	9
Degeneration; myofiber	5	.	.	0	1	.	.	1
.... minimal	5	.	.	0	1	.	.	1
Infiltration, mononuclear cell	1	.	.	1	0	.	.	0
.... minimal	1	.	.	1	0	.	.	0
NERVE, OPTIC
Examined	10	0	0	10	10	0	0	10
No Visible Lesions	10	.	.	10	10	.	.	10
NERVE, SCIATIC
Examined	10	0	0	10	10	0	0	10
No Visible Lesions	10	.	.	10	10	.	.	10
OVARY
Examined	.	.	.	.	10	0	1	10
No Visible Lesions	.	.	.	.	10	.	1	10
PANCREAS
Examined	10	0	0	10	10	0	0	10
No Visible Lesions	10	.	.	10	10	.	.	9
Atrophy; acinar	0	.	.	0	0	.	.	1
.... minimal	0	.	.	0	0	.	.	1
SKIN
Examined	10	0	0	10	10	0	0	10
No Visible Lesions	10	.	.	10	10	.	.	10
SMALL INTESTINE,
DUODENUM
Examined	10	0	0	10	10	0	0	10
No Visible Lesions	10	.	.	10	10	.	.	10
SMALL INTESTINE, ILEUM
Examined	10	0	0	10	10	0	0	10
No Visible Lesions	10	.	.	10	10	.	.	10
SMALL INTESTINE,
JEJUNUM
Examined	10	0	0	10	10	0	0	10
No Visible Lesions	10	.	.	10	10	.	.	10
SPINAL CORD
Examined	10	0	0	10	10	0	0	10
No Visible Lesions	10	.	.	10	10	.	.	10
SPLEEN
Examined	10	1	0	10	10	0	0	10
No Visible Lesions	1	1	.	3	0	.	.	0
Pigment	9	0	.	6	10	.	.	10
.... minimal	8	0	.	6	9	.	.	9
.... mild	1	0	.	0	1	.	.	1
Extramedullary hematopoiesis	3	0	.	2	6	.	.	5
.... minimal	2	0	.	0	4	.	.	4
.... mild	1	0	.	2	2	.	.	1
STOMACH
Examined	10	0	1	10	10	10	10	10
No Visible Lesions	8	.	1	5	7	10	8	4
Hyperplasia; basal cell; glandular	0	.	0	1	0	0	0	1
.... minimal	0	.	0	1	0	0	0	1
Hyperplasia; squamous cell;	0	.	0	0	0	0	1	3
non-glandular
.... minimal	0	.	0	0	0	0	1	2
.... mild	0	.	0	0	0	0	0	1
Vacuolation; epithelial, limiting	0	.	0	0	2	0	0	2
ridge
.... minimal	0	.	0	0	2	0	0	2
Erosion; non-glandular	0	.	0	0	0	0	0	1
.... minimal	0	.	0	0	0	0	0	1
Infiltration, lymphocytic;	2	.	0	3	1	0	1	1
submucosal, glandular
.... minimal	2	.	0	3	1	0	1	1
Infiltration, lymphocytic; limiting	0	.	0	0	0	0	0	1
ridge
.... minimal	0	.	0	0	0	0	0	1
Infiltration, mixed cell;	1	.	0	1	0	0	0	1
submucosal, glandular
.... minimal	1	.	0	1	0	0	0	1
TESTIS
Examined	10	0	0	10	.	.	.	.
No Visible Lesions	6	.	.	8	.	.	.	.
Degeneration/atrophy; tubular	4	.	.	2	.	.	.	.
.... minimal	2	.	.	2	.	.	.	.
.... mild	1	.	.	0	.	.	.	.
.... marked	1	.	.	0	.	.	.	.
THYMUS
Examined	10	1	0	10	10	0	0	10
No Visible Lesions	10	0	.	10	10	.	.	10
Hemorrhage; acute	0	1	.	0	0	.	.	0
.... minimal	0	1	.	0	0	.	.	0
TRACHEA
Examined	10	0	0	10	10	0	0	10
No Visible Lesions	10	.	.	10	10	.	.	10
URINARY BLADDER
Examined	10	0	0	10	10	0	0	10
No Visible Lesions	10	.	.	10	10	.	.	10
UTERUS
Examined	.	.	.	.	10	2	0	10
No Visible Lesions	.	.	.	.	10	2	.	10
VAGINA
Examined	.	.	.	.	10	0	0	10
Diestrus	.	.	.	.	5	.	.	7
Estrus	.	.	.	.	2	.	.	3
Proestrus	.	.	.	.	3	.	.	0

Table 28: microscopic pathology (males & females) - end of recovery

Removal Reason(s): RECOVERY EUTHANASIA Summary: Incidence Number of Animals:	Male 0 1000 mg/kg/ mg/kg/ day day Group Group 1 4		Female 0 1000 mg/kg/ mg/kg/ day day Group Group 1 4
	5	5	5	5
KIDNEY
Examined	0	1	0	1
No Visible Lesions	.	0	.	1
Cyst; cortical	.	1	.	0
.... mild	.	1	.	0
LARGE INTESTINE, RECTUM
Examined	5	5	.	.
No Visible Lesions	5	5	.	.
LIVER
Examined	1	0	.	.
Atrophy	1	.	.	.
.... minimal	1	.	.	.
LUNG
Examined	.	.	1	0
Hemorrhage; acute, alveolar	.	.	1	.
.... minimal	.	.	1	.
STOMACH
Examined	0	1	5	5
No Visible Lesions	.	1	4	3
Vacuolation; epithelial, limiting ridge	.	0	1	1
.... minimal	.	0	1	1
Erosion; glandular	.	0	0	1
.... minimal	.	0	0	1
THYMUS
Examined	.	.	0	1
Hemorrhage; acute	.	.	.	1
.... minimal	.	.	.	1
UTERUS
Examined	.	.	2	1
No Visible Lesions	.	.	2	1

Table 29: clinical observations (males & females) - Main

Observation Type: All Types From Day 1 (Start Date) to 93 (Start Date)	0 mg/kg/day Group 1	Male 100 300 mg/kg/day mg/kg/day Group 2 Group 3		1000 mg/kg/day Group 4	0 mg/kg/day Group 1	Female 100 300 mg/kg/day mg/kg/day Group 2 Group 3		1000 mg/kg/day Group 4
Retching
Number of Animals Affected	0	1	0	6	0	1	3	9
Number of Times Recorded	0	1	0	8	0	1	10	13
% of Affected Animals	0	10	0	40	0	10	30	60
First to Last seen	-	44 - 44	-	43 - 48	-	44 - 44	44 - 49	43 - 48
Salivation
Number of Animals Affected	0	0	10	15	0	0	4	15
Number of Times Recorded	0	0	96	1223	0	0	25	1205
% of Affected Animals	0	0	100	100	0	0	40	100
First to Last seen	-	-	12 - 91	7 - 91	-	-	12 - 89	7 - 92
Breathing, Deep
Number of Animals Affected	0	0	0	0	0	0	1	0
Number of Times Recorded	0	0	0	0	0	0	2	0
% of Affected Animals	0	0	0	0	0	0	10	0
First to Last seen	-	-	-	-	-	-	91 - 92	-
Breathing, Labored
Number of Animals Affected	0	0	0	0	0	0	0	1
Number of Times Recorded	0	0	0	0	0	0	0	1
% of Affected Animals	0	0	0	0	0	0	0	7
First to Last seen	-	-	-	-	-	-	-	81 - 81
Breathing, Abnormal Sounds
Number of Animals Affected	0	3	3	10	1	1	5	6
Number of Times Recorded	0	7	10	36	11	3	15	20
% of Affected Animals	0	30	30	67	7	10	50	40
First to Last seen	-	9 - 91	8 - 91	8 - 91	61 - 90	74 - 76	42 - 92	3 - 91
Breathing, Shallow
Number of Animals Affected	0	2	0	1	0	0	0	1
Number of Times Recorded	0	6	0	1	0	0	0	1
% of Affected Animals	0	20	0	7	0	0	0	7
First to Last seen	-	89 - 91	-	89 - 89	-	-	-	89 - 89
Hunched Posture
Number of Animals Affected	0	0	1	1	0	0	0	1
Number of Times Recorded	0	0	13	2	0	0	0	1
% of Affected Animals	0	0	10	7	0	0	0	7
First to Last seen	-	-	23 - 35	28 - 29	-	-	-	81 - 81
Fur, Erected
Number of Animals Affected	0	0	2	4	0	0	1	1
Number of Times Recorded	0	0	7	9	0	0	2	1
% of Affected Animals	0	0	20	27	0	0	10	7
First to Last seen	-	-	23 - 35	42 - 67	-	-	91 - 92	81 - 81
Fur, Loss
Number of Animals Affected	0	0	1	0	0	0	0	0
Number of Times Recorded	0	0	75	0	0	0	0	0
% of Affected Animals	0	0	10	0	0	0	0	0
First to Last seen	-	-	26 - 92	-	-	-	-	-
Skin, Lesion, Dorsal Cervical
Number of Animals Affected	0	0	1	0	0	0	0	0
Number of Times Recorded	0	0	9	0	0	0	0	0
% of Affected Animals	0	0	10	0	0	0	0	0
First to Last seen	-	-	12 - 29	-	-	-	-	-
Skin, Scab, Dorsal Cervical
Number of Animals Affected	0	0	1	0	0	0	0	0
Number of Times Recorded	0	0	8	0	0	0	0	0
Skin, Scab, Dorsal Cervical (Continued...)
% of Affected Animals	0	0	10	0	0	0	0	0
First to Last seen	-	-	18 - 25	-	-	-	-	-
Ploughing
Number of Animals Affected	0	0	1	10	0	0	0	12
Number of Times Recorded	0	0	2	33	0	0	0	36
% of Affected Animals	0	0	10	67	0	0	0	80
First to Last seen	-	-	43 - 44	43 - 47	-	-	-	43 - 47

Table 30: clinical observations (males & females) - Recovery

Observation Type: All Types From Day 92 (Start Date) to 121 (Start Date)	Male 0 1000 mg/kg/day mg/kg/day Group 1 Group 4		Female 0 1000 mg/kg/day mg/kg/day Group 1 Group 4
Breathing, Labored
Number of Animals Affected	0	0	0	1
Number of Times Recorded	0	0	0	1
% of Affected Animals	0	0	0	20
First to Last seen	-	-	-	106 - 106
Breathing, Abnormal Sounds
Number of Animals Affected	0	0	0	1
Number of Times Recorded	0	0	0	4
% of Affected Animals	0	0	0	20
First to Last seen	-	-	-	106 - 121
Fur, Erected
Number of Animals Affected	0	0	1	0
Number of Times Recorded	0	0	1	0
% of Affected Animals	0	0	20	0
First to Last seen	-	-	121 - 121	-

Conclusions:

A 90-day repeated dose toxicity study was performed according to OECD TG 408 and in accordance with GLP principles. In conclusion, administration of the substance by once daily oral gavage was tolerated in Wistar Han rats at levels up to 1000 mg/kg bw/day. There were no adverse test material related alterations observed.
Based on these results, the No Observed Adverse Effect Level (NOAEL) was considered to be at least 1000 mg/kg bw/day in males and females. This result is read across to the registered substance, the rationale is attached in Section 13.

Executive summary:

A 90-day repeated dose toxicity study was performed according to OECD TG 408 and in accordance with GLP principles.

The objectives of this study were to determine the potential toxicity of the substance, when given orally by gavage for 90 days to Wistar Han rats and to evaluate the potential reversibility of any findings. In addition, a No Observed Adverse Effect Level (NOAEL) was evaluated. Ten animals per sex per dose received 0 (vehicle), 100, 300 or 1000 mg/kg bw/day. An additional 5 animals per sex per dose in the recovery group received 0 or 1000 mg/kg bw/day.

Chemical analyses of formulations were conducted on Weeks 1, 6 and 12 on Groups 1-3 to assess accuracy and homogeneity. Formulation analyses confirmed that formulations were prepared accurately and homogenously for all occasions. The following parameters and end points were evaluated in this study: mortality, clinical signs, functional observation tests, body weights, food consumption, ophthalmology, stage of estrous, clinical pathology parameters (hematology, coagulation, and clinical chemistry), macroscopic examination, organ weights and microscopic examination.

No mortality occurred during the study.
At 100 mg/kg bw/day, abnormal breathing sounds, shallow breathing and retching were noted in males and/or females, which were considered to be non-adverse.
At 300 mg/kg bw/day, non-adverse abnormal breathing sounds, deep breathing, retching, hunched posture, erected fur were noted in males and/or females. Histopathological changes consisted of non-adverse squamous cell hyperplasia in the stomach in females, which was often accompanied by hyperkeratosis.
At 1000 mg/kg bw/day, abnormal breathing sounds, labored breathing, shallow breathing, retching, hunched posture and erected fur were noted in males and/or females and were considered to be non-adverse. Non-adverse decreased total movements and ambulations were noted for females. Hematological findings comprised non-adverse decreased red blood cell count and red blood cell distribution width and increased mean corpuscular volume and mean corpuscular hemoglobin in males. Red blood cell count was recovered, while red blood cell distribution width, mean corpuscular volume and mean corpuscular hemoglobin were not recovered at the end of the Recovery Period. It is noted that all reported changes on Hematology parameters were within historical control for this rat strain at the site. Nonadverse clinical chemistry findings consisted of an increase in triglyceride concentration in males, which partly recovered at the end of the Recovery Period. Hormone analysis showed an increased T4 levels in females compared to the concurrent control group however, the mean values still remained within the historical control range and levels recovered at the end of the Recovery Period. Furthermore, no histopathological changes were observed in the thyroid. Overall, the observed increase in the study was considered non-adverse and not toxicologically relevant. Histopathological changes consisted of non-adverse squamous cell hyperplasia often accompanied by hyperkeratosis in the stomach in females and goblet cell hyperplasia of the rectum in a few males. In addition, a higher kidney and liver weight was noted in females. As kidney and liver weights remained within the historical control data and changes were no longer observed after recovery and in the absence of concomitant histopathological changes, the effects on organ weights were considered non-adverse. All histopathological and organ weight changes were fully recovered at the end of the Recovery Period.

No test material-related changes were noted in any of the remaining parameters investigated in this study (i.e., mortality, body weight, food consumption, ophthalmoscopy, coagulation and macroscopic pathology).
In conclusion, administration of the substance by once daily oral gavage was tolerated in Wistar Han rats at levels up to 1000 mg/kg bw/day. There were no adverse test material related alterations observed.
Based on these results, the No Observed Adverse Effect Level (NOAEL) was considered to be at least 1000 mg/kg bw/day in males and females.

This result is read across to the registered substance, the rationale is attached in Section 13.

Endpoint conclusion

Endpoint conclusion:: no adverse effect observed
Dose descriptor:: NOAEL
: 1 000 mg/kg bw/day
Study duration:: subchronic
Species:: rat
Quality of whole database:: The study was performed with a substance analogue, according to OECD/EC guidance and GLP principles (Klimisch 1 study). The rationale to read across the data is attached in Section 13.

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion

Endpoint conclusion:: no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion

Endpoint conclusion:: no study available

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion

Endpoint conclusion:: no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion

Endpoint conclusion:: no study available

Additional information

The data for repeated dose toxicity are read across from a substance analogue. The rationale to read across the data is attached in Section 13.

Treatment with Amphoacetates C12-C14 in a 90-day repeated dose toxicity study was performed according to OECD TG 408 and in accordance with GLP principles. The objectives of this study were to determine the potential toxicity of the substance, when given orally by gavage for 90 days to Wistar Han rats and to evaluate the potential reversibility of any findings. In addition, a No Observed Adverse Effect Level (NOAEL) was evaluated.

Records with supporting studies on structural analogues are included for information purposes.

Justification for classification or non-classification

Based on the available data, the registered substance is not classified for repeated dose exposure according to CLP Regulation (EC) No. 1272/2008.

Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice.
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