Pentapotassium bis(peroxymonosulphate) bis(sulphate)

Endpoint summary

Description of key information

KMPS triple salt was tested in repeated dose toxicity studies for oral and inhalation toxicity.

 

Oral administration

KMPS triple salt was tested in a 14-days subacute study and in a 13-weeks subchronic study in rats. The NOAEL observed in both studies was 200 mg/kg bw/day.

 

Inhalation

KMPS triple salt was tested in a 14-days subacute inhalation study in rats. The NOAEC observed in this study was 1.4 mg/m³ air.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2001-04-09 - 2002-03-19

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

This study was selected as the key study because the information provided for the hazard endpoint is sufficient for the purpose of classification and labelling and/or risk assessment. Due to the high mortality the dose of 1000 mg/kg bw/day was reduced to 600 mg/kg bw/day. Therefore, the LOAEL is difficult to derive but assigned to 600 mg/kg bw/d for precautionary reasons.

Qualifier:

according to guideline

Guideline:

OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)

Version / remarks:

guideline 408 “Repeated dose 90-day oral toxicity study in rodents” (May

Deviations:

no

GLP compliance:

yes

Limit test:

no

Species:

rat

Strain:

other: Crl:CD(SD) IGS BR

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River (UK) Ltd., Margate, Kent, UK
- Age at study initiation: approx. 6 weeks
- Weight at study initiation: 168 – 215 g (males); 141 – 180 g (females)

Route of administration:

oral: gavage

Vehicle:

other: Deionised water

Details on oral exposure:

The subchronic toxicity of KMPS triple salt was assessed following oral administration to male and female Crl:CD(SD) IGS BR rats for a period of 13 weeks by gavage in water as the vehicle. Dose levels administered were 0, 25, 200 and 1000 mg/kg bw/day (dose volume: 10 mL/kg bw/day). Following 4 weeks of treatment at 1000 mg/kg bw/day, this dosage level was reduced to 600 mg/kg bw/day due to 4 unscheduled deaths and adverse clinical signs.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The stability in deionised water was confirmed at nominal concentrations of 1 mg/mL and 100 mg/mL, during ambient temperature.

Duration of treatment / exposure:

91 days

Frequency of treatment:

Once daily, 7 days per week

Dose / conc.:

0 mg/kg bw/day (nominal)

Dose / conc.:

25 mg/kg bw/day (nominal)

Remarks:

corresponding to 2.5 mg/mL

Dose / conc.:

200 mg/kg bw/day (nominal)

Remarks:

corresponding to 20 mg/mL

Dose / conc.:

1 000 mg/kg bw/day (nominal)

Remarks:

corresponding to 100 mg/mL; due to 4 unscheduled deaths and adverse clinical signs the high dosage level was reduced to 600 mg/kg bw/day after 4 weeks of treatment

No. of animals per sex per dose:

10 animals/group/sex (40 animals per sex in total)

Control animals:

yes, concurrent vehicle

Details on study design:

Clinical signs of toxicity and mortality/viability were observed daily. Body weights, body weight gains and food consumption was recorded on a weekly basis. A complete haematological and clinical-chemical examination was as well as an ophthalmoscopic examination performed on day 90 and during the Week 12, respectively. Functional and observational battery and motor activity assessments were performed before initiation and during the 12th week of treatment. A shortened battery was performed during Weeks 1 11 and Week 13. On termination of the study, organ weights were determined and a macroscopic as well as microscopic examination of all organs was performed.

Positive control:

no

Observations and examinations performed and frequency:

OBSERVATIONS: regularly throughout the study
- Clinical signs. Yes; at least twice daily; a detailed physical examination of each animal was performed weekly
- Mortality. Yes, twice daily

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

FOOD CONSUMPTION: Yes
- Weekly

WATER CONSUMPTION: Yes
- Time schedule for examinations: Daily monitoring by visual appraisal was maintained throughout the dosing period.

OPHTHALMOSCOPIC EXAMINATION: Yes
Performed on all animals following instillation of tropicamide solution before commencement of treatment and during the week 12 of treatment.

HAEMATOLOGY: Yes
- Time schedule for collection of blood: on day 90
- How many animals: all animals
- Parameters checked:
Haematocrit, haemoglobin concentration, red blood cell count (RBC), mean cell haemoglobin concentration (MCHC), MCV, MCH, total and differential leukocyte count (neutrophils, basophils, eosinophils, lymphocytes, monocytes, large unstained cells (LUC), platelet count, prothrombin time, partial thromboplastin time

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: on day 90
- How many animals: all animals
- Parameters checked: sodium, potassium, glucose, urea, creatinine, total protein, albumin, alanine aminotransferase, aspartate aminotransferase, Albumin/Globulin ratio (A/G ratio), alkaline phosphatase, cholesterol

URINALYSIS: No

Sacrifice and pathology:

ORGAN WEIGHTS
yes
organs:
adrenals, brain, epididymides, heart, kidneys, liver, ovaries, spleen, testes, thymus, uterus with cervix

GROSS AND HISTOPATHOLOGY
yes
all dose groups
The macroscopic appearance of the tissue of all rats was recorded and samples of tissues were preserved:
adrenals, aorta, brain, caecum, colon, duodenum, epididymides, eyes, femur (with joint), head (including nasal cavity, paranasal sinuses and nasopharynx), heart, ileum, jejunum, kidneys, liver, lung (and bronchi), lymph nodes (mandibular and mesenteric), mammary area, oesophagus, ovaries, pancreas, pituitary gland, prostate gland, rectum, salivary glands, spinal cord, pituitary, prostate, rectum, salvary glands, sciatic nerves, seminal vesicles, skin, spinal cord, spleen, sternum, stomach, testes, thymus, thyroid, parathyroid, trachea, urinary bladder, uterus, (with cervix), other macroscopically abnormal tissue.

Microscopic examination of the prepared slides (from all above indicated tissues) was carried out for all terminal animals. Macroscopic abnormalities were processed and examined for any animal. Stomach was assessed for all dosage groups.

Other examinations:

Functional and observational battery and motor activity assessments were performed during the study at app. the same time of day before initiation of treatment and during the 12th week of treatment (monitoring by using a Coulbourn Infra-Red Activity Monitoring System).
A shortened battery was performed during Weeks 1 11 and Week 13. The battery comprised of 3 sets of observations. The first set was performed when initially handling the animal. The second set of observations was performed in the test arena and the third set comprised handling/specific testing of the animal.
1. Observations in the hand: Ease of removing the animal from the cage, reactivity to handling, occurrence of convulsions, tremors, twitches, salvation/lachrymation, piloerection, vocalisation on handling etc.
2. Observation in the arena: occurrence of convulsions, tremors, twitches, activity counts, rearing counts, assessment of gait/posture etc.
3. Sensory reactivity: approach response, touch response, auditory startle response, tail pinch response, grip strength

Statistics:

The following sequence of statistical tests was used for bodyweight, haematology, blood chemistry, organ weight and functional observational battery data:
If 75 % of the data were the same value, then a frequency analysis was applied. Treatment groups were compared using a Mantel test for a trend in proportions and also pairwise Fisher’s Exact tests for each dose against the control values
If Bartlett’s test for variance homogeneity was not significant at the 1 % level, then parametric analysis was applied. If the F1 test for monotonicity of dose-response was not significant at the 1 % level, William’s test for a monotonic trend was applied. If the F1 test was significant, suggesting that the dose-response was not monotone, Dunnett’s test was performed instead.
If Bartlett’s test was significant at the 1 % level, then logarithmic and square-root transformations were tried. If Bartlett’s test was still significant, then non-parametric tests were applied. If the H1 test for monotonicity of dose-response was not significant at the 1 % level, Shirley's’test for a monotonic trend was applied. If the H1 test was significant, suggesting that the dose-response was not monotone, Dunn’s test was performed instead.
Where appropiate, analysis of covariance was used in place of analysis of variance in the above sequence.

Clinical signs:

effects observed, treatment-related

Mortality:

mortality observed, treatment-related

Body weight and weight changes:

effects observed, treatment-related

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Food efficiency:

effects observed, treatment-related

Water consumption and compound intake (if drinking water study):

no effects observed

Ophthalmological findings:

no effects observed

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

not examined

Behaviour (functional findings):

no effects observed

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

effects observed, treatment-related

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Histopathological findings: neoplastic:

no effects observed

Details on results:

Tables mentioned below are presented under "Remarks on results including tables and figures" and under "Overall remarks"

OBSERVATIONS
Clinical signs
• Post-dose salivation was noted for all animals receiving 1000/600 mg/kg bw/day, with incidence of post-dose salivation also noted among animals receiving 200 mg/kg bw/day.
• During Week 4, piloerection, abnormal gait gasping and hunched posture were noted on occasions for a few animals receiving 1000 mg/kg bw/day. These findings were generally noted from 1 2 hours after dosing but no longer evident the following morning.
• Noisy respiration was noted for 5/10 males and 4/10 females receiving 1000 mg/kg bw/day and one animal receiving 200 mg/kg bw/day during the first 4 weeks of treatment. Following a reduction of the dose level from 1000 to 600 mg/kg bw/day, noisy respiration was noted for 1 male in Weeks 8 and 12 only.
• Incidences of wet coat, paddling of forepaws and piloerection were noted among animals receiving 600 mg/kg bw/day during Week 12and 13.
• Incidental findings of vocalisation and hair loss were noted in all groups.
Mortality
One male (no. 17M; 1000 mg/kg bw/day) was humanely sacrificed in week 3. Two males and one female (no. 14M, 15M ad 64F; 1000 mg/kg bw/day) were also sacrificed due to adverse clinical signs in week 4. Necrotic and inflammatory lesions of the stomach and small intestine were considered to have been contributory to death.

BODY WEIGHT/BODY WEIGHT GAIN
After 4 weeks of treatment, bodyweight gain was lower than that of the concurrent controls for all treated groups of males. The difference from controls was statistically significant for all groups but not dosage related for those animals receiving 25 or 200 mg/kg bw/day. Group mean bodyweight gain by all treated groups of females was however marginally higher than that of controls. Please refer to tables A6.4.1/01-1 and A6.4.1/01-2.

FOOD CONSUMPTION
• Food intake was reduced among males receiving 1000 mg/kg bw/day from Weeks 1 4, continuing to a slightly lesser degree when those animals received 600 mg/kg bw/day from Weeks 5-13.
• Food consumption among males receiving 25 or 200 mg/kg bw/day was slightly less than that by controls but no dose relationship was apparent and the degree of difference from control was minimal.
• Consumption by females at all three dosages was higher than controls to a similar degree and these variations are not thought to be treatment related.
Food conversion efficiency was calculated (please refer to table A6.4.1/01-3):
• The efficiency for animals receiving 25 or 200 mg/kg bw/day was similar to that of controls.
• Food conversion efficiency for animals receiving 600 mg/kg bw/day (previously 1000 mg/kg bw/day) continued to be inferior to that of controls for the period Week 5 13.
The reduction in food utilisation efficiency for animals receiving 1000 mg/kg bw/day and then 600 mg/kg bw/day indicate that the reduced body weight gain was not entirely attributable to reduced food intake and is suggestive of a toxic effect.

WATER CONSUMPTION
No changes in water consumption were observed.

OPHTHALMOSCOPIC EXAMINATION
The incidence and distribution of findings noted in Week 12 fall within background ranges for this age and strain


BLOOD ANALYSIS
Haematology
Week 13 examination:
Please refer to Table A6.4.1/01-4.
There were no adverse treatment-related changes in haematology. The following statistically significant changes in haematology parameters were considered to be non-adverse because of the magnitude or direction of change.
• Eosinophils were minimally increased in males dosed with 1000 mg/kg bw/day. This change was due primarily to one rat with increased WBC as well. Increased eosinophils were possibly related to treatment.
• WBC, basophils, and LUCs were minimally decreased in females dosed with 200 or 1000/600 mg/kg bw/day. These changes were possibly related to treatment. Due to the very small magnitude of response and the lack of dose-response between 200 and 1000/600 mg/kg bw/day these changes were considered to be non-adverse.
• Platelets were increased in females dosed with 1000/600 mg/kg bw/day. This change was considered treatment related. However, minimally increased platelets have no adverse consequences.
• Activated partial thromboplastin time (APTT) was minimally shortened in males dosed with 1000/600 mg/kg bw/day after 13 weeks. Shortened APTT, although likely related to treatment due to the consistency of change, was considered non-adverse because minimal decreases in coagulation times are not associated with toxicity.
• Prothrombin time (PT) was prolonged in males dosed with 200 or 1000/600 mg/kg bw/day. However, all individual values were within the reference interval.

Clinical chemistry
Week 13 examination:
Please refer to Table A6.4.1/01-5
There were no adverse treatment related changes in clinical chemistry parameters.
• ALT was minimally decreased in males dosed with 1000/600 mg/kg bw/day. This was unlikely to be due to treatment due to lack of a dose response.
• Creatinine was minimally decreased in males dosed with 1000/600 mg/kg bw/day. This was not considered to be treatment related due to the magnitude of the change. A minimal creatinine decrease is also not associated with toxicity.
• Glucose was increased in males dosed with 1000/600 mg/kg bw/day. This was not considered to be treatment related. Glucose concentrations of individual rats dosed with 25 or 200 mg/kg bw/day were more variable than those in rats dosed with 1000 mg/kg bw/day. All glucose concentrations of rats dosed with 1000/600 mg/kg bw/day were less than the mean of the reference interval.
• Potassium was decreased in males of all dose groups. However, potassium concentrations in individual rats were equal or greater than the 50th percentile of the reference interval. Therefore, although this change was likely related to treatment, it was not considered adverse.
• Sodium was decreased in females dosed with 1000/600 mg/kg bw/day. Although this change was likely related to treatment, it was not considered adverse due to the magnitude of the change.
• Total protein was minimally decreased in males and females dosed with 1000/600 mg/kg bw/day. This was not considered adverse because the magnitude of change was small. However, this change may be secondary to loss of fluid rich in protein into the gastrointestinal tract. Therefore, although this change was not considered adverse, it was secondary to adverse effects observed histologically.


URINANALYSIS
Not performed


SACRIFICE AND PATHOLOGY
Organ weights
Please refer to table A6.4.01/016.
• There was a significantly higher bodyweight adjusted group mean liver weight for males and females receiving 1000/600 mg/kg bw/day in comparison to controls. This may be treatment related but in the absence of any histopathological change this finding is not considered adverse. No histological changes were observed in the liver to explain the increased organ weight.
• The bodyweight adjusted group mean adrenal weight was also significantly higher for males receiving 1000/600 mg/kg bw/day and the bodyweight adjusted mean brain weights were statistically significantly higher in all treated groups of males. In the absence of dose related trends, consistency between the sexes or related histopathological changes, these differences are not considered to be related to treatment.
• In addition, an increased relative organ weight (see above) might be explainable by reduced absolute body weights.

Gross and histopathology
Macroscopic examination of the 4 decedents:
Please refer to table A6.4.01/01-7.
Stomach: congested in 2/3 males and 1 female
Forestomach: thickened in all decedent animals; oedematous in 1/3 males
Limiting ridge and forestomach: thickened/depression in 1/3 males
Antrum: pale area/white nodules in 1/3 males
Lymph nodes: mandibular, enlarged in 1 female
Lumbar: enlarged in 1/3 males
Caecum: distended in 2/3 males
Duodenum: congested in 1/3 males
Pancreas: dark in 2/3 males
Spleen: pale in 1 female
Abnormal G.I.tract in 1/3 males and distension of G.I.tract in 2/3 males and 1 female.

Macroscopic observations performed on survivors at termination revealed changes to the stomach:
Please refer to table A6.4.1/01-8.
- Forestomach
Thickening of the limiting ridge in 7/7 males and 8/9 females receiving 1000/600mg/kg bw/day and 1/10 females receiving 200 mg/kg bw/day
Generalised thickening in 3/7 males and 1/9 females, with roughening in 4/7 males and depressions in 2/7 males and 2/9 females receiving 1000/600 mg/kg bw/day
All other findings at termination were considered to fall within the background range of macroscopic changes.
Microscopic examination of survivors at termination:
Treatment related effects were observed in the stomach only in the high dose group
In the forestomach inflammation, oedema and haemorrhage in the mucosal and submucosal areas, epithelial necrosis and ulceration (less pronounced in females), epithelial hyperplasia (including the limiting ridge) was noted.
In few cases submucosal inflammation was observed in the glandular stomach in male and female high dose group compared to controls.

Microscopic examination of the Decedents:
Please refer to table A6.4.1/01-9.
In all cases, necrotic and inflammatory lesions of the stomach and, to a lesser extent, the small intestine were considered to be factors contributory to death. The principal findings were epithelial necrosis and subepithelial inflammation and, to a lesser extent, in the glandular region of stomach and villous atrophy and necrosis and also epithelial hyperplasia in the small intestine. Also considered to be associated with the clinical condition of the animals was apotosis, seen in various lymphoid tissues including spleen, mesenteric lymph node, mandibular lymph node and Peyer’s patch in the ileum.


OTHER
FOB and MA examinations:
Treatment with Oxone Monopersulfate Compound for 13 weeks was not associated with any behavioural changes which are considered to be indicative for neuotoxicity.

Dose descriptor:

LOAEL

Effect level:

600 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: based on the changes in the nonglandular stomach

Key result

Dose descriptor:

NOAEL

Effect level:

200 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: absence of effects

Key result

Critical effects observed:

no

Table 1: 90-day gavage study with KMPS triple salt in rats – group mean bodyweight (g)

Week	Control	25 mg/kg bw/day	200 mg/kg bw/day	1000/600 mg/kg bw/day
-	males
0	196.5	188.4	189.7	191.6
1	255.3	242.3	245.5	242.6
2	316.5	296.2	297.1	289.9
3	366.3	334.5	339.2	323.2
4	404.7	367.5	372.0	339.8
5	430.7	389.2	398.0	363.9
6	448.6	406.4	418.5	382.1
7	465.7	414.9	435.3	398.4
8	488.4	434.0	451.9	408.0
9	504.2	450.0	465.6	422.4
10	518.9	461.3	479.7	431.4
11	537.7	473.5	492.5	442.1
12	546.3	483.3	504.6	449.1
13	548.3	487.2	506.8	453.1
-	females
0	157.4	161.2	160.6	163.4
1	183.5	191.4	191.3	194.8
2	207.6	215.9	216.0	217.9
3	224.4	236.2	236.6	239.2
4	237.3	250.2	247.6	253.4
5	250.5	259.0	258.3	265.4
6	257.8	267.7	268.6	273.3
7	264.7	277.6	277.4	278.8
8	269.6	285.6	281.4	283.4
9	278.7	287.5	286.2	292.3
10	282.6	295.1	291.0	293.9
11	288.3	302.7	296.5	295.5
12	287.0	303.9	300.0	298.2
13	288.8	299.0	298.7	298.6

Table 2: 90-day gavage study with KMPS triple salt in rats – group mean bodyweight gain

Week	Control	25 mg/kg bw/day	200 mg/kg bw/day	1000/600 mg/kg bw/day
-	males
0-4	208.2	179.1*	182.3*	147.8**
% of control	-	86	88	71
4-13	143.6	119.7	134.8	113.3*
% of control	-	83	94	79
0-13	351.8	298.8*	317.1*	261.1**
% of control	-	85	90	74
-	females
0-4	79.9	88.9	87.0	89.5
% of control	-	111	109	112
4-13	51.3	48.9	51.0	45.2
% of control	-	95	99	88
0-13	131.4	137.8	138.0	134.7
% of control	-	105	105	102

                          *P< 0.05; **P <0.01 in comparison with controls (Williams’ test)

 

Table 3: Food conversion efficiency – group mean values (%)

Dosage (mg/kg bw/day)	Male				Female
	0	15	200	1000/ 600	0	15	200	1000/ 600
Weeks	-				-
1-4	25.2	23.6	23.6	20.3	15.4	15.7	15.8	15.9
5-13	8.1	7.3	8.0	7.0	4.2	3.8	4.1	3.7
1-13	13.5	12.5	12.9	11.1	7.6	7.5	7.6	7.6

 

Table 4: Haematology

Dosage (mg/kg bw/day)	Males
	Eosinophils x10-9/L		Prothrombin time (sec.)		Activated partial thromboplastin time (sec.)
0	-		-		-
Mean	0.18		12.7		20.0
SD	0.051		0.6		1.49
n	10		10		10
25	-		-		-
Mean	0.21		13.3		20.3
SD	0.077		0.42		1.79
n	10		10		10
200	-		-		-
Mean	0.19		13.5*		19.2
SD	0.058		0.93		1.83
n	10		10		10
1000/600	-		-		-
Mean	0.30*		13.7**		16.9**
SD	0.208		1.04		1.58
n	7		7		7
Dosage (mg/kg bw/day)	Females
	WBC x10-9/L	Basophils x10-9/L		LUC x10-9/L		Plt x10-9/L
0	-	-		-		-
Mean	10.14	0.03		0.26		1014
SD	2.730	0.014		0.054		115.6
n	10	10		10		10
25	-	-		-		-
Mean	8.83	0.02		0.27		955
SD	2.602	0.008		0.097		314.3
n	10	10		10		10
200	-	-		-		-
Mean	6.88*	0.01**		0.19**		1074
SD	1.453	0.005		0.058		132.7
n	10	10		10		10
1000/600	-	-		-		-
Mean	8.59*	0.02**		0.20**		1217++
SD	2.492	0.010		0.071		139.9
n	9	9		9		9

*      p<0.05 in comparison with controls (Williams’ test)                               WBC: White blood cell count
**     p<0.01 in comparison with controls (Williams’ test)                               LUC: Large unstained cells
++     p<0.01 in comparison with controls (Shirley’s test)                                Plt: Platelet count

SD   Standard deviation

 

Table 5: Blood chemistry

Dosage (mg/kg bw/day)	Males
	ALT U/L	Creatinine µmol/L	Glucose mmol/L	K mmol/L	Total Protein g/L
0	-	-	-	-	-
Mean	44	50	5.29	4.4	71
SD	2.5	2.9	0.505	0.21	1.3
n	10	10	10	10	10
25	-	-	-	-	-
Mean	50	50	5.99	4.1*	70
SD	16.3	1.7	0.927	0.24	2.7
n	10	10	10	10	10
200	-	-	-	-	-
Mean	50	50	5.60	4.1*	71
SD	6.1	2.5	1.025	0.28	3.6
n	9	9	9	9	9
1000/600	-	-	-	-	-
Mean	39	47*	6.38**	3.9**	67**
SD	3.6	1.9	0.166	0.08	2.6
n	7	7	7	7	7

*      P<0.05 in comparison with controls (Williams’ test)                              ALT: Alanine aminotransferase
**     P<0.01 in comparison with controls (Williams’ test)                              K: Potassium

 

Table 5 (continued): Blood chemistry 

Dosage (mg/kg bw/day)	Females
	Na mmol/L	Total Protein g/L	Albumin g/L
0	-	-	-
Mean	139	77	40
SD	1.4	4.8	2.2
n	10	10	10
25	-	-	-
Mean	139	77	40
SD	1.3	3.7	1.9
n	10	10	10
200	-	-	-
Mean	138	75	39
SD	0.8	4.5	2.5
n	10	10	10
1000/600	-	-	-
Mean	137*	72*	37*
SD	1.3	3.6	2.3
n	9	9	9

*      p<0.05 in comparison with controls (Williams’ test)                      Na: Sodium

 

Table 6: Organ weights – group mean values (g) for animals killed after 13 weeks of treatment

Dosage (mg/kg bw/day)	Males
	Terminal Body weight	Brain	Adrenals	Liver
0	-	-	-	-
Mean (unadjusted)	542.6	2.15	0.063	20.49
SD	72.4	0.10	0.014	3.87
Mean (adjusted)	-	2.09	0.055	18.13
n	10	10	10	10
25	-	-	-	-
Mean (unadjusted)	483.0	2.15	0.058	19.12
SD	40.4	0.07	0.010	3.70
Mean (adjusted)	-	2.17*	0.061	19.94
n	10	10	10	10
200	-	-	-	-
Mean (unadjusted)	505.3	2.16	0.059	20.02
SD	32.2	0.08	0.012	2.95
Mean (adjusted)	-	2.15*	0.058	19.66
n	10	10	10	10
1000/600	-	-	-	-
Mean (unadjusted)	447.6	2.12	0.068	19.94
SD	40.9	0.10	0.014	2.72
Mean (adjusted)	-	2.19**	0.077**	22.65**
n	7	7	7	7

*       p<0.05 in comparison with controls (Williams’ test)

**      p<0.01 in comparison with controls (Williams’ test)

 

Table 6 (continued): Organ weights – group mean values (g) for animals killed after 13 weeks of treatment 

Dosage (mg/kg bw/day)	Females
	Terminal Body weight	Liver
0	-	-
Mean (unadjusted)	286.2	10.53
SD	24.9	1.18
Mean (adjusted)	-	10.80
n	10	10
25	-	-
Mean (unadjusted)	296.5	10.59
SD	17.0	1.15
Mean (adjusted)	-	10.48
n	10	10
200	-	-
Mean (unadjusted)	295.9	11.34
SD	39.9	1.86
Mean (adjusted)	-	11.25
n	10	10
1000/600	-	-
Mean (unadjusted)	295.5	12.13*
SD	19.3	0.98
Mean (adjusted)	-	12.06**
n	9	9

*       p<0.05 in comparison with controls (Williams’ test)
**      p<0.01 in comparison with controls (Williams’ test)

 

Table 7: Macropathology –group distribution of findings for animals killed or dying during the study

Organ	Males 1000/600 mg/kg bw/day	Females 1000/600 mg/kg bw/day
Caecum          No. examined          distended	3 2	1 0
Pancreas          No. examined          dark	3 2	1 0
Stomach          No. examined          Forestomach thickened          Limiting Ridge, thickened          Forestomach, roughened          Forestomach, depression(s)          congested          Antrum, white nodule(s)          Antrum, pale area(s)          Forestomach oedematous	3 3 1 1 1 2 1 1 1	1 1 0 0 0 1 0 0 0
Miscellaneous          No. examined          Abnormal contents (G.I. tract)          Distension G.I. tract	3 1 2	1 0 1

 

Table 8: Macroscopic observations performed at termination

Dosage (mg/kg bw/day)	Males
	0	25	200	1000/600
Organ	-	-	-	-
Stomach          No. examined          Forestomach, thickened          Limiting ridge, thickened          Forestomach roughened          Forestomach depression(s)          Antrum, white nodule(s)          Cyst(s) Dark area(s)	10 0 0 0 0 2 1 0	10 0 0 0 0 1 0 0	10 0 0 0 0 2 0 0	7 3 7 4 2 1 0 1
Skin          No. examined          Hairloss	10 2	10 1	10 1	7 1
Dosage (mg/kg bw/day)	Females
	0	25	200	1000/600
Organ	-	-	-	-
Stomach          No. examined          Forestomach, thickened          Limiting ridge, thickened          Forestomach depression(s)          Antrum, white nodule(s)          Cyst(s)         Forestomach raised area(s)	10 0 0 0 1 0 0	10 0 0 0 0 0 0	10 0 1 0 4 0 0	9 1 8 2 1 1 1
Skin          No. examined          Hairloss	10 6	10 7	10 3	9 4

 

Table 9: Histopathology –group distribution of findings for animals killed or dying during the study

Organ	Males 1000/600 mg/kg bw/day	Females 1000/600 mg/kg bw/day
Stomach          No. examined          Epithelial hyperplasia-nonglandular region          Submucosal oedema-nonglandular region          Submucosal inflammation-nonglandular region          Epithelial hyperplasia-glandular region          Epithelial necrosis-nonglandular region          Ectopic nonglandular epithelium in glandular          Mucosa, focal          Subepithelial inflammation-glandular region          Epithelial hyperplasia-limiting ridge          Subepithelial inflammation-nonglandular region          Mucosal inflammation-nonglandular region          Mcosal inflammtion-glandular region          Subepithelial oedema-nonglandular region	3 3 3 3 3 3 3 3 1 1 3 1 2 2	1 1 1 1 1 1 1 1 0 0 1 0 0 0
Thymus          No. examined          Apotosis-Cortex          Apotosis Medulla	3 3 2	1 1 0
Necrosis and inflammation of stomach and intestine	3/3	1/1

Conclusions:

The study and the conclusions which are drawn from it fulfill the quality criteria (validity, reliability, repeatability). Treatment of rats with KMPS triple salt at a combined dosage of 1000/600 mg/kg bw/day for 13 weeks resulted in pathological changes of the stomach. Effects were severe and resulted in the death of 4 animals. Following reduction of the high dose level to 600 mg/kg bw/day there were no more deaths but changes in the nonglandular stomach persisted. There were no toxic effects noted at 200 or 25 mg/kg bw/day and the NOAEL is considered to be 200 mg/kg bw/day. Changes in the stomach of the decedents are considered to be a direct local effect following bolus administration and the oxidative reaction mechanisms of KMPS triple salt at the site of first contact.
No classification and labelling regarding repeated dose toxicity is necessary according to Regulation 1272/2008/EC (CLP).

Executive summary:

Materials and methods

The subchronic toxicity of KMPS triple salt was assessed following oral administration to male and female Crl:CD(SD) IGS BR rats for a period of 13 weeks by gavage in water as the vehicle. Dose levels administered were 0, 25, 200 and 1000 mg/kg bw/day (dose volume: 10 ml/kg bw/day). Following 4 weeks of treatment at 1000 mg/kg bw/day, this dosage level was reduced to 600 mg/kg bw/day due to 4 unscheduled deaths and adverse clinical signs.

Clinical signs of toxicity and mortality/viability were observed daily. Body weights, body weight gains and food consumption was recorded on a weekly basis. A complete haematological and clinical-chemical examination was as well as an ophthalmoscopic examination performed on day 90 and during the Week 12, respectively. Functional and observational battery and motor activity assessments were performed before initiation and during the 12th week of treatment. A shortened battery was performed during Weeks 1 11 and Week 13. On termination of the study, organ weights were determined and a macroscopic as well as microscopic examination of all organs was performed.

 

Results and discussion

Pathological changes resulting from the administration of KMPS triple salt at 1000 mg/kg bw/day was marked, culminating in the death of 4 rats during Week 3 or 4 of treatment. The dose level was reduced to 600 mg/kg bw/day for the surviving animals of this level from Week 5 to termination (Week 13). In all 4 decedent animals, necrotic and inflammatory lesions of the stomach and the small intestine were considered to factors contributory to death. Which is considered to be a direct local effect following bolus administration and the oxidative reaction mechanisms of KMPS at the site of first contact.

Histopathological change was evident in animals receiving 1000/600 mg/kg bw/day for 13 weeks. The target organ was identified as the stomach with inflammatory, degenerative and hyperplastic changes mainly in the nonglandular stomach, the principal findings of which were inflammation, epithelial necrosis and ulceration, and epithelial hyperplasia in the nonglandular stomach. Macropathological changes were also apparent.

The pathological findings seen in the nonglandular area of the stomach are considered to represent damage due to the physical nature of the test substance and as such not considered to be associated with systemic toxicity. The changes in the gut were not of sufficient severity to affect nutrient availability or disturb general homeostasis, when animals were given 600 mg/kg bw/day.

No pathological changes related to treatment were identified in animals receiving 200 mg/kg bw/day. The complete absence of any adverse effect of treatment at this dose level indicates a steep dose-response curve between 200 and 1000 mg/kg bw/day for stomach pathology. The nature of lesions suggests that treatment at 600 mg/kg bw/day was potentiating damage initially caused by treatment at 1000 mg/kg bw/day.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

NOAEL

200 mg/kg bw/day

Study duration:

subchronic

Species:

rat

Quality of whole database:

reliable guideline study

Repeated dose toxicity: inhalation - systemic effects

Link to relevant study records

Reference

Endpoint:

short-term repeated dose toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1980-09-22 - 1981-08-12

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Remarks:

The fact, that only male rats were used, is not considered to have compromised the study results, as male rats were shown to be the more sensitive sex in the acute inhalation toxicity study (please refer to A6.1.3/01, IUCLID section 7.2.2). This study was selected as the key study because the information provided for the hazard endpoint is sufficient for the purpose of classification and labelling and/or risk assessment.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 412 (Subacute Inhalation Toxicity: 28-Day Study)

Version / remarks:

No guideline available at time of study conduct

Deviations:

yes

Remarks:

Only male rats were used , particle size not analysed, no individual animal data available, food consumption not investigated

GLP compliance:

no

Remarks:

GLP was not obligatory at the time of the study conduct; laboratories in the U.S.A. are not certified by any governmental agency, but are subject to official inspections. The study was performed according to good scientific practise.

Limit test:

no

Species:

rat

Strain:

other: Crl:CD

Sex:

male

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Not stated
- Age at study initiation: 8 weeks
- Weight at study initiation: 230 – 266 g

Route of administration:

inhalation

Type of inhalation exposure:

head only

Vehicle:

other: unchanged (no vehicle); dilution with clean air only

Remarks on MMAD:

MMAD / GSD: not indicated

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE

- System of generating particulates/aerosols: Dust aerosol atmospheres of Oxone were generated with a 3 stage glass generator composed of a dust reservoir, cyclone generator, and elutriator.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Analytical concentrations are given under "Doses/concentrations"

Duration of treatment / exposure:

2 weeks, 6 hours per day

Frequency of treatment:

5 days/week

Dose / conc.:

0 mg/L air

Dose / conc.:

0.001 mg/L air (nominal)

Remarks:

0.0014 mg/L (analytical)

Dose / conc.:

0.01 mg/L air (nominal)

Remarks:

0.0101 mg/L (analytical)

Dose / conc.:

0.05 mg/L air (nominal)

Remarks:

0.0431 mg/L (analytical)

No. of animals per sex per dose:

10/group

Control animals:

yes, concurrent no treatment

Details on study design:

not indicated

Positive control:

Not applicable

Observations and examinations performed and frequency:

Observations
- Clinical signs: yes
- Mortality: yes

Body weight: yes (daily)

Food consumption: No

Water consumption: No

Ophthalmoscopic examination: yes

Haematology: Yes, blood samples were taken after the 10th and 13th observation day. Measurements included erythrocyte count, haemoglobin concentration, mean corpuscular volume, platelet count, leukocyte count, and relative number of neutrophils, lymphocytes, eosinophils, monocytes, and basophils. The haematocrit, mean corpuscular haemoglobin, and mean corpuscular haemoglobin concentration were calculated from these data.

Clinical Chemistry: Yes, blood serum measurements included alkaline phosphatase, glutamic-pyruvic transaminase, glutamic-oxalacetic acid, urea nitrogen, creatinine, and total protein.

Urinalysis: Yes, after the 9th exposure and the 12th observation day all test animals were individually housed in stainless steel metabolism cages for overnight (16 hours) urine collection.
The measurement included urine volume, osmolality, pH, and tests for sugar, blood, protein, bilirubin, urobilinogen, and acetone. The urine colour and appearance were noted and sediment from pooled specimens was examined macroscopically.

Sacrifice and pathology:

Yes, after the 10th exposure, 5 rats from each level were selected at random and sacrificed for gross and histopathological examination.

Remaining rats were sacrificed on the 13th observation day for an identical examination.
Organs weighed at necropsy: thymus, spleen, heart, lungs, kidneys, testes

Organs examined: ear pinna, skin thymus, mediastinal tissue, spleen, bone marrow (sternum), heart, trachea, lungs, oesophagus, stomach, small intestines (duodenum, jejum, and ileum), large intestines (caecum and colon), liver, kidneys, testes, epididymides, thyroids, adrenals, brain, eyes

Other examinations:

not indicated

Statistics:

LSD and Dunnett test was used as statistical model.

Clinical signs:

effects observed, treatment-related

Mortality:

mortality observed, treatment-related

Body weight and weight changes:

effects observed, treatment-related

Food consumption and compound intake (if feeding study):

not examined

Food efficiency:

not specified

Water consumption and compound intake (if drinking water study):

not examined

Ophthalmological findings:

effects observed, treatment-related

Haematological findings:

effects observed, treatment-related

Clinical biochemistry findings:

effects observed, treatment-related

Urinalysis findings:

effects observed, treatment-related

Behaviour (functional findings):

not specified

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

effects observed, treatment-related

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Histopathological findings: neoplastic:

no effects observed

Details on results:

CLINICAL SIGNS
During the exposure:
The predominant clinical observations were characteristic of eye irritation (please refer to point 4.5).
A red discharge from the noses of control and test rats, and slight lung noise in high dose level rats was observed during exposure.
Post-exposure:
The 13 day observation period allowed only partial recovery from these ocular effects. Most of the clinical signs were still present but their severity had decreased.

MORTALITY
No unscheduled deaths were reported.

BODY WEIGHT AND WEIGHT GAIN
There was a sporadic weight loss in all groups (including controls), with significant weight depression in the intermediate and high levels during the latter part of the exposure period (Please refer to table 1, which is presented under “Remarks on results including tables and figures”).


OPHTHALMOSCOPIC EXAMINATION
No effects were noted in low dose level rats (0.0014 mg/L). Intermediate and high level rats exhibited dose dependent alopecia around the eye, conjunctival swelling, severe opacity, corneal ulceration and haemorrhage, corneal vascularisation, and clear discharge. In general, a dry, crusty scab formed around the eyes, keeping them closed unless forced open. One control rat had a cloudy, glazed right eye.

HAEMATOLOGY
After 10 exposures, rats at the high dose level had higher than normal leukocyte counts, with 3/10 rats having counts outside of the norm established by controls.

CLINICAL CHEMISTRY
High dose level rats tended to have elevated serum glutamic-oxalacetic acid, glutamic-pyruvic transaminase, and depressed levels in alkaline phosphatase, and serum creatinine.
After a 13-day observation period serum glutamic oxalacetic acid transaminase, creatinine, and total protein levels were statistically lower than controls, however, these effects were not considered clinically significant. Low levels in total protein and glutamic oxalacetic acid transaminase are not of toxicological relevance due to the absence of a treatment-related effect for total protein during treatment and due to a non-consistent direction of change for glutamic oxalacetic acid transaminase.
Furthermore, changes seen on glutamic-pyruvic transaminase and alkaline phosphatase were shown to be completely reversible during the recovery period.

URINALYSIS
Rats at the high dose level had an elevated nitrogen activity as well as depressed levels in urine pH.


ORGAN WEIGHTS
Analysis of mean absolute organ weight after 10 exposures and the 13-day observation period did not reveal any dose-dependent effects. Organ/body weight ration analysis likewise did not reveal any dose related responses (Please refer to table 2, which is presented under “Remarks on results including tables and figures”).


GROSS PATHOLOGY
No gross findings were observed in control or low dose level rats. Intermediate and high dose level rats generally had ocular discharge, swollen eyelids with hair loss, and a cloudy appearance of the cornea.

HISTOPATHOLOGY:
Microscopic lesions attributable to the test compound were limited to the eyes and eyelids of rats exposed at the 2 highest exposure levels. These changes included: blepharitis, keratitis, corneal vascularisation, iritis, exflammatory exudate in the anterior and posterior chambers of the eye, haemorrhage mainly in the vitreous body of the eye and degeneration of the lens (cataract). These findings were equal in severity in the 2 high test groups but higher in incidence in the high level. The 13 day observation period allowed only slight recovery of these lesions and it is not expected that complete recovery would occur.

Dose descriptor:

LOAEL

Effect level:

0.01 mg/L air (analytical)

Sex:

male

Basis for effect level:

other: based on ocular effects

Dose descriptor:

LOAEL

Effect level:

2.73 other: mg/kg bw/d (assuming a respiratory volume of 86.4 L/rat/d and a body weight of 320 g)

Sex:

male

Basis for effect level:

other: based on ocular effects

Key result

Dose descriptor:

NOEL

Effect level:

0.001 mg/L air (analytical)

Sex:

male

Basis for effect level:

other: based on histological findings in the eyes at the next higher dose level.

Dose descriptor:

NOEL

Effect level:

0.38 other: mg/kg bw/d (assuming a respiratory volume of 86.4 L/rat/d and a body weight of 320 g)

Sex:

male

Basis for effect level:

other: based on histological findings in the eyes at the next higher dose level

Key result

Critical effects observed:

no

Table 1: Summary: Body weight (g)

 

Test Day	Dose (mg/L)
-	0	0.0014	0.0101	0.0431
1	249.70	246.90	251.80	249.56
5	237.60	243.60	237.90	240.89
10	259.30	265.70	258.80	247.44#
12	259.70	264.10	243.80#	235.56#
15	278.00	285.00	271.00	256.25+
25	327.60	337.60	329.20	321.00

 

#  Significantly different (P0.05) from control group by Dunnett test and LSD.

+  Significantly different (P0.05) from control group by LSD.

 

 

Table 2:  Organ/Body weight mean relative data of rats sacrificed after 13 day observation period.

Organ	Dose (mg/L)
-	0	0.0014	0.0101	0.0431
Heart	0.3451	0.3320	0.3287	0.3526
Lungs	0.6509	0.6132	0.6263	0.5565
Liver	3.7396	3.7591	3.8695	3.9592
Spleen	0.2508	0.2525	0.2112	0.2020
Kidney	0.7931	0.7655	0.8019	0.7929
Testis	1.0050	0.9866	0.9647	0.9665
Thymus	0.2205	0.2200	0.2088	0.1960

Conclusions:

The study and the conclusions which are drawn from it fulfill the quality criteria (validity, reliability, repeatability).
14-days LOAEC: 0.0101 mg/L (10.1 mg/m³) corresponding to 2.73 mg/kg bw/d (assuming a respiratory volume of 86.4 L/rat/d and a body weight of 320 g)
14-days NOAEC: 0.0014 mg/L (1.4 mg/m³) corresponding to 0.38 mg/kg bw/d (assuming a respiratory volume of 86.4 L/rat/d and a body weight of 320 g), based on histological findings in the eyes at the next higher dose level.
No classification and labelling in view of repeated dose inhalation toxicity is necessary according to Regulation 1272/2008/EC (CLP).

Executive summary:

Materials and methods

In a 14-day inhalation toxicity study (6 hours per day, 5 days per week), 10 male Crl:CD rats per group were exposed to dust aerosol of KMPS triple salt at exposure levels of 0, 0.0014, 0.0101 and 0.0431 mg KMPS triple salt/L (head only exposure). Animals were examined for clinical signs and mortality. Body weight was examined daily. Haematology and clinical chemistry was performed after the 10th exposure and the 13th observation day. All animals were subjected to gross pathology. Selected organs were weighed.

 

Results and discussion

During the exposure period clinical observations were severe ocular irritation at mid and high dose levels. These included: alopecia around eyes, conjunctival swelling, weeping, severe opacity, and corneal vascularisation, ulceration, and haemorrhaging. In general, a dry crusty scab formed around the eyes keeping them closed unless forced open. Also noted during exposures was a red nasal discharge from both control and test rats and slight lung noise in high level rats. Sporadic weight depression was recorded in the two highest test levels during the latter part of the exposure period. The 13 day observation period allowed only partial recovery from these ocular effects. Most of the clinical signs continued but were less severe.

Following the 10th exposure, rats at the high dose level had elevated leukocyte counts as well as elevated serum glutamic oxalactic acid, glutamic pyruvic transaminase, and urea nitrogen activities. This group also had depressed levels in urine pH, alkaline phosphatase, and serum creatinine. No other clinical pathologic changes were seen in this group. No corresponding changes in haematology or clinical chemistry were evident at the low and mid dose level.

After the observation period, serum glutamic oxalactic acid, creatitine, and total protein levels were statistically lower than controls.

Gross pathology showed significant findings in the ocular area only at the mid and high dose level. These findings included ocular discharge, swollen eyelids with hair loss, and a cloudy appearance of the cornea. Histologic changes attributable to the test compound were also noted in the eyes and blespharitis, keratitis, corneal vascularisation, iritis, exflammatory exudate in the anterior and posterior chambers of the eye, haemorrhage (mainly in the vitreous body of the eye), and degeneration of the lens (cataract). These changes were equal in severity at the mid and high dose level but the incidences were higher at the high dose level. No ocular changes were observed in rats of the low level.

In a 13-days observation period allowed these eye lesions only slightly recovered and complete reversal is not considered likely to occur.

Organ and body weight at the high dose level noted only a significant decrease in body weight on test days 10-16 and at the intermediate dose level on day 12. All rats gained weight at parallel rates during the observation period. Analysis of mean absolute organ weight and organ/body weight ratios did not reveal any dose dependent changes (Please refer to tables 1 and 2, which are presented under “Remarks on results including tables and figures”).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEC

1.4 mg/m³

Study duration:

subacute

Species:

rat

Quality of whole database:

relibale as the study was conducted comparable to OECD Guideline 412, only males were exposed but this is not considered to have compromised the study results, as male rats were shown to be the more sensitive sex in the acute inhalation toxicity study (please refer to A6.1.3/01, IUCLID section 7.2.2)

Repeated dose toxicity: inhalation - local effects

Link to relevant study records

Reference

Endpoint:

short-term repeated dose toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1980-09-22 - 1981-08-12

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Remarks:

The fact, that only male rats were used, is not considered to have compromised the study results, as male rats were shown to be the more sensitive sex in the acute inhalation toxicity study (please refer to A6.1.3/01, IUCLID section 7.2.2). This study was selected as the key study because the information provided for the hazard endpoint is sufficient for the purpose of classification and labelling and/or risk assessment.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 412 (Subacute Inhalation Toxicity: 28-Day Study)

Version / remarks:

No guideline available at time of study conduct

Deviations:

yes

Remarks:

Only male rats were used , particle size not analysed, no individual animal data available, food consumption not investigated

GLP compliance:

no

Remarks:

GLP was not obligatory at the time of the study conduct; laboratories in the U.S.A. are not certified by any governmental agency, but are subject to official inspections. The study was performed according to good scientific practise.

Limit test:

no

Species:

rat

Strain:

other: Crl:CD

Sex:

male

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Not stated
- Age at study initiation: 8 weeks
- Weight at study initiation: 230 – 266 g

Route of administration:

inhalation

Type of inhalation exposure:

head only

Vehicle:

other: unchanged (no vehicle); dilution with clean air only

Remarks on MMAD:

MMAD / GSD: not indicated

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE

- System of generating particulates/aerosols: Dust aerosol atmospheres of Oxone were generated with a 3 stage glass generator composed of a dust reservoir, cyclone generator, and elutriator.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Analytical concentrations are given under "Doses/concentrations"

Duration of treatment / exposure:

2 weeks, 6 hours per day

Frequency of treatment:

5 days/week

Dose / conc.:

0 mg/L air

Dose / conc.:

0.001 mg/L air (nominal)

Remarks:

0.0014 mg/L (analytical)

Dose / conc.:

0.01 mg/L air (nominal)

Remarks:

0.0101 mg/L (analytical)

Dose / conc.:

0.05 mg/L air (nominal)

Remarks:

0.0431 mg/L (analytical)

No. of animals per sex per dose:

10/group

Control animals:

yes, concurrent no treatment

Details on study design:

not indicated

Positive control:

Not applicable

Observations and examinations performed and frequency:

Observations
- Clinical signs: yes
- Mortality: yes

Body weight: yes (daily)

Food consumption: No

Water consumption: No

Ophthalmoscopic examination: yes

Haematology: Yes, blood samples were taken after the 10th and 13th observation day. Measurements included erythrocyte count, haemoglobin concentration, mean corpuscular volume, platelet count, leukocyte count, and relative number of neutrophils, lymphocytes, eosinophils, monocytes, and basophils. The haematocrit, mean corpuscular haemoglobin, and mean corpuscular haemoglobin concentration were calculated from these data.

Clinical Chemistry: Yes, blood serum measurements included alkaline phosphatase, glutamic-pyruvic transaminase, glutamic-oxalacetic acid, urea nitrogen, creatinine, and total protein.

Urinalysis: Yes, after the 9th exposure and the 12th observation day all test animals were individually housed in stainless steel metabolism cages for overnight (16 hours) urine collection.
The measurement included urine volume, osmolality, pH, and tests for sugar, blood, protein, bilirubin, urobilinogen, and acetone. The urine colour and appearance were noted and sediment from pooled specimens was examined macroscopically.

Sacrifice and pathology:

Yes, after the 10th exposure, 5 rats from each level were selected at random and sacrificed for gross and histopathological examination.

Remaining rats were sacrificed on the 13th observation day for an identical examination.
Organs weighed at necropsy: thymus, spleen, heart, lungs, kidneys, testes

Organs examined: ear pinna, skin thymus, mediastinal tissue, spleen, bone marrow (sternum), heart, trachea, lungs, oesophagus, stomach, small intestines (duodenum, jejum, and ileum), large intestines (caecum and colon), liver, kidneys, testes, epididymides, thyroids, adrenals, brain, eyes

Other examinations:

not indicated

Statistics:

LSD and Dunnett test was used as statistical model.

Clinical signs:

effects observed, treatment-related

Mortality:

mortality observed, treatment-related

Body weight and weight changes:

effects observed, treatment-related

Food consumption and compound intake (if feeding study):

not examined

Food efficiency:

not specified

Water consumption and compound intake (if drinking water study):

not examined

Ophthalmological findings:

effects observed, treatment-related

Haematological findings:

effects observed, treatment-related

Clinical biochemistry findings:

effects observed, treatment-related

Urinalysis findings:

effects observed, treatment-related

Behaviour (functional findings):

not specified

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

effects observed, treatment-related

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Histopathological findings: neoplastic:

no effects observed

Details on results:

CLINICAL SIGNS
During the exposure:
The predominant clinical observations were characteristic of eye irritation (please refer to point 4.5).
A red discharge from the noses of control and test rats, and slight lung noise in high dose level rats was observed during exposure.
Post-exposure:
The 13 day observation period allowed only partial recovery from these ocular effects. Most of the clinical signs were still present but their severity had decreased.

MORTALITY
No unscheduled deaths were reported.

BODY WEIGHT AND WEIGHT GAIN
There was a sporadic weight loss in all groups (including controls), with significant weight depression in the intermediate and high levels during the latter part of the exposure period (Please refer to table 1, which is presented under “Remarks on results including tables and figures”).


OPHTHALMOSCOPIC EXAMINATION
No effects were noted in low dose level rats (0.0014 mg/L). Intermediate and high level rats exhibited dose dependent alopecia around the eye, conjunctival swelling, severe opacity, corneal ulceration and haemorrhage, corneal vascularisation, and clear discharge. In general, a dry, crusty scab formed around the eyes, keeping them closed unless forced open. One control rat had a cloudy, glazed right eye.

HAEMATOLOGY
After 10 exposures, rats at the high dose level had higher than normal leukocyte counts, with 3/10 rats having counts outside of the norm established by controls.

CLINICAL CHEMISTRY
High dose level rats tended to have elevated serum glutamic-oxalacetic acid, glutamic-pyruvic transaminase, and depressed levels in alkaline phosphatase, and serum creatinine.
After a 13-day observation period serum glutamic oxalacetic acid transaminase, creatinine, and total protein levels were statistically lower than controls, however, these effects were not considered clinically significant. Low levels in total protein and glutamic oxalacetic acid transaminase are not of toxicological relevance due to the absence of a treatment-related effect for total protein during treatment and due to a non-consistent direction of change for glutamic oxalacetic acid transaminase.
Furthermore, changes seen on glutamic-pyruvic transaminase and alkaline phosphatase were shown to be completely reversible during the recovery period.

URINALYSIS
Rats at the high dose level had an elevated nitrogen activity as well as depressed levels in urine pH.


ORGAN WEIGHTS
Analysis of mean absolute organ weight after 10 exposures and the 13-day observation period did not reveal any dose-dependent effects. Organ/body weight ration analysis likewise did not reveal any dose related responses (Please refer to table 2, which is presented under “Remarks on results including tables and figures”).


GROSS PATHOLOGY
No gross findings were observed in control or low dose level rats. Intermediate and high dose level rats generally had ocular discharge, swollen eyelids with hair loss, and a cloudy appearance of the cornea.

HISTOPATHOLOGY:
Microscopic lesions attributable to the test compound were limited to the eyes and eyelids of rats exposed at the 2 highest exposure levels. These changes included: blepharitis, keratitis, corneal vascularisation, iritis, exflammatory exudate in the anterior and posterior chambers of the eye, haemorrhage mainly in the vitreous body of the eye and degeneration of the lens (cataract). These findings were equal in severity in the 2 high test groups but higher in incidence in the high level. The 13 day observation period allowed only slight recovery of these lesions and it is not expected that complete recovery would occur.

Dose descriptor:

LOAEL

Effect level:

0.01 mg/L air (analytical)

Sex:

male

Basis for effect level:

other: based on ocular effects

Dose descriptor:

LOAEL

Effect level:

2.73 other: mg/kg bw/d (assuming a respiratory volume of 86.4 L/rat/d and a body weight of 320 g)

Sex:

male

Basis for effect level:

other: based on ocular effects

Key result

Dose descriptor:

NOEL

Effect level:

0.001 mg/L air (analytical)

Sex:

male

Basis for effect level:

other: based on histological findings in the eyes at the next higher dose level.

Dose descriptor:

NOEL

Effect level:

0.38 other: mg/kg bw/d (assuming a respiratory volume of 86.4 L/rat/d and a body weight of 320 g)

Sex:

male

Basis for effect level:

other: based on histological findings in the eyes at the next higher dose level

Key result

Critical effects observed:

no

Table 1: Summary: Body weight (g)

 

Test Day	Dose (mg/L)
-	0	0.0014	0.0101	0.0431
1	249.70	246.90	251.80	249.56
5	237.60	243.60	237.90	240.89
10	259.30	265.70	258.80	247.44#
12	259.70	264.10	243.80#	235.56#
15	278.00	285.00	271.00	256.25+
25	327.60	337.60	329.20	321.00

 

#  Significantly different (P0.05) from control group by Dunnett test and LSD.

+  Significantly different (P0.05) from control group by LSD.

 

 

Table 2:  Organ/Body weight mean relative data of rats sacrificed after 13 day observation period.

Organ	Dose (mg/L)
-	0	0.0014	0.0101	0.0431
Heart	0.3451	0.3320	0.3287	0.3526
Lungs	0.6509	0.6132	0.6263	0.5565
Liver	3.7396	3.7591	3.8695	3.9592
Spleen	0.2508	0.2525	0.2112	0.2020
Kidney	0.7931	0.7655	0.8019	0.7929
Testis	1.0050	0.9866	0.9647	0.9665
Thymus	0.2205	0.2200	0.2088	0.1960

Conclusions:

The study and the conclusions which are drawn from it fulfill the quality criteria (validity, reliability, repeatability).
14-days LOAEC: 0.0101 mg/L (10.1 mg/m³) corresponding to 2.73 mg/kg bw/d (assuming a respiratory volume of 86.4 L/rat/d and a body weight of 320 g)
14-days NOAEC: 0.0014 mg/L (1.4 mg/m³) corresponding to 0.38 mg/kg bw/d (assuming a respiratory volume of 86.4 L/rat/d and a body weight of 320 g), based on histological findings in the eyes at the next higher dose level.
No classification and labelling in view of repeated dose inhalation toxicity is necessary according to Regulation 1272/2008/EC (CLP).

Executive summary:

Materials and methods

In a 14-day inhalation toxicity study (6 hours per day, 5 days per week), 10 male Crl:CD rats per group were exposed to dust aerosol of KMPS triple salt at exposure levels of 0, 0.0014, 0.0101 and 0.0431 mg KMPS triple salt/L (head only exposure). Animals were examined for clinical signs and mortality. Body weight was examined daily. Haematology and clinical chemistry was performed after the 10th exposure and the 13th observation day. All animals were subjected to gross pathology. Selected organs were weighed.

 

Results and discussion

During the exposure period clinical observations were severe ocular irritation at mid and high dose levels. These included: alopecia around eyes, conjunctival swelling, weeping, severe opacity, and corneal vascularisation, ulceration, and haemorrhaging. In general, a dry crusty scab formed around the eyes keeping them closed unless forced open. Also noted during exposures was a red nasal discharge from both control and test rats and slight lung noise in high level rats. Sporadic weight depression was recorded in the two highest test levels during the latter part of the exposure period. The 13 day observation period allowed only partial recovery from these ocular effects. Most of the clinical signs continued but were less severe.

Following the 10th exposure, rats at the high dose level had elevated leukocyte counts as well as elevated serum glutamic oxalactic acid, glutamic pyruvic transaminase, and urea nitrogen activities. This group also had depressed levels in urine pH, alkaline phosphatase, and serum creatinine. No other clinical pathologic changes were seen in this group. No corresponding changes in haematology or clinical chemistry were evident at the low and mid dose level.

After the observation period, serum glutamic oxalactic acid, creatitine, and total protein levels were statistically lower than controls.

Gross pathology showed significant findings in the ocular area only at the mid and high dose level. These findings included ocular discharge, swollen eyelids with hair loss, and a cloudy appearance of the cornea. Histologic changes attributable to the test compound were also noted in the eyes and blespharitis, keratitis, corneal vascularisation, iritis, exflammatory exudate in the anterior and posterior chambers of the eye, haemorrhage (mainly in the vitreous body of the eye), and degeneration of the lens (cataract). These changes were equal in severity at the mid and high dose level but the incidences were higher at the high dose level. No ocular changes were observed in rats of the low level.

In a 13-days observation period allowed these eye lesions only slightly recovered and complete reversal is not considered likely to occur.

Organ and body weight at the high dose level noted only a significant decrease in body weight on test days 10-16 and at the intermediate dose level on day 12. All rats gained weight at parallel rates during the observation period. Analysis of mean absolute organ weight and organ/body weight ratios did not reveal any dose dependent changes (Please refer to tables 1 and 2, which are presented under “Remarks on results including tables and figures”).

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

NOAEC

1.4 mg/m³

Study duration:

subacute

Species:

rat

Quality of whole database:

study conducted similar to OECD Guideline

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

KMPS triple salt was tested for repeated dose toxicity by oral administration and inhalation.

 

Repeated dose oral toxicity studies
14-days study:

The repeated dose oral toxicity of KMPS triple salt was investigated in a range finding study (similar or equivqlent to OECD Guideline 407 – Repeated 28-Day Oral Toxicity in Rodents) in male and female CD rats by daily administration of dose levels of 0, 50, 400 and 1000 mg/kg bw/day via gavage for a period of 14 days. Dose levels were based on a acute oral toxicity study (Doc. D_K1.Oral.Gav.R.DPT 567/012267/AC; IUCLID section 7.2.1). In the 14-day study, animals had free access to food and drinking water during the study. Clinical signs and mortality/viability were observed at regular intervals following dosing each day. Body weights recorded daily during treatment Week 1 and before necropsy.
On termination of the study a macroscopic examination of all animals was performed.
In the 14-day dose-range-finding study, there were no treatment related effects observed on the recorded parameters.

13-weeks study:
The subchronic toxicity of KMPS triple salt was assessed in a 90-day study according OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity in Rodents). Oral administration to male and female Crl:CD(SD) IGS BR rats for a period of 13 weeks was performed by gavage in water as the vehicle. Dose levels administered were 0, 25, 200 and 1000 mg/kg bw/day (dose volume: 10 ml/kg bw/day). Following 4 weeks of treatment at 1000 mg/kg bw/day, this dosage level was reduced to 600 mg/kg bw/day due to 4 unscheduled deaths and adverse clinical signs.
Histopathological change was evident in animals receiving 1000/600 mg/kg bw/day for 13 weeks.
The target organ was identified as the stomach with inflammatory, degenerative and hyperplastic changes mainly in the nonglandular stomach, the principal findings of which were inflammation, epithelial necrosis and ulceration, and epithelial hyperplasia in the nonglandular stomach.
No pathological changes related to treatment were identified in animals receiving 200 mg/kg bw/day.
Therefore, the NOAEL was determined as 200 mg/kg bw/d.

Repeated dose inhalation
The repeated dose inhalation toxicity was tested in male rats in a study similar or equivalent to OECD Guideline 412 (Repeated Dose Inhalation Toxicity: 28/14-Day). Despite some deviations to the guideline (only male rats were used, particle size was not analysed, no individual animal data available, food consumption not investigated), the study was considered valid.
In a 14-day inhalation toxicity study (6 hours per day, 5 days per week), 10 male Crl:CD rats per group were exposed to dust aerosol of KMPS triple salt at exposure levels of 0, 0.0014, 0.0101 and 0.0431 mg KMPS triple salt/L (head only exposure). Animals were examined for clinical signs and mortality. Body weight was examined daily. Haematology and clinical chemistry was performed after the 10th exposure and the 13th observation day. All animals were subjected to gross pathology. Selected organs were weighed.
The following results were observed in this study:
14-days LOAEC: 0.0101 mg/L (10.1 mg/m³) corresponding to 2.73 mg/kg bw/d (assuming a respiratory volume of 86.4 L/rat/d and a body weight of 320 g)
14-days NOAEC: 0.0014 mg/L (1.4 mg/m³) corresponding to 0.38 mg/kg bw/d (assuming a respiratory volume of 86.4 L/rat/d and a body weight of 320 g), based on histological findings in the eyes at the next higher dose level.

 

Local effects were restricted to the eyes

Gross pathology showed significant findings in the ocular area only at the mid and high dose level. These findings included ocular discharge, swollen eyelids with hair loss, and a cloudy appearance of the cornea. Histologic changes attributable to the test compound were also noted in the eyes and blespharitis, keratitis, corneal vascularisation, iritis, exflammatory exudate in the anterior and posterior chambers of the eye, haemorrhage (mainly in the vitreous body of the eye), and degeneration of the lens (cataract). These changes were equal in severity at the mid and high dose level but the incidences were higher at the high dose level. No ocular changes were observed in rats of the low level.

Justification for classification or non-classification

Classification, Labelling, and Packaging Regulation (EC) No 1272/2008

The available experimental test data are reliable and suitable for classification purposes under Regulation (EC) No 1272/2008. As a result the substance is not considered to be classified for repeated dose toxicity under Regulation (EC) No 1272/2008, as amended for the seventeenth time in Regulation (EU) 2021/849.