Reaction mass of calcium dihydroxide and calcium peroxide

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

23/03/2018 - 23/04/2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.3 (Acute Toxicity (Dermal))

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

fixed dose procedure

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Calcium peroxide

EC Number:

215-139-4

EC Name:

Calcium peroxide

Cas Number:

1305-79-9

Molecular formula:

CaO2

IUPAC Name:

calcium peroxide

Constituent 2

Reference substance name:

Calcium dihydroxide

EC Number:

215-137-3

EC Name:

Calcium dihydroxide

Cas Number:

1305-62-0

Molecular formula:

CaH2O2

IUPAC Name:

calcium dihydroxide

Test material form:

solid: particulate/powder

Test animals

Species:

rat

Strain:

Wistar

Remarks:

Crl: WI(Han)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 10-11 weeks old.
- Weight at study initiation: 192 to 212 g.
- Housing: On arrival, animals were group housed (up to 5 animals of the same sex together) in polycarbonate cages (Makrolon MIV type; height 18 cm.) and following assignment to the study, animals were individually housed in polycarbonate cages (Makrolon MIII type; height 18 cm.) containing sterilized sawdust as bedding material (Lignocel S 8-15, JRS - J.Rettenmaier & Söhne GmbH + CO. KG, Rosenberg, Germany) equipped with water bottles.
- Diet: Pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany) ad libitum.
- Water: tap-water from public distribution system ad libitum.
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 - 21
- Humidity (%): 43 - 51
- Air changes (per hr): at least 10
- Photoperiod (hrs dark / hrs light): 12 / 12

Administration / exposure

Type of coverage:

other: non-occlusive

Vehicle:

CMC (carboxymethyl cellulose)

Details on dermal exposure:

TEST SITE
- Area of exposure: at least 10% of the body surface area
- % coverage: at least 10% of the body surface area
- Type of wrap if used: non-occlusive porous gauze dressing (50x50 mm2 non-woven swab, 4-layer patch, MEDISTOCK) secured in position with a strip of surgical adhesive tape (50 mm wides hypoallergenic micropore™ adhesive tape from 3M).

REMOVAL OF TEST SUBSTANCE
- Washing: the gauze dressings were removed and the treated area was rinsed with distilled water.
- Time after start of exposure: 24 h

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 10 mL/kg bw
- Concentration (if solution): 0.2 g/ml
- Constant volume or concentration used: yes
- For solids, paste formed: no

VEHICLE
- Amount(s) applied (volume or weight with unit): 10 mL
- Concentration (if solution): 1% aqueous CMC
- Lot/batch no. (if required): Genfarma, Zaandam (NL)
- Rationale: selected based on the results of trial preparation of formulations.

Duration of exposure:

24 h

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

5 males and 5 females per dose

Control animals:

yes

Remarks:

control group referred to historical values.

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: daily observations, the animals were weighed on day 0 (just before administering the test item) then on day 2, day 7, and day 14.
- Necropsy of survivors performed: yes. Observed organs: oesophagus, stomach, duodenum, jejunum, ileum, caecum, colon, rectum, spleen, liver, thymus, trachea, lungs, heart, kidneys, urinary bladder, testicles, adrenals, pancreas. As no abnormalities were observed, no microscopic examinations were performed.
- Other examinations performed: clinical signs, body weight. Histopathological examination was not performed because no macroscopic anomalies were observed in any organ.

Results and discussion

Preliminary study:

One animal was dosed at 1000 mg/kg in order to select the dose causing no mortality or significant toxicity to be used in the main study. Based on the results, one additional animal was dosed at 1000 mg/kg 48h after the first. Based on the results of the range finding study, two animals were dosed at 2000 mg/kg.

Effect levels

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred during the study.

Clinical signs:

other: At 2000 mg/kg, fissures, scales, scabs, general erythema and/or white staining were seen in the treated skin-area of the animals between Days 2 and 15. Scabs and/or focal erythema were noted on the back of two animals between Days 8 and 15. Chromodacryor

Gross pathology:

No abnormalities were found at macroscopic post mortem examination of the animals.

Any other information on results incl. tables

Table 1 MORTALITY DATA

TEST DAY HOURS AFTER TREATMENT	1 0	1 2	1 4	2	3	4	5	6	7	8	9	10	11	12	13	14	15
FEMALES 1000 MG/KG	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
FEMALES 2000 MG/KG	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
FEMALES 2000 MG/KG	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-

Table 2 CLINICAL SIGNS

TEST DAY HOURS AFTER TREATMENT	MAX GRADE	1 0	1 2	1 4	2	3	4	5	6	7	8	9	10	11	12	13	14	15
FEMALES 1000 MG/KG
ANIMAL 1
Skin / fur
White staining (Treated skin)	(1)	-	-	-	1	-	-	-	-	-	-	-	-	-	-	-	-	-
Secretion / excretion
Chromodacryorrhoea (Snout)	(3)	-	1	1	-	-	-	-	-	-	-	-	-	-	-	-	-	-
FEMALES 2000 MG/KG
ANIMAL 2
Skin / fur
Erythema focal (Back)	(4)	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	1
Fissures (Treated skin)	(3)	-	-	-	-	-	1	1	-	-	-	-	-	-	-	-	-	-
Scales (Treated skin)	(3)	-	-	-	-	-	-	-	1	1	-	-	-	-	-	-	-	-
Scabs (Back)	(3)	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	1	-
White staining (Treated skin)	(1)	-	-	-	1	-	-	-	-	-	-	-	-	-	-	-	-	-
Various
Red (Treated skin)	(1)	-	-	-	-	-	1	1	1	1	-	-	-	-	-	-	-	-
FEMALES 2000 MG/KG
ANIMAL 3
Skin / fur
General erythema (Treated skin)	(4)	-	-	-	-	1	1	1	1	1	-	-	-	-	-	-	-	-
Scabs (Back)	(3)	-	-	-	-	-	-	-	-	-	1	1	1	1	1	1	1	1
White staining (Treated skin)	(1)	-	-	-	1	-	-	-	-	-	-	-	-	-	-	-	-	-
Secretion / excretion
Chromodacryorrhoea (Nose)	(3)	-	1	1	-	-	-	-	-	-	-	-	-	-	-	-	-	-
ANIMAL 4
Skin / fur
General erythema (Treated skin)	(4)	-	-	-	-	1	1	1	1	1	1	1	1	1	1	1	-	-
Scabs (Treated skin)	(3)	-	-	-	-	-	-	-	-	-	1	1	1	1	1	1	1	1
White staining (Treated skin)	(1)	-	-	-	1	-	-	-	-	-	-	-	-	-	-	-	-	-
Secretion / excretion
Chromodacryorrhoea (Nose)	(3)	-	1	1	-	-	-	-	-	-	-	-	-	-	-	-	-	-
- = Sign not observed

 

Table 3 BODY WEIGHTS (GRAM)
SEX/DOSE LEVEL	ANIMAL	DAY 1	DAY 8	DAY 15
FEMALES 1000 MG/KG
	1	200	199	218
	MEAN	200	199	218
	ST.DEV.	---	---	---
	N	1	1	1
FEMALES 2000 MG/KG
	2	192	192	211
	MEAN	192	192	211
	ST.DEV.	---	---	---
	N	1	1	1
FEMALES 2000 MG/KG
	3	207	180	224
	4	212	185	218
	MEAN	210	183	221
	ST.DEV.	4	4	4
	N	2	2	2

Table 4 MACROSCOPIC FINDINGS
ANIMAL	ORGAN	FINDING	DAY OF DEATH
FEMALES 1000 MG/KG
1		No  findings noted	Scheduled necropsy
			Day 15 after treatment
FEMALES 2000 MG/KG
2		No  findings noted	Scheduled necropsy
			Day 15 after treatment
FEMALES 2000 MG/KG
3		No  findings noted	Scheduled necropsy
			Day 15 after treatment
4		No  findings noted	Scheduled necropsy
			Day 15 after treatment

Table 5 IRRITATION
SEX/DOSE LEVEL	ANIMAL	24 Hours		48 Hours		72 Hours
		Erythema	Oedema	Erythema	Oedema	Erythema	Oedema
FEMALES 1000 MG/KG
	1	0	0	0	0	0	0
FEMALES 2000 MG/KG
	2	0	0	0	0	0	0
FEMALES 2000 MG/KG
	3	0	0	0	0	0	0
	4	0	0	0	0	0	0

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Remarks:

EU criteria.

Conclusions:

The dermal LD50 value of the test item in Wistar rats was established to exceed 2000 mg/kg body weight.

Executive summary:

To determine the acute dermal toxicity of the test item in rats, a limit test was performed, according to OECD 402 (GLP study). Initially, the test item was administered to single female Wistar rats by a single dermal application at 1000 mg/kg and 2000 mg/kg body weight for 24 hours in a range finding study.  Based on the results, the main study was performed by dosing two females at 2000 mg/kg.  All animals were subjected to daily observations and weekly determination of body weight for 14 days.  Macroscopic examination was performed on the day of death or after terminal sacrifice (Day 15). No mortality ocurred at any dose tested. No significant signs of systemic toxicity were noted at 1000 mg/kg. At 2000 mg/kg, fissures, scales, scabs, general erythema and/or white staining were seen in the treated skin-area of the animals between Days 2 and 15.  Scabs and/or focal erythema were noted on the back of two animals between Days 8 and 15. Chromodacryorrhoea of the nose was noted for two animals on Day 1. Based on the study results, the dermal LD50 value of the test item in Wistar rats was established to exceed 2000 mg/kg body weight.