Sulfates of potassium, sodium and calcium, byproduct from fermentation

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 7 April 2017 to 2 June 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

yes

Remarks:

Relative humidity value (28%) outside the expected range of 30-70% was recorded at one time. This deviation was considered to have no impact on the outcome of the study.

GLP compliance:

yes (incl. QA statement)

Test type:

standard acute method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

calcium dipotassium disodium trisulfate

Molecular formula:

SO42-, K+, Na+, Ca2+

IUPAC Name:

calcium dipotassium disodium trisulfate

Test material form:

solid

Remarks:

light to beige solid

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: EXP LIberica Lote Al16503047
- Expiration date of the lot/batch: 30 November 2019
- Purity test date: 1 February 2017

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Controlled room temperature (15-25 deg Celsius, below 70% relative humidity) protected from light and humidity
- Stability under test conditions: not applicable
- Solubility and stability of the test substance in the solvent/vehicle: not applicable

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
The test item was ground to a fine powder before the application. Sufficient water was used to dampen the test material to ensure good contact with the skin.

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories, Research Models and Services, Germany
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: Young adult rats
- Weight at study initiation: Between 219 g and 266 g
- Fasting period before study: No
- Housing: Type II. polypropylene/polycarbonate
- Diet (e.g. ad libitum): ssniff® SM R/M "Autoclavable complete diet for rats and mice breeding and maintenance", ad libitum
- Water (e.g. ad libitum): tap water from the municipal supply, as for human consumption from a 500 mL bottle, ad libitum
- Acclimation period: 5 Days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20.4–24.7°C
- Humidity (%): 28–61%
- Air changes (per hr): 15-20 air exchanges/hour
- Photoperiod (hrs dark / hrs light): 12 hours daily, from 6.00 a.m. to 6.00 p.m.

IN-LIFE DATES: From: 6 April 2017 To: 25 April 2017

Administration / exposure

Type of coverage:

semiocclusive

Vehicle:

water

Details on dermal exposure:

TEST SITE
- Area of exposure: back of the animals
- % coverage: approximately 10% area of the total body surface
- Type of wrap if used: Sterile gauze pads maintened with adhesive hypoallergenic plaster.The entire trunk was wrapped with semi occlusive plastic wrap for 24 hours

REMOVAL OF TEST SUBSTANCE
- Washing (if done): washing with water at body temperature
- Time after start of exposure: 24 hours

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg/kg bw
- Concentration (if solution): not applicable
- Constant volume or concentration used: adjusted to animal bodyweight
- For solids, paste formed: yes

Duration of exposure:

24 hours

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

5 animals per sex per dose were used

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Clinical observations were performed on the day of treatment at 1 and 5 hours after application of the test item and once each day for 14 days thereafter. Observations included the skin and fur, eyes and mucous membranes, the respiratory, circulatory, autonomic and central nervous system, somatomotor activity and behaviour pattern. Particular attention was directed to observation of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma. The body weights were recorded on Day 0 (before the test item administration) and on Days 7 and 14 (before necropsy). Adverse skin reactions at the site of application were recorded daily following the removal of the dressing.
- Necropsy of survivors performed: yes

Results and discussion

Effect levels

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

The test item did not cause mortality at the dose level of 2000 mg/kg bw.

Clinical signs:

other: There were no systemic clinical signs noted in any animal throughout the study

Gross pathology:

There was no evidence of any gross macroscopic changes at a dose level of 2000 mg/kg bw.

Other findings:

No adverse local dermal signs were observed after treatment with the test item or during the 14 days observation period.

Any other information on results incl. tables

TABLE 1:            Clinical Observations

 

DOSE LEVEL: 2000mg/kg bw                                                                                           SEX:MALE

Cage No.

Animal No.

Observations

Observation days

Frequency

0

1

2

3

4

5

6

7

8

9

10

11

12

13

14

1h

5h

1

7970

Symptom Free

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

16/16

2

7971

Symptom Free

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

16/16

3

7972

Symptom Free

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

16/16

4

7973

Symptom Free

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

16/16

5

7974

Symptom Free

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

16/16

 

DOSE LEVEL: 2000mg/kg bw                                                                                      SEX:FEMALE

Cage No.

Animal No.

Observations

Observation days

Frequency

0

1

2

3

4

5

6

7

8

9

10

11

12

13

14

1h

5h

6

7985

Symptom Free

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

16/16

7

7986

Symptom Free

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

16/16

8

7987

Symptom Free

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

16/16

9

7988

Symptom Free

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

16/16

10

7989

Symptom Free

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

16/16

 

Remarks:         += present

h =hour(s)               Treatment day = Day0

Frequency of observation = number of occurence of observation / total number of observations

 

TABLE 2:            Body Weight Data

 

DOSE LEVEL: 2000mg/kgbw                                                                              SEX:MALE

Cage No.	Animal No.	Body weight (g) Days			Body Weight Gain (g)
		0	7	14	0-7	7-14	0-14
1	7970	253	310	369	57	59	116
2	7971	238	272	307	34	35	69
3	7972	242	305	361	63	56	119
4	7973	241	285	339	44	54	98
5	7974	234	273	313	39	40	79
Mean:		241.6	289.0	337.8	47.4	48.8	96.2
Standard deviation:		7.1	17.7	27.7	12.2	10.6	22.1

 

DOSE LEVEL: 2000mg/kgbw                                                                         SEX:FEMALE

Cage No.	Animal No.	Body weight (g) Days			Body Weight Gain (g)
		0	7	14	0-7	7-14	0-14
6	7985	264	276	298	12	22	34
7	7986	266	272	277	6	5	11
8	7987	256	270	279	14	9	23
9	7988	234	238	246	4	8	12
10	7989	219	257	260	38	3	41
Mean:		247.8	262.6	272.0	14.8	9.4	24.2
Standard deviation:		20.5	15.5	19.8	13.6	7.4	13.3

 

TABLE 3:            Macroscopic Findings

 

DOSE LEVEL: 2000mg/kgbw                                                                                      SEX:MALE

Cage No.	Animal No.	Necropsy Date/ Necropsy Day	External Observations	Internal Observations	Organ/ Tissue
1	7970	25 April 2017 Day 14	No external observations	No internal observations	Not applicable
2	7971	25 April 2017 Day 14	No external observations	No internal observations	Not applicable
3	7972	25 April 2017 Day 14	No external observations	No internal observations	Not applicable
4	7973	25 April 2017 Day 14	No external observations	No internal observations	Not applicable
5	7974	25 April 2017 Day 14	No external observations	No internal observations	Not applicable

 

DOSE LEVEL: 2000mg/kgbw                                                                                  SEX:FEMALE

 

Cage No.	Animal No.	Necropsy Date/ Necropsy Day	External Observations	Internal Observations	Organ/ Tissue
6	7985	25 April 2017 Day 14	No external observations	No internal observations	Not applicable
7	7986	25 April 2017 Day 14	No external observations	No internal observations	Not applicable
8	7987	25 April 2017 Day 14	No external observations	No internal observations	Not applicable
9	7988	25 April 2017 Day 14	No external observations	No internal observations	Not applicable
10	7989	25 April 2017 Day 14	No external observations	No internal observations	Not applicable

 

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The acute dermal median lethal dose (LD50) of the test item Sulfates of potassium, sodium and calcium, by-product from fermentation was found to be greater than 2000 mg/kg body weight in male and female Crl:WI rats. According to the CLP criteria, the test item was not classified for Acute Dermal Hazard.

Executive summary:

The purpose of this GLP compliant study was to assess the potential Acute Toxicity of the test substance Sulfates of potassium, sodium and calcium, by-product from fermentation, when applied dermally on Wistar rats, perfomed according to OECD 402 method.

A limit test  was  carried  out  at  2000 mg/kg  body  weight  (bw)  in  both sexes (5 rats/sex). The test item was powdered and was applied as a single dermal 24- hour exposure followed by a 14-day observation period.

Clinical observations were performed on all animals at 1 and 5 hours  after dosing and daily for 14 days thereafter. Body weight was measured prior to dosing on Day 0 and on Days 7 and 14 (before necropsy). Gross macroscopic examination was performed on all animals at the end of the 2-week observation period (Day 14).

Test item did not cause mortality at the dose level of 2000 mg/kg bw.

There were no systemic clinical signs noted in any animal throughout the study.

No adverse local dermal signs were observed after treatment with the test item or during the 14 days observation period.

There were no treatment related effects on body weight or body weight gain during the observation period.

There was no  evidence  of  any  macroscopic  changes  at  a  dose  level  of  2000 mg/kg bw.

The  acute  dermal  median  lethal  dose  (LD50)   of   the   test   item   Sulfates of potassium, sodium and calcium, by-product from fermentation was found to be greater than 2000 mg/kg body weight in male and female Crl:WI rats. According to the CLP criteria, the test item was not classified for Acute Dermal Hazard.