Barium bis(dihydrogenorthophosphate)

Endpoint summary

Description of key information

Skin irritation (OECD 402 including dermal irriation scoring), rat: not irritating

Eye irritation (OECD 437), in vitro: Eye Dam. Cat. 1 (H318)

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin corrosion: in vitro / ex vivo

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

the study does not need to be conducted because an acute toxicity study by the dermal route does not indicate skin irritation up to the relevant limit dose level (2 000 mg/kg body weight)

Reason / purpose for cross-reference:

data waiving: supporting information

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

7 October 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage)

Version / remarks:

adopted July 2013

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Hess. Ministerium für Umwelt, Klimaschutz, Landwirtschaft und Verbraucherschutz, Wiesbaden, Germany

Species:

cattle

Strain:

not specified

Details on test animals or tissues and environmental conditions:

SOURCE OF COLLECTED EYES
- Source: Schlachthof Aschaffenburg, Aschaffenburg, Germany
- Characteristics of donor animals: at least 9 months old
- Storage, temperature and transport conditions of ocular tissue: The isolated eyes were stored in HBSS containing 1% (v/v) Penicillin/Streptomycin (100 units/mL penicillin and 100 μg/mL streptomycin) in the cooled slaughterhouse until transportation on the same morning to the laboratory using a Styrofoam box. The corneae were isolated on the same day after delivery of the eyes.
- Indication of any existing defects or lesions in ocular tissue samples: no
- Indication of any antibiotics used: streptomycin/penicilline

Vehicle:

physiological saline

Controls:

yes, concurrent vehicle

yes, concurrent positive control

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied: 0.75 mL
- Concentration: 20%

VEHICLE
- Amount(s) applied: 0.75 mL
- Concentration (if solution): saline (0.9% NaCl in deionised water)

Duration of treatment / exposure:

240 minutes

Duration of post- treatment incubation (in vitro):

none

Number of animals or in vitro replicates:

3

Details on study design:

SELECTION AND PREPARATION OF CORNEAS: All eyes were carefully examined macroscopically for defects. Those presenting defects such as vascularization, pigmentation, opacity and scratches were discarded. The cornea was carefully removed from the eye using scalpel and rounded scissors. A rim of about 2 mm of tissue (sclera) was left for stability and handling of the isolated cornea. The corneae were directly used in the BCOP test on the same day.

QUALITY CHECK OF THE ISOLATED CORNEAS: The basal opacity of all corneae was recorded. Each cornea with a value of the basal opacity > 7 was discarded.

NUMBER OF REPLICATES: 3

NEGATIVE CONTROL USED: none

SOLVENT CONTROL USED: Saline (0.9% NaCl in deionised water)

POSITIVE CONTROL USED: 10% (w/v) Benzalkonium chloride in 0.9% (w/v) NaCl (saline)

APPLICATION DOSE AND EXPOSURE TIME: The test item was tested as a 20% suspension (w/v) in saline. 0.75 mL were applied on each surface of the cornea and incubated for 240 minutes.

TREATMENT METHOD: closed chamber

POST-INCUBATION PERIOD: no

REMOVAL OF TEST SUBSTANCE
- Number of washing steps after exposure period: the test item or control items, respectively, were rinsed off from the application side with saline


METHODS FOR MEASURED ENDPOINTS:
- Corneal opacity: measured with opacitometer
- Corneal permeability: passage of sodium fluorescein dye measured with the aid of microtiter plate reader at 490 nm (OD490)

SCORING SYSTEM: In Vitro Irritancy Score (IVIS)
The following formula is used to determine the IVIS of the negative control:
IVIS = opacity value + (15 x OD490 value)
The following formula is used to determine the IVIS of the positive control and the test item:
IVIS = (opacity value – opacity value mean negative control) + (15 x corrected OD490 value)
The mean IVIS value of each treated group is calculated from the IVIS values.

DECISION CRITERIA: decision criteria as indicated in the OECD 437 was used;
IVIS <= 3: no category
IVIS > 3 and >= 55: no prediction can be made
IVIS > 55: serious eye damage according to CLP/GHS (Cat 1)

Irritation parameter:

in vitro irritation score

Run / experiment:

240 minutes

Value:

131.94

Vehicle controls validity:

valid

Remarks:

IVIS = 1.43

Negative controls validity:

not examined

Positive controls validity:

valid

Remarks:

IVIS = 122.17

Remarks on result:

positive indication of irritation

Other effects / acceptance of results:

OTHER EFFECTS:
- Visible damage on test system: no

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: yes
- Acceptance criteria met for positive control: yes
- Range of historical values if different from the ones specified in the test guideline: The historical data of the IVIS of the negative control spaced in a range of 0.73 – 2.04 with a mean IVIS of 1.32. The historical data of the IVIS of the positive control spaced in a range of 98.49 – 167.85 with a mean IVIS of 122.01.

Table 1: Results after 240 minutes incubation time

Test Group	Opacity value = Difference (t240-t0) of Opacity		Permeability at 490 nm (OD490)		IVIS	Mean IVIS	Proposedin vitroIrritancy Score
		Mean		Mean
Negative Control	1	0.33	0.074	0.073	2.11	1.43	No Category
	0		0.073		1.10
	0		0.072		1.08
Positive Control	118.67*		0.043*		119.31	122.17	Category 1
	124.67*		0.098*		126.14
	119.67*		0.092*		121.05
Test Item	124.67*		0.050*		125.42	131.94	Category 1
	127.67*		0.036*		128.21
	141.67*		0.036*		142.21

* corrected values

Interpretation of results:

other: Category 1 based on CLP GHS criteria (EU criteria according to Regulation (EC) No. 1272/2008)

Conclusions:

In conclusion, according to the current study and under the experimental conditions reported, Barium bis(dihydrogenorthophosphate) is considered to induce serious eye damage (CLP/GHS Cat 1).

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irreversible damage)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin:

According to Annex VII of the Reach Regulation (EC) 1272/2008 no skin irritation study has to be conducted if an acute toxicity study by the dermal route does not indicate skin irritation up to the limit dose level (2 000 mg/kg bw). The available acute toxicity study by dermal route up to the limit dose of 2000 mg/kg bw in rats (LPT, 2017) with the test substance did not show any skin irritation throughout the observation period. Thus it is conlcuded, that the test substance is not a skin irritant.

Eye:

The eye irritation potential of the test substance was determined in a bovine corneal opacity and permeability test (BCOP test) according to OECD Guideline 437 and in compliance with GLP (Envigo, 2017). After a first opacity measurement of the fresh bovine corneae, 750 µL of a 20% suspension (test substance in saline) was applied directly to the epithelial surface of three cattle corneae in an incubation chamber in horizontal position for 240 min at 32 ± 1°C. After the incubation phase the test substance was rinsed from the corneae. Afterwards, opacity was measured a second time. In addition, the permeability of the corneae was determined by measuring spectrophotometrically the transfer of sodium fluorescein after incubation in a horizontal position for 90 ± 10 min at 32 ± 1°C. The results of the opacity and permeability measurement of the test substance were used to calculate an in vitro irritation score (IVIS) of 131.94. With the negative control saline neither an increase of opacity nor permeability of the corneae could be observed. The mean IVIS of the negative control was 1.43. The mean IVIS of the positive control (benzalkonium chloride) was 122.17. Based on the results, the test substance was damaging to the eye under the conditions of the test.

Justification for classification or non-classification

The available data on skin irritation does not meet the criteria for classification according to Regulation (EC) 1272/2008, and is therefore conclusive but not sufficient for classification. The available data on eye irritation meets the criteria for classification according to Regulation (EC) 1272/2008, and is therefore classfied as Eye Dam. Cat. 1 (H318).