Manganese antimony titanium buff rutile

Endpoint summary

Description of key information

oral
LD50 rat > 5000 mg/kg bw (standardized test protocol; BASF 1985a, 1985 b);

dermal
not relevant

inhalation
LC50 > 2.199 mg/L for both sexes.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Comparable to guideline study with acceptable restrictions (purity unknown, non-GLP)

Principles of method if other than guideline:

BASF Test
The study was conducted according to an internal BASF method which in principle is comparable to the OECD Guideline 401.
A test group consisting of 5 animals/sex was treated by single gavage with an aqueous solution of the test substance; the concentration of test substance in vehicle (w/v) was 50% and the application volume was 10 ml/ kg bw.
The animals were observed for mortality and for clinical symptoms of toxicity. They were weighed prior treatment and thereafter on day 3, day 7 and day 13 post-treatment. At the end of the observation period of 14 days, the surviving animals were sacrificed for the purpose of necropsy.

GLP compliance:

no

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Dr. Thomae GmbH, Biberach, Germany
- Mean weight at study initiation: 169 g (males) and 171 g (females)
- Fasting period before study: 16 h
- Housing: 5 per cage (Stainless steel wire mesh cages)
- Diet: Kliba Labordiaet ad libitum
- Water: tap water ad libitum
- Acclimation period: at least 1 week


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24
- Humidity (%): 30-70
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: gavage

Vehicle:

CMC (carboxymethyl cellulose)

Details on oral exposure:

VEHICLE
- Concentration in vehicle: Test substance formulation with 0.5% aqueous carboxymethyl cellulose
- Justification for choice of vehicle: Aqueous formulation corresponds to the physiological medium


MAXIMUM DOSE VOLUME APPLIED: 10 ml

Doses:

5000 mg/kg

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: observations at least once each workday, weighing at d0, d3, d7 and d 13
- Necropsy of survivors performed: no

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Remarks on result:

other: 1% confidence level

Sex:

male/female

Dose descriptor:

other: NOAEL

Effect level:

>= 5 000 mg/kg bw

Mortality:

none

Clinical signs:

other: no abnormalities observed

Gross pathology:

no abnormalities observed

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Quality of whole database:

Two valid almost identical limit tests were available, which were both performed according to a standardized internal method which is in principle comparable to the OECD TG 401 but did not follow GLP requirements (BASF 1985a, 1985b).

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2021-01-27 to 2021-03-23

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

At least three concentrations or a limit test should be used in the traditional protocol, only one concentration (2.199 mg/L) was used in the study. A sighting study was not performed. Oxygen and carbon dioxide concentration were not measured. The relative humidity should be in the range of 30-70 %, throughout the exposure, the relative humidity was 15.8 ± 0.8 %. Method of randomisation of the animals and the pressure difference in the inhalation chamber were not stated.

Qualifier:

according to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Version / remarks:

2009

Deviations:

yes

Remarks:

please refer to the field `Rationale for reliability incl. deficiencies´.

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1300 (Acute inhalation toxicity)

Version / remarks:

1998

Qualifier:

according to guideline

Guideline:

other: COMMISSION REGULATION (EU) No 260/2014

Version / remarks:

2014

GLP compliance:

yes (incl. QA statement)

Remarks:

signed, 2020-04-03

Test type:

traditional method

Limit test:

no

Specific details on test material used for the study:

- Appearance, physical state, colour: solid, brown
- Storage conditions: room temperature

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 Laboratories, Research Models and Services, Germany GmbH, Sandhofer Weg 7, 97633 Sulzfeld, Germany
- Females nulliparous and non-pregnant: yes
- Age at study initiation: young adult ; males: 61 - 65 d; females 75 - 79 d
- Weight at study initiation: males: 248.0 - 257.8 g, mean 253.7 ± 5.1 g; females: 196.4 - 219.4, mean 212.5 ± 10.2 g
- Housing: Single housing (if the animals showed clinical signs and findings) or up to 5 animals (caged in groups, if the animals were free from clinical signs and findings)
- Historical data: not specified
- Diet (ad libitum): Mouse and rat maintenance diet, GLP, 12 mm pellets, Granovit AG, Kaiseraugst, Switzerland
- Water (ad libitum): tap water
- Acclimation period: at least 5 d

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 – 24
- Humidity (%): 45 – 65
- Air changes (per hr): 5
- Photoperiod (hrs dark / hrs light): 12 (6.00 p.m. - 6.00 a.m.) / 12 (6.00 a.m. - 6.00 p.m.)

Route of administration:

inhalation: dust

Type of inhalation exposure:

nose only

Vehicle:

clean air

Mass median aerodynamic diameter (MMAD):

>= 1.11 - <= 1.12 µm

Geometric standard deviation (GSD):

>= 2.41 - <= 2.68

Remark on MMAD/GSD:

please refer to the field `Any other information on results incl. tables´

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: nose-only inhalation system INA 10 (glass-steel construction, BASF SE): the animals were restrained in glass tubes and their snouts projected into the inhalation system.
- Exposure chamber volume: V≈ 34 L
- Method of holding animals in test chamber: the animals were restrained in glass tubes and their snouts were projected into the inhalation system (nose-only inhalation system INA 10).
- Source and rate of air (airflow): 1.5 m³/h (compressed air, from a central air-conditioning system), exhaust air flow: 1.3 m³/h.
- Method of conditioning air: cold (15 °C) air passes through an activated charcoal filter, is adjusted to room temperature of 20 to 24 °C and passes through a second particle filter (H13 (HEPA) Camfil Farr, Germany). The generated conditioned air was used to generate inhalation atmospheres.
- Method of compressioning air: air is filtered by an inlet air strainer and introduced into the oil-free compressor (HT 6, Josef Mehrer GmbH & Co KG, Germany). After passing through a second ultra-filter (SMF 5/3, 108 mm, Donalson), the compressed air (15 bar) is stored in a storage of 1500 or 5000 L. The compressed air is conducted to the laboratories via pipes, where the pressure is reduced to 5 - 6 bar. In the laboratory, the compressed air can be taken as required.
- System of generating particulates/aerosols: Equipment: balance (Mettler AT250), Flat-Tray Feeder (ZD 5 FB-C-1M-50, Three-Tec GmbH, Seon, Switzerland), Aerosol mixing tube (stainless steel; BASF SE, Ludwigshafen, Germany)
Generation technique: the test substance was dosed unchanged, and the dust aerosol was generated inside the inhalation system with the above-mentioned dust generator and compressed air. The concentrations were adjusted by varying the feed of the double concave screw.
- Method of particle size determination: the particle size was determined with a 7 stages cascade impactor (29.5 µm, 18.2 µm, 8.5 µm, 5.5 µm, 2.8 µm, 1.2 µm). MMADs and GSD were calculated based on the collected [mg] and cumulative [%] masses at each stage.
Equipment: Stack Sampler Mark III (Andersen), vacuum pump (Millipore), sampling probe (internal diameter 6.9 mm), limiting orifice 3 L/min, balance: Sartorius MSA6.6S-000-DF.
The impactor was assembled with preweighed glass-fiber collecting discs, equipped with a backup particle filter, and connected to the vacuum pump. Two samples were taken from the breathing zone of the animals. After sampling the impactor was taken apart. The collecting discs and the backup particle filter were re-weighed.
- Treatment of exhaust air: the exhaust air was filtered and conducted into the exhaust air of the building. The exposure system was located inside an exhaust cabin in an air-conditioned laboratory.
- Temperature, humidity, pressure in air chamber: supply air: 1.5 ± 0.0 m³/h; exhaust air 1.3 ± 0.0 m³/h; test substance flow 21.0 g/h; temperature: 23.4 ± 0.2 °C; relative humidity 15.8 ± 0.8 %.

TEST ATMOSPHERE
- Brief description of analytical method and equipment used: Vacuum pump (Millipore)
Sampling devices: filtration equipment with probe, internal diameter: 7 mm, (Millipore); filter: MN 85/90 BF (d = 47 mm); equipment: Balance Sartorius MSA6.6S-000-DF
Sampling position: immediately adjacent to the animals' noses at a separate spare port
Sampling flow: 3 L/min
Sampling velocity: 1.25 m/s
Sampling volume: 15 L
Sampling frequency: 4 samples at about hourly intervals
Preweighed filters were placed into the filtration equipment. By means of the vacuum pump metered volumes of the dust were drawn through the filter. For each sample the dust concentration in mg/L was calculated from the difference between the preweight of the filter and the weight of the filter after sampling, with reference to the sample volume of the inhalation atmospheres. Mean and standard deviation were calculated for the concentration from the results of the individual measurements.
- Samples taken from breathing zone: yes
- Time needed for equilibrium of exposure concentration before animal exposure: at least 30 min.

TEST ATMOSPHERE
- Particle size distribution: the particle size distribution at different passages were reported in the field `Any other information on results incl. tables´, the D10, D50 and D90 vales were note determined,
- MMAD (Mass median aerodynamic diameter) / GSD (Geometric st. dev.): two measurements: MMAD 1: 1.11 µm, GSD 1: 2.41; MMAD 2: 1.13 µm, GSD 2 : 2.68

Analytical verification of test atmosphere concentrations:

yes

Remarks:

see above `Details on inhalation exposure´

Duration of exposure:

4 h

Concentrations:

The following mean analytical concentrations were obtained:
- Mean: 2.199 mg/L
- Standard deviation: 0.244
- Nominal concentration: 14.0 mg/L

No. of animals per sex per dose:

Five per sex

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations: once during the pre- exposure day (d -1) and from day 0 – day 14, hourly observation during exposure
- Frequency of weighing: day -1, 0, 1, 3 and 7, and before the sacrifice of the animals at the end of the observation period
- Necropsy of survivors performed: yes , at the end of the observation period the animals were sacrificed with CO2-inhalation in a chamber with increasing concentration over time, and were subjected to gross-pathological examination
- Mortality: check for dead or moribund animal was made twice each workday and once on Saturdays, Sundays and on public holidays.

Statistics:

CALCULATIONS OF LC50 AND PARTICLE SIZE DISTRIBUTION
In this study, only one concentration was tested, at which no animals died. The result belonged to the type ”LC50 greater than”. For the statistical analysis the Binomial-Test [1] was performed. The calculation of the particle size distribution was carried out in the inhalation laboratory on the basis of mathematical methods for evaluating particle measurements [2].

[1] Steel R.G.D. , Torrie J.H. (1984): Principles and procedures of statistics a biometrical approach. McGraw-Hill
[2] DIN 66141: Darstellung von Korngrößenverteilungen, DIN 66161: Partikelgrößenanalyse (Beuth-Vertrieb GmbH, Berlin 30, FRG und Köln 1, FRG)

Preliminary study:

not specified

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 2.199 mg/L air (analytical)

Based on:

test mat.

Exp. duration:

4 h

Remarks on result:

other: St.dev.: 0.244 mg/L

Remarks:

the LC50 value is statistically significant to a level of 99 %.

Sex:

male

Dose descriptor:

LC50

Effect level:

> 2.199 mg/L air (analytical)

Based on:

test mat.

Exp. duration:

4 h

Remarks on result:

other: St.dev.: 0.244 mg/L

Remarks:

the LC50 value is statistically significant to a level of 95 %.

Sex:

female

Dose descriptor:

LC50

Effect level:

> 2.199 mg/L air (analytical)

Based on:

test mat.

Exp. duration:

4 h

Remarks on result:

other: St.dev.: 0.244 mg/L

Remarks:

the LC50 value is statistically significant to a level of 95%

Mortality:

No mortality was observed in male and female animals during the study period.

Clinical signs:

other: Please refer to the field `Any other information on results incl. tables´

Body weight:

The mean body weights of the animals decreased slightly on the first post-exposure observation day but increased thereafter. This is a typical finding for this test design.

Gross pathology:

No gross pathological findings were detected during the necropsy in the animals at the termination of the post-exposure period.

Particle size distribution, MMAD & GSD

 

Table1: Particle size analysis 1

Size range [µm]	Collected mass [mg]	Cumulative mass [%]
<1.2	6.717	32.82
1.2-1.8	12.138	92.11
2.8-5.5	1.404	98.97
5.5-8.5	0.113	99.53
8.5-18.2	0.002	99.54
18.2-29.5	0.095	100
<29.5	0	100

MMAD: 1.11 µm, GSD: 2.41

Cumulative concentration: 1.36 mg/L

Calculated mass below 1 µm: 45.5 %

Calculated mass below 3 µm: 87.2 %

 

Table 2: Particle size analysis 2

Size range [µm]	Collected mass [mg]	Cumulative mass [%]
<1.2	8.055	32.29
1.2-1.8	14.526	90.52
2.8-5.5	1.810	97.78
5.5-8.5	0.278	98.89
8.5-18.2	0	98.89
18.2-29.5	0.277	100
<29.5	0	100

MMAD: 1.13 µm, GSD: 2.68

Cumulative concentration: 1.66 mg/L

Calculated mass below 1 µm: 45.0 %

Calculated mass below 3 µm: 83.8 %

[MMAD (mass median aerodynamic diameter) is the calculated aerodynamic diameter which divides the size distribution in half when measured by mass. Geometrical standard deviation (GSD) is the ratio of the estimated 84 percentile to the 50 percentile and indicates the slope of the cumulative particle size distribution curve.]

 

Clincal signs:

Clinical signs of toxicity in animals comprised accelerated respiration, piloerection, substance contaminated and discolored fur substance like. Findings were observed from hour 1 of exposure through study day 1. No clinical signs and findings were observed from study day 2 onwards.

 

Table 3: Duration of signs

	Male animals	Female animals
Fur, substance-contaminated	d0 – d1	d0
Fur, discolored, substance-like	d1	d1
Respiration, accelerated	h1 – d1	h1 – d0
Piloerection	d0	d0

 

Interpretation of results:

study cannot be used for classification

Conclusions:

Under the current study conditions, the LC50 value was > 2.199 mg/L in male and female Wistar rats after a 4-hour inhalation exposure to the dust aerosol of Pigment Yellow 164 (Manganese antimony titanium buff rutile).

Executive summary:

An acute inhalation toxicity test was performed according to the OECD guideline 403 (2009) with some deviations. Five male and five female rats were exposed to a dust aerosol of manganese antimony titanium buff rutile for 4 hours. At least three concentrations or a limit test should be used according to the guideline, but only one concentration (2.199 mg/L) was tested in the study. A sighting study was not performed. Oxygen and carbon dioxide concentration during the exposure were not measured. The relative humidity should be in the range of 30-70 %, throughout the exposure, the relative humidity was 15.8 ± 0.8 %. The method of randomisation of the animals and the pressure difference in the inhalation chamber were not stated. Futhermore, a second device for the particle measurement was not used. The test animals were observed for 14 days. No mortality was observed in male and female animals during the study period. Clinical signs of toxicity in animals comprised accelerated respiration, piloerection, substance contaminated and discoloured fur substance like. Findings were observed from hour 1 of exposure through study day 1. No clinical signs and findings were observed from study day 2 onwards. The mean body weights of the animals decreased slightly on the first post-exposure observation day but increased thereafter. No gross pathological findings were detected during the necropsy in the animals at the termination of the post-exposure period.

Under the current study conditions, the LC50 value was > 2.199 mg/L in male and female Wistar rats after a 4-hour inhalation exposure to the dust aerosol of Pigment Yellow 164 (Manganese antimony titanium buff rutile).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LC50

Value:

> 2.199 mg/L air

Physical form:

inhalation: dust / mist

Quality of whole database:

The study cannot be used for classification based on the fact, that the Category 4 in the CLP Regulation (1272/2008) goes up to a concentration of 5 mg/L and not only up to the tested concentration.

Additional information

Oral

For the oral exposure pathway, two valid almost identical limit tests were available, which were both performed according to a standardized internal method which is in principle comparable to the OECD TG 401 but did not follow GLP requirements (BASF 1985a, 1985b). Since there were no mortalities, clinical signs or necropsy findings observed in male and female rats dosed with 5000 mg/kg bw after the 14 d observation period, the acute oral LD50 is > 5000 mg/kg bw and the LD0 is >= 5000 mg/kg bw in rats.

Inhalation

Five male and five female rats were exposed to a dust aerosol of manganese antimony titanium buff rutile for 4 hours at a mean concentration of 2.199 mg/L. The test animals were observed for 14 days. No mortality was observed in male and female animals during the study period. Clinical signs of toxicity in animals comprised accelerated respiration, piloerection, substance contaminated and discoloured fur substance like. Findings were observed from hour 1 of exposure through study day 1. No clinical signs and findings were observed from study day 2 onwards. The mean body weights of the animals decreased slightly on the first post-exposure observation day but increased thereafter. No gross pathological findings were detected during the necropsy in the animals at the termination of the post-exposure period.

Under the current study conditions, the LC50 value was > 2.199 mg/L in male and female Wistar rats after a 4-hour inhalation exposure to the dust aerosol of Pigment Yellow 164 (Manganese antimony titanium buff rutile).

The study cannot be used for classification based on the fact, that the Category 4 in the CLP Regulation (1272/2008) goes up to a concentration of 5 mg/L and not only up to the tested concentration.

 

Dermal

No data were available for acute dermal toxicity. The acute oral toxicity study did not reveal any systemic toxicity up to the limit dose, in addition the dermal irritation study in rabbits did not reveal any local or systemic intolerances. Therefore, the dermal exposure pathway is assessed as not relevant.

Justification for classification or non-classification

In the available acute oral toxicity studies no mortality was observed up to the limit dose of 5000 mg/kg bw. Hence, no classification required in accordance with EU CLP or GHS.

 

In the available acute inhalation toxicity study no mortality was observed up to the concentration of 2.199 mg/L. Only very mild treatment-related effects were observed up to 1 day post exposure, which were judged typical for this type of study and test item. Due to the complete lack of signs of toxicity following oral and inhalation exposure, it is anticipated that acute inhalation toxicity testing up to the limit concentration of 5mg/L would not have yielded a different outcome, i.e. no systemic of local effects after 4 hrs acute inhalation. The lack of local reactivity to biological membranes (such as the respiratory epithelium) of manganese antimony titanium buff rutile is also corroborated by the complete lack of irritating properties in skin and eye irritation testing in rabbits. Finally, the lack of acute inhalation toxicity for manganese antimony titanium buff rutile is in agreement with the results obtained for other members of the inorganic pigments category substances.

 

Acute dermal toxicity was not assessed in an in vivo test. The acute oral toxicity study did not reveal any systemic toxicity up to the limit dose, in addition the dermal irritation study in rabbits did not reveal any local or systemic intolerances. It is therefore anticipated that manganese antimony titanium buff rutile is not acutely toxic via the dermal route.