mancozeb (ISO); manganese ethylenebis(dithiocarbamate) (polymeric) complex with zinc salt  

Endpoint summary

Description of key information

In the acute oral toxicity study no adverse effects have been observed and a discriminating dose > 2000 mg/kg bw was determined for rats.

The substance was tested for inhalation toxicity in rats in a 4 h head-only inhalation study according to OECD TG 403.

The acute inhalation median lethal concentration (4hr LC50) value to rat was found to be more than the maximum achieved concentration of the test item dust aerosol viz.,1.766 mg/L of air at breathing zone.

The acute dermal median lethal dose (LD50) of the test item to Wistar rat was found to be more than 2000 mg/kg body weight.

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

Study period:

1997-10-29 to 1997-11-12

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Version / remarks:

24th February 1987

GLP compliance:

yes (incl. QA statement)

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: Breeding facility, Jai Research Foundation, India
- Weight at study initiation: 93.0 g - 118.0 g
- Fasting period before study: 17 hours prior to dosing
- Housing: Group-housed (5 animals of same sex per cage) in polypropylene rat cage
- Diet: Rat pellet feed (Amrut brand) ad libitum (MF: Nav Maharashtra Chakan Oil Mills Limited, 43, Shaniwar Peth, Pune - 411 030, Maharashtra, India)
- Water: Pure drinking water filtered through Aquaguard water filter system ad libitum
- Acclimation period: 5 days prior to dosing

ENVIRONMENTAL CONDITIONS
- Temperature (°C): between 20 ± 2.0 and 22 ± 1.8
- Humidity (%): 76 ± 6.0
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

VEHICLE
- Amount of vehicle: 10 mL/kg bw

Doses:

0 and 2000 mg/kg bw

No. of animals per sex per dose:

5

Control animals:

yes

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Deaths and signs of toxicity were recorded 1, 2 and 3 hours after administration and subsequently once daily for 14 days. Individual body weights were recorded prior to dosing and on day 7 and 14.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality was observed in 2000 mg/kg body weight dose group and control group.
The oral LD50 was determined as more than 2000 mg/kg in both sexes.

Clinical signs:

other: No toxicity symptoms were observed in any group.

Gross pathology:

After termination of experiment all animals from control as well as treated group were sacrificed and subjected to thorough gross pathological examination. External examination of carcass did not reveal gross lesions of significance.
In control group, lung showed mild congestion with pinpoint haemorrhages in one female animal and diffused pinpoint haemorrhages in another female animal.
In treated group, lung showed gross pathology such as diffused focal emphysema (1 male animal), mild congestion (1 female animal) and diffused ecchymotic congestion (1 female animal). Whereas, gross pathology in liver showed patchy congestion in one male animal and two female animals. Congested spleen was observed in one male animal and peritoneal adhesion in another male animal was observed in this study
In conclusion, the gross lesion recorded in lung in animal of control group taken as incidental. Whereas, in treated group the vascular pathology in lung and liver showed widespread lesions when compared with control group. Peritoneal adhesion were also recorded in one animal of this group. The severity and incidence of vascular changes in lung, liver, spleen and mesentery in treated group is a result of toxic effect of test substance.

Interpretation of results:

GHS criteria not met

Conclusions:

The acute oral LD50 of the test item was determined to be > 2000 mg/kg bw in rats.

Executive summary:

This study was performed to assess the acute oral toxicity of the test item to Wistar rat . The method followed was as per the guidelines of the Organization for Economic Cooperation and Development (OECD) for testing of chemicals No.401 (adopted 24th February 1987) "Acute Oral Toxicity". Two groups of rats comprising 5 males and 5 females per group were randomly selected. One group was kept as control and another group were subjected to a single oral dose of the test item at the dose level of 2000 mg/kg body weight and observed for 14 days. No mortality was observed in control and 2000 mg/kg body weight dose group. All the animals that survived at the end of the treatment period were subjected to gross pathological examination. The acute oral median lethal dose (LD50) of the test item to the rat was found to be more than 2000 mg/kg body weight.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating dose

Value:

> 2 000 mg/kg bw

Quality of whole database:

according to OECD TG 401

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:

1997-10-23 to 1997-11-07

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Version / remarks:

12th May 1981

GLP compliance:

yes (incl. QA statement)

Test type:

traditional method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Breeding facility, Jai Research Foundation, India
- Weight at study initiation: 112.0 g - 148.0 g
- Housing: Group-housed (5 animals of same sex per cage) in solid floor polypropylene rat cage with clean paddy husk bedding
- Diet: Rat pellet feed (Amrut brand) ad libitum (MF: Nav Maharashtra Chakan Oil Mills Limited, 43, Shaniwar Peth, Pune-411 030, Maharashtra, India)
- Water: Pure drinking water filtered through Aquaguard water filter system ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): between 21 ± 1.38 and 24 ± 1.38
- Humidity (%): 66 ± 5.44
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

other: inhalation: dust aerosol

Type of inhalation exposure:

head only

Vehicle:

air

Remark on MMAD/GSD:

The size of generated respirable particle was found in between 6 to 10 microns because 94.90% of the particles were collected at the Effective Cut-off Diameter (particle size 6 to 10 microns) . Therefore the particle size distribution curve was not plotted and the mass median aerodynamic diameter could not be evaluated.

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: Dynamic inhalation equipment consisting of an inhalation chamber, air compressor, gas flow meter, dust generator system, rotameter, Andersen personal cascade impactor, Humitherm, Oxygen monitor and rat confinement cages.
- Exposure chamber volume:
Dynamic inhalation equipment: 3 main parts: inlet, exposure and outlet chambers. Each part is 30 cm in height and 30 cm in internal diameter. The total capacity of the chamber is 63.5 L
- Method of holding animals in test chamber: in rat confinement cages accommodated in the port-holes of the inhalation chamber.
- Source and rate of air (airflow): air inflow rate: 20 L/min and outflow rate 21.04 L/min
- Method of conditioning air:
- System of generating particulates/aerosols:
The test item was used as such and loaded into powder filling funnel once every 30 minutes during the exposure period. The regulated quantity of the test item fell on the vibrating funnel through the rotary wheel feeder which in turn filled uniform cavities (holes) on the circular rotating dosage disc. The test item from the cavities was sucked with the help of compressed air venturi dust aerosol ejector and aerosolised. The test item dust aerosol thus generated was directed into the nose only inhalation exposure chamber with the conducting pipe. Uniform distribution of the test item aerosol inside the inhalation chamber was ensured with help of placing a baffle plate at the end of conducting pipe.
- Method of particle size determination: by using Andersen Personal Cascade Impactor (6 stage)
- Temperature, humidity, oxygen concentration in air chamber: The mean chamber temperature and relative humidity during the exposure were 24 ± 0.95°C and 65.6 ± 7.80%, respectively. The mean chamber oxygen concentration was 20.9 ± 0.08%.

TEST ATMOSPHERE
A known quantity of the test item was used as such and loaded in to powder filling funnel. The nominal concentration of the test item was calculated using the following formula:

N.C = (Test substance carrying capacity of dosage disc (mg) / Air flow rate through dust generator venturi (L)) x ( r.p.m of dosage disc) = mg/L air

The test substance carrying capacity of dosage disc was found to be 471.5 mg at 1 revolution per minute (r.p.m) of the dosage disc. The the test item was evaluated at the nominal concentration of 23.57 mg/L air. To measure the breathing zone concentration, a measured volume of air was drawn from the inhalation chamber at the level of breathing zone every one hour of exposure and determined by the gravimetric method.

- Time needed for equilibrium of exposure concentration before animal exposure : 10 min


TEST ATMOSPHERE
The size of generated respirable particle was found in between 6 to 10 microns because 94.90% of the particles were collected at the Effective Cut-off Diameter (particle size 6 to 10 microns). Therefore the particle size distribution curve was not plotted and the mass median aerodynamic diameter could not be evaluated.

Analytical verification of test atmosphere concentrations:

no

Duration of exposure:

4 h

Concentrations:

0 and 1.766 mg/L air

No. of animals per sex per dose:

5

Control animals:

yes

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: The toxicity symptoms and mortality were observed at 1, 2, 3 and 4 h during exposure and once daily up to 14 days from the day of the exposure. Individual body weights were recorded prior to dosing and on day 7 and 14.
- Necropsy of survivors performed: yes
- Clinical signs including body weight : The toxic symptom observed in the treated group during exposure was abdominal breathing and nasal irritation in treated group only. The animals in both treated and control groups showed a gain in body weight during the study.
- Other examinations performed: clinical signs, body weight

Key result

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 1.766 mg/L air

Based on:

test mat.

Exp. duration:

4 h

Mortality:

There was no mortality in any group.

Clinical signs:

other: abdominal breathing and nasal irritation in treated group

Body weight:

The animals in both treated and control groups showed a gain in body weight during the study.

Gross pathology:

At the end of the experimental period the animals from the control and treated groups were sacrificed by cervical dislocation and subjected to thorough necropsy examination.
External examination of carcasses of both control and treated groups did not reveal any external findings of significance.
In control group, vascular pathology in lung revealed focal emphysema (1 male), mild congestion (1 female) and necrotic patches (1 female) while kidney revealed mild paleness in one male animal.
In treated group, lung revealed emphysema in six animal (3 male + 3 female), haemorrhages in 3 animals (2 male + 1 female). Liver showed congestion in 2 female animals. The vascular pathology in kidney showed cystic foci. Whereas, adrenal in two animals (1 male + 1 female) revealed slightly hyperaemia.
In sum, it may be concluded that the lesions recorded in lung and kidney of control group were having mild intensity of toxic effect and.it may considered as incidental. Whereas, in treated group, vascular lesions met in lung, liver, kidney and adrenal showed effect of the toxic substance by inhalation route as evidenced by the result of this study.

Interpretation of results:

GHS criteria not met

Conclusions:

The acute inhalation toxicity (LC50) value to rat was found to be more than the maximum achieved concentration of the test item dust aerosol viz.,1.766 mg/L of air at breathing zone.

Executive summary:

This study was undertaken to assess the acute inhalation toxicity (LC50) of the test item to rat. The method followed was as per the guidelines of Organization for Economic Co-operation and Development (OECD) for testing of chemicals No. 403 (adopted 12th May 1981) "Acute Inhalation Toxicity".

This study was carried out using inhalation equipment (head only exposure) supplied by Bio-Tox Instrumentations International, New Delhi, India. Two groups of rats each comprising 5 males and 5 females were taken up for the study. One group was exposed to the maximum achieved concentration the test item dust aerosol viz., 1.766 mg/L of air and another control group was exposed to only compressed air without test item dust for a duration of 4 h continuously and observed for 14 days. No mortality was observed in both the groups. The surviving animals were subjected to necropsy 14 days after the exposure. Animals sacrificed at termination of the study revealed mild toxicity by inhalation route at the concentration level tested.

The acute inhalation toxicity (LC50) value to rat was found to be more than the maximum achieved concentration of the test item dust aerosol viz., 1.766 mg/L of air at breathing zone.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating conc.

Value:

> 1.766 mg/L air

Physical form:

other: dust aerosol

Quality of whole database:

according to TG 403

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1997-10-29 to 1997-11-12

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Version / remarks:

24th February 1987

GLP compliance:

yes (incl. QA statement)

Test type:

fixed dose procedure

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Breeding facility, Jai Research Foundation - 396 108, India
- Weight at study initiation: 202.0 g - 297.0 g
- Housing: individually in polypropylene rat cages
- Diet: Rat pellet feed (Amrut brand) ad libitum (MF: Nav Maharashtra Chakan Oil Mills Limited, 43, Shaniwar Peth, Pune - 411 030, Maharashtra, India)
- Water: Pure drinking water filtered through Aquaguard water filter system ad libitum
- Acclimation period: 5 days prior to the application

ENVIRONMENTAL CONDITIONS
- Temperature (°C): between 20 ± 2.0 and 22 ± 1.8
- Humidity (%): 76 ± 6.0
- Photoperiod (hrs dark / hrs light): 12/12

Type of coverage:

semiocclusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE
- Area of exposure: dorso-lumbar area of the trunk
- % coverage: ca. 10 %
- Type of wrap: porous gauze dressing (not more than 8-ply) and a non-irritating tape

REMOVAL OF TEST SUBSTANCE
- Washing: residual test substance was removed using wet cotton.
- Time after start of exposure: 24 h

TEST MATERIAL
- The test substance was moistened with distilled water.
- For solids, paste formed: no

Duration of exposure:

24 h

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

5

Control animals:

yes, concurrent no treatment

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: observations: 1 hour, 2 hours and 3 hours after the application and thereafter once a day; weighing: prior to the application and after 7 and 14 days.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality was observed in 2000 mg/kg body weight dose group and control group.

Clinical signs:

other: No toxicity symptoms were observed in any group.

Gross pathology:

After termination of experiment, all animals from control as well as treated group were sacrificed by over dose of chloroform and gross pathological examination was carried out. External examination of carcasses did not reveal gross lesion of significance.
In control group, lung showed focal emphysema in two male animals and mild congestion with mottling of liver in one male animal. Whereas, no any internal gross lesions were observed any of the female animal.
In treated group, lung showed mild congestion in three male animals and four female animals. Whereas, liver of one male animal showed mottling and hydrometra in one female animal. In conclusion, the mild gross lesions recorded in lung and liver in animals of control group were taken as incidental. Whereas, in treated group out of ten animals seven animals showed vascular pathology in lung, liver and hydrometra in one female. This clearly points to the mild toxic effect of the test substance with dose induced lesions.

Interpretation of results:

GHS criteria not met

Conclusions:

The acute dermal median lethal dose (LD50) of the test item to Wistar rat was found to be more than 2000 mg/kg body weight.

Executive summary:

This study was performed to assess the acute dermal toxicity (LD50) of the test item to Wistar rat. The study was conducted in compliance with Good Laboratory Practice following the guidelines of Organisation for Economic Co-operation and Development (OECD) for Testing of Chemicals No. 402 (adopted 24 February 1987) "Acute Dermal Toxicity".

Two groups of rats having 5 males and 5 females per group were randomly selected. One group was kept as control and another group was given a single dose of the test item by dermal application at the dose level of 2000 mg/kg body weight and observed for fourteen days. No mortality was observed in the any group. All the animals that survived at the end of the treatment period were subjected to gross pathological examination. For both the sexes acute dermal median lethal dose (LD50) of the test item to the rat was found to be more than 2 000 mg/kg body weight.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating dose

Value:

> 2 000 mg/kg bw

Quality of whole database:

according to OECD TG 402

Additional information

Acute oral

An acute oral toxicity study was performed in rats. The study was performed as GLP study according to OECD guideline 401. Five rats/sex received a single oral gavage treatment with the test item in distilled water at 2000 mg/kg bw (10 mL/kg bw) and were then observed for 14 days. Animals were observed for 1, 2 and at 3 hours after the administration and thereafter once a day. Clinical signs were recorded at each observation. The animals were weighed prior to the administration and on day 7 and 14 day. A gross necropsy was performed at the termination of the experiment. No mortality, clinical signs or unusual changes in body weight were observed. Gross necropsy revealed no abnormality in any rats of either sex. The acute oral LD50 was determined to be > 2000 mg/kg bw.

Acute inhalation

An acute inhalation toxicity study was performed in male and female Wistar rats. This study was undertaken to assess the acute inhalation toxicity (LC50) of the test item to rat. The method followed was as per the guidelines of Organization for Economic Co-operation and Development (OECD) for testing of chemicals No. 403 (adopted 12th May 1981) "Acute Inhalation Toxicity".

This study was carried out using inhalation equipment (head only exposure) supplied by Bio-Tox Instrumentations International, New Delhi, India. Two groups of rats each comprising 5 males and 5 females were taken up for the study. One group was exposed to the maximum achieved concentration the test item dust aerosol viz., 1.766 mg/L of air and another control group was exposed to only compressed air without test item dust for a duration of 4 h continuously and observed for 14 days. No mortality was observed in both the groups. The surviving animals were subjected to necropsy 14 days after the exposure. Animals sacrificed at termination of the study revealed mild toxicity by inhalation route at the concentration level tested.

The acute inhalation toxicity (LC50) value to rat was found to be more than the maximum achieved concentration of the test item dust aerosol viz., 1.766 mg/L L of air at breathing zone.

Acute dermal

An acute dermal toxicity study was performed with the test item in Wistar rats. The study was conducted in compliance with Good Laboratory Practice following the guidelines of Organisation for Economic Co-operation and Development (OECD) for Testing of Chemicals No. 402 (adopted 24 February 1987) "Acute Dermal Toxicity". Two groups of rats having 5 males and 5 females per group were randomly selected. One group was kept as control and another group was given a single dose of the test item by dermal application at the dose level of 2000 mg/kg body weight and observed for fourteen days. No mortality was observed in the any group. All the animals that survived at the end of the treatment period were subjected to gross pathological examination. For both the sexes acute dermal median lethal dose (LD50) of the test item to the rat was found to be more than 2000 mg/kg body weight.

Justification for classification or non-classification

The following conclusion is taken from the Renewal Assessment Report prepared according to the Commission Regulation (EU) N° 1107/2009, Volume 1, Dec 2018, of Mancozeb:

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

The acute toxicity of the test item in mammals was evaluated previously in the original DAR (2000) in studies that were considered to be acceptable. No new studies have been submitted by the MTF. Agria have submitted new acute toxicity studies conducted with their source of the test item. These were generated to support registration of Agria's products in other regulatory regimes outside of the EU (Brazil). The results obtained are comparable to the EU-agreed endpoints (Review Report, SANCO/4058/2001-rev. 4.4, July 2009).

The test item is of low acute toxicity by the oral (LD50 > 5000 mg/kg bw), dermal (LD50 > 2000 mg/kg bw) and inhalation (4hr- LC50 > 5 mg/L) route. This is consistent with the current harmonised classification of the test item.