4,4'-methylenedianiline

Toxicological Summary

Workers - Hazard via inhalation route

Systemic effects

Long term exposure

Hazard assessment conclusion:

DMEL (Derived Minimum Effect Level)

Value:

0.015 mg/m³

Most sensitive endpoint:

carcinogenicity

Route of original study:

Oral

DNEL related information

DNEL derivation method:

other: Risk-based concept for carcinogenic substances by the German Committee for Hazardous Substances (AGS)

Overall assessment factor (AF):

3 125

Dose descriptor starting point:

LOAEL

Value:

9 mg/kg bw/day

Modified dose descriptor starting point:

T25

Value:

16.5 mg/m³

Explanation for the modification of the dose descriptor starting point:

Neoplastic liver nodules in a chronic drinking water study in male rats were identified as the most critical systemic effect. A linear increase in the dose range tested was observed, resulting in a LOAEL of 9 mg/kg bw at which 13 out of 50 male rats were affected (12 nodules, 1 carcinoma), compared to 1 out of 50 in the control group.

 

Adaptation of starting point:

Net incidence at 9 mg/kg/d = (13-1)/50 = 0.24 = 24 %

T25_{oral, rat}= 9 mg/kg bw * 25%/24% = 9.375 mg/kg bw

Route to route extrapolation:   

The bioavailability of MDA after oral application is at least 90 % (see rational) and therefore as a conservative approach similar bioavailabilities on the oral and inhalation route of exposure need to be anticipated.

 

T25_{inhal., human}=   T25_{oral, rat}* (1/sRV_rat) * (ABS_{oral rat}/ABS_{human inhal.}) * (sRV_human/RV_worker)

T25_{inhal., human}=   9.375 mg/kg bw* (1/0.38 m³/d) 100%/100% * (6.7/10) = 16.5 mg/m³.

 (sRV:                       standard respiratory volume (for rat 0.38 m³/kg/8h or 1.15 m³/kg/24h))

Acute/short term exposure

Hazard assessment conclusion:

hazard unknown but no further hazard information necessary as no exposure expected

DNEL related information

Local effects

Long term exposure

Hazard assessment conclusion:

no hazard identified

Acute/short term exposure

Hazard assessment conclusion:

no hazard identified

DNEL related information

Workers - Hazard via dermal route

Systemic effects

Long term exposure

Hazard assessment conclusion:

DMEL (Derived Minimum Effect Level)

Value:

0.004 mg/kg bw/day

Most sensitive endpoint:

carcinogenicity

Route of original study:

Oral

DNEL related information

DNEL derivation method:

other: Risk-based concept for carcinogenic substances by the German Committee for Hazardous Substances (AGS)

Overall assessment factor (AF):

12 500

Dose descriptor starting point:

LOAEL

Value:

9 mg/kg bw/day

Modified dose descriptor starting point:

T25

Value:

18.75 mg/kg bw/day

Explanation for the modification of the dose descriptor starting point:

Neoplastic liver nodules in a chronic drinking water study in male rats were identified as the most critical systemic effect. A linear increase in the dose range tested was observed, resulting in a LOAEL of 9 mg/kg bw at which 13 out of 50 male rats were affected (12 nodules, 1 carcinoma), compared to 1 out of 50 in the control group.

 

Adaptation of starting point:

Net incidence at 9 mg/kg/d = (13-1)/50 = 0.24 = 24 %

T25_{oral, rat}= 9 mg/kg bw * 25%/24% = 9.375 mg/kg bw

 

Route to route extrapolation:   

Basic assumptions :  100% bioavailability oral

50% bioavailability dermal in rats (El-Hawari et al, 1986).

T25_{dermal, rat}= 9.375 mg/kg bw / 0.5 = 18.75 mg/kg bw

Acute/short term exposure

Hazard assessment conclusion:

hazard unknown but no further hazard information necessary as no exposure expected

DNEL related information

Local effects

Long term exposure

Hazard assessment conclusion:

high hazard (no threshold derived)

Acute/short term exposure

Hazard assessment conclusion:

high hazard (no threshold derived)

Workers - Hazard for the eyes

Local effects

Hazard assessment conclusion:

no hazard identified

Additional information - workers

Rationale for choice of toxicological key values:

The main target organs of MDA´s carcinogenic and non-carcinogenic effects in rodents are the liver and the thyroid gland. Also in humans liver effects following MDA exposure were documented. Moreover MDA is a skin sensitizer in humans.

 

Dermal Absorption:

Following dermal application (2 mg/kg bw onto 2 cm² application area) MDA was well absorbed in rats (50% in 96 h), though to a minor extend in guinea pigs (29% in 96 h) and monkeys (21% in 168 h) (El-Hawari et al., 1986). The transport process exhibited saturable transport kinetics and significant amounts of MDA were recovered in the skin of the application area at the end of the observation periods. In an occlusive in vitro dermal penetration assay resorption rates of 33% were determined with isolated human skin and 13% with isolated rat skin during an observation period of 72 h (Hotchkiss et al., 1993). These resorbtion rates decreased significantly when the application area was not occluded. Like in the in vivo assay major amounts of radioactivity were recovered in the skin surface. As a worst-case approach, it is assumed that 50% of MDA is absorbed via the skin in humans, which is in line with the proceeding by RAC (RAC/32/2015/11 rev 1, "Application for authorisation: establishing a reference dose response relationship for carcinogenicity of technical MDA").

 

Acute toxicity:

Following single oral ingestion MDA exhibits severe irreversible liver toxicity with a distinct interspecies difference in severity.

In rats gross pathological liver and eye effects were noticeable from dose levels of 400 mg/kg bw (BASF AG, 1961, 1965). Microscopically hepatocellular necrosis was observed in male rats treated with 100 mg/kg bw MDA and markers of liver injury (e.g. serum bilirubin, liver weight) were already altered at dose level as low as 25-75 mg/kg bw. These observations were supported by Dugas et al. (2001) demonstrating that a single oral dose of 25 mg/kg (female rat) or 50 mg/kg bw (male rat) is resulting in bile duct injury, and an increase in AP, GGT, ALT and bilirubin in serum.

As expected for an aromatic diamine cats were shown to be the most susceptible species for systemic MDA liver toxicity, followed by dogs, rats and rabbits (Oettel and Hofmann, 1961). In an acute gavage study performed with cats (Oettel & Hoffmann, 1961), animals at the lowest dose of 10 mg/kg were without clinical signs, though hematological, renal and hepatic disturbances were observed. Significant macroscopic liver effects (icterus) were observed with 25 mg/kg bw and higher dose levels additionally affected the kidneys and the eyes. Severe, irreversible mydriasis, followed by blindness was observed in almost all cats from dose levels of 25 mg/kg bw. Similar effects on the eyes were observed in dogs (from 100 mg/kg bw) and rabbits (at 300 mg/kg bw).

 

Repeated dose toxicity:

Subchronic uptake of MDA by rats via the drinking water resulted in irreversible hemotoxic effects, irreversible hyperplasia of small biliary ducts and stimulation of the follicular epithelium in the thyroids (Ciba-Geigy, 1982). The LOEL was identified as 7.5 mg/kg bw. With respect to organ toxicity female animals were more susceptible than males and rats were more susceptible than mice.

 

Carcinogenicity:

The point of departure for the DMEL derivation were liver tumors in rats observed in a chronic drinking water study with MDA hydrochloride (NTP, 1983). This tumor type was identified as the most critical regarding incidence and dose levels. Oral uptake of 9 mg/kg bw resulted in a significant increase in neoplastic liver nodules (LOAEL 9 mg/kg bw/d) with male rats being more susceptible than female rats and rats generally being more susceptible than mice. Additionally one carcinoma was observed in each male dose group. The thyroid tumors in rats and the liver tumors in mice cannot be conclusively evaluated with respect to their human relevance.

No data is available to evaluate carcinogenicity on the inhalation route of exposure and only a chronic study of very low reliabilitiy is available for the dermal route of exposure. Though, confirming the liver as primary target of liver neoplasms.

The mechanism of carcinogenicity is yet not fully understood. Genotoxic and/or secondary mechanisms (e.g. thyroid stimulation following glucuronidation in the liver) can be postulated. Based on this uncertainty, a DMEL needs to be derived for the safety assessment based on the conservative assumption of a genotoxic mechanism of liver carcinogenesis and, thus, a linear dose-response curve.

 

Toxicity to reproduction:

The reliability of the little data available for reproductive toxicology of MDA is low and does not allow the derivation of a reliable dose descriptor. However, the hazard and safety assessment of MDA with respect to reproductive toxicology is superimposed by its classification as a cat. 2 carcinogen.

 

Details:

The following DNELs / DMELs were not derived:

• oral exposure: In principle ingestion is not an anticipated route of exposure in an industrial setting, since general workplace hygiene yield to avoid any oral ingestion. Particularly for MDA as cat2 carcinogen the low occupational exposure limits applied prohibit from any oral ingestion at the workplace.

• acute effects: occupational exposure limits for carcinogens (DMEL for MDA, cat. 2 carcinogen) are established as 8 h time weighed averages, a ceiling factor for acute peak exposures is not envisioned. Acute occupational exposure limits are designed to e.g. prevent from local irritation (not relevant for MDA, see above) or adverse systemic effects. In the case of MDA the derivation of a DNEL for acute-systemic-effects is resulting in significantly higher risk numbers and would therefore be misleading with respect to the derived DMEL

Derivation of DMELs:

The “Large assessment factor approach” for the derivation of a DMEL (see ECHA guidance chapter R8, appendix R.8-7) was followed.

Long-term exposure – systemic effects – inhalation DMEL:

The following assessment factors were used:

•  Allometric scaling rat → man:       1 (Allometric scaling does not need to be applied in cases where doses in experimental animal studies are expressed as concentrations (mg/m³)).
• remaining differences:                    2.5 (covering e.g. differences in toxicodynamics)
• intra-species differences:               5 (for workers)
• point of comparison:                     10 (not having a NOEL)
• nature of carcinogenic process:      10 (genotoxic carcinogen)
• T25 instead of BMDL10:              2.5

Adaptation for differences in worker and experimental exposure conditions:

Workers exposure is 5 days a week, 48 weeks a year and 40 years in an average lifetime of 75. This results in a correction factor of 2.8 (7/5 days/week * 52/48 weeks/year * 75/40 years/life).

 

DMEL_{inhal., worker} = 16.5 mg/m³ / 3125 *2.8 = 14.8 µg/m³

Due to the conservative nature of the DMEL derivation, systemic organ toxicity is sufficiently covered by the DMEL (DNEL derivation not presented).

Note: According to Directive(EU) 2019/130 of the European Parliament and of the Council of 16 January 2019 amending Directive 2004/37/EC on the protection of workers from the risks related to exposure to carcinogens or mutagens at work, a limit value for MDA of 0.08 mg/m³ (8 hours) was established, including a skin notation indicating the possibility of significant uptake through the skin. This value represents a binding occupational exposure limit in the EU and is to be adopted as a minimum standard for MDA by the Member States by 20 February 2021. The presented DMEL is more than five times lower than the binding EU occupational exposure limit and is therefore considered sufficiently conservative.

Long-term exposure – systemic effects – dermal DMEL:

The following assessment factors were used:

• Allometric scaling rat → man:         4
• remaining differences:                    2.5 (covering e.g. differences in toxicodynamics)
• intra-species differences:               5 (for workers)
• point of comparison:                      10 (not having a NOEL)
• nature of carcinogenic process:       10 (genotoxic carcinogen)
• T25 instead of BMDL10:               2.5

Adaptation for differences in worker and experimental exposure conditions:

Workers exposure is 5 days a week, 48 weeks a year and 40 years in an average lifetime of 75.

This results in a correction factor of 2.8 (7/5 days/week * 52/48 weeks/year * 75/40 years/life). 

DMEL_{dermal, worker} = 18.75 mg/kg bw / 12500 *2.8 = 4.2 µg/kg bw

Due to the conservative nature of the DMEL derivation, systemic organ toxicity is sufficiently covered by the DMEL (DNEL derivation not presented).

The following DMELs/DNELs were not derived:

Acute/short-term exposure – systemic effects – dermal DNEL

Not quantifiable; see above

 

Acute/short-term exposure – systemic effects – inhalation DNEL

Not quantifiable; see above

 

Acute/short-term exposure – local effects – dermal DNEL

Not quantifiable; see above

 

Acute/short-term exposure – local effects – inhalation DNEL

Not quantifiable; see above

Long-term exposure – local effects – dermal DNEL

Not quantifiable; see above

 

Long-term exposure – local effects – inhalation DNEL

Not quantifiable; see above

General Population - Hazard via inhalation route

Systemic effects

Long term exposure

Hazard assessment conclusion:

hazard unknown but no further hazard information necessary as no exposure expected

Acute/short term exposure

Hazard assessment conclusion:

hazard unknown but no further hazard information necessary as no exposure expected

DNEL related information

Local effects

Long term exposure

Hazard assessment conclusion:

hazard unknown but no further hazard information necessary as no exposure expected

Acute/short term exposure

Hazard assessment conclusion:

hazard unknown but no further hazard information necessary as no exposure expected

DNEL related information

General Population - Hazard via dermal route

Systemic effects

Long term exposure

Hazard assessment conclusion:

hazard unknown but no further hazard information necessary as no exposure expected

Acute/short term exposure

Hazard assessment conclusion:

hazard unknown but no further hazard information necessary as no exposure expected

DNEL related information

Local effects

Long term exposure

Hazard assessment conclusion:

hazard unknown but no further hazard information necessary as no exposure expected

Acute/short term exposure

Hazard assessment conclusion:

hazard unknown but no further hazard information necessary as no exposure expected

General Population - Hazard via oral route

Systemic effects

Long term exposure

Hazard assessment conclusion:

hazard unknown but no further hazard information necessary as no exposure expected

Acute/short term exposure

Hazard assessment conclusion:

hazard unknown but no further hazard information necessary as no exposure expected

DNEL related information

General Population - Hazard for the eyes

Local effects

Hazard assessment conclusion:

hazard unknown but no further hazard information necessary as no exposure expected

Additional information - General Population

It is possible to calculate consumer DN(M)EL long term, (dermal and inhalation route-systemic), by increasing the assessment factor for intra-species differences from 5 to 10. Though, since no exposure of the general population is supported by the CSA, no DMELs are presented.