2-(4-tert-butylphenyl)-6-cyano-5-[bis(ethoxycarbonylmethyl)carbamoyloxy]-1H-pyrrolo[1,2-b][1,2,4] triazole-7-carboxylic acid 2,6-di-tert-butyl-4-methylcyclohexylester

Endpoint summary

Description of key information

Oral (OECD 423), rat: LD50 = 5000 mg/kg bw (limit test)
Inhalation (OECD 403), rat: LD50 > 5.05 mg/L (limit test)
Dermal (OECD 402), rat: LD50 > 2000 mg/kg bw (limit test)

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:

22 Jul - 08 Aug 2003

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP - guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), 12 Nousan, Notification No. 8147, November 2000, including the most recent partial revisions

Deviations:

no

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

yes

Species:

rat

Strain:

other: Wistar Crl:(WI)BR

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland, Sulzveld, Germany
- Age at study initiation: Approximately 11 weeks
- Weight at study initiation: 344 g (mean, males) and 227 g (mean, females)
- Fasting period before study: Food was withheld overnight (for a maximum of 20 hours) prior to dosing until 3-4 hours after administration of the test substance
- Housing: Group housing of 3 animals per sex per cage in labelled Macrolon cages (type IV; height 18 cm) containing purified sawdust as bedding material (SAWI, Jelu Werk, Rosenberg, Germany)
- Diet: Standard pelleted laboratory animal diet (from Altromin (code VRF 1), Lage, Germany), ad libitum
- Water: Tap water, ad libitum
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17.2-24.5
- Humidity (%): 44-83
- Air changes (per hr): approximately 15
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: gavage

Vehicle:

propylene glycol

Details on oral exposure:

VEHICLE
- Amount of vehicle (if gavage): 10 mL/kg bw

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bw

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: According to OECD 423, if the acute oral toxicity of a substance is expected to be low, a limit test may be performed with the highest dose level of 2000 mg/kg bw

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

Step 1: 3 males
Step 2: 3 females

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Observations of mortality were made twice daily; observations of clinical signs were made several times on day 1 and daily thereafter, graded according to a severity scale of 1-4; weighing was performed weekly on day 1 (pre-administration), 8 and 15
- Necropsy of survivors performed: Yes, all gross pathological changes were recorded

Sex:

male/female

Dose descriptor:

LD50

Effect level:

5 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: LD50 cut-off according to OECD 423

Mortality:

There was no mortality during the 14-day study period.

Clinical signs:

Uncoordinated movements were noted in all males 2-4 hours after adminstration, lasting until day 2 in 2/3 animals. One male had a hunched posture from 2 hours after dosing until day 3. No clinical signs were observed in the females.

Body weight:

There were no effects on body weight.

Gross pathology:

Necropsy and histopathological examination revealed no substance-related findings.

Table 1: Mortality and clinical signs

Dose [mg/kg bw]	Toxicological results*	Duration of clinical signs	Time of death	Mortality (%)
Males
2000	0/3/3	1 h - 3 days	-	0
LD50 > 2000 mg/kg bw
Females
2000	0/0/3	-	-	0
LD50 > 2000 mg/kg bw

                                                                                           

* first number = number of dead animals                                 

 second number = number of animals with clinical signs         

 third number = number of animals used                               

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

CLP: not classified
DSD: not classified

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

5 000 mg/kg bw

Quality of whole database:

The study was performed under GLP conditions and in accordance with standardised guidelines. The study was therefore assigned a reliability score of 1 in accordance with the principles defined by Klimisch et al. (1997). The quality of the dataset is therefore considered to be high.

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:

08 August 2013 - 28 August 2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Study conducted in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results. The study report was conclusive, done to valid guidelines and the study was conducted under GLP conditions.

Qualifier:

according to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.2 (Acute Toxicity (Inhalation))

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1300 (Acute inhalation toxicity)

Deviations:

no

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
- Stain: CRL:(WI) rats (SPF).
- Age at study initiation: Young adult rats, 9 - 12 weeks old.
- Weight at study initiation: males: 350 - 458 g; females: 211 - 288 g.
- Housing: Grouped by sex; 5 for the main test and 2 for the sighting group. Cages were Type III solid floor cages with stainless steel mesh lids.
- Diet: Complete feed for rats and mice, ad libitum.
- Water: Tap water, ad libitum.
- Acclimation period: 35 days (sighting group), 13 days (main group).

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 °C
- Humidity (%): 30 – 70 %
- Air changes (per hr): 15 – 20 air charges/hour
- Photoperiod (hrs dark / hrs light): 12 hours of light daily, from 6.00 a.m. to 6.00 p.m.

Route of administration:

inhalation: aerosol

Type of inhalation exposure:

nose only

Vehicle:

air

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: TSE Rodent Exposure System. This system comprises of two, concentric anodised aluminium chambers and a computer control system incorporating pressure detectors and mass flow controllers. Fresh aerosol from the generation system was constantly supplied to the inner plenum (distribution chamber) of the exposure system from where, under positive pressure, it was distributed to the individual exposure ports. Atmosphere generation was therefore dynamic.
- Exposure chamber volume: 3.85 L (inner plenum)
- Equilibration period: Sighting test 16 minutes; main test 13 minutes.
- Method of holding animals in test chamber: Each rat was individually held in a tapered, polycarbonate restraining tube fitted onto a single tier of the exposure chamber. Only the nose of each animal was exposed to the test atmosphere.
- Rate of air: Mean airflow into the inner plenum measured 15.0 L/min. Air flow was at least 0.7 L/min through each port.
- Method of conditioning air: Compressed air was supplied by an oil-free compressor and passed through a suitable filter system prior to introduction to the dust generator.
- System of generating particulates/aerosols: Due to the granular nature of the material, prior to use it was ground using a Ball Mill (‘Mixer Mill MM 400’). The test material was aerosolised using a dust generator (Wright’s Dust Feed Systems) located at the top of the exposure chamber.
- Method of particle size determination: The particle size of the test atmosphere was determined three times during the exposure period using a 7-stage impactor of Mercer style. Such devices employ an inertial separation technique to isolate particles in the discrete aerodynamic size ranges. Samples were taken from an unoccupied exposure port.
The collection substrates and the backup filter were weighed before and after sampling and the weight of test material, collected at each stage, calculated by this difference.
The total amount collected for each stage was used to determine the cumulative amount below each cut-off point size. In this way, the proportion (%) of aerosol less than 0.55, 0.96, 1.55, 2.11, 3.56, 6.66 and 10.55 μm was calculated.
From these data, using software supplied with the impactor, the Mass Median Aerodynamic Diameter (MMAD) and Geometric Standard Deviation (GSD) were calculated. In addition, the proportion (%) of aerosol less than 4 μm (considered to be the inhalable portion) was determined.
- Treatment of exhaust air: After passing through the animals’ breathing zone, used aerosol entered the outer cylinder from where it was exhausted through a suitable filter system.
- Conditions in test chamber:
> Sighting Group: Mean temperature: 24.5 °C (range 24.3 – 24.7 °C); Mean relative humidity: 60.2 % (range 58.8 – 74.7 %).
> Main Group: Mean temperature: 24.7 °C (range 24.0 – 25.0 °C); Mean relative humidity: 62.4 % (range 61.5 – 70.8 %).

TEST ATMOSPHERE
- Brief description of analytical method used: Aerosol concentrations were measured gravimetrically.
The test atmosphere was sampled at regular intervals during each exposure period. Samples were taken from an unoccupied exposure port by pulling a suitable, known volume of test atmosphere through weighed GF10 glass fibre filters. The difference in the pre- and post-sampling weights, divided by the volume of atmosphere sampled, was equal to the actual achieved test atmosphere concentration.
The nominal concentration was calculated by dividing the mass of test material disseminated into the chamber by the total volume of air that went through the chamber during the same period.
- Samples taken from breathing zone: yes
- See Table 1 for test atmosphere concentrations for the main test.

TEST ATMOSPHERE
- The percentage of inhalable fraction (% < 4 µm) in the sighting test was 61.2 % and 61.9 % in the main test.
- MMAD (Mass median aerodynamic diameter) / GSD (Geometric st. dev.):
> Sighting test: MMAD = 3.11 µm; GSD = 2.42
> Main test: MMAD = 3.00; GSD = 2.56
- See Table 2 for particle size distribution for the main test.

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

4 h

Concentrations:

Mean achieved concentration sighting test: 5.07 ± 0.12 mg/L (nominal concentration 15.18 mg/L)
Mean achieved concentration main test: 5.05 ± 0.17 mg/L (nominal concentration 12.88 mg/L)

No. of animals per sex per dose:

Sighting test: 2 per sex per dose
Main test: 5 per sex per dose

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Morbidity/Mortality: Animals were observed hourly during exposure, one hour after exposure and then twice daily during the 14 day post exposure observation period.
- Clinical signs: All animals were observed hourly during exposure, as soon as practically possible following removal from restraint at the end of exposure, one hour after exposure and subsequently once daily for 14 days.
- Bodyweight: Individual bodyweights were recorded prior to treatment on the day of exposure (Day 0) and on Days 1, 3, 7 and 14.
- Necropsy of survivors performed: Yes. At the end of the 14 day observation period, the animals were euthanised by exsanguination under anaesthesia and gross macroscopic examination was performed. All animals were subject to a gross necropsy which included a detailed examination of the abdominal and thoracic cavities. Special attention was given to the respiratory tract for macroscopic signs of irritancy or local toxicity.

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 5.05 mg/L air (analytical)

Based on:

test mat.

Exp. duration:

4 h

Remarks on result:

other: Mean achieved concentration

Mortality:

No mortality was observed in either the sighting or main tests.

Clinical signs:

other: Wet fur and fur staining were commonly recorded mostly on the day of exposure and in some occasions several days after exposure. These observations were considered to be related to the restraint and exposure procedures and, in isolation, were considered n

Body weight:

Normal bodyweight gain was noted during the observation period for all animals.

Gross pathology:

No treatment related gross lesions were noted.

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Under the conditions of the test, no death or overt signs of toxicity occurred in a group of five male and five female rats exposed to a mean achieved atmosphere of 5.05 mg/L for four hours. The LC50 was therefore determined to be greater than 5.05 mg/L.

Executive summary:

The acute toxicity of the test material was determined using Wistar strain rats in an acute inhalation toxicity study. The study was performed under GLP conditions and in accordance with the standardised guidelines OECD 403, EU Method B.2 and EPA OPPTS 870.1300.

A single sighting exposure was performed prior to the main study with 2 males and 2 females at the target concentration of 5 mg/L.

In the main study group, 10 (5 male and 5 female) rats were exposed to the mean achieved concentration of 5.05 mg/L. The animals were exposed for 4 hours using a nose-only exposure system, followed by a 14 day observation period. Aerosol concentrations were measured gravimetrically. The particle size distribution of the test aerosol was determined regularly during the exposure period. Clinical observations and bodyweights were recorded throughout the study, and at the end of the observation period the animals were euthanised and subjected to a gross examination post mortem.

In the main study the MMAD was 3.00 µm with a GSD of 2.56. The inhalable fraction (% < 4 µm) was 61.9 %.

Under the conditions of the test, no death or overt signs of toxicity occurred in a group of ten rats exposed to a mean achieved atmosphere of 5.05 mg/L for four hours. As no deaths occurred at 5.06 mg/L no further testing was required. The LC50 was therefore determined to be greater than 5.05 mg/L.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Quality of whole database:

The study was performed under GLP conditions and in accordance with standardised guidelines. The study was therefore assigned a reliability score of 1 in accordance with the principles defined by Klimisch et al. (1997). The quality of the dataset is therefore considered to be high.

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:

21 January 2004 - 04 February 2004

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Study conducted in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results. The study report was conclusive, done to valid guidelines and the study was conducted under GLP conditions.

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.3 (Acute Toxicity (Dermal))

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1200 (Acute Dermal Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: JMAFF, 12 Nousan, Notification No. 8147 (2000)

Deviations:

no

GLP compliance:

yes

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
- Strain: Wistar strain Crl:(Wl) BR (outbred, SPF-Quality).
- Age at study initiation: Young adult animals (approx. 10 weeks old) were selected.
- Weight at study initiation: Mean value for males on day 1 of the study was 347 g; mean value for females on day 1 of the study was 246 g. Body weight variation did not exceed ± 20 % of the sex mean.
- Housing: Individually housed in labelled Macrolon cages (type III, height 15 cm) containing purified sawdust as bedding material.
- Diet: Standard pelleted laboratory animal diet, ad libitum.
- Water: tap water, ad libitum.
- Acclimation period: At least 5 days before start of treatment under laboratory conditions.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.0 ± 3.0 °C (actual range 18.6 – 22.9 °C)
- Humidity (%): 30 – 70 % (actual range 38 – 80 %)
- Air changes (per hr): Approximately 15 air charges per hour.
- Photoperiod (hrs dark / hrs light): 12 hours of artificial fluorescent light and 12 hours of darkness per day.

Type of coverage:

occlusive

Vehicle:

propylene glycol

Details on dermal exposure:

TEST SITE
- Area of exposure: One day before exposure (day -1) an area of approximately 5 x 7 cm on the back of the animal was clipped.
- % coverage: 10 % of the total body surface (approximately 25 cm² for males and 18 cm² for females).
- Type of wrap if used: The formulation was held in contact with the skin with a dressing, consisting of a surgical gauze patch (Surgy 1D) successively covered with aluminium foil and a Coban elastic bandage. A piece of Micropore tape was additionally used for fixation of the bandages in females only.

REMOVAL OF TEST SUBSTANCE
- Washing: The dressing was removed and the skin cleaned of residual test material using water.
- Time after start of exposure: 24 hours.

PREPARATION OF SOLUTION
- The formulation (w/w) was prepared within 4 hours prior to dosing. Homogeneity was accomplished to a visually acceptable level. Adjustment was made for specific gravity of the vehicle, which was 1.036.

APPLICATION
- Volume applied: 10 mL/kg bw.

Duration of exposure:

A single application for 24 hours.

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

5 per sex per dose

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Animals were observed for mortality/viability twice daily. Body weights were measured on Days 1 (pre-administration), 8 and 15.
- Observations of clinical signs were carried out at periodic intervals on the day of dosing (day 1) and once daily thereafter, until day 15. The time of onset, degree and duration were recorded and the symptoms graded according to fixed scales:
Maximum grade 4: grading slight (1) to very severe (4)
Maximum grade 3: grading slight (1) to severe (3)
Maximum grade 1: presence is scored (1).
- Necropsy of survivors performed: yes. At the end of the observation period (day 15), all animals were sacrificed by asphyxiation using an oxygen/carbon dioxide procedure and subjected to necropsy. Descriptions of all internal macroscopic abnormalities were recorded.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred.

Clinical signs:

Hunched and flat posture, chromodacryorrhoea (snout) and ptosis were noted among the animals. The animals had recovered from the symptoms between days 3 and 7. Scales, scabs and erythema were seen on the treated skin area among the animals during the observation period.

Body weight:

The changes noted in body weight gain in males and females were within the range expected for rats used in this type of study and were therefore considered not indicative of toxicity.

Gross pathology:

No abnormalities were found at macroscopic post mortem examination of the animals.

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Under the conditions of the test, no death or signs of treatment-related toxicity occurred in five male and five female rats exposed to the test material at 2000 mg/kg bw. The LC50 was therefore determined to be greater than 2000 mg/kg bw.

Executive summary:

The acute toxicity of the test material was determined using Wistar strain rats in an acute dermal toxicity test performed under GLP conditions and in accordance with the standardised guidelines OECD 402, EU Method B.3, EPA OPPTS 870.1200 and JMAFF 12 Nousan, Notification No. 8147.

Ten rats (5 male and 5 female) were exposed to the test material in an occlusive fashion by dermal application at a limit dose of 2000 mg/kg bw in propylene glycol for 24 hours (dose volume 10 mL/kg bw). Animals were subjected to daily observations and weekly determination of body weight. Macroscopic examination was performed after terminal sacrifice.

Under the conditions of the test, no death or signs of treatment related toxicity occurred in the rats. The LC50 was therefore determined to be greater than 2000 mg/kg bw.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Quality of whole database:

The study was performed under GLP conditions and in accordance with standardised guidelines. The study was therefore assigned a reliability score of 1 in accordance with the principles defined by Klimisch et al. (1997). The quality of the dataset is therefore considered to be high.

Additional information

Oral Toxicity

In an acute oral toxicity study performed according to OECD guideline 423, the limit dose of 2000 mg/kg bw 2-(4-tert-butylphenyl)-6-cyano-5-[bis(ethoxycarbonylmethyl)carbamoyloxy]-1 H-pyrrolo[1 ,2-b][1 ,2,4]triazole-7-carboxylic acid-2,6-di-tert-butyl-4-methyl-cyclohexyl ester (UC-141) was administered to male and female rats by gavage (Hooiveld, 2003). No mortality was observed, and there were no effects on body weight during the 14 day observation period. No substance-related clinical signs were observed. The necropsy and gross pathological examination did not reveal any treatment-related effects. According to the acute toxic class method described in the OECD guideline 423, if there is no mortality following administration of 2000 mg/kg bw, the LD50 cut-off limit is 5000 mg/kg bw. Therefore, the LD50 is considered to be 5000 mg/kg bw for male and female rats.

Inhalation Toxicity

The acute toxicity of the test material was determined using Wistar strain rats in an acute inhalation toxicity study (Nagy, 2013) performed under GLP conditions and in accordance with the standardised guidelines OECD 403, EU Method B.2 and EPA OPPTS 870.1300.

A single sighting exposure was performed prior to the main study with 2 males and 2 females at the target concentration of 5 mg/L.

In the main study group, 10 (5 male and 5 female) rats were exposed to the mean achieved concentration of 5.05 mg/L. The animals were exposed for 4 hours using a nose-only exposure system, followed by a 14 day observation period. Aerosol concentrations were measured gravimetrically. The particle size distribution of the test aerosol was determined regularly during the exposure period. Clinical observations and bodyweights were recorded throughout the study, and at the end of the observation period the animals were euthanised and subjected to a gross examination post mortem.

In the main study the MMAD was 3.00 µm with a GSD of 2.56. The inhalable fraction (% < 4 µm) was 61.9 %.

Under the conditions of the test, no death or overt signs of toxicity occurred in a group of ten rats exposed to a mean achieved atmosphere of 5.05 mg/L for four hours. As no deaths occurred at 5.06 mg/L no further testing was required. The LC50 was therefore determined to be greater than 5.05 mg/L. 

Dermal Toxicity

The acute toxicity of the test material was determined using Wistar strain rats in an acute dermal toxicity test (Hooiveld, 2004) performed under GLP conditions and in accordance with the standardised guidelines OECD 402, EU Method B.3, EPA OPPTS 870.1200 and JMAFF 12 Nousan, Notification No. 8147.

Ten rats (5 male and 5 female) were exposed to the test material in an occlusive fashion by dermal application at a limit dose of 2000 mg/kg bw in propylene glycol for 24 hours (dose volume 10 mL/kg bw). Animals were subjected to daily observations and weekly determination of body weight. Macroscopic examination was performed after terminal sacrifice.

Under the conditions of the test, no death or signs of treatment related toxicity occurred in the rats. The LC50 was therefore determined to be greater than 2000 mg/kg bw.

Justification for selection of acute toxicity – oral endpoint
Only one study is available.

Justification for selection of acute toxicity – inhalation endpoint
Only one study is available.

Justification for selection of acute toxicity – dermal endpoint
Only one study is available.

Justification for classification or non-classification

Oral Toxicity

The available data on the acute oral toxicity of the test substance does not meet the criteria for classification according to Regulation (EC) 1272/2008 or Directive 67/548/EEC, and is therefore conclusive but not sufficient for classification.

Inhalation Toxicity

The available data on the acute inhalation toxicity of the test substance does not meet the criteria for classification according to Regulation (EC) 1272/2008 or Directive 67/548/EEC, and is therefore conclusive but not sufficient for classification.

Dermal Toxicity

The available data on the acute dermal toxicity of the test substance does not meet the criteria for classification according to Regulation (EC) 1272/2008 or Directive 67/548/EEC, and is therefore conclusive but not sufficient for classification.