Reaction product of Chloro[29H,31H-phthalocyaninato(2-)-N29,N30,N31,N32]aluminium, sulphuric acid and caustic soda

Endpoint summary

Description of key information

The NOAEL, based on a 90 -day oral study, is 1000 mg/kg diet, which corresponds to 68.7 mg/kg bw for males and 71.8 mg/kg bw for females.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)

GLP compliance:

no

Remarks:

Study pre-dates GLP regulations

Limit test:

no

Specific details on test material used for the study:

- Name as used in study report: FAT 60'149/B
- Batch number: Partie 1
- Purity: 70%

Species:

rat

Strain:

other: Tif: RAIf (SPF)

Remarks:

F3-hybrid of RII 1/Tif x RII 2/Tif

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Animal Production Stein, CIBA-GEIGY LTD
- Age at study initiation: approx. 4 weeks
- Weight at study initiation: 47-109 g; initial mean group body weight: 91-94 g, males, 85-87 g females
- Fasting period before study: none
- Housing: in groups of 5 in Macrolon cages type 4
- Diet: Pelleted, certified standard diet Nafag No. 890 Tox, ad libitum
- Water: ad libitum
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2
- Humidity (%): 55 ± 10
- Air changes (per hr): 15-17
- Photoperiod (hrs dark / hrs light): 14/10

Route of administration:

oral: feed

Vehicle:

unchanged (no vehicle)

Analytical verification of doses or concentrations:

yes

Remarks:

concentrations 95-102% of nominal value

Details on analytical verification of doses or concentrations:

Four samples taken and analysed during the study in weeks 1-3, 4-5, 6-8 and 9-13.

Duration of treatment / exposure:

3 months, daily (90 days)

Frequency of treatment:

Continuous

Dose / conc.:

0 ppm

Remarks:

0 mg/kg diet

Dose / conc.:

1 000 ppm

Remarks:

1000 mg/kg diet

Dose / conc.:

3 000 ppm

Remarks:

3000 mg/kg diet

Dose / conc.:

10 000 ppm

Remarks:

10000 mg/kg diet

No. of animals per sex per dose:

20

Control animals:

yes, plain diet

Positive control:

no

Observations and examinations performed and frequency:

- Mortality and symptoms were observed daily
- Body weight: weekly
- Food and water consumption: weekly
- Mean food conversion calculated according to the following formula: weekly food consumption (g) / midweek body weight (g) * 1000/7 (unit: g food/kg body weight/day)
- Hearing test: at the beginning and towards the end of the treatment period

Laboratory investigations:
Laboratory investigations were carried out in 10 rats of each sex and group after completion of:
- month 1, week 4
- month 2, week 9
- month 3, week 13
To reduce the biological variability due to circadian rhythms, blood sampling for haematology and blood chemistry was between the hours of 0700 and 0900 a.m. For blood chemistry measurements food was witheld for about 20 hours prior to blood removal.
The site of blood removal was the orbital sinus and microhaematocrit glass capillary tubes were used. Blood samples from each animal with the respective anticoagulant (EDTA for performing the complete blood count, 3.8% Sodium citrate for coagulation testing and Heparin for blood chemistry measurements) were aliquoted into individual vials. No anaesthesia was used to restrain the animals. All blood collection was by manual restraint only. Urine for analysis was collected overnight. The individual rats were housed in special metabolism cages. Food and water was withheld during the time of urine collection. The parameters measured were:
- haematology: Haemoglobin, Erythrocytes, Mean Corpuscular Volume, Mean Corpuscular Heamaglobin, Haematocrit, Leucocytes Total Count, Leucocytes Differential Count, Reticulocytes, Inclusion Bodies (Heinz Bodies), Thrombocytes, Partial Thromboplastin Time
- blood chemistry: Glucose, Urea, Total Bilirubin, Creatinine, Total Protein, Albumin, Globulin, A/G Ratio, Sodium, Potassium, Chloride, Calcium, Phosphorus Inorganic, Asp. Aminotransferase, Ala. Aminotransferase, Lactate dehydrogenase, Alkaline Phosphate
- urinalysis: Colour, Specific Gravity, pH, Protein, Glucose, Ketone, Blood, Bilirubin, Urobilinogen, Urine Sediment

Sacrifice and pathology:

At the end of the test period all control and test animals were bled under ether anaesthesia and subjected to detailed autopsy. Liver,adrenals, brain, thymus, heart, kidneys and gonads were weighed. The following organs and tissues were preserved in 10% neutral formalin: brain (cerebrum, cerebellum, brain stem), spinal cord, eyes, lacrimal glands, pituitary, salivary glands, heart, aorta, thymus, thyroid, lungs, trachea, spleen, bone marrow (sternum), lymph nodes (axillary and mesenteric), sciatic nerve, oesophagus, stomach, small and large intestine, adrenal glands, pancreas, liver, kidneys, urinary bladder, ovaries, testes, seminal vesicle, epididymis, prostate, uterus, skin (mammary aera), skeletal muscle, organs and tissues showing macroscopical changes.
After the fixation organ samples from each control and test rat were taken, embedded in paraplast, sectionned at 3-5um, stained with haematoxylin and 3-5 µm, stained with haematoxylin and eosin and subjected to microscopic examination.

Statistics:

For each time point and parameter a uni-variate statistical analysis was conducted. Due to the routine manner of the analysis system, parameter free methods were applied. Each treated group was compared to the control group in respect of dispersion and displacement. In addition a trend test was applied considering all groups.

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Food efficiency:

no effects observed

Water consumption and compound intake (if drinking water study):

no effects observed

Ophthalmological findings:

not examined

Haematological findings:

effects observed, treatment-related

Description (incidence and severity):

A slight tendency to anaemia, reflected by a decreased number of Erythrocytes, a decrease of the Haemoglobin concentration and a decrease of the Haematocrit was observed in male groups 3 and 4 (3000 and 10 000 ppm) at weeks 4, 9 and 13 of the study. The males of these two dosage groups also showed increased numbers of Reticulocytes at weeks 4 and 9 of test. Though these chemges are only slight they are considered to be treatment related.

Clinical biochemistry findings:

effects observed, non-treatment-related

Description (incidence and severity):

At week 4 an increase in Glucose and Creatinine concentration was observed in all treated male groups. Since the concentrations of Glucose and Creatinine were relatively low in the respective control group at this point in time the increase is considered to be unrelated to the treatment.
Furthermore the values of Glucose and Creatinine were within the normal range at week 9 and 13 of treatment in the control group as well as in the treated male groups.

Urinalysis findings:

no effects observed

Behaviour (functional findings):

not examined

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

Significantly increased liver weights in male groups 3 and 4 (3000 and 10 000 ppm) and in female group 4 (10 000 ppm) were observed. Male group 4 (10 000 ppm) showed a slight decrease of thymus weight and ratios. However microscopical examination of these organs showed no histopathological abnormalities which could be attributed to the treatment.

Gross pathological findings:

effects observed, treatment-related

Description (incidence and severity):

Macroscopical examination at necropsy showed discoloration of the digestive tube in treated groups 3 and 4 (3000 and 10 000 ppm) and additionally of some abdominal organs in male and female groups 4 (10 000 ppm). No abnormalities were seen in these organs and tissues upon histopathological examination and microscopical appearance was comparable to that of the controls.

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

no effects observed

Other effects:

no effects observed

Description (incidence and severity):

Hearing tests performed at the beginning and towards the end of the application period revealed no treatment related effects on the auditory perception.

Key result

Dose descriptor:

NOAEL

Effect level:

1 000 mg/kg diet

Based on:

test mat.

Sex:

male/female

Basis for effect level:

gross pathology

haematology

organ weights and organ / body weight ratios

Remarks on result:

other: NOAEL of 1000 mg/kg diet corresponds to daily intake of 68.7 mg/kg bw for males and 71.8 mg/kg for females

Key result

Critical effects observed:

yes

Lowest effective dose / conc.:

3 000 mg/kg diet

System:

haematopoietic

Organ:

blood

Treatment related:

yes

Dose response relationship:

yes

Relevant for humans:

yes

Key result

Critical effects observed:

yes

Lowest effective dose / conc.:

3 000 mg/kg diet

System:

hepatobiliary

Organ:

liver

Treatment related:

yes

Dose response relationship:

yes

Relevant for humans:

yes

Critical effects observed:

yes

Lowest effective dose / conc.:

3 000 mg/kg diet

System:

gastrointestinal tract

Organ:

pharynx

Treatment related:

yes

Dose response relationship:

yes

Relevant for humans:

yes

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

NOAEL

68.7 mg/kg bw/day

Study duration:

subchronic

Species:

rat

System:

other: haematopoietic and hepatobilliary

Organ:

blood

liver

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

In an 28 -days palpability study in rats, with daily administration via diet, male and female rats were administered doses of 0, 1000, 3000 or 10 000 ppm (= mg/kg feed) (CIBA-GEIGY, 1980). No death occurred during the 28 days test period. No clinical symptoms and no signs of local and/or systemic toxicity were observed. The mean food consumption of all treated male and female rats was comparable to the controls. The mean bodyweight gain of all treated males and females was comparable to the controls. The mean food conversion of all male and female treated rats was comparable to the controls. Ophthalmic inspections made during the application period revealed no evidence of a reaction to treatment.

In a reliable oral subchronic study in rats, with 3 months daily administration via diet, male and female rats were administered doses of 0, 1000, 3000 and 10 000 ppm (= mg/kg feed) (CIBA-GEIGY 1982). The mean body weight gain of all treated male and female groups was similar to that of the respective control groups. The mean food consumption values of all treated male and female groups were similar to that of the respective control groups. The mean water consumption values of all treated male and female groups were similar to that of the respective control groups. The mean food conversion in treated animals was similar to that of the respective control groups. No death occurred during the 3 months test period. No clinical symptoms emd no signs of local and/or systemic toxicity were observed. Hearing tests performed at the beginning and towards the end of the application period revealed no treatment related effects on the auditory perception. A slight tendency to anaemia, reflected by a decreased number of erythrocytes, a decrease of the haemoglobin concentration and a decrease of the haematocrit was observed in male groups 3 and 4 (3000 and 10 000 ppm) at weeks 4, 9 and 13 of the study. The males of these two dosage groups also showed increased numbers of reticulocytes at weeks 4 and 9 of test. Though these changes are only slight they are considered to be treatment related. At week 4 an increase in glucose and creatinine concentration was observed in all treated male groups. Since the concentrations of glucose and creatinine were relatively low in the respective control group at this point in time the increase is considered to be unrelated to the treatment. Furthermore the values of glucose and creatinine were within the normal range at week 9 and 13 of treatment in the control group as well as in the treated male groups. The findings in the urine were unremarkable. Significantly increased liver weights in male groups 3 and 4 (3000 and 10 000 ppm) and in female group 4 (10 000 ppm) were observed. Male group 4 (10 000 ppm) showed a slight decrease of thymus weight and ratios. However microscopical examination of these organs showed no histopathological abnormalities which could be attributed to the treatment. Macroscopical examination at necropsy showed discoloration of the digestive tube in treated groups 3 and 4 (3000 and 10 000 ppm) and additionally of some abdominal organs in male and female groups 4 (10 000 ppm). No abnormalities were seen in these organs and tissues upon histopathological examination and microscopical appearance was comparable to that of the controls. Based on this study, the NOAEL of the substance is 1000 mg/kg diet, which corresponds to 68.7 mg/kg bw for males and 71 mg/kg bw for females.

In a two month skin depigmentation study with concentrations up to 0.3% of the substance, no depigmentation of the skin of Guinea pigs was observed (CIBA-GEIGY, 1980).

Justification for classification or non-classification

Though there are effects that are considered to be treatment related (slight tendency to anaemia, increased liver weight) and which lead to the derivation of a No Observed Adverse Effect Level, the effects are not very severe and occur at 3000 ppm in diet (above 100 mg/kg bw). No macroscopical changes, other than discoloration, nor abnormalities upon histopathology were observed. It is therefore concluded that the effects do not require classification for Specific Target Organ Toxicity - Repeated Exposure according to the criteria of the CLP Regulation (EC) No 1272/2008.