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EC number: 805-523-5 | CAS number: 9025-52-9
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
- Melting point / freezing point
- Boiling point
- Density
- Particle size distribution (Granulometry)
- Vapour pressure
- Partition coefficient
- Water solubility
- Solubility in organic solvents / fat solubility
- Surface tension
- Flash point
- Auto flammability
- Flammability
- Explosiveness
- Oxidising properties
- Oxidation reduction potential
- Stability in organic solvents and identity of relevant degradation products
- Storage stability and reactivity towards container material
- Stability: thermal, sunlight, metals
- pH
- Dissociation constant
- Viscosity
- Additional physico-chemical information
- Additional physico-chemical properties of nanomaterials
- Nanomaterial agglomeration / aggregation
- Nanomaterial crystalline phase
- Nanomaterial crystallite and grain size
- Nanomaterial aspect ratio / shape
- Nanomaterial specific surface area
- Nanomaterial Zeta potential
- Nanomaterial surface chemistry
- Nanomaterial dustiness
- Nanomaterial porosity
- Nanomaterial pour density
- Nanomaterial photocatalytic activity
- Nanomaterial radical formation potential
- Nanomaterial catalytic activity
- Endpoint summary
- Stability
- Biodegradation
- Bioaccumulation
- Transport and distribution
- Environmental data
- Additional information on environmental fate and behaviour
- Ecotoxicological Summary
- Aquatic toxicity
- Endpoint summary
- Short-term toxicity to fish
- Long-term toxicity to fish
- Short-term toxicity to aquatic invertebrates
- Long-term toxicity to aquatic invertebrates
- Toxicity to aquatic algae and cyanobacteria
- Toxicity to aquatic plants other than algae
- Toxicity to microorganisms
- Endocrine disrupter testing in aquatic vertebrates – in vivo
- Toxicity to other aquatic organisms
- Sediment toxicity
- Terrestrial toxicity
- Biological effects monitoring
- Biotransformation and kinetics
- Additional ecotoxological information
- Toxicological Summary
- Toxicokinetics, metabolism and distribution
- Acute Toxicity
- Irritation / corrosion
- Sensitisation
- Repeated dose toxicity
- Genetic toxicity
- Carcinogenicity
- Toxicity to reproduction
- Specific investigations
- Exposure related observations in humans
- Toxic effects on livestock and pets
- Additional toxicological data
Endpoint summary
Administrative data
Description of key information
It is concluded that oral administration of trehalase, batch PPT42347 to Han Wistar rats at doses up to 100% of trehalase, batch PPT42347 for 13 weeks was well-tolerated and did not cause any adverse change. The no-observed adverse-effect level (NOAEL) was therefore considered to be 100% of the trehalase, batch PPT42347 (equivalent to 1312 mg TOS/kg body weight/day).
Key value for chemical safety assessment
Repeated dose toxicity: via oral route - systemic effects
Link to relevant study records
- Endpoint:
- sub-chronic toxicity: oral
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 01-12-2016 to 16-08-2017, (study protocol amended on 01-08-2017)
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)
- Version / remarks:
- 1998
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Limit test:
- no
- Species:
- rat
- Strain:
- Wistar
- Details on species / strain selection:
- RccHan™:WIST rat.
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: Envigo RMS Limited
- Females nulliparous and non-pregnant: Yes
- Age at study initiation: 41-47 days
- Weight at study initiation: 121 to 173 g for males; 113-148 g for females
- Fasting period before study: None
- Housing: 5 animals of the same sex per cage
- Diet: Teklad 2014C Diet ad libitum (removed overnight before blood sampling for hematology or blood chemistry).
- Water: Ad libitum. Potable water from the public supply via polycarbonate bottles with sipper tubes.
- Acclimation period: 11 days
ENVIRONMENTAL CONDITIONS
- Temperature : 20-24°C
- Humidity : 40-70 % RH
- Photoperiod (hrs dark / hrs light): 12/12
IN-LIFE DATES: From: 18 January 2016 To: 18 April 2016 - Route of administration:
- oral: gavage
- Vehicle:
- water
- Remarks:
- reverse osmosis water
- Details on oral exposure:
- PREPARATION OF DOSING SOLUTIONS:
The test enzyme was provided deep-frozen in a number of containers. The day before use, the required numbers of test material containers were transferred from the freezer to nominally 4ºC storage to allow them to thaw. On the day of formulation the test material containers were inverted 10 times and gently magnetically stirred. The required amount of test material was measured out and the residue returned to the freezer as quickly as practicable. The required volume of vehicle was added to the pre-measured material and gently mixed by magnetic stirring and inversion to avoid any potential damage to the protein and ensure homogeneity of the dose preparation. Formulations were retained refrigerated (2-8°C) overnight (when prepared the day before) or at ambient temperature.
VEHICLE
- Concentration in vehicle: 10, 33 and 100%, corresponding to 112, 368 and 1116 mg total organic solids (TOS) /kg bw/day.
- Amount of vehicle (if gavage): constant volume 10 mL/kg body weight.
- Purity: Reverse osmosis water - Analytical verification of doses or concentrations:
- yes
- Details on analytical verification of doses or concentrations:
- Dose samples were analysed according to GLP.
Stability of formulations at 10 and 100%, when retained for up to 24 hours at refrigerated (nominally 5°C) or ambient temperature was demonstrated b y Novozymes A/S. Samples of each formulation prepared for administration in Weeks 1, 6 and 13 of treatment were analysed for achieved concentration of the test substance. A total of 6 x 10 mL samples were taken from the middle of each formulation and all samples were frozen at approximately -20°C upon completion of sampling. Three samples from each formulation/occasion were subsequently dispatched (deep frozen on dry ice) to the Principal Investigator responsible for formulation analysis. All remaining samples were retained (at approximately -20°C) as a contingency but were discarded after finalization of the formulation analysis report since no further analysis was required. - Duration of treatment / exposure:
- 13 weeks
- Frequency of treatment:
- Daily
- Dose / conc.:
- 131 mg/kg bw/day (actual dose received)
- Remarks:
- Total organic substance
- Dose / conc.:
- 433 mg/kg bw/day (actual dose received)
- Remarks:
- Total organic substance
- Dose / conc.:
- 1 312 mg/kg bw/day (actual dose received)
- Remarks:
- Total organic substance
- No. of animals per sex per dose:
- 10
- Control animals:
- yes, concurrent vehicle
- Details on study design:
- - Dose selection rationale: The test enzyme was not anticipated to cause any significant findings, based on the results obtained from studies with similar materials. The highest dose (100%) is the maximum practical dose and represents administration of the enzyme, as received, at a volume dosage of 10 mL/kg body weight. The lower doses were selected using a ratio of approximately 3.3 between doses.
- Positive control:
- Not included
- Observations and examinations performed and frequency:
- CAGE SIDE OBSERVATIONS: Yes
- Time schedule: At least once daily
DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Animals were inspected visually at least twice daily for evidence of ill-health or reaction to treatment.
BODY WEIGHT: Yes
- Time schedule for examinations: The weight of each animal was recorded one week before treatment commenced, on the day that treatment commenced (Week 0), weekly throughout the study and before necropsy.
FOOD CONSUMPTION:
- The weight of food supplied to each cage, that remaining and an estimate of any spilled was recorded for the week before treatment started and for each week throughout the treatment period.
WATER CONSUMPTION:
- Time schedule for examinations: Daily by visual observation.
OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: Before treatment started and during week 12.
- Dose groups that were examined: All animals before treatment. Control and highest dose group after treatment.
HAEMATOLOGY: Yes
- Time schedule for collection of blood: Week 13
- Anaesthetic used for blood collection: Yes (isoflurane anaesthesia)
- Animals fasted: Yes, overnight
- How many animals: From all animals
- Parameters checked:
Haematocrit (Hct)
Haemoglobin (Hb)
Erythrocyte count (RBC)
Mean cell haemoglobin (MCH)
Mean cell haemoglobin concentration (MCHC)
Mean cell volume (MCV)
Total white cell count (WBC)
Platelet count (Plt)
Prothrombim time (PT)
Activated partial thromboplastin time (APTT)
Differential WBC count:
Neutrophils (N)
Lymphocytes (L)
Eosinophils (E)
Basophils (B)
Monocytes (M)
Large unstained cells (LUC)
CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: Week 13
- Animals fasted: Yes, overnight
- How many animals: From all animals
- Parameters checked:
Alkaline phosphatase (ALP)
Alanine aminotransferase (ALT)
Aspartate aminotransferase (AST)
Urea
Creatinine (Creat)
Glucose (Gluc)
Total cholesterol (Chol)
Triglycerides (Trig)
Sodium (Na)
Potassium (K)
Total protein (Total Prot)
Albumin (Alb)
Albumin/globulin ratio (A/G Ratio)
URINALYSIS: No
NEUROBEHAVIOURAL EXAMINATION: yes
- Time schedule for examinations: During week 12
- Dose groups that were examined: all
- Battery of functions tested: sensory activity / grip strength / motor activity / : Yes - Sacrifice and pathology:
- GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes - Other examinations:
- FAECAL ANALYSIS: No
Hematology, Bone Marrow: Yes
Weight of individual organs: Yes
- Time schedule for collection of organs: At necropsy - Statistics:
- The main tests used were Dunnett’s test, Shirley’s test, Williams’ test. Significant differences between the groups compared were expressed at the 5% (p<0.05) or 1% (p<0.01) level.
- Clinical signs:
- no effects observed
- Mortality:
- mortality observed, non-treatment-related
- Description (incidence):
- One incidental and non-treatment related death occurred in this study. One Control female was killed for welfare reasons in Week 9 after displaying decreased activity, laboured breathing, gasping, distended abdomen and piloerect coat. The macroscopic examination indicated that the cause of the respiratory distress and abdominal distension to be the presence of gaseous distension of the stomach and intestinal tract, though the histopathological examination suggested that the condition of the animal was due to lesions reported in the kidneys (moderate pelvic dilation).
- Body weight and weight changes:
- no effects observed
- Food consumption and compound intake (if feeding study):
- no effects observed
- Food efficiency:
- not examined
- Water consumption and compound intake (if drinking water study):
- no effects observed
- Ophthalmological findings:
- no effects observed
- Haematological findings:
- effects observed, non-treatment-related
- Description (incidence and severity):
- All inter-group differences from control, including those attaining statistical significance, were minor, lacked dose relationship or were confined to one sex and were therefore attributed to normal biological variation. Such differences included the small but statistically significant decrease of mean cell hemoglobin concentration in males given 100% trehalase but since the extent of the difference from controls was small and at this dose only a single animal had a marginally lower value than concurrent control and all individual values were within the background range it was attributed to normal biological variation.
In males given 33 or 100%, a statistically significant increase in monocyte counts, but this was attributable mainly to a low group mean value for controls caused by the individual values of 5/10 animals being below the background range, with almost all individual values for males at 33% (9/10 animals) or 100% (8/10 animals) trehalase being within the background range and, furthermore, the individual values that were outside the background range were within the concurrent control range. - Clinical biochemistry findings:
- effects observed, treatment-related
- Description (incidence and severity):
- All inter-group differences from control, including those attaining statistical significance, were minor, lacked dose-relationship or were confined to one sex and could all, therefore, be attributed to normal biological variation. Slight but statistically significant increases of plasma creatinine occurred at all doses in males, and also in females receiving 100% trehalase, batch PPT42347, but the extent of the differences from controls was small in each case and all individual values were within the background range.
There was also a slight but statistically significant decrease of potassium concentrations in females given 100% trehalase, with the majority of animals having values that were just below the background range, however those values were generally within the concurrent control range (there were many individual values for females across the groups, including controls, that had plasma potassium concentrations that were also below the background range). Consequently, and also in the absence of any other findings that were indicative of an effect on kidney function in this study, these differences were attributed to normal biological variation and were therefore not related to treatment. - Urinalysis findings:
- not examined
- Behaviour (functional findings):
- no effects observed
- Immunological findings:
- not examined
- Organ weight findings including organ / body weight ratios:
- no effects observed
- Gross pathological findings:
- no effects observed
- Neuropathological findings:
- no effects observed
- Histopathological findings: non-neoplastic:
- no effects observed
- Histopathological findings: neoplastic:
- no effects observed
- Key result
- Dose descriptor:
- NOAEL
- Effect level:
- >= 1 312 other: mg TOS/kg bw/day
- Based on:
- other: total organic solids (TOS)
- Sex:
- male/female
- Basis for effect level:
- other: Trehalase was well-tolerated and did not cause any adverse change.
- Critical effects observed:
- no
- Conclusions:
- It is concluded that oral administration of trehalase, batch PPT42347 to Han Wistar rats at doses up to 100% of trehalase, batch PPT42347 for 13 weeks was well-tolerated and did not cause any adverse change. The no-observed adverse-effect level (NOAEL) was therefore considered to be 100% of the trehalase, batch PPT42347 (equivalent to 1312 mg TOS/kg body weight/day).
- Executive summary:
The objective of this study was to assess the systemic toxic potential of trehalase, batch PPT42347 when administered orally by gavage to Han Wistar rats for 13 weeks. Three groups, each comprising ten males and ten females, received doses of 10, 33 or 100% of Trehalase, batch PPT42347 (equivalent to 131, 433 and 1312 mg TOS/kg bw/day). A similarly constituted control group received the vehicle (reverse osmosis water) at the same volume-dose (10 mL/kg body weight) as the treated groups.
During the study, clinical condition, detailed physical and arena observations, sensory reactivity, grip strength, motor activity, body weight, food consumption, visual water consumption, ophthalmic examination, hematology (peripheral blood), blood chemistry, organ weight, macropathology and histopathology investigations were undertaken.
There were no deaths associated with trehalase, batch PPT42347. The general appearance and behavior, sensory reactivity responses, grip strength and motor activity were not affected by treatment during the study. A control female was killed for welfare reasons in Week 9, but this was clearly an incidental death. There was no effect of treatment on body weight, food consumption or (visually assessed) water consumption. There were no treatment-related ophthalmic findings. The hematology and blood chemistry investigations during Week 13 did not identify any treatment-related findings. Organ weights were unaffected by treatment and there were no treatment-related macroscopic or histopathological findings.
It is concluded that oral administration of trehalase, batch PPT42347 to Han Wistar rats at doses up to 100% of trehalase, batch PPT42347 for 13 weeks was well-tolerated and did not cause any adverse change. The no-observed adverse-effect level (NOAEL) was therefore considered to be 100% of the trehalase, batch PPT42347 (equivalent to 1312 mg TOS/kg body weight/day).
Reference
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed
- Dose descriptor:
- NOAEL
- 1 312 mg/kg bw/day
- Study duration:
- chronic
- Species:
- rat
- Quality of whole database:
- Toxicological data has been generated within the enzyme producing industry during the last 40 years. Substantial documentation on the safety of the production strains has been generated, and the enzyme test materials are thoroughly characterized. High quality studies for all relevant endpoints for in vivo studies as well as in vitro studies show that industrial enzymes from well-known and well characterized production strains have very similar safety profiles across the catalytic activities. Read-across can therefore be applied for the majority of toxicological endpoints. The database can therefore be considered of high quality.
Repeated dose toxicity: inhalation - systemic effects
Link to relevant study records
- Endpoint:
- sub-chronic toxicity: inhalation
- Data waiving:
- exposure considerations
- Justification for data waiving:
- other:
Reference
Repeated dose toxicity: inhalation - local effects
Link to relevant study records
- Endpoint:
- sub-chronic toxicity: inhalation
- Data waiving:
- exposure considerations
- Justification for data waiving:
- other:
Reference
Repeated dose toxicity: dermal - systemic effects
Link to relevant study records
- Endpoint:
- short-term repeated dose toxicity: dermal
- Data waiving:
- study scientifically not necessary / other information available
- Justification for data waiving:
- other:
Reference
Repeated dose toxicity: dermal - local effects
Link to relevant study records
- Endpoint:
- short-term repeated dose toxicity: dermal
- Data waiving:
- study scientifically not necessary / other information available
- Justification for data waiving:
- other:
Reference
Additional information
It is concluded that oral administration of trehalase, batch PPT42347 to Han Wistar rats at doses up to 100% of trehalase, batch PPT42347 for 13 weeks was well-tolerated and did not cause any adverse change. The no-observed adverse-effect level (NOAEL) was therefore considered to be 100% of the trehalase, batch PPT42347 (equivalent to 1312 mg TOS/kg body weight/day).
The repeated dose inhalation and dermal toxicity studies were waived.
- The dermal study was waived because of the low likelihood of absorption of enzymes through the skin due to the physico-chemical properties of the enzyme protein.
- The inhalation study was waived because exposure is too low to exert any toxicity. Potential exposure by inhalation to an amount of enzyme that toxicologically relevant is unrealistic, due to the formulation of enzymes and the stringent work practices, enforced because of the risk of sensitisation by inhalation.
Justification for classification or non-classification
Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice.
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