lysophospholipaseIUBMB 3.1.1.5

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

September 23, 2013 - November 21, 2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2013

Report date:

2013

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

Adopted 17 December 2001.

Deviations:

no

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Active enzyme protein of lysophospholipase ( (EC no.618-333-0, CAS no. 9001-85-8, EC name Lysophospholipase, Enzyme class no 3.1.1.5 )

Molecular formula:

Not available

IUPAC Name:

Active enzyme protein of lysophospholipase ( (EC no.618-333-0, CAS no. 9001-85-8, EC name Lysophospholipase, Enzyme class no 3.1.1.5 )

Constituent 2

Reference substance name:

Protein as a constituent of enzyme deriving from the fermentation or extraction process

Molecular formula:

Not available

IUPAC Name:

Protein as a constituent of enzyme deriving from the fermentation or extraction process

Constituent 3

Reference substance name:

Carbohydrates constituent of enzyme deriving from the fermentation or extraction process

Molecular formula:

Not available.

IUPAC Name:

Carbohydrates constituent of enzyme deriving from the fermentation or extraction process

Constituent 4

Reference substance name:

Inorganic salts as a constituent of enzyme deriving from the fermentation or extraction process

Molecular formula:

Not available.

IUPAC Name:

Inorganic salts as a constituent of enzyme deriving from the fermentation or extraction process

Constituent 5

Reference substance name:

Lipids as a constituent of enzyme deriving from the fermentation or extraction process

Molecular formula:

Not available.

IUPAC Name:

Lipids as a constituent of enzyme deriving from the fermentation or extraction process

Test material form:

liquid

Details on test material:

- Lot/batch No.: PPW35424
- Expiration date of the lot/batch: 01 July 2023
- Stability under test conditions: The test material is stable for at least 24 hours at room temperature
- Storage condition of test material: Frozen (-20°C)

Test animals

Species:

rat

Strain:

other: CD (Crl:CD ‘SD’) albino rats

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River (UK) Ltd
- Females nulliparous and non-pregnant: Yes
- Age at study initiation: 8 - 12 weeks
- Weight at study initiation: 221 to 238 g
- Fasting period before study: overnight fasting prior to dosing
- Housing: Barriered rodent facility with control of temperature, humidity and lighting. The cages were solid bottomed polycarbonate cages with a stainless steel mesh lid. Softwood bark-free fibre were provided as bedding. Aspen chew block and plastic shelter for environmental enrichment.
- Diet: standard rodent diet (Rat and Mouse No. 1 Maintenance Diet) ad libitum
- Water: ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 to 23°C
- Humidity (%): 40-70%
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 2013-10-01 To: 2013-10-15

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

Undiluted test material for the top dose.

Doses:

Dose volume was 16 mL/kg bodyweight (equivalent to 2085 g TOS/kg bodyweight).

No. of animals per sex per dose:

6 (female only)

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Observations for clinical signs of effect: soon after dosing, and at frequent intervals for the remainder of Day 1. On subsequent days, animals were observed once in the morning and again at the end of the experimental day (with the exception of Day 15 - morning only). The weight of each rat was recorded on Days 1 (prior to dosing), 8 and 15.
- Necropsy of survivors performed: All animals were subject to a macroscopic examination which consisted of opening the cranial, thoracic and abdominal cavities. The macroscopic appearance of all examined organs was recorded.

Statistics:

No

Results and discussion

Effect levels

Key result

Sex:

female

Dose descriptor:

LD0

Effect level:

>= 2 085 mg/kg bw

Based on:

other: Total Organic Solids (TOS)

Mortality:

No animals died during the study.

Clinical signs:

No animals died and no sign of toxicity or ill health was seen at the routine physical examination.

Body weight:

No effect was observed on the body weights.

Gross pathology:

Effects on organs:
Macroscopic examination of animals killed on Day 15 of the observation period did not reveal any treatment-related findings.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

No signs of toxicity were observed in any of the rats treated with a single oral dose of 2085 mg total organic solids/kg body weight for a period of 14 days.

Executive summary:

The objective of the study was to assess the acute toxicity of lysophospholipase when administered by gavage as a single oral dose to one group six female rats followed by an observation period of 14 days.

The study was conducted in accordance with the OECD Guideline No 423, “Acute Oral Toxicity – Acute Toxic Class method”. The design of the limit test was used. The test item was supplied as a liquid ready to use. The dose volume administered was 2085 mg/kg body weight, based on the Total Organic Solids (TOS) content of the test substance.

A group of three fasted female rats received a single oral gavage dose of the test substance. As results at this dose level indicated the acute lethal oral dose of the test substance to be greater than 2085 mg TOS/kg body weight, in compliance with the study guidelines, a further group of three fasted females was similarly dosed at 2085 mg TOS/kg body weight to complete the study.

There were no deaths during the study. There were no clinical signs of reaction to treatment throughout the study. All animals were considered to have achieved satisfactory body weight gains throughout the study. No abnormalities were noted in any animal at the macroscopic examination at study termination on Day 15.

In conclusion, the acute median lethal oral dose (LD50) to rats of lysophospholipase, batch PPW35424 was demonstrated to be greater than 2085 mg TOS/kg body weight.