Table of Contents
ToggleAcute Toxicity Studies - Study Design of OECD Guideline 425: Acute oral toxicity – up and down procedure
The OECD Guideline 425 outlines the study design for the Acute Oral Toxicity – Up-and-Down Procedure, another method for assessing the potential harm of a substance following a single oral administration. Here’s a breakdown of its key aspects:
Objectives:
- Estimate the median lethal dose (LD50) of a substance, which is the dose at which 50% of the test animals die.
- Classify the substance according to its acute oral toxicity based on the GHS criteria.
- Provide initial information for further safety testing, if necessary.
Test System:
- Healthy young adult animals of a single sex (usually female) are utilized.
- Rodents (rats or mice) are the most common choices, but other species may be used in exceptional cases.
Dose Progression:
- The test starts with a dose level estimated to be below the LD50.
- Animals are dosed one at a time, with the dose for the next animal determined based on the previous animal’s response:
- If the previous animal survives, the dose for the next animal is increased by a predetermined factor (e.g., 3.2 times).
- If the previous animal dies, the dose for the next animal is decreased by the same factor.
- This up-and-down procedure continues until a statistically valid estimate of the LD50 is obtained.
Observation Period:
- Similar to the fixed dose method, animals are observed for at least 14 days after dosing.
- Detailed observations are made at least hourly during the first 24 hours and at specific intervals thereafter.
Endpoints:
- Mortality and time to death.
- Signs of toxicity, including gross behavioral changes, tremors, convulsions, salivation, and respiratory distress.
- Body weight changes.
- Clinical observations, including physical appearance, skin and fur condition, eyes, mucous membranes, and excreta.
- Necropsy findings (at termination of the study).
Data Analysis:
- Statistical methods are used to calculate the LD50 and its confidence interval based on the observed responses.
- Classification of the substance is based on the estimated LD50 and GHS criteria.
Benefits:
- Compared to the fixed dose method, this procedure requires fewer animals and provides a more precise estimate of the LD50.
- It avoids administering unnecessarily high doses that cause severe suffering.
Limitations:
- Requires more complex statistical analysis compared to the fixed dose method.
- May not be suitable for substances with very steep dose-response curves.
- Still relies on animal use, raising ethical concerns.
Additional Resources:
- OECD Test Guideline 425: https://www.oecd-ilibrary.org/environment/test-no-425-acute-oral-toxicity-up-and-down-procedure_9789264071049-en
- Acute Oral Toxicity Testing – Guidance Document Number 24, National Toxicology Program: https://ntp.niehs.nih.gov/sites/default/files/iccvam/suppdocs/feddocs/oecd/oecd-gd24.pdf
- Globally Harmonized System of Classification and Labelling of Chemicals (GHS): https://unece.org/DAM/trans/danger/publi/ghs/ghs_rev06/English/ST-SG-AC10-30-Rev6e.pdf