When this test applies
ISO 10993-1:2025 flags systemic toxicity for devices with prolonged (24 h to 30 days) body contact. The 14-day repeated dose is the subacute study that fits that window: it covers cumulative exposure that a single acute dose cannot, without committing to the 28-day study that long-term and permanent contact usually calls for.
When to run it
After cytotoxicity, sensitization and irritation are clear, and only once the material and its manufacturing process are locked — a formulation change invalidates the study. Run chemical characterisation first where you can: an ISO 10993-17 toxicological risk assessment built on good extractables data often addresses the subacute endpoint without an animal study. Confirm the duration against your actual contact duration before commissioning; if contact is long-term, the 28-day dual route study (GRK-4004-04) is the one regulators will expect.
What a pass tells you
That across 14 consecutive days of dosing there were no toxicologically significant differences between the test and control groups in clinical signs, body weight, food consumption, haematology, clinical chemistry, organ weights or histopathology — assessed per ISO 10993-11.
What it does not tell you
Nothing about exposure beyond 14 days; the 28-day study is the next tier and chronic toxicity is separate again. Nothing about local effects at an implant site, genotoxicity or sensitization. And it tests what the two vehicles extract: a constituent neither the polar nor the non-polar extract recovers is not assessed here.
Method
Testing per ISO 10993-11:2017. Two extracts are prepared at 18 mL per extract — 0.9% sodium chloride (polar) and sesame oil, NF (non-polar) — giving at least 28 discrete dose portions, one of each extract on each of the 14 days. Dosing is by systemic injection in the rat (preferred) or mouse against concurrent controls, with routine body weights, feed consumption, haematology, clinical chemistry, gross pathology, organ weights and histopathology at termination. Extraction ratio follows ISO 10993-12: 6 cm²/mL for material under 0.5 mm thick, 3 cm²/mL at 0.5 mm or thicker, or 0.1 to 0.2 g/mL where surface area is indeterminate.
Where it runs
This method is internationally sourced (OUS). The laboratory is ISO/IEC 17025 accredited and GLP certified, and the report is issued under Groenakker’s ISO 13485:2016 certified quality system, in Groenakker’s report template or in your document template on request at no extra cost.
Usually ordered alongside
Acute systemic injection (GRK-4004-02) is normally run first or in the same campaign. Material-mediated pyrogenicity (GRK-4004-01) is a separate endpoint where blood or CSF contact makes it relevant. Chemical characterisation with an ISO 10993-17 assessment can reduce or remove the need for this study, and the 28-day dual route study (GRK-4004-04) replaces it where contact is long-term.
