Overview
EN ISO 14183:2008 (ISO 14183:2005) specifies a validated high-performance liquid chromatographic (HPLC) method for the determination of monensin, narasin and salinomycin in animal feeding stuffs, supplements (dry and liquid) and mineral premixtures. The method uses reverse‑phase HPLC with post‑column derivatization (vanillin) and spectrophotometric detection at 520 nm. It is not applicable to pharmaceutical drug premixes; lasalocid and semduramicin cannot be determined by this method.
Key topics and technical requirements
- Scope: Applicable to animal feeds, supplements and mineral premixtures; excludes drug premixes.
- Extraction: Methanol/water (90 + 10) with mechanical shaking for 1 hour, then filtration.
- Chromatography: Reverse‑phase HPLC with post‑column derivatization using vanillin (daily‑prepared reagent) and detection at 520 nm.
- Confirmation: Suspect trace positives or unexpected ionophores can be confirmed by hexane extraction or post‑column derivatization with dimethylaminobenzaldehyde (DMAB).
- Reference standards & preparation: Requires authenticated monensin, narasin and salinomycin standards; stock standards prepared in neutralized methanol for certain analytes (narasin). Concentrations and component composition must be recorded.
- Limits of quantitation (LOQ): Approximately 1 mg/kg for monensin and 2 mg/kg for salinomycin and narasin. Users may validate lower LOQs for their laboratory.
- Safety & quality: Use HPLC‑grade solvents, protect standards from light, handle toxic ionophore standards in a fume hood and follow appropriate PPE.
- Reference for sampling: ISO 6498:1998 (preparation of test samples) is a normative reference cited in the standard.
Applications and users
EN ISO 14183:2008 is intended for:
- Feed testing and analytical laboratories performing routine or regulatory analysis of ionophores in feeds.
- Quality control teams at feed manufacturers and premix producers monitoring monensin, narasin and salinomycin levels.
- Regulatory authorities and compliance labs verifying feed composition, safety and labeling claims.
Practical advantages include established extraction and HPLC confirmation procedures, defined LOQs, and guidance on standard preparation and detection-facilitating reproducible, comparable results across laboratories.
Related standards
- ISO 14183:2005 - original international text adopted as EN ISO 14183:2008.
- ISO 6498:1998 - preparation of test samples for animal feeding stuffs.
- CEN/TC 327 - European technical committee responsible for animal feeding stuffs methods.
Keywords: EN ISO 14183:2008, ISO 14183:2005, monensin, narasin, salinomycin, HPLC, post‑column derivatization, animal feeding stuffs, limit of quantitation.
