ISO 14797:1999

Overview

ISO 14797:1999 specifies a validated high-performance liquid chromatography (HPLC) method for the determination of furazolidone in animal feeding stuffs and premixtures. The method covers quantitative analysis for animal feeds with furazolidone levels from 25 mg/kg to 5 000 mg/kg and premixtures with a mass fraction up to 20%. Furazolidone is a nitrofuran veterinary chemotherapeuticum; this standard supports accurate measurement for quality control and regulatory testing.

Key topics and technical requirements

• Scope and ranges: Applicable to feeding stuffs (25–5 000 mg/kg) and premixtures (up to 20% m/m).
• Sample preparation: Follow ISO 6498 for test sample preparation; grinding to 1 mm and representative sampling are emphasized.
• Extraction: Use acetonitrile:methanol (1:1 v/v) as the extraction solvent; animal feeds are pre-wetted with water prior to extraction.
• Clean-up: Short aluminum oxide column purification for feed extracts; premixtures are diluted directly.
• HPLC conditions: Reverse-phase chromatography with a mobile phase of sodium acetate buffer and acetonitrile (filtered and degassed). Typical parameters in the standard include:
  • Flow rate: 0.6 mL/min
  • Injection volume: 20 µL
  • Detection: UV at 365 nm (diode array detector may be used for confirmation)
  • Column: silica bonded C18 packing (e.g., 200 mm × 3 mm, 5 µm)
• Standards and calibration: Stock (≈250 µg/mL) and working solutions (≈5 µg/mL and 12.5 µg/mL) are used. Linearity and reproducibility criteria are specified (peak variance <5%).
• Confirmation: Identity confirmation by co‑chromatography or diode array detection with criteria such as retention time within ±5% and spectral purity checks (difference spectra within specified tolerances).
• Safety: Furazolidone is light‑sensitive and toxic - work in the absence of daylight, use fume hood, and follow protective measures.

Applications

• Routine quality control of veterinary premixtures and compounded feeds.
• Regulatory and compliance testing for furazolidone residues in feed materials.
• Contract analytical laboratories performing HPLC-based feed analysis.
• Research on nitrofuran distribution and stability in feed matrices.

Who should use ISO 14797

• Feed manufacturers and premixture producers for product verification.
• Food and feed testing laboratories and quality assurance teams.
• Regulatory agencies and inspection bodies setting compliance limits.
• Contract analytical laboratories offering trace-level HPLC testing.

Related standards

• ISO 6498:1998 - Preparation of test sample (normative reference cited by ISO 14797).

Keywords: ISO 14797, furazolidone, HPLC, animal feeding stuffs, premixtures, nitrofuranes, feed analysis, analytical method, UV detection.