ISO 15141:2018 PDF

Overview

ISO 15141:2018 specifies a validated analytical procedure for the determination of ochratoxin A (OTA) in cereals and cereal products using high performance liquid chromatography (HPLC) with immunoaffinity column cleanup and fluorescence detection. The standard defines sample preparation, extraction, cleanup, HPLC conditions, calibration, identification and confirmation steps. The limit of quantification (LOQ) is 0.2 μg/kg; the method detection limit depends on sample matrix and instrument performance. ISO 15141:2018 replaces earlier ISO 15141-1/2 editions and refines extraction and purification principles.

Key Topics and Technical Requirements

• Sample preparation and extraction
  • Typical sample mass: 25 g (12.5 g for light samples such as wheat bran).
  • Extraction solvent: acetonitrile–water (60:40, v/v); shake 30 min or blend 3 min; centrifuge or filter.
• Dilution and cleanup
  • Dilution into phosphate buffered saline (PBS) and centrifugation/filtration options.
  • Immunoaffinity column containing anti‑OTA antibodies for selective cleanup (example product cited; equivalents allowed).
  • Column flow ~1–2 drops/s; wash with specified washing solution and water; elute with 1.5 ml methanol/acetic acid (98:2) and evaporate.
• HPLC and detection
  • Reverse‑phase column (example: C18, 150 × 4.6 mm, 5 μm) for baseline resolution of OTA.
  • Mobile phase: acetonitrile/water/glacial acetic acid (48/51/1, v/v/v), degassed.
  • Fluorescence detection with optional confirmation by derivatization (boron trifluoride in methanol).
• Calibration and performance
  • Calibration solutions covering 0.05–1.0 ng per 100 μl injection.
  • Precision data and interlaboratory results are included; laboratories must verify standard solution stability and instrument suitability.
• Safety
  • OTA is nephrotoxic and a probable carcinogen - follow appropriate handling and decontamination procedures.

Applications and Users

ISO 15141:2018 is intended for:

• Food testing and quality control laboratories analyzing cereals, flours, breakfast cereals, and related products.
• Regulatory agencies enforcing mycotoxin limits and import/export control.
• Agricultural processors and cereal manufacturers implementing HACCP and product safety monitoring.
• Method developers and proficiency-test organisers who require a standardized, HPLC‑based OTA procedure.

Benefits include high selectivity from immunoaffinity cleanup, robust quantification at low μg/kg levels, and harmonized methodology to support compliance, risk assessment, and international trade.

Related Standards

• ISO 24333 (sampling guidance for cereals)
• ISO 3696 (water for analytical laboratory use)
• Previous: ISO 15141-1 and ISO 15141-2 (replaced by this edition)

Keywords: ISO 15141:2018, ochratoxin A, HPLC method, immunoaffinity column, fluorescence detection, cereals testing, LOQ 0.2 μg/kg.