Liquid chromatographic method development for anabolic androgenic steroids using a monolithic column Application to...

Area of Science:

• Analytical Chemistry
• Food Safety
• Veterinary Science

Background:

• Anabolic androgenic steroids (AASs) are prohibited growth promoters in animal feed.
• Accurate detection methods are crucial for ensuring food safety and regulatory compliance.

Purpose of the Study:

• To develop and validate an isocratic High-Performance Liquid Chromatography (HPLC) method for screening AASs in finishing pig feed.
• To establish reliable analytical procedures for detecting specific AAS compounds used as growth promoters.

Main Methods:

• Utilized reversed-phase HPLC with UV detection on a Chromolith RP-18e column.
• Optimized mobile phase composition (ultrapure water:acetonitrile), flow-rate (3mL/min), and temperature (40°C) for optimal separation.
• Implemented a sample preparation protocol involving acetonitrile leaching, saponification, and solid-phase extraction (SPE) using Nexus cartridges.

Main Results:

• Achieved baseline resolution of target AASs within approximately 15 minutes.
• Demonstrated high extraction efficiencies (83-96%) for the analyzed compounds.
• Validated method performance according to European Commission Decision 2002/657/EC, with decision limits (CCα) and detection capabilities (CCβ) reported.

Conclusions:

• The developed HPLC method is suitable for screening anabolic androgenic steroids in pig feed.
• The validated method provides reliable and sensitive detection of AASs, supporting food safety regulations.
• Successfully applied the method to analyze various animal feed samples, confirming its practical utility.