Development and validation of solvent free elution procedures for the isolation of mycotoxins by immunoaffinity

Inhaltsverzeichnis (Table of Contents)

• Introduction and Scope of the work
• Theoretical Part
  • Fungi, moulds and mycotoxins
  • Major Groups of mycotoxins occuring in food and feed
  • Food Safety Aspects and Regulations
  • Mycotoxin-Analysis
  • Immunoaffinity Clean-Up
  • Immunosorbents and Antibodies
  • Supports for Immunosorbents
  • Bonding density
  • Selective Extractions and Cross-Reactivity
  • Capacity
  • Immuno-based applications for Mycotoxins
  • Determination of Mycotoxins
  • Principles of reversed-phase high performance liquid chromatography (RP-HPLC)
  • Individual Mycotoxins
  • Liquid chromatography / mass spectrometry (LC-MS) as universal detector for multi-toxin extracts
  • Results and Discussion
  • Exploratory Experiments with Deoxynivalenol
  • Background
  • The Surveillance procedure
  • pH solutions
  • Heated elution procedures
  • Statistical analysis
  • Zearalenone
  • Mixtures of water with organic solvents
  • Unspecific bindings
  • Recombination of antibodies
  • Interfering Peaks
  • Statistical analysis
  • Aflatoxins
  • Statistical Analysis
  • Aflatoxin B1
  • Aflatoxin B2
  • Aflatoxin G1
  • Aflatoxin G2
  • Ochratoxin A
  • Statistical analysis
  • Fumonisins
  • Statistical analysis
  • T-2 and HT-2 toxin
  • Technical
  • Alternative heating procedures
  • Self-built heating Block
  • Microwave
  • Soldering Rod
  • Oscillating circuit
  • Electrical operated heating
  • (Semi-) Automation
  • Summary

  Zielsetzung und Themenschwerpunkte (Objectives and Key Themes)

  The thesis aims to develop and validate solvent-free elution procedures for the isolation of mycotoxins using immunoaffinity techniques. The focus is on providing a safe and reliable method for mycotoxin analysis, which is crucial for ensuring the safety of food and feed products.

  • Development of solvent-free elution procedures for mycotoxin isolation
  • Validation of the developed procedures
  • Optimization of immunoaffinity clean-up techniques
  • Exploration of alternative heating methods for elution
  • Evaluation of the impact of solvent-free elution on mycotoxin analysis

  Zusammenfassung der Kapitel (Chapter Summaries)

  The thesis begins with a comprehensive introduction to the importance of mycotoxin analysis in food safety and the challenges associated with current methods. It then delves into the theoretical aspects of mycotoxins, their classification, and the analytical techniques commonly employed for their determination.

  The subsequent chapters present the results of the research, focusing on the development and validation of solvent-free elution procedures for various mycotoxins, including deoxynivalenol, zearalenone, aflatoxins, ochratoxin A, fumonisins, and T-2 and HT-2 toxin. The results are presented in detail, including statistical analysis and discussions on the factors influencing elution efficiency and potential interferences.

  The final chapter explores alternative heating procedures for the elution process, aiming to improve the efficiency and automation of the developed methods.

  Schlüsselwörter (Keywords)

  Mycotoxin analysis, immunoaffinity, solvent-free elution, food safety, HPLC, LC-MS, deoxynivalenol, zearalenone, aflatoxins, ochratoxin A, fumonisins, T-2 and HT-2 toxin, heating procedures, automation.

  Frequently Asked Questions

  What are mycotoxins and why are they dangerous?

  Mycotoxins are toxic compounds produced by fungi (moulds). They can contaminate food and feed, causing serious health problems in humans and animals, ranging from acute poisoning to long-term effects like cancer.

  What is immunoaffinity clean-up in mycotoxin analysis?

  It is a selective purification technique using antibodies to bind specific mycotoxins from a complex sample, allowing for more accurate detection and measurement.

  Why develop solvent-free elution procedures?

  Traditional methods use organic solvents which are often toxic and environmentally harmful. Solvent-free procedures (e.g., using heat) are safer, greener, and can be easier to automate.

  What heating methods were explored for elution?

  The research explored alternative heating methods such as self-built heating blocks, microwaves, and electrical heating to release the toxins from the antibodies without using solvents.

  Which mycotoxins were tested in this study?

  The study focused on major toxins including Deoxynivalenol, Zearalenone, Aflatoxins, Ochratoxin A, Fumonisins, and T-2/HT-2 toxins.