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IN VITRO TOXICOLOGICAL STUDIES AND REAL TIME ANALYSIS OF RESIDUES IN FOOD

Objective

1.Primary objective:

To provide rapid food quality measurement techniques for detection and safety assessment of xenobiotics in food in order to satisfy consumer and/or industrial needs within the European Community.

2. Secondary objectives:

- To develop chromatographic/mass spectrometric methods for trace analysis of food contaminants, such that real time monitoring becomes possible;

- To develop and validate a set of rapid, low cost in-vitro test systems for the screening of the toxic potency of food contaminants;

- To integrate these methods to design suitable monitoring systems for the food industry;

- To evaluate and harmonize analytical and toxicological test methods for safety evaluation procedures at the European level.
Rapid and in vitro toxicological and analytical test systems for screening food contaminants and natural toxins are under development. Such tests will provide rapid food quality measurement techniques for detection and safety assessment.
The following aspects with respect to the development of real time analytical methodology were identified for the Concerted Action 'In Vitro Toxicological Studies and Real Time Analysis of Residues in Food':
method development for sample introduction into a tandem monitoring system;
automation of sample introduction;
sample pretreatment procedures to ensure minimum handling and rapid throughput;
back up analysis by chromatography;
production of reference materials.

With respect to the development of cell culture methodology the Concerted Action identified the following aspects:
isolation and characterization of cell organelles, primary cellcultures, and cell lines;
biotransformation profiles of xenobiotics;
characterization of Cyt P-450 isoenzymes;
adduct formation between xenobiotics and cellular macromolecules;
animal species and sex dependency of metabolic pathways;
mode of action and molecular mechanisms of toxicity of xenobiotics;
in vivo validation in target and test animals.

In a series of workshops scientific papers were presented and the subjects ranged from instrumental approaches to residue analysis to detailed analytical procedures for defined classes of compounds or individual xenobiotics. The available analytical instrumentation seemed sufficient for sensitive analysis of individual compounds and therefore efforts to improve sample preparation and pretreatment were considered with high priority. Papers were presented on the isolation and maintenance of various in vitro toxicological systems and how these systems are being used for toxicology problems. Research into the use of in vitro techniques showed clearly the advantages and limitations of these systems for risk assessment purposes. As regards th e level of external flora, there are a large number of yeasts, from 5E7 to E10/g. The presence of bacteria resistant to salt, such as Micrococaccae and Brevibacterium were also found.
The project attempts to develop rapid and in vitro toxicological and analytical test systems for screening of foodcontaminants and natural toxins. There is a strong need for such tests given the large number of food contaminants and the costs of traditional protocol safety testing. Moreover the food industry has to meet nowadays demands of consumers which tend to be more and more stringent with respect to the presence of contaminants in food. For that purpose this Action has brought together scientists from eleven EC and EC affiliated countries, with a common interest in the safety of residues of foodcontaminants, but coming from different scientific disciplines, namely toxicology and analytical chemistry.

The following aspects were identified for this Concerted Action:

1. Development of Real-Time Analytical Methodology

- method development for sample introduction into tandem MS;

- automation of sample introduction;

- sample pretreatment procedures to ensure minimum handling and rapid throughput;

-back up analysis by chromatography;

- production of reference materials.

2. Development of Cell Culture Methodology:

- isolation and characterization of cell organelles, primary cellcultures, and cell lines;

- biotransformation profiles of xenobiotics;

- characterization of Cyt P-450 isoenzymes;

- adduct formation between xenobiotics and cellular macromolecules;

- animal species and sex dependency of metabolic pathways;

- mode of action and molecular mechanisms of toxicity of xenobiotics;

- in-vivo validation in target and test animals.

Coordinator

State Institute for Quality Control of Agricultural Products
Address
45 Bornsesteeg, Po Box 230
6700 AE Wageningen
Netherlands