Table 8.

Main steps in preparing food samples for mycotoxin analysis.

Sample Preparation	Importance	Procedure	Challenges
Sampling	Accurate mycotoxin analysis relies on proper sampling, as uneven distribution in food can cause incorrect results.	The process begins with collecting food samples from different parts of a batch to account for variability in mycotoxin contamination. To form a composite sample, random sub-samples are collected and combined for solid foods like grains, nuts, and cereals. Mixing ensures even distribution before sampling for liquids, milk, or fruit juices [100].	Heterogeneous mycotoxin contamination complicates sampling, requiring larger samples for bulk goods like grains to minimize errors.
Homogenization	After sampling, food must be homogenized to ensure even distribution of mycotoxins, particularly in solid or semi-solid foods [101].	Homogenization involves grinding or blending the sample into a fine, uniform consistency. Equipment such as mills or blenders reduces the particle size of solid foods like grains or nuts. Mixing ensures consistency in liquids.	Prevent contamination during homogenization, as excessive grinding heat can degrade sensitive mycotoxins.
Extraction	The extraction aims to separate mycotoxins from the food matrix into a solvent, isolating them from interfering compounds like proteins, fats, and carbohydrates for easier analysis.	The solvent choice depends on the food type and mycotoxin analyzed. Common organic solvents like methanol and acetonitrile effectively dissolve mycotoxins. The homogenized food is mixed with the solvent and agitated for solid samples, while liquid samples require filtration or centrifugation to remove debris before analysis [102].	The selection of an appropriate solvent system is crucial. It must efficiently extract mycotoxins while minimizing the coextraction of other food components that may interfere with the analysis.
Cleanup	Following extraction, a cleanup step is often necessary to remove unwanted compounds from the extract, such as fats, sugars, and proteins, which can interfere with the sensitivity and accuracy of detection methods.	Cleanup methods vary by analytical technique and food matrix:Solid-Phase Extraction (SPE): extracts pass through an adsorbent column that binds unwanted substances while allowing mycotoxins to pass.Immunoaffinity Columns (IAC): use antibodies to selectively bind and isolate mycotoxins, which are then eluted with a solvent.Liquid–Liquid Partitioning: separates mycotoxins based on solubility in two immiscible phases, typically an organic solvent and water [103].	Cleanup must be optimized for different food types, as excessive removal of matrix components can result in the loss of mycotoxins, reducing the sensitivity of the analysis.
Concentration	After extraction and cleanup, mycotoxin levels may be too low for accurate detection. Concentration improves these levels, making quantification easier.	Concentration is typically achieved by evaporating the solvent used during extraction, leaving behind a more concentrated sample of mycotoxins. This is usually performed under reduced pressure or using rotary evaporation to avoid degradation of the mycotoxins [98].	Avoid over-concentration, as it can cause matrix effects or unwanted compound precipitation that interferes with analysis.