Table 6.

Primary techniques used for mycotoxin analysis.

Method	Description
Chromatography	Chromatography is one of the most widely used techniques for analyzing mycotoxins due to its high sensitivity, accuracy, and ability to separate and identify multiple mycotoxins simultaneously. Two common forms of chromatography used in mycotoxin analysis are high-performance liquid chromatography (HPLC) and gas chromatography–mass spectrometry (GC-MS) [84].
	Principle	Application	Advantage
High-Performance Liquid Chromatography (HPLC)	HPLC involves the separation of mycotoxins based on their interaction with a stationary phase (usually a column) and a mobile phase (usually a solvent). The different affinities of mycotoxins for the stationary phase allow them to be separated, detected, and quantified [85].	HPLC is commonly used to analyze aflatoxins, fumonisms in various foods, including cereals, nuts, and dairy products. It is highly effective when coupled with fluorescence or UV detection methods, which enhance sensitivity for specific mycotoxins.	HPLC offers high resolution, accuracy, and the ability to detect low levels of mycotoxins. It is widely accepted in regulatory testing and can be used for routine food safety monitoring.
Gas Chromatography–Mass Spectrometry (GC-MS)	GC-MS vaporizes mycotoxin samples, separates them via gas chromatography, and identifies them by mass spectrometry. Mycotoxins are derivatized to ensure volatility [86].	GC-MS is beneficial for the detection of volatile mycotoxins like patulin. It is susceptible and specific, making it suitable for detecting trace levels of mycotoxins in complex food matrices.	GC-MS provides high specificity and sensitivity, making it the gold standard for detecting mycotoxins like patulin in fruit juices.
Spectrometry	Mass spectrometry (MS) is often combined with chromatography to improve the sensitivity and specificity of mycotoxin detection. MS measures the mass-to-charge ratio of ionized mycotoxin molecules, providing precise molecular identification and quantification.
Mass Spectrometry (MS)	MS works by ionizing chemical compounds and measuring the mass-to-charge ratio of the resulting ions. Coupled with HPLC or GC, it allows for separating and identifying mycotoxins based on their mass [87].	HPLC-MS and GC-MS are widely used to analyze various mycotoxins, including aflatoxins, ochratoxins, and fumonisins. These techniques are valuable in multi-mycotoxin analysis, where several toxins may exist in a single sample.	MS provides high accuracy and detects multiple mycotoxins at low concentrations, which is crucial for regulatory testing and detailed mycotoxin profiling in food products.
Immunoassays	Immunoassays are rapid, sensitive, and cost-effective techniques for detecting mycotoxins in food. They rely on antibodies’ specific binding to mycotoxins and are suitable for quickly screening large numbers of samples [88].
Enzyme-Linked Immunosorbent Assay (ELISA)	ELISA is based on antibodies binding to mycotoxins, followed by an enzyme–substrate reaction that produces a detectable signal, usually colorimetric or fluorescent. The intensity of the signal corresponds to the concentration of mycotoxins in the sample [89].	ELISA is commonly used to detect aflatoxins, ochratoxns, zearalenone, and fumonisms in food products such as grains, nuts, and milk. It is often employed for routine screening in food industries and regulatory bodies.	ELISA is a quick, affordable method for mycotoxin detection but may lack the specificity of chromatographic techniques due to cross-reactivity.
Lateral Flow Immunoassay (LFIA)	LFIA is similar to ELISA but uses a test strip format. Mycotoxin–antibody interactions produce a visible line or signal on the test strip, indicating the presence of mycotoxins [90].	LFIA is used for rapid, on-site testing of mycotoxins in agricultural products. It is commonly applied to detect aflatoxins, fumonisins, and zearalenone in grains, nuts, and animal feed.	LFIA is a portable, quick method for mycotoxin detection, ideal for field testing, though less precise than lab methods.
Hyperspectral Analysis	Utilizes the spectral signature of materials across a wide range of wavelengths to identify and quantify mycotoxin contamination [91].	Sorting and detecting mycotoxin presence in grains, nuts, and other food products.	Non-destructive, rapid analysis, can be applied in real-time sorting, and high-throughput screening.
Immuno-detection	Employs specific antibodies that bind to mycotoxins, allowing their detection through various methods (e.g., ELISA, lateral flow assays) [90].	Food safety testing, monitoring mycotoxin levels in processed and raw food products.	High specificity and sensitivity, can detect low concentrations of mycotoxins, and suitable for various matrices.