Environmental Analysis & Electrochemistry
The role of analytical technologies in detecting mycotoxin and tropane alkaloid food contamination amidst climate change
Holly Lee, PhD, Staff Scientist, Food Market, SCIEX
As climate change continues to reshape our world, its impact on food safety is becoming a growing concern. One of the most pressing issues in agriculture and food production is the increased prevalence of mycotoxin contamination. Mycotoxins are toxic secondary metabolites produced by fungi, particularly species of Aspergillus, Fusarium, and Penicillium, which thrive in warm and humid conditions.[1]
Rising global temperatures and shifting precipitation patterns have created an environment that fosters fungal growth, leading to increased occurrences of mycotoxin contamination in crops.
These contaminants pose serious health risks, including carcinogenic, nephrotoxic, hepatotoxic, and immunosuppressive effects. In response, the food industry and regulatory agencies must strengthen food safety testing measures to mitigate risks associated with mycotoxins. Moreover, with climate change facilitating the spread of new plant pathogens, farmers are compelled to adjust their pesticide strategies, either by increasing application frequency or adopting novel pesticide formulations. This creates an additional concern: plants are often weakened by periods of extreme weather, making them more susceptible to pests and diseases and necessitating increased pesticide use.[2] At the same time, mycotoxin-producing fungi are fl ourishing under these altered environmental conditions.
Beyond mycotoxins, another growing concern in food safety is the presence of tropane alkaloids, such as scopolamine and hyoscyamine. These naturally occurring toxins, found in certain invasive plant species such as Datura stramonium (Jimson weed), pose serious health risks when inadvertently co-harvested with food crops.[3] The increase in erratic climate patterns has led to the unintentional spread of tropane alkaloid-containing plants into agricultural fi elds, raising the risk of contamination in grain and seed-based products. Addressing these risks requires advanced analytical techniques that can effectively detect and quantify both mycotoxins and tropane alkaloids in food matrices.
Furthermore, masked mycotoxins - modifi ed forms of mycotoxins that escape conventional detection methods like immunoassay-based techniques - are emerging as a critical challenge.[1,4] These metabolites are transformed by plants or microorganisms and can revert to their toxic parent compounds during digestion, posing a hidden health risk. Advanced analytical techniques like mass spectrometry (MS) enable targeted and nontargeted screening to achieve comprehensive coverage of both these compounds and their parent analogues, while also providing the sensitivity required for their quantitation.
Regulatory agencies, such as the European Union (EU), Food and Agriculture Organization (FAO), and the World Health Organization (WHO), have imposed strict maximum residue levels (MRLs) to ensure consumer safety, particularly for baby foods and staple grains.[5,6] Key regulated mycotoxins include afl atoxins (e.g., AFB1), which are highly carcinogenic, ochratoxins (OTA), patulin, fumonisins (e.g., FB1, FB2), zearalenone (ZEN), deoxynivalenol (DON), ergot sclerotia and alkaloids, T-2 and HT-2 toxins, and trichothecenes.[5]
Ultra-sensitive and selective detection methods are necessary to accurately identify and quantify mycotoxins, masked mycotoxins, and tropane alkaloids in complex food matrices at levels that are compliant with those established by regulatory agencies. These analytical methods include advanced applications, workfl ows, and instruments, with cutting-edge MS leading the way.
Analytical approaches to detect mycotoxins and tropane alkaloids
The selection of an appropriate method depends on factors, such as sensitivity, specifi city, throughput, cost, and regulatory compliance. Conventional approaches have distinct advantages and inherent limitations. These include bioanalytical methods that rely on immuno-recognition or receptor binding, such as enzyme-linked immunosorbent assay (ELISA), dipsticks, lateral fl ow devices, and immuno-sensors. Immunoassay- based techniques, like ELISA and lateral fl ow assays, are cost-effective, rapid, and ideal for high-throughput and fi eld applications. They enable on-site food testing but are prone to cross-reactivity, which may lead to false positives. Their inability to differentiate between masked and free mycotoxins also limits their effectiveness, making them more suitable for preliminary rapid screening.[7,8]
Other techniques, such as thin-layer chromatography (TLC), may also be used, provided that the signals they generate directly correspond to the mycotoxins of interest.[5,6] As a low-cost and easy-to-use technique, TLC is valuable in resource-limited settings for initial screenings.
Figure 1: Extracted ion chromatogram (XIC) showing chromatographic separation of mycotoxins in a wine matrix, with all tested mycotoxin compounds overlaid, and using SCIEX OS 3.4 software for data processing, including the peak-to-peak signal-to-noise and MQ4 integration algorithm.[11] Credit: SCIEX.
However, its low sensitivity, semi-quantitative nature, and time-consuming process make it unsuitable for regulatory routine testing without confi rmatory analysis.[7,9] Advancements in miniaturised MS and biosensors enable real-time, onsite monitoring. Handheld devices utilising near-infrared and Raman spectroscopy provide non- destructive detection of mycotoxins and tropane alkaloids, enhancing food safety interventions.[10,11,12,13]
Chromatographic approaches, such as high-performance liquid chromatography (HPLC) and gas chromatography (GC), are often integrated with detection methods like fl uorescence detection (FLD) and mass spectrometry (MS and MS/MS) to enhance orthogonal separation of different compounds. HPLC-FLD reliably quantifi es mycotoxins with high sensitivity but is limited to fl uorescent analytes. Its limited detection of masked mycotoxins and tropane alkaloids, along with the need for complex derivatisation, adds to its drawbacks.[7,9]
Ambient MS enables rapid and onsite screening of food contaminants with minimal preparation. However, it lacks the sensitivity and quantitative performance of LC- MS/MS and may be prone to matrix interferences.[7,10] GC-MS excels in detecting volatile and thermally stable compounds, particularly tropane alkaloids. However, it requires derivatisation for non-volatile mycotoxins, has longer analysis times, and is less sensitive to polar mycotoxins.[7,9] Regarded as the gold standard for mycotoxin quantitation, LC-MS/MS offers high specifi city, sensitivity, and multi-residue detection capability, but high costs and the requirements of highly trained personnel and complex sample preparation may present challenges for widespread adoption.[7,11]
The advantages of the latest mass spectrometry technologies
Effective chromatographic separation is crucial for the accurate quantifi cation of mycotoxins through LC-MS/MS analysis. HPLC helps ensure that structurally similar mycotoxins are well separated (see Figure 1). Advanced workfl ows in toxin analysis integrate streamlined approaches, such as the Quick Easy Cheap Effective Rugged Safe (QuEChERS) sample preparation protocol, solid-phase extraction (SPE) purifi cation technique, and optimised gradient elution methods.[11] These approaches help minimise matrix interferences and enhance analyte retention and peak resolution, leading to more accurate quantitation of mycotoxins and tropane alkaloids. Additionally, optimised targeted workfl ows, such as multiple reaction monitoring (MRM), improves specifi city by further minimising matrix interferences, resulting in high signal-to noise ratios.
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