Using tandem mass spectrometry to quantify mycotoxin and ensure food safety
The security and stability of global food networks are gravely threatened by mycotoxin contamination. These potentially harmful secondary metabolites, which are produced by fungus, have an impact on a variety of foods, such as cereals, fruits, and spices. Furthermore, a large portion of crops—roughly 25%—are poisoned, making the issue pervasive.1.
Even though mycotoxins are already common (Figure 1), the contamination issue is expected to worsen. Weather variations, such rising temperatures, create the cozy conditions required to alter the growth and prevalence of toxins in different parts of the planet. In addition to increasing the levels of contamination that already exist, global warming raises the possibility that new strains and combinations will emerge.
Global rules are in place to guarantee that food and crops are fit for eating in order to protect human health in this ever-changing environment. Products are thoroughly analyzed to make sure they don’t surpass permissible mycotoxin levels in order to comply with this legislation. To guarantee correct quantification and consequently promote food safety, sensitive and precise procedures are essential. Can liquid chromatography with tandem mass spectrometry (LC-MS/MS) be the “comprehensive answer”?
Contamination by mycotoxin: An increasing issue
Exposure to mycotoxin can happen when food items are directly consumed that are contaminated or when animals consume feed that is contaminated. This indicates that mycotoxins can be harmful to livestock and people alike. The hazards are not the same for humans and cattle. In humans, mycotoxin ingestion is linked to illnesses like cancer and weakened immune systems; in livestock, the primary effects are diminished liver function and difficulties in reproducing.
Mycotoxin use has extremely worrying health implications, but it can also have severe economic effects. Breeding difficulties might result in a significant loss of livestock output and, consequently, revenue. The FDA calculates that losses from mycotoxin contamination for all crops amount to more than $932 million per year.Six And what about the businesses and vendors themselves? Reputation of the brand is in jeopardy. It is more crucial than ever to keep an eye on and control the maximum levels of mycotoxin in food, since there is so much at risk and even more losses are probably in store given global warming.
Reducing the risk of mycotoxin
To guarantee food safety, strict laws governing the highest levels of allowable mycotoxin in food are in place. Mycotoxin level standards are established by the Joint Expert Committee on Food Additives (JEFCA), an international initiative consisting of the Food and Agriculture Organization (FAO) and the World Health Organization (WHO). These are typically quite low, though they can differ based on the nation and the particular toxin. For instance, the maximum allowable levels of aflatoxin for food are 0.5–15 µg/kg, whereas the permissible levels of ochratoxin A are 1–50 µg/kg for food and 100–1,000 µg/kg for animal feed.
Our knowledge of mycotoxins and the analytical methods’ sensitivity are both rapidly advancing. The maximum permitted amounts of mycotoxins in food are changing as a result of these developments and worries about the rise in mycotoxin prevalence. For instance, the Canadian Food Inspection Agency (CFIA) started revising its rules pertaining to animal feed only a year ago.
Sensitive procedures are available, however it might be difficult to analyze and quantify mycotoxins in order to comply with laws. Sample preparation for analysis can be very difficult, and different approaches work well for different matrices and target analytes. The complexity of the situation is increased by modifications to maximum levels and the potential for newly emerging and modified/conjugated toxins, necessitating enlarged panels and greater analytical costs.
These days, analysts may easily and confidently analyze mycotoxins with increased confidence because of the advancements in LC-MS techniques.
LC-MS/MS: A combined method for analyzing mycotoxins
The use of LC-MS-based analytical methods in the food testing sector is becoming more and more common. Mycotoxin analysis is much enhanced when the method is combined with tandem mass spectrometry, such as LC-MS/MS, because of the higher sensitivity and improved structural understanding of the analyte. Most crucially, target mycotoxin chemicals in complex food matrices can be selectively and quantitatively determined using LC-MS/MS, which streamlines analysis.
Over the past ten years, a great deal of work has been done to advance analysis by gradually switching from single class to multiclass protocols. These protocols guarantee the identification of a large number of compounds using a limited number of techniques and are an affordable means to enhance the effectiveness and efficiency of the current official residue control system. In order to meet the demanding testing requirements in analytical laboratories, LC-MS/MS provides high-throughput capabilities and multi-residue protocols, allowing for multi-group screening approaches that cover analyte groups such pesticides and veterinary medications.8 Simpler and more ubiquitous sample extraction tools, including the Quick, Easy, Cheap, Effective, Rugged, and Safe (QuEChERs) sample preparation kits, contribute to these capabilities.
With many benefits, LC-MS/MS is the best method for finding mycotoxins at low concentrations in challenging matrices. Additionally, because of its versatility, it may be expanded to cover new targets. Additionally, because of its high resolution potentialities and software compatibility, it can screen chemicals that are unknown.
48 regulated mycotoxins that are of interest to quantify
In the EU, 48 mycotoxins are presently controlled or are being considered for regulation in the future, which can be very difficult for panels that are already in place. A created panel that analyzed all 48 mycotoxins in a single run was reported in one study that illustrated a method for quantifying these mycotoxins in grain-based meals.
A mass spectrometer and an ultra-high-performance LC (UHPLC) device were part of the setup. Following the extraction of the cereals using acetonitrile and water, the samples were reconstituted by evaporating them. Table 1 displays the results for the eleven mycotoxins that are currently covered by law; all of these compounds’ MRLs were determined with good precision and repeatability.
The limits of quantification (LOQs) for every controlled substance under investigation were less than the maximum residue limits (MRLs) that had been set. On five replicates at the LOD, the scientists also discovered good precision and reproducibility. The precision (in percentage coefficient of variation, percent CV) and intra-assay and inter-assay accuracy (in percent bias) were also measured and found to be below 20 percent, indicating the method’s acceptability for mycotoxin analysis.
The limits of quantification (LOQs) for every controlled substance under investigation were less than the maximum residue limits (MRLs) that had been set. On five replicates at the LOD, the scientists also discovered good precision and reproducibility. The precision (in percentage coefficient of variation, percent CV) and intra-assay and inter-assay accuracy (in percent bias) were also measured and found to be below 20 percent, indicating the method’s acceptability for mycotoxin analysis.
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Comply with present laws and expand to meet future demands
Because of the rising levels of contamination and the emergence of new strains, it is more crucial than ever to monitor mycotoxins in food. Mycotoxin levels must be effectively monitored in order to guarantee food safety, reduce food waste, and preserve brand reputation.
Because LC-MS/MS provides accurate, precise, and dependable mycotoxin level readings, it helps analysts comply with increasingly strict standards both today and in the future. Furthermore, the method can keep up with the increasing needs for quantitation because to its multiresidue capabilities. Its adaptability will make it possible to detect unknown substances in the future, together with high resolution and the right compound identification software. By utilizing these tactics, analysts, producers, and regulators are able to maintain the required high standards of food quality, even in the face of emerging obstacles.