Broad-specific monoclonal antibody based IACs purification coupled UPLC-MS/MS method for T-2 and HT-2 toxin determination in maize and cherry samples

Abstract

In this work, broad-specific monoclonal antibody based immunoaffinity columns (IACs) coupled to ultra-high performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) for purification and determination of T-2 and HT-2 toxin in maize and cherry samples. Based on broad-specific anti-T-2 monoclonal antibody, the prepared IACs could be used for both T-2 and HT-2 purification with cross reactivity values at 100% for T-2 and 108% for HT-2. Column capacity was 3.14 μg mL −1 gel for T-2 toxin, and 3.20 μg mL −1 gel for HT-2 toxin. After 10 cycles of usage at intervals of 2 days, column capacity decreased to 0.85 μg mL −1 gel for T-2 and 1.22 μg mL −1 gel for HT-2 toxin. The developed method exhibited high sensitive results with limit of detection (LOD) lower than 0.35 μg kg −1 and 1.42 μg kg −1 .

Solid‐phase extraction techniques based on nanomaterials for mycotoxin analysis: An overview for food and agricultural products

“…For in-house fabricated IACs, antibodies coupled to solid supports have been packed in empty SPE columns. Com-monly, monoclonal antibodies against aflatoxins, DON, OTA, ZEN, T-2, and sterigmatocystin (STER) were used [71][72][73]. Contrary to widely commercialized IACs, AACs have been mainly prepared in laboratories to analyze aflatoxins and OTA [74,75].…”

Immunoaffinity Extraction and Alternative Approaches for the Analysis of Toxins in Environmental, Food or Biological Matrices

“…To immobilize Abs on a solid sorbent, the most common approach is with regard to their covalent bonding, which is often achieved by coupling an accessible amino group of the Abs with a support that contains reactive groups such as epoxy [ 93 ] or aldehyde [ 97 ] or groups that can be activated using glutardialdehyde [ 71 , 100 , 101 ], carbonyldiimidazole, cyanogen bromide (CNBr) or N-hydrosuccinimide (NHS). Some activated supports are commercially available such as NHS- [ 70 ] or CNBr- [ 69 , 76 , 84 , 85 , 86 , 87 , 88 , 91 , 94 , 95 , 96 , 103 , 112 ] activated Sepharose or glutardialdehyde activated silica [ 74 , 77 , 82 ]. Non-covalent binding can also be used to couple Abs to the sorbent.…”

“…Reported or calculated dilution factor values are mainly between 2 and 10. It is worthwhile noticing that a residual amount of solvent in the extract can sometimes be necessary to ensure the complete solubilization of the toxins [ 74 , 77 , 84 , 85 , 86 , 94 , 95 , 96 ]. Moreover, some authors suggested to optimize the extraction conditions of the toxins from the sample not only regarding the final extraction recovery of the toxins but also by studying the effect of the nature of the extraction solvent on the final selectivity measured by the removal of the interfering peak in the final chromatogram.…”

“…Regarding the ISs prepared in laboratories, softer elution conditions are indicated in Table 2 , such as the use of a water–acetonitrile or water–methanol mixture, to favor the reuse of the ISs. However, it is worthwhile to notice that the use of pure methanol does not prevent the reuse of ISs [ 86 , 88 , 94 , 95 ]. The reusability of ISs was not so much studied even for laboratory-made ISs, but some works demonstrated that ISs can be reused 5 [ 86 , 88 ], 6 [ 70 ], 8 [ 96 ] or even more than 60 times [ 90 ] without observing a decrease in the extraction recoveries.…”

“…This approach was also proposed by different groups to evaluate the clean-up effect of ISs [ 38 , 60 , 65 , 68 , 80 ]. Indeed, it was considered as necessary for the simultaneous quantification of HT-2 and T-2 toxins in maize and cherry samples [ 94 ], of several toxins in cereals [ 48 , 54 , 58 , 85 ], feed samples [ 84 ], or urine [ 62 ]. In return, matrix match calibration was studied and considered as not necessary for the analysis of a mix of mycotoxins in cereals [ 55 , 61 ] or SMC [ 31 ] in various samples thus allowing the use of the much simpler external calibration method.…”