One sample multi-point calibration curve as a novel approach for quantitative LC-MS analysis: The quantitation of six aflatoxins in milk and oat-based milk as an example

Integrating a Multiple Isotopologue Reaction-Monitoring Technique and LC-MS/MS for Quantitation of Small Molecules: Ten Mycotoxins in Cereals as an Example

“…As depicted in Figure A, we delve into the preparation of the MSCC strategy, wherein six blank matrix samples are meticulously prepared, each infused with varying concentrations of the target analyte Figure B vividly illustrates the procedural workflow of the OSCC strategy, which simplifies the process to a single analyte concentration addition to a blank matrix sample. , In stark contrast to the MSCC, which relies solely on quantifying the analyte using the MIRM channel with the highest abundance, the OSCC harnesses the potential of multiple MIRM channels associated with the analyte. In this study, a quantitative method combining the OSCC approach with the SIL-IS for ten prevalent mycotoxins found in cereals was devised and evaluated, which represents a significant advancement in our analytical capabilities.…”

“…Several strategies have been proposed to eliminate sample dilution, such as extending the calibration curve range or using fewer calibration points. , However, these approaches may result in inaccurate quantitation, particularly for samples at low concentration levels. In our proposed calibration strategy, based on the MIRM channels, sample dilution for samples outside the calibration curve range is obviated by monitoring the MIRM with proper intensity. ,, It is important to note that the dilution factor requires further evaluation. In this study, a series of dilution factors, including 2 ULOQ, 4 ULOQ, 8 ULOQ, 10 ULOQ, 15 ULOQ, and 20 ULOQ, were evaluated, with the results presented in Figure D.…”

“…In our proposed calibration strategy, based on the MIRM channels, sample dilution for samples outside the calibration curve range is obviated by monitoring the MIRM with proper intensity. 15,21,36 It is important to note that the dilution factor requires further evaluation. In this study, a series of dilution factors, including 2 ULOQ, 4 ULOQ, 8 ULOQ, 10 ULOQ, 15 ULOQ, and 20 ULOQ, were evaluated, with the results presented in Figure 3D.…”

“…The primary contributing factor to this performance differential is attributed to the OSCC strategy's requirement for a single concentration and a single injection analysis. 15,20,21,36 This streamlined approach significantly mitigates operational and analytical errors compared with the multiple concentration samples and multiple injection analyses mandated by the MSCC. Consequently, the OSCC strategy exhibits enhanced performance.…”

“…Over the past two decades, liquid chromatography coupled with triple quadrupole mass spectrometry (LC-MS/MS) and high-resolution mass spectrometry have attracted widespread popularity for mycotoxin determination due to their inherent specificity, sensitivity, and accuracy. ,, These techniques have emerged as the “gold standard” for multimycotoxin quantitative analysis. , Since mass spectrometry lacks inherent calibration, diverse calibration strategies have been developed to enhance accuracy and precision . Consequently, in the pursuit of a dependable analytical approach, the creation of calibration curves stands out as a pivotal phase. − The present state of research methods predominantly concentrated on individual mycotoxins or specific mycotoxin types, employing the conventional multisample external calibration curve (MSCC). , This approach involves introducing six nonzero concentrations into a blank matrix to perform the MSCC, enabling the back-calculation of the analyte’s concentration of interest . Nevertheless, when confronted with the simultaneous quantification of an array of compounds, the MSCC method proves laborious and time-intensive, primarily due to the extensive preparation of working standard solutions. , The one-point calibration (OPC) method simplifies the procedure by requiring just a single measurement point; its drawback is evident, that is, a lack of a second calibration point to characterize the instrument’s dynamic response adequately. , Typically, the OPC method is more suitable when the sample’s analyte concentration closely aligns with the value of the concentrations generated in the OPC.…”