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

Preparing of calibration curves are critical steps for accurate quantitative LC-MS bioanalysis. Traditional multi -sample external calibration curve (MSCC) is labor-intensive and prone to error. In this study, a novel strategy of one sample multi-point calibration curve (OSCC) using multiple isotopologue reaction monitoring (MIRM) was proposed and validated using LC-MS for the quantitation of six aflatoxins in milk and oat-based milk samples. The developed MIRM-OSCC methodology is comprehensively validated and the results indicated that the established method exhibits good performance in selectivity, sensitivity, accuracy and precision. Furthermore, the OSCC could realize sample dilution by monitoring the MIRM channel with less intensity for samples beyond the upper limit of quantification, without the need of sample dilution, which improves the assay throughput. Considering the advantages of excluding the MSCC preparation and sample dilution in OSCC, this strategy can be widely applied in various fields such as drugs, food safety and environmental analysis.

Automated summary

This study proposed and validated a novel strategy of one sample multi-point calibration curve (OSCC) using multiple isotopologue reaction monitoring (MIRM) for quantitative LC-MS bioanalysis. The developed OSCC methodology exhibited good performance in selectivity, sensitivity, accuracy, and precision, and could realize sample dilution by monitoring the MIRM channel. The advantages of excluding the preparation of multi-sample external calibration curve (MSCC) and sample dilution in OSCC make this strategy widely applicable in various fields.