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.

校准曲线的准备是准确定量 LC-MS 生物分析的关键步骤。传统的多样本外部校准曲线 (MSCC) 工作量大且容易出错。在本研究中，提出了一种使用多重同位素体反应监测 (MIRM) 的单一样品多点校准曲线 (OSCC) 新策略，并使用 LC-MS 验证了牛奶和燕麦奶样品中六种黄曲霉毒素的定量分析。所开发的 MIRM-OSCC 方法得到了全面验证，结果表明所建立的方法在选择性、灵敏度、准确度和精密度方面表现出良好的性能。此外，对于超出定量上限的样品，OSCC可以通过监测强度较低的MIRM通道实现样品稀释，无需稀释样品，这提高了化验吞吐量。考虑到在 OSCC 中排除 MSCC 制备和样品稀释的优势，该策略可广泛应用于药物、食品安全和环境分析等各个领域。