淡豆豉炮制过程中黄曲霉毒素含量的动态变化规律分析

目的: 检测淡豆豉炮制过程中不同时间点样本的黄曲霉毒素(AFTs)含量,明确AFTs的动态变化规律及其产生的关键时间点。方法: 按实验室前期已建立的规范炮制工艺制备淡豆豉,获取淡豆豉炮制过程中不同时间点的样本;采用超高效液相色谱-串联质谱法(UPLC-MS/MS)检测各样本中4种AFTs 黄曲霉毒素B₁(AFB₁),AFB₂,AFG₁,AFG₂]含量。结果: 4种黄曲霉毒素在选定的质量浓度范围内与峰面积线性关系良好(R²均大于0.99),重复性相对标准偏差(RSD)0.4%~0.9%,精密度RSD 0.7%~2.6%,稳定性RSD 0.8%~1.74%,加样回收率介于95.09%~107.20%(RSD 3.14%~12.71%);AFB₁含量在淡豆豉整个炮制过程中呈先上升后下降的趋势,在"再闷"第6天时达最高值6.95 μg·kg^-1,再闷第12天后各样本均未检测出AFTs。结论: 本研究建立了简单、快速、灵敏度高且适用于淡豆豉中4种AFTs检测的UPLC-MS/MS法;淡豆豉炮制过程中AFTs含量呈动态变化,表明淡豆豉炮制中生物拮抗作用自然存在,从安全性角度证实淡豆豉炮制中"再闷"环节的重要性和"再闷"时间的合理性。

Analysis of dynamic change law of aflatoxin content in processing of Sojae Semen Praeparatum

OBJECTIVE To determine the key producing time point of aflatoxins (AFTs) by detecting the content of AFTs at different time points in the processing of Sojae Semen Praeparatum (SSP).METHODS SSP was prepared according to the standard processing process established by our laboratory in the early stage, and samples were obtained at different time points during the processing of SSP. Ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was used to detect the contents of four AFTs aflatoxin B1 (AFB1), AFB2, AFG1, AFG2] in each sample.RESULTS The four aflatoxins had good linear relationships with the peak areas within the selected mass concentration ranges (R²＞0.99). The relative standard deviation (RSD) of repeatability was 0.4%-0.9%. The precision RSD was 0.7%-2.6%. The stable RSD was 0.8%-1.74%, and the sample recovery rate was between 95.09% and 107.20% (RSDs of 3.14%-12.71%). The AFB₁ content showed a trend of first increasing and then decreasing during the whole processing process of SSP, reaching the highest value of 6.95 μg·kg^-1 on the 6th day of secondary fermentation, and no AFT was detected in each sample after the 12th day of secondary fermentation.CONCLUSION In this study, a simple, rapid and highly sensitive UPLC-MS/MS method was established for the detection of four AFTs in SSP. The contents of AFTs change dynamically during the processing of SSP, and the secondary fermentation in the processing of SSP is important, and the time of this stage is crucial. This study may help us understand secondary fermentation from the perspective of safety.