目的 探讨淡豆豉炮制过程中环境因素对黄曲霉毒素（AFs）含量的影响。方法 依据《中国药典》2020年版记载的淡豆豉制法，设置不同的温度、湿度、是否水封“再闷”环境条件下的6个组进行自然炮制，检测各组炮制至不同时间豆粒的温度、水分活度（AW）、含氧量；采用超高效液相色谱-串联质谱法（UPLC-MS/MS）检测各组在“黄衣上遍”第3天（F3）、第6天（F6）和“再闷”第6天（Z6）、第12天（Z12）样品中AFB₁、AFB₂、AFG₁、AFG₂含量，分析淡豆豉自然炮制中影响AFs产生的主要环境因素；模拟淡豆豉自然炮制，通过纯培养考察炮制原料、“再闷”环境（低含氧量）对产毒黄曲霉菌标准株（简称：产毒株）产AFB₁的影响，实时荧光定量PCR（qRT-PCR）法检测产毒株产毒关键基因（aflR、aflD、aflM、aflO、aflQ） mRNA相对表达量。结果 自然炮制过程中湿度与AFs含量呈显著性相关（r＝0.435，P＜0.05），且组6（湿度为90%）中F6、Z6、Z12的AFs含量均最高（分别为159.11、45.43、8.78 μg·kg^-1）；纯培养实验中，产毒株在黑豆基质上比马铃薯葡萄糖肉汤（PDB）培养基上的产毒量减少95.31%及以上，在密封罐内培养比罐外培养的产毒量减少99.93%及以上，均以aflM、aflO、aflQ mRNA表达下调更显著（P＜0.001）。结论 淡豆豉自然炮制过程中湿度是影响AFs含量的主要因素，炮制原料、“再闷”环境可显著降低产毒株的产毒量。

Objective The effects of environmental factors on the content of aflatoxins (AFs) in the processing of Sojae Semen Preaparatum (SSP) were investigated. Methods According to the 2020 edition of Chinese Pharmacopoeia, six groups were naturally processed under different temperature, humidity, and whether they were sealed with water or not in the process of the “secondary fermentation” stage. Conventional methods were used to detect the temperature, water activity (aw), and oxygen content of each group at different processing times. The contents of AFs (AFB₁, AFB₂, AFG₁, AFG₂) in the samples of each group on the 3rd day (F3) and the 6th day (F6) of the "yellow cladding" stage, the 6th day (Z6) and the 12th day (Z12) of the "secondary fermentation" stage were detected by method of ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The main environmental factors affecting the production of AFs in natural processing of SSP were analyzed. Simulating the natural processing of SSP, the effects of processing raw materials and the environment of the "secondary fermentation" stage on the production of AFB₁ by the standard strain of Aspergillus flavus (abbreviated as toxigenic strain) were investigated through pure culture. Real -time quantitative PCR (RTqPCR) was used to detect the mRNA expression levels of the key genes aflR, aflD, aflM, aflO and aflQ of toxigenic strains. Results There was a significantly positive correlation between humidity and AFs content during natural processing (r = 0.435, P < 0.05), and the AFs content of F6, Z6 and Z12 in group 6 (humidity of 90%) was the highest (159.11, 45.43 and 8.78 μg·kg^-1, respectively). In pure culture experiments, the toxin amount produced by toxigenic strain was reduced by 95.31% or more on black bean substrate compared to PDB medium; and by 99.93% or more in sealed jar compared to out of jar culture. The mRNA expression of aflM, aflO, and aflQ was significantly inhibited (P < 0.001). Conclusion Humidity was the main factor affecting the content of AFs during the natural processing of SSP. Processing raw material and the environment of the "secondary fermentation" stage can significantly reduce the amount of toxic production.

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国家自然科学基金资助项目（82360772）;国家自然科学基金资助项目（82060709）;国家自然科学基金资助项目（82060699）;南昌大学香樟育才计划;江西省自然科学基金资助项目（20242BAB26173）;江西省自然科学基金资助项目（20232BAB206161）