Zhao Lingguo, Lv Ronggui, Song Jiang, Chen Xi, Lei Lei, Zeng Weixian, She Miaowen, Li Dabing, Yu Xiaxia, Liu Zhanguo, Wang Xiao, Liu Yong
Department of Physical and Chemical Laboratory, Center for Disease Prevention and Control of Baoan District, Guangdong, Shenzhen, China.
Department of Critical care medicine, Shenzhen Hospital, Southern Medical University, Guangdong, Shenzhen, China.
Front Chem. 2025 Jan 28;12:1386635. doi: 10.3389/fchem.2024.1386635. eCollection 2024.
A UPLC-MS/MS method with one-step extraction and simultaneous determination of bongkrekic acid (BA) and dehydroacetic acid (DHA) in rice noodles has been established in this study for the first time. The extraction solvent consisted of a mixture of methanol, ammonia, and water. Electrospray ionization negative ion mode (ESI-) was used for multiple reaction detection and external standard method was used for quantitative determination. The method demonstrated a good linearity range of 0-50 ng/mL with a correlation coefficient (r) of 0.9998 for BA, and showed a linearity range of 0∼500 ng/mL with r of 0.9993 for DHA. The limits of detection (LOD) were determined to be 0.1 μg/kg for BA and 0.6 μg/kg for DHA. Additionally, the method exhibited good recovery of 90.1%∼105.4% for BA and 80.4%∼102% for DHA. Meanwhile, the method also exhibited good precision with RSD of 0.4%∼7.5% for BA, and with RSD of 4.5%∼11.4% for DHA. Furthermore, the established method was sucessfully applied to the analysis of rice noodles from two production enterprises. BA was detected in one rice noodle sample and DHA was detected in 7 rice noodle samples in the enterprises A which was the manufacturer of food consumed by the poisoned patient. To investigate the cause of death of the poisoned patient, the toxigenic bacterium was isolated and cultured to produce BA. The results showed that the established method could also be used to qualitatively screen BA and its isomer isobongkrekic acid (IBA) in GVC enrichment broth, potato glucose agar and rice sample, which facilitates a rapid and accurate judgement of whether the isolated bacterium strain produces BA. With high sensitivity and accuracy, and simple sample pretreatment, the proposed method shows potential for safety assessment of the entire production process of rice-derived products, addressing a significant public health concern.
本研究首次建立了一种超高效液相色谱-串联质谱法（UPLC-MS/MS），用于一步提取并同时测定米粉中的米酵菌酸（BA）和脱氢乙酸（DHA）。提取溶剂由甲醇、氨水和水的混合物组成。采用电喷雾电离负离子模式（ESI-）进行多反应监测，并采用外标法进行定量测定。该方法对BA显示出良好的线性范围，为0 - 50 ng/mL，相关系数（r）为0.9998；对DHA显示出的线性范围为0∼500 ng/mL，r为0.9993。确定BA的检测限（LOD）为0.1 μg/kg，DHA的检测限为0.6 μg/kg。此外，该方法对BA的回收率良好，为90.1%∼105.4%，对DHA的回收率为80.4%∼102%。同时，该方法精密度良好，BA的相对标准偏差（RSD）为0.4%∼7.5%，DHA的RSD为4.5%∼11.4%。此外，所建立的方法成功应用于对两家生产企业的米粉分析。在中毒患者食用的食品生产企业A的一个米粉样品中检测到了BA，在7个米粉样品中检测到了DHA。为调查中毒患者的死因，分离并培养了产毒细菌以产生BA。结果表明，所建立的方法还可用于定性筛查广口瓶富集肉汤、马铃薯葡萄糖琼脂和大米样品中的BA及其异构体异米酵菌酸（IBA），这有助于快速准确判断分离出的菌株是否产生BA。该方法具有高灵敏度和准确性，且样品前处理简单，在大米衍生产品的整个生产过程安全评估方面显示出潜力，解决了一个重大的公共卫生问题。
