超声波-固相萃取-高效液相色谱法测定毒蘑菇中肽类毒素  

Determination of peptide toxins in poisonous mushroom by ultrasonic solid phase extraction high performance liquid chromatography

作　　者：许莹[1] 李春盛[1] 郭维静[1] XU Ying;LI Chun-sheng;GUO Wei-jing(Dalian City Center for Disease Control and Prevention,Dalian,Liaoning,116021,China)

机构地区：[1]大连市疾病预防控制中心,辽宁大连116021

出　　处：《预防医学论坛》2020年第10期783-786,共4页Preventive Medicine Tribune

摘　　要：目的利用超声波提取、固相萃取分离技术相结合,对大连本地野生蘑菇进行肽类毒素排查。方法 2019年利用超声波细胞破壁提取蘑菇样品中毒素成分,通过固相萃取(SPE)纯化分离蕈毒素,对野生蘑菇中a鹅膏毒肽(a-amanitin)和鬼笔毒肽(phalloidin)进行提取和纯化,再用高效液相色谱进行定性与定量分析;并对大连本地野生蘑菇进行肽类毒素排查。结果利用超声波提取、固相萃取结合,有效地分离提取出蘑菇样品中的a-amanitin、phalloidin两种毒素。对照标样,保留时间一致,在改变流动相极性后各组分的保留时间仍然保持一致,可以实现对蘑菇样品中a-amanitin、phalloidin毒素的定性和定量。在15种大连本地野生蘑菇中,利用本方法成功地从一种蘑菇中检测到了a-amanitin(1.26 mg/kg)。结论鉴于误食毒菇中毒事件的特点,本研究在检验周期、提取率、方法简便化方面都有了较大改善,为肽类毒素中毒提供了很好的技术支持。Objective To investigate the peptide toxins of wild mushrooms in Dalian city by using the combination of ultrasonic extraction and solid phase extraction separation technology. Methods The toxin components in mushroom samples were extracted by ultrasonic cell wall breaking in 2019.The fumigating toxin was purified and separated by solid phase extraction(SPE),and the a-amanitin and phalloidin in wild mushroom were extracted and purified,followed by qualitative and quantitative analysis by high performance liquid chromatography.And the peptide toxoids of wild mushrooms in Dalian city were investigated. Results The combination of ultrasonic extraction and solid phase extraction was used to effectively separate and extract the toxins-amanitin and phalloidin from the mushroom sample.Compared with the standard sample,the retention times were consistent,and the retention times of each component were still consistent after the polarity of the mobile phase was changed,thus realizing the qualitative and quantitative determination of the toxins-amanitin and phalloidin in the mushroom sample.In 15 kinds of wild mushrooms native to dalian,amanitin(1.26 mg/kg) was successfully detected from one mushroom by this method. Conclusion In view of the characteristics of poisoning events caused by ingestion of poisonous mushrooms,this study has greatly improved the inspection cycle,extraction rate and simplification of the method,which provides a good technical support for peptide poisoning.

关 键 词：毒蘑菇 a鹅膏毒肽 鬼笔毒肽 固相萃取 超声波 高效液相色谱 

分 类 号：R155.54[医药卫生—营养与食品卫生学]