QuEChERS—超高效液相—三重四极杆串联质谱测定白茅根中16种真菌毒素(1)
[摘要] 该文建立了一种同时检测白茅根中16种真菌毒素的QuEChERS(Quick， Easy， Cheap， Effective， Rugged， Safe)超高效液相色谱串联质谱定量分析方法。白茅根以1%乙酸-乙腈超声提取，QuEChERS方法净化，用甲醇和含0.01%甲酸水溶液为流动相进行梯度洗脱，超高效液相色谱仪应用Agilent Eclipse Plus C18色谱柱，优化选择在电喷雾离子源正离子模式下以多反应监测方式检测。结果表明，16种真菌毒素在各自线性范围内线性关系良好，相关系数r在0.996 2～1.000，定量限为0.03～186.68 μg·kg-1，回收率为 60.28%～129.2%，RSD为 0.29%～11%。该方法净化效果好、灵敏度高、重复性好，适用于基质成分复杂的白茅根中多种真菌毒素的定量检测。
[关键词] 真菌毒素； QuEChERS法； 超高效液相-三重四级杆联用质谱； 白茅根
[Abstract] A method for the simultaneous determination of sixteen mycotoxins in cogon rootstalk was developed using ultra-performance liquid chromatography coupled with triple quadropole mass spectrometry(UPLC-QqQ-MS/MS). The samples were extracted with acetonitrile contained 1% acetic acid and purified by QuEChERS method. The separation was performed on an Agilent Eclipse Plus C18 column by gradient elution using methanol and 0.01% aqueous formic acid as mobile phase. The targeted compounds were detected in MRM mode by mass spectrometry with electrospray ionization(ESI)source operated in positive ionization mode. The linear relationships of the sixteen mycotoxins were good in their respective linear ranges. The correlation coefficients(r)ranged among 0.996 2-1.000. The LOQs of the sixteen mycotoxins were between 0.03 and 186.68 μg·kg-1. The average recoveries ranged from 60.28% to 129.2% with relative standard deviations(RSDs)within 0.29%-11%. The results demonstrated that the proposed method was sensitive and accurate， and suitable for the mycotoxins quantification in cogon rootstalk.
, 百拇医药
[Key words] mycotoxin； QuEChERS； UPLC-QqQ-MS/MS； cogon rootstalk
白茅根為禾本科植物白茅Imperata cylindrical Beauv. var. major(Mees)C. E. Hubb.的新鲜或干燥根茎，具有凉血止血，清热利尿的功效 [1]。在食品、保健食品和中药中有广泛的应用。真菌毒素是真菌产生的次级代谢产物，产毒菌种主要分为曲霉菌毒素(如黄曲霉毒素、赭曲霉毒素等)和镰刀菌毒素(如T-2 毒素、HT-2 毒素、脱氧雪腐镰刀菌烯醇、玉米赤霉烯酮等)等[2]。动物试验和流行病学的调查结果表明，许多真菌毒素可在体内积累后产生致癌、致畸、致突变、类激素中毒，白细胞缺乏症等，对机体造成永久性损害[3]。随着对真菌毒素危害的深入研究，公众逐渐认识到其对社会经济和人类健康造成的严重威胁，各国政府对药品、食品中真菌毒素的分布及检测也都予以了极大的关注，欧美等国家对此设定了严格的限量规定[4]。真菌毒素检测方法的研究与开发一直是热点和难点，实现快速、准确地同时测定多种毒素是一个长期的技术挑战。QuEChERS(Quick，Easy，Cheap，Effective，Rugged，Safe)法是利用吸附剂填料与基质中的杂质相互作用，吸附杂质从而达到除杂净化的目的。QuEChERS方法回收率较高，精确度和准确度高[5]，可分析的真菌范围广，分析速度快，溶剂使用量少[6]，污染小，价格低廉且不使用含氯化物溶剂，操作简便且装置简单[7]。本文采用QuEChERS-超高相色谱-串联三重四极杆质谱同时测定白茅根中黄曲霉毒素B1，B2，G1，G2，M1，M2(AFB1，AFB2，AFG1，AFG2，AFM1，AFM2)，伏马毒素B1，B2(FB1，FB2)，赭曲霉毒素A(OTA)，T-2毒素，HT-2毒素，ST毒素，脱氧雪腐镰刀菌烯醇(DON)，玉米赤霉烯酮(ZEN)，3-乙酰脱氧雪腐镰刀菌烯醇(3-ACE)，15-乙酰脱氧雪腐镰刀菌烯醇(15-ACE)等16种真菌毒素，该方法快速、准确，适用于真菌毒素多成分含量测定。, 百拇医药(范妙璇 董娇娇 王京辉 郭洪祝 陈有根 傅欣彤)
[关键词] 真菌毒素； QuEChERS法； 超高效液相-三重四级杆联用质谱； 白茅根
[Abstract] A method for the simultaneous determination of sixteen mycotoxins in cogon rootstalk was developed using ultra-performance liquid chromatography coupled with triple quadropole mass spectrometry(UPLC-QqQ-MS/MS). The samples were extracted with acetonitrile contained 1% acetic acid and purified by QuEChERS method. The separation was performed on an Agilent Eclipse Plus C18 column by gradient elution using methanol and 0.01% aqueous formic acid as mobile phase. The targeted compounds were detected in MRM mode by mass spectrometry with electrospray ionization(ESI)source operated in positive ionization mode. The linear relationships of the sixteen mycotoxins were good in their respective linear ranges. The correlation coefficients(r)ranged among 0.996 2-1.000. The LOQs of the sixteen mycotoxins were between 0.03 and 186.68 μg·kg-1. The average recoveries ranged from 60.28% to 129.2% with relative standard deviations(RSDs)within 0.29%-11%. The results demonstrated that the proposed method was sensitive and accurate， and suitable for the mycotoxins quantification in cogon rootstalk.
, 百拇医药
[Key words] mycotoxin； QuEChERS； UPLC-QqQ-MS/MS； cogon rootstalk
白茅根為禾本科植物白茅Imperata cylindrical Beauv. var. major(Mees)C. E. Hubb.的新鲜或干燥根茎，具有凉血止血，清热利尿的功效 [1]。在食品、保健食品和中药中有广泛的应用。真菌毒素是真菌产生的次级代谢产物，产毒菌种主要分为曲霉菌毒素(如黄曲霉毒素、赭曲霉毒素等)和镰刀菌毒素(如T-2 毒素、HT-2 毒素、脱氧雪腐镰刀菌烯醇、玉米赤霉烯酮等)等[2]。动物试验和流行病学的调查结果表明，许多真菌毒素可在体内积累后产生致癌、致畸、致突变、类激素中毒，白细胞缺乏症等，对机体造成永久性损害[3]。随着对真菌毒素危害的深入研究，公众逐渐认识到其对社会经济和人类健康造成的严重威胁，各国政府对药品、食品中真菌毒素的分布及检测也都予以了极大的关注，欧美等国家对此设定了严格的限量规定[4]。真菌毒素检测方法的研究与开发一直是热点和难点，实现快速、准确地同时测定多种毒素是一个长期的技术挑战。QuEChERS(Quick，Easy，Cheap，Effective，Rugged，Safe)法是利用吸附剂填料与基质中的杂质相互作用，吸附杂质从而达到除杂净化的目的。QuEChERS方法回收率较高，精确度和准确度高[5]，可分析的真菌范围广，分析速度快，溶剂使用量少[6]，污染小，价格低廉且不使用含氯化物溶剂，操作简便且装置简单[7]。本文采用QuEChERS-超高相色谱-串联三重四极杆质谱同时测定白茅根中黄曲霉毒素B1，B2，G1，G2，M1，M2(AFB1，AFB2，AFG1，AFG2，AFM1，AFM2)，伏马毒素B1，B2(FB1，FB2)，赭曲霉毒素A(OTA)，T-2毒素，HT-2毒素，ST毒素，脱氧雪腐镰刀菌烯醇(DON)，玉米赤霉烯酮(ZEN)，3-乙酰脱氧雪腐镰刀菌烯醇(3-ACE)，15-乙酰脱氧雪腐镰刀菌烯醇(15-ACE)等16种真菌毒素，该方法快速、准确，适用于真菌毒素多成分含量测定。, 百拇医药(范妙璇 董娇娇 王京辉 郭洪祝 陈有根 傅欣彤)