目的: 建立固相萃取液质联用法同时测定黄精、麦冬中10种真菌毒素(伏马毒素B1、B2、 B3,黄曲霉毒素B1、B2、G1、G2,T-2毒素、赭曲霉毒素A,玉米赤霉烯酮)。方法: 在黄精、麦冬药材粉末中加入70%甲醇水溶液,涡旋及震荡提取,固相萃取柱 (HLB)净化,正离子模式以0.01%甲酸水溶液(含0.5 mmol·L-1乙酸胺)和乙腈-甲醇(50∶50)混合溶液为流动相;负离子模式以0.1%甲酸水溶液-乙腈为流动相,经C18色谱柱梯度洗脱分离,采用电喷雾(ESI)离子源、多反应监测(MRM)及正负离子模式采集数据,以基质匹配标准曲线外标法进行定量。结果: 10种真菌毒素线性关系良好,相关系数(r)均≥0.99,麦冬和黄精的方法检测限为0.20~4.00 μg·kg-1和0.33~0.67 μg·kg-1,低、中、高水平加样回收率分别为72.3%~130.8%和69.7%~122.7%;RSD分别为0.50%~7.7%和0.13%~8.7%。22批黄精样品中有一批检出玉米赤霉烯酮,检出值1.48 μg·kg-1,检出率为5.0%;20批麦冬样品中有14批检出伏马毒素B2,检出值为4.99~378.99 μg·kg-1,检出率为70.0%。结论: 该方法简便、快速,实用性强,适用于黄精、麦冬中10种真菌毒素的检测。
Objective: To establish a high-performance liquid chromatography (HPLC) coupled with electrospray ionization (ESI) tandem triple quadrupole mass spectrometry (QQQ/MS) method for the simultaneous determination of multiple residues of ten mycotoxins (fumonisin B1, fumonisin B2, fumonisin B3, aflatoxin G1, aflatoxin G2, aflatoxin B1, aflatoxin B2, T-2 toxin, ochratoxin A and zearalenone) in Polygonati Rhizoma and Ophiopogonis Radix by solid-phase extraction. Methods: Samples were extracted with 70% methanol solution by vortex and oscillation. The concentrated liquid supernatant was purified by a hydrophilic-lipophilic balance (HLB) solid-phase extraction (SPE) column. HPLC separation was performed on an Agilent Eclipse plus C18 column. 0.01% formic acid (containing 0.5 mmol·L-1 ammonium acetate) and acetonitrile-methanol (50∶50) solution was used as mobile phase in positive mode. 0.1% acetic acid-acetonitrile was used as the mobile phase in negative mode. MS/MS was adopted for multiple reaction monitoring (MRM) in the positive/negative mode. The quantification was achieved using matrix-matched standard calibrations as the external standard. Results: Under the optimized conditions, the calibration curve showed good linearity with correlation coefficients above 0.99. The limits of detection (LODs) were 0.20-4.0 μg·kg-1(Ophiopogonis Radix) and 0.33-0.67 μg·kg-1 (Polygonati Rhizoma). The recoveries of 10 mycotoxins at three levels were in the range of 72.3%-130.8%(Ophiopogonis Radix)and 69.7%-122.7%(Polygonati Rhizoma),and the relative standard deviation(RSDs) were 0.50%-7.7%(Ophiopogonis Radix)and 0.13%-8.7% (Polygonati Rhizoma). The result showed that among 14 batch of Polygonati Rhizoma samples, zearalenone was detected in one batch of sample, with a detection value of 1.48 μg·kg-1 and the detection rate of 5.0%. Amang 20 batches of Ophiopogonis Radix samples, fumonisin B2 were detected in 14 batches of samples, with a detection value of 4.99-378.99 μg·kg-1 and the detection rate of 70.0%. Conclusion: This experiment provides a simple, rapid, and practical method that is suitable for the quantitative analysis of 10 kinds of fungal mycotoxin in Ophiopogonis Radix and Polygonati Rhizoma. Some mycotoxin residues in Ophiopogonis Radix and Polygonati Rhizoma have certain health risks and should arouse our attention more.
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