改进通过式固相萃取柱-超高效液相色谱-串联质谱法测定谷物及制品中脱氧雪腐镰刀菌烯醇及衍生物的含量

Determination of Deoxynivalenol and Its Derivatives in Cereals and Its Products by Ultra-High Performance Liquid Chromatography-Tandem Mass Spectrometry with Modified Pass-Through Solid Phase Extraction Column

开发、设计一种改进通过式固相萃取柱,用于谷物及其制品中脱氧雪腐镰刀菌烯醇及其4种衍生物的前处理。称取固体样品2.00 g(液体样品5.00 g),加入400μL 1.0 mg·L^(-1)内标混合溶液,振荡混匀,静置30 min,再加入20.0 mL 84%(体积分数)乙腈溶液(液体样品加入15.0 mL乙腈),涡旋振荡20 min,离心5 min。移取约8 mL上清液,用改进通过式固相萃取柱[填料为160 mg C_(18)、300 mg石墨化碳黑、100 mg氨丙基净化剂(NH_(2))和300 mg硅藻土的混合物]处理,取5.0 mL滤液,于40℃氮吹至干后,加入1.0 mL水,超声30 s,涡旋30 s,过0.22μm微孔滤膜。采用超高效液相色谱-串联质谱法测定其中脱氧雪腐镰刀菌烯醇及其4种衍生物的含量,内标法定量。结果表明:经改进通过式固相萃取柱处理后,样品溶液澄清透明,并且5种目标物的基质效应减少;5种目标物标准曲线的线性范围为10~2 000μg·L^(-1),检出限为(3S/N)为5.0μg·kg^(-1);对小麦粉、大米、玉米、啤酒、白酒等基质进行3个浓度水平的加标回收试验,回收率为74.6%~106%,测定值的相对标准偏差(n=6)均不大于9.1%;方法用于162份样品分析,小麦粉、玉米、大米、啤酒中均检出脱氧雪腐镰刀菌烯醇,衍生物中仅检出3-乙酰化脱氧雪腐镰刀菌烯醇和雪腐镰刀菌烯醇。

A modified pass-through solid phase extraction column was developed and designed for the pretreatment of deoxynivalenol and its 4 derivatives in cereals and its products.The solid sample(2.00 g) or liquid sample(5.00 g) was taken,and 400 μL of 1.0 mg·L^(-1) internal standard mixed solution was added.The mixture was shaken,mixed well,and settled for 30 min.Then 20.0 mL of 84%(volume fraction) acetonitrile solution was added into the solid sample or 15.0 mL of acetonitrile was added into the liquid sample,and the mixed solution was swirled for 20 min and centrifuged for 5 min.About 8 mL of the supernatant obtained was treated with the modified pass-through solid phase extraction column(a mixture composed of 160 mg of C_(18),300 mg of graphitized carbon black,100 mg of aminopropyl purifier(NH_(2)) and 300 mg of diatomite as filler),and 5.0 mL of filtrate was taken and blown to dryness by nitrogen at 40 ℃.Then 1.0 mL of water was added into the residue,and the solution was treated by ultrasound for 30 s,swirled for 30 s,and filtrated by 0.22 μm microporous filter membrane.Deoxynivalenol and its 4 derivatives were determined by ultra-high performance liquid chromatography-tandem mass spectrometry,with internal standard method for quantification.As shown by the results,the sample solution was clear and transparent after treatment with the modified pass-through solid phase extraction column,and the matrix effect of the 5 targets was significantly reduced.The linear range of the standard curves of 5 targets was in the range of 10-2 000 μg·L^(-1),and detection limit(3S/N) was 5.0 μg·kg^(-1).Test for recovery was made on wheat flour,rice,corn,beer and liquor,giving results in the range of 74.6%-106%,and RSDs(n=6) of the determined values were not more than 9.1%.The proposed method was applied to analysis of 162 samples,and deoxynivalenol was detected in wheat flour,corn,rice and beer,but only 3-acetylated deoxynivalenol and nivalenol were detected among the 4 derivatives.