超高效液相色谱-串联质谱法检测食用菌中的游离棉酚

Determination of Free Gossypol in Edible Fungi by Ultra-high Performance Liquid Chromatography-Tandem Mass Spectrometry

目的:建立超高效液相色谱-串联质谱法定量测定食用菌中游离棉酚含量的分析方法。方法:样品经丙酮-盐酸(0.5 mol·L^(-1))-水(13∶1∶6,v/v/v)混合溶液振荡离心,通过超高效液相色谱C18反向色谱柱分离,以乙腈和0.1%甲酸(含2 mmol·L^(-1)乙酸铵)为流动相进行梯度洗脱,采用电喷雾负离子的多反应离子监测模式,基质匹配标准溶液外标法定量。结果:棉酚在0.25~25.00μg·L^(-1)质量浓度范围内呈现出良好的线性关系,3个添加水平下的平均回收率在86.1%~99.8%,相对标准偏差为2.3%~8.2%(n=6),方法的检出限(S/N≥3)均为1μg·kg^(-1),定量限(S/N≥10)均为3μg·kg^(-1)。结论:该方法简单、快速、可靠,具有良好的灵敏度、回收率和精密度,适用于食用菌中痕量游离棉酚的大批量快速检测及定量分析。

Objective:An ultra-high performance liquid chromatography-tandem mass spectrometry method was developed for the quantitative determination of free gossypol in edible fungi.Method:The sample was extracted with acetone-hydrochloric acid(0.5 mol·L^(-1))-water(13∶1∶6,v/v/v),shaken and centrifuged,and separated by an ultrahigh performance liquid chromatography C18 reverse chromatographic column,using gradient elution with a mobile phase of acetonitrile and 0.1%formic acid in water(containing 2 mmol·L^(-1) ammonium acetate).Detected using an electrospray ionization source in the negative ion multiple reaction monitoring(MRM)mode,and quantified by an external standard method using a matrix-match standard solution.Result:The results showed that gossypol showed a good linear relationship in the mass concentration range of 0.25~25.00μg·L^(-1),the average recoveries from edible fungi matrix spiked at the three different levels ranged from 86.1%to 99.8%,with the relative standard deviation between 2.3%and 8.2%(n=6).The method limits of detection(LODs,S/N≥3)is 1μg·kg^(-1)and the limit of quantification(LOQs,S/N≥10)is 3μg·kg^(-1).Conclusion:The method is simple,rapid,reliable,has good sensitivity,recovery rate and precision,and could meet the requirements for rapid detection and quantitative analysis of trace amounts of free gossypol in edible fungi.