摘要
目的:建立免疫亲和柱净化-HPLC柱后光化学衍生法同时测定何首乌中4种黄曲霉毒素AflaG1、G2、B1、B2的含量。方法:样品经80%乙腈-水溶液提取后,采用含2%的吐温-20溶液稀释,月旭Aflatoxin(B1、B2、G1、G2)3 mL免疫亲和柱净化,Ultimate^(■)ODS-3(4.6 mm×250 mm,5μm)色谱柱分离,以甲醇-乙腈-水为流动相进行等度洗脱,流速为1.0 mL/min,柱温为30℃,进样量为15μL,荧光检测器激发波长为360nm,发射波长为450nm的条件进行测定。结果:所发展的方法能够对生何首乌中黄曲霉毒素AflaB1、B2、G1、G2进行准确检测并定量,其线性范围相关系数均大于0.999,回收率为93.6%-101.8%,RSD小于10%。结论:所建立的方法能同时检测生何首乌中黄曲霉毒素B1、B2、G1、G2,通过在稀释和淋洗阶段加入吐温-20,有效去除杂质干扰,保证了免疫亲和柱的高回收率,使黄曲霉毒素的检测变得更高效准确。
Objectives:To establish an immunoaffinity column purification-HPLC photochemical post-column derivatization method for the simultaneous determination of four aflatoxins G1,G2,B1 and B2 in polygonum multiflorum thunb.Methods:Samples were extracted with 80%acetonitrile aqueous solution,the extractions were purified by Welchrom Aflatoxin(B1,B2,G1,G2)immune-affinity column after dilution with 2%Tween-20 solution.The separation was performed on a Ultimate^(■)ODS-3(4.6 mm×250 mm、5μm)column with methanolacetonitrile-water as the mobile for isocratic elution at the flow rate of 1.0 mL/min.The column temperature was 30℃and the injection volume was 15μL.The excitation wavelength of the fluorescence detector was 360 nm and the emission wavelength was 450 nm.Results:Aflatoxin B1,B2,G1,G2 in polygonum multiflorum thunb can be accurately detected and quantified by the developed method.The correlation coefficient of all aflatoxins was greater than 0.999,and the recovery rates were from 93.6%to 101.8%,RSD was below 10%.Conclusion:The established method is stable and effective,which can simultaneously detect aflatoxin B1,B2,G1,G2 in raw polygonum multiflorum thunb.By adding Tween-20 in the dilution and rinsing processes,the interference of impurities can be effectively eliminated and the high recovery of the immunoaffinity column can be ensured,which makes the detection of aflatoxins more efficient and accurate.
作者
任兴发
邹天骏
袁凯旭
屠炳芳
张维冰
REN Xingfa;ZOU Tianjun;YUAN Kaixu;TU Bingfang;ZHANG Weibing(East China University of Science and Technology,Shanghai,200237;Welch technology(Shang Hai)Co.,LTD,Shanghai,201600)
出处
《生命科学仪器》
2023年第2期94-100,共7页
Life Science Instruments
关键词
何首乌
黄曲霉毒素
高效液相色谱法
光化学衍生
免疫亲和
Prediction of pulmonary nodules
CT image
Three dimensional voxel reconstruction
Thermonuclear characteristics
作者简介
任兴发(1983-),男,山西河曲人,在读博士研究生,研究方向:生物医药,E-mail:y16220019@mail.ecust.edu.cn;通信作者:张维冰(1964-),男,安徽合肥人,教授,博士,研究方向:色谱理论及应用,E-mail:weibingzhang@ecust.edu.cn
