2种HPLC-柱后衍生化-荧光检测法测定陈皮中黄曲霉毒素的比较  

Comparison of two post-column derivatization systems for HPLC determination of aflatoxins in Citri Reticulatae Pericarpium

作　　者：沙东旭[1] Uwe Gasser 孙苓苓[1] 张满来[1] 侯峰[1] 康强[1] 王维宁[1] 

机构地区：[1]辽宁省食品药品检验所,沈阳110023 [2]Kneipp-werke Kneipp-Mittel-Zentrale GmbH&Co.KG,Bad Wtirishofen,Germany

出　　处：《药物分析杂志》2013年第8期1367-1371,共5页Chinese Journal of Pharmaceutical Analysis

摘　　要：目的:对陈皮中黄曲霉毒素测定的2种柱后衍生化方法(碘衍生化和电化学衍生化)进行比较。方法:样品经过免疫亲和柱净化,HPLC-柱后衍生化-荧光检测器检测。采用Diamonsil ODS C18(5μm,4.6 mm×200 mm)色谱柱,荧光检测,激发波长360 nm,发射波长450 nm。柱后衍生化系统(1)碘衍生化:以甲醇-乙腈-水(25∶18∶57)为流动相,流速0.8 mL.min-1;衍生化溶液为0.05%碘溶液,流速0.3 mL.min-1,衍生化反应温度70℃;(2)电化学衍生化:以甲醇-乙腈-水(20∶20∶60)为流动相(每1 L流动相中加入溴化钾120 mg和4 mol.L-1硝酸350μL),流速1.0 mL.min-1。结果:碘衍生化,黄曲霉毒素G2、B2在2～36 pg,黄曲霉毒素G1、B1在2～120 pg范围内线性关系良好;电化学衍生化,黄曲霉毒素G2、B2在1.2～36 pg,黄曲霉毒素G1、B1在2～120 pg范围内线性关系良好,r>0.9997;回收率在60%～101%之间。结论:2种衍生化方法的测定结果比较接近,且电化学衍生化方法较灵敏,操作简单、分析速度快,也适用于陈皮中黄曲霉毒素的测定。Objective： To evaluate two post-column derivatization systems（derivatization with iodine,and with electrochemically generated bromine） for HPLC determination of aflatoxins in Citri Reticulatae Pericarpium.Methods： The samples were cleaned up on immunoaffinity columns and analyzed by HPLC with post-column derivatization and fluorescence detection.For chromatographic separation,a Diamonsil ODS C18（5 μm,4.6 mm × 200 mm） column was employed.The detection was observed by fluorescence with the excitation wavelength at 360 nm and the emission wavelength at 450 nm.For the post-column derivatization system with iodine（I2）,the separation was carried out on the mobile phase of methanol-acetonitrile-water（25∶ 18∶ 57） with a flow rate of 0.8 mL.min-1,the iodine 0.05% solution was used as the derivatization reagent which was delivered at a flow rate of 0.3 mL.min-1 and the reaction coil was maintained at 70 ℃.For derivatization with electrochemically generated bromine（KOBRA）,the mobile phase consisted of a mixture of methanol-acetonitrile-water（20∶ 20∶ 60） with a flow rate of 1.0 mL.min-1.To each liter of the mobile phase was added 120 mg potassium bromide and 350 μL of 4 mol.L-1 nitric acid.Results： Good linear relationships were obtained within the ranges of 1.2-36 pg for aflatoxin G2 and B2,and 2-120 pg for aflatox in G1 and B1,respectively,r 0.9997.The recoveries were between 60%-101%.Conclusion： The results of two post-column derivatization systems are equivalent.However,KOBRA has higher sensitivity,easier operating procedure,and less analytical time,and it is also applicable to the determination of aflatoxins in Citri Reticulatae Pericarpium.

关 键 词：黄曲霉毒素 陈皮 柱后衍生化 碘衍生化 电化学衍生化 高效液相色谱 中药安全监测 

分 类 号：R917[医药卫生—药物分析学]