采用 SPE 和 N-乙酰-l-半胱氨酸衍生化后，HPLC-DAD-MS 同时测定植物样品中的单个异硫氰酸酯。
Simultaneous determination of individual isothiocyanates in plant samples by HPLC-DAD-MS following SPE and derivatization with N-acetyl-l-cysteine.
机构信息
Department of Food Chemistry, Technology and Biotechnology, Faculty of Chemistry, Gdańsk University of Technology, Narutowicza 11/12, 80-233 Gdańsk, Poland.
Department of Organic Chemistry, Faculty of Chemistry, Gdańsk University of Technology, Narutowicza 11/12, 80-233 Gdańsk, Poland.
出版信息
Food Chem. 2017 Jan 1;214:587-596. doi: 10.1016/j.foodchem.2016.07.125. Epub 2016 Jul 21.
The procedure for the isothiocyanates (ITCs) determination that involves derivatization with N-acetyl-l-cysteine (NAC) and separation by HPLC was developed. Prior to derivatization, plant ITCs were isolated and purified using solid-phase extraction (SPE). The optimum conditions of derivatization are: 500μL of isopropanolic eluate obtained by SPE combined with 500μL of derivatizing reagent (0.2M NAC and 0.2M NaHCO3 in water) and reaction time of 1h at 50°C. The formed dithiocarbamates are directly analyzed by HPLC coupled with diode array detector and mass spectrometer if required. The method was validated for nine common natural ITCs. Calibration curves were linear (R(2)⩾0.991) within a wide range of concentrations and limits of detection were below 4.9nmol/mL. The recoveries were in the range of 83.3-103.7%, with relative standard deviations <5.4%. The developed method has been successfully applied to determine ITCs in broccoli, white cabbage, garden cress, radish, horseradish and papaya.
建立了涉及用 N-乙酰-L-半胱氨酸 (NAC) 衍生化和 HPLC 分离的异硫氰酸酯 (ITC) 的测定方法。在衍生化之前，使用固相萃取 (SPE) 对植物 ITC 进行分离和纯化。衍生化的最佳条件为：SPE 得到的 500μL 异丙醇洗脱液与 500μL 衍生化试剂（水中 0.2M NAC 和 0.2M NaHCO3）结合，并在 50°C 下反应 1 小时。如果需要，形成的二硫代氨基甲酸盐可直接通过 HPLC 与二极管阵列检测器和质谱仪进行分析。该方法针对九种常见的天然 ITC 进行了验证。校准曲线在较宽的浓度范围内呈线性（R(2)⩾0.991），检测限低于 4.9nmol/mL。回收率在 83.3-103.7%范围内，相对标准偏差<5.4%。所开发的方法已成功应用于测定西兰花、白菜、豆瓣菜、萝卜、辣根和木瓜中的 ITC。
Zotero 插件