柱前衍生化高效液相检测法测定桑叶中的1-脱氧野尻霉素  

Determination of 1-deoxynojirimycin in Morus alba L. Leaves by High Performance Liquid Chromatography with Precolumn Derivatization

作　　者：张旻[1] 吴联[1] 周乐[2] 

机构地区：[1]西北农林科技大学生命科学学院,陕西杨凌712100 [2]西北农林科技大学理学院,陕西杨凌712100

出　　处：《西北农业学报》2008年第6期258-261,共4页Acta Agriculturae Boreali-occidentalis Sinica

基　　金：国家自然科学基金(30571402;30771454)

摘　　要：建立了2，4-二硝基氟苯（DNFB）柱前荧光衍生-高效液相色谱法测定桑叶中1-脱氧野尻霉素（DNJ）的方法。桑叶干粉经索式提取器除去部分色素后加入0.1%的冰醋酸甲醇溶液，在水浴超声条件下（50°C，5min）重复提取2次，活性炭脱色后用DNFB进行衍生，DNJ反应生成荧光产物，然后用高效液相-紫外检测器测定。流动相为乙腈-0.1%醋酸甲醇（25∶75，V/V），流速1.0 mL/min，柱温10℃。线性范围为0.16-8.14μg/mL，（相关系数r=0.9994），检出限为0.03μg/mL。试验测得桑叶中DNJ的平均含量为0.30%，平均回收率为96.8%，RSD为2.0%（n=6）。A rapid and reliable method has been developed, suitable for assays of a large number of mulberry leaf for 1-deoxynojirimycin （DNJ）. Dried leaf was extracted with fresh chloroform in Soxhlet extractor to remove pigments, and then DNJ in the dried leaf was double extracted in 0.1 acetic acid in methanol, assisted by ultrasound for 5 min at 50℃ for each time. After that active car bon was used to the pooled extract and vortexed for 30 s to remove residual pigments. Finally, the extract was derivatized with 2,4-Dinitro-1-fluorobenzene （DNFB）, and analyzed by high performance liquid chromatography equipped with ultraviolet detector. Mobile phase consisted of acetonitrile-0.1 acetic acid in methanol （25 ： 75, V/V ） with a flow rate of 1.0 mL/min. The linear calibration curve was established with the concentration range from 0.16 ug/ mE to 8.14 ug/mL, r=0. 9994 of the correlation coefficient, and 0.03 ug/mL （30） of the detective limit. The results showed that the content of DNJ in leaf of Morusalba L. was 0. 30% , the average recovery was 96.8% , and RSD was 2. 0% （n=6）.

关 键 词：桑叶 1-脱氧野尻霉素(DNJ) 高效液相色谱法(HPLC) 柱前衍生化 

分 类 号：R284[医药卫生—中药学]