荧光显微技术与HPLC联用法快速检测虎杖组织器官中白藜芦醇的分布与含量

Fluorescence Microscopy and HPLC Assay for Rapid Detection of Distribution and Content of Resveratrol in Polygonum cuspidatum

目的:建立荧光显微技术与HPLC法联用,快速检测不同生长期虎杖组织器官中白黎芦醇的分布与含量。方法:采用序贯试验设计冷冻切片的条件,荧光显微镜观察;超声波辅助提取所切材料中的白黎芦醇,HPLC检测其含量。结果:冷冻切片的条件为阿拉伯胶浓度先20%后40%,浸渍时间20%为5～6 h、40%为2～5 min,冷冻室温度-5℃,厚度为15μm;白藜芦醇主要积累在虎杖各器官、组织与细胞的纤维和纤维素中,其含量在根状茎中髓>木质部>韧皮部>周皮;器官间则是根状茎的芽>根状茎>地上茎>叶;根茎中白藜芦醇的含量随年龄增长而增加。结论:荧光显微镜观察结果与HPLC检测结果一致,说明该方法简便快捷,结果可靠,为虎杖最佳采收期与收购品质的确定提供了快速可靠的检测方法。

Objective：To establish fluorescence microscopy combined with HPLC method for rapid detection the distribution and content of resveratrol tissues in different growth stages of Polygonum cuspidatum.Methods：Used sequential experiment to design conditions of frozen and observe of the section by fluorescence microscopy;Resveratrol was extracted by ultrasonic-assisted extraction and its content was detected by HPLC.Results：The results showed that frozen condition for concentration of gum Arabic was from 20%（dipping time was 5~6 h） to 40%（2~5min）,the freezer temperature was-5 ℃,and the thickness was 15 μm.Resveratrol in polygonum cuspidatum was mainly accumulated in the organs,tissues and cells of fiber and cellulose,its content in rhizomes declined as the following sequence：spinal cordxylemphloemperiderm;Its content declined in organ as the following sequence：buds rhizomesground stemleaves;The content of resveratrol in root increased with age.Conclusion：The results of fluorescence microscopic observation is in accordance with the HPLC results,indicating that the method is simple,fast and reliable,and provides a fast and reliable detection method for the determination of optimum harvesting period of Polygonum cuspidatum and acquisition of quality.