高效液相層析法及毛細管電泳法測定蕎麥茶及印度蕎麥種子中之芸香苷   全文下載

Determination of Rutin in Buckwheat Tea and Fagopyrum tataricum Seeds by High Performance Liquid Chromatography and Capillary Electrophoresis

本研究開發簡單，快速且靈敏度高的高效液相層析法（HPLC）和毛細管電泳法（CE）檢測芸香?。高效液相層析法使用Chromolith RP 18管柱分離芸香?，偵測波長356 nm。移動相組成為甲醇：水（50：50，v/v）之10 mM pH值4.1醋酸緩衝液。以訊雜比為3之偵測極限23.91 ng/mL。芸香?滯留時間、波鋒面積及高度之相對標準差（n = 5）分別為0.199、0.832和0.522%。毛細管電泳法使用bare fused silica管柱分離芸香?。pH 9.4的硼酸緩衝液為背景電解質，檢測波長208 nm。注射時間5秒，電源20 kV，偵測極限55.67 ng/mL。芸香?滯留時間、波鋒面積及高度之相對標準差 （n = 5）分別為0.377、0.923和2.446%。兩種方法都可應用於蕎麥茶，印度蕎麥種子中之芸香?檢驗。蕎麥茶樣品基質於HPLC和CE注射前，以SEP - PAK C18移除。高效液相層析法檢測芸香?之結果與毛細管電泳法一致。

Simple, fast and very sensitive high performance liquid chromatography (HPLC) and capillary electrophoresis (CE) methods for the determination of rutin were developed. The separation of rutin by HPLC was performed using a Chromolith RP 18 column with detection wavelength at 356 nm. Composition of mobile phase was methanol : water (50 : 50, v/v) in 10 mM acetate buffer at pH 4.1. The limit of detection (at a signal-to-noise ratio of 3) was 23.91 ng/mL. Relative standard deviations of retention time, peak area and peak height (n = 5) for rutin were 0.199, 0.832 and 0.522%, respectively. For CE method, rutin was separated on a bare fused silica column. Borate buffer at pH 9.4 was used as the background electrolyte and detected wavelength was 208 nm. The limit of detection (at a signal-to-noise ratio of 3) was 55.67 ng/mL by using a 5 s injection time and +20 kV power supply. Relative standard deviations of retention time, peak area and peak height (n = 5) were 0.377, 0.923 and 2.446%, respectively. Both methods were applied to determine rutin in buckwheat tea and Fagopyrum tataricum seeds. Sample matrix in buckwheat tea was removed by using Sep-Pak C18 prior injection to HPLC and CE. The results for rutin determination obtained by the HPLC method agreed well with those from CE method.