Determination Of Higenamine By Reversed Phase High Performance Liquid Chromatography With Precolumn Derivatization

There is an active ingredient of Higenamine in many Chinese herbal medicine.Shuch as Gnetum parvifolium,embryo of Nelumbo nucifera, Aconite root and Asarum. Higenamine is an effective component of cardiac in Aconite root of Chinese herbal medicine. A sensitive method for determination of higenamine based on a pre-column derivatization reaction with a fluorescent derivatization reagent,1-{[(9H-Fluoren-9-ylmethoxy)carbonyl]oxy}pyrrolidine-2,5-dione (Fmoc-Osu) has been established. The fluorescent derivatization conditions is at 25℃for 50 min with the presence of Boric acid buffer solution PH=8.5.The condition of determination isλex=265nm andλem=315nm. The method limits of detection (S/N=3) was 5 ng/mL. The linear concentration range of higenamine was 0.05～20μg/mL. The best method of sample treatment was established.The established method was applied for the first time successfully to the determination of higenamine in embryo of Nelumbo nucifera, Aconite root and North Asarum, Chinese herbal medicine. High sensitivity, good stability, high selectivity, simple and rapid analysis methods are available. This article specifically content as follows:Chapter one:At first, the research progress of precolumn derivatization high performance liquid chromatography were simply presented. The Second, Physiological pharmacological effects and analysis method of higenamine was introduced. At last, The objective significance and contents of experiment research were summarized.Chapter two:The fluorescent derivatization reagent was used to derivatize at precolumn with higenamine. All kinds of condition of derivatization was examed to find affact to the derivatization reaction.And types and proportion of mobile phase was groped.The best condition of derivatization and chromatograph were finded.lt was separated by HPLC and detected by fluorescence detector. The method of for determination of higenamine based on a pre-column derivatization reaction with a fluorescent derivatization reagent has been established.And the established method was verified by Precision and stability.Chapter three:The efficacy of embryo of Nelumbo nucifera, Aconite and North Asarum were introduced.The best method of sample treatment was established. The established method was applied successfully to the determination of higenamine in embryo of Nelumbo nucifera, Aconite root and North Asarum, a Chinese herbal medicine. And the established method was verified by spike sample.