RP-HPLC法同时测定藏药手掌参中3种抗AD活性成分的含量

目的]建立同时测定藏药手掌参中天麻素、dactylorhin A和militarine的含量测定方法.方法]采用反相高效液相色谱法（RP-HPLC）,以ZORBAX SB-C18（4.6 mm×250 mm,5 μm）色谱柱;进样10μl;乙腈-浓度0.5％的磷酸水梯度洗脱（0 min,7％;10 min,15％;20 min,20％;25 min,23％;30 min,40％;35 min,40％）;流速为0.7 ～ 1.0 ml/min;柱温30℃;检测波长222 nm.结果]在35 min内完成对3种抗AD活性成分的分析,各成分之间均达到基线分离.天麻素、dactylorhin A和militarine的线性范围分别为：0.031 34～1.003 00mg/ml（R=0.999 8）、0.038 06 ～1.218 00 mg/ml（R =0.999 6）、0.039 28 ～1.257 00 mg/ml（R=0.999 6）;平均加样回收率（n=6）分别为98.88％、99.91％和100.48％.测定得青藏高原15个不同地区不同种属的手参中3种抗AD活性成分的含量,以互助变异手参的含量最高.结论]该方法建立的HPLC分析方法简单准确,能快速测定手参中有效成分的含量,青藏高原不同地区不同种属手参中3种抗AD活性成分含量差异较大.

Quantitative Analysis of Three Anti-Alzheimer＇s Active Constituents in Tibetan Herb Gymnadenia conopsea by RP-HPLC

Objective] To determine the content of gastrodin,dactylorhin A and militarine in Tibetan herb Gymnadenia conopsea.Method]The sample were separated on the column of ZORBAX SB-C18 （4.6 mm × 250 mm,5 μm） which eluted with Acetonitrile（A） and water（B,0.5％ phosphoric acid）.The sample size was 10 μl.The initial condition was 7％ A,chsnhrf to 15％ A in 10 min,to 20％ A in 10 min,to 23％ in 5 min,to 40％ in 5 min with detected wavelength at 222 nm,flow rate was from 0.7 to 1 ml/min,column temperature at 30 ℃.Result] Three active constituents were base-isolated.The linear ranges of gastrodin,dactylorhin A and militarine were 0.031 34-1.003mg/ml （R =0.999 8）,0.038 06-1.218 mg/ml （R =0.999 6）,0.039 28-1.257 mg/ml （R =0.999 6） respectively ; the average recoveries（n =6） were 98.88％,99.91％ and 100.48％ respectively.The content of three Anti-Alzheimer＇ s active constituents in Tibetan herb Gymnadenia conopsea from fifteen different areas in Tibetan Plateau was determined.Conclusion] The method is easy and precise which can be used for quantitative analysis of active constituents in Gymnadenia conopsea rapidly.The content of three active constituents changed at a great degree in different species and area.