李耀利;尚明英;耿长安;张雪梅;陈纪军;蔡少青;
LI Yao-li1,2,SHANG Ming-ying2,GENG Chang-an1,ZHANG Xue-mei1,CHEN Ji-jun1,CAI Shao-qing2(1.State Key Laboratory of Phytochemistry and Plant Resources in West China,Kunming Institute of Botany, Chinese Academy of Sciences,Kunming 650204,China; 2.State Key Laboratory of Natural and Biomimetic Drugs,School of Pharmaceutical Sciences, Peking University,Beijing 100191,China)

• 1:中国科学院昆明植物研究所植物化学与西部植物资源持续利用国家重点实验室
• 2:北京大学药学院天然药物与仿生药物国家重点实验室

摘要(Abstract)：

目的:建立1种用于测定青叶胆及习用品药材中獐牙菜苦苷、龙胆苦苷、獐牙菜苷、芒果苷和红白金花内酯含量的HPLC方法,并研究5种成分在青叶胆及习用品药材中的分布。方法:Thermo BDS Hypersil C18色谱柱(4.6 mm×250 mm,5μm),流动相0.1%磷酸溶液-甲醇,梯度洗脱,流速0.7～1.0 mL.min-1,柱温32℃,检测波长为250,260,225 nm。结果:5种成分的分离度良好,标准曲线在检测范围内均呈良好线性,r=0.999 7～0.999 9,平均回收率97.03%～102.7%,RSD为1.8%～6.2%。对8种30份青叶胆测定结果显示8个青叶胆品种中5种成分的有无及含量存在差异;弥勒獐牙菜、丽江獐牙菜等獐牙菜苦苷含量高达34.47～118.05 mg.g-1;西南獐牙菜中龙胆苦苷含量最高,紫红獐牙菜中獐牙菜苷的含量较高;根据獐牙菜苦苷及龙胆苦苷与獐牙菜苷含量之和的比较可以鉴别小青叶胆和大青叶胆;根据龙胆苦苷与5成分总和的比值可以区别西南獐牙菜。结论:本研究建立的方法操作简单、结果准确,具有较好的重复性和稳定性,可用于青叶胆类药材的鉴定及质量评价。
Objective: To develop an HPLC method for simultaneous determination of swertiamarin,gentiopicroside,sweroside,mangiferin,erythrocentaurin,and to detect these five constituents in eight Qingyedans derived from Swertia mileensis,S.cincta,S.patens,S.punicea,S.delavayi,S.nervosa,S.macrosperma and S.yunnanensis.Method: The separation was carried out on a Thermo BDS Hypersil C18(4.6 mm×250 mm,5 μm) column eluted with mobile phase of water containing 0.1% phosphoric acid and methanol(B) in gradient program(0-10 min,18%-20% B;10-30 min,20%-35% B;30-35 min,35%-60% B).The column temperature was 32 ℃,and the detection wavelength was set at 250,260,225 nm.The flow rate was 0.7 mL· min-1 from 0 to 30 min,and be increased to 1.0 mL· min-1 in 35 min.Result: The five compounds were well separated.The linear response ranges of swertiamarin,gentiopicroside,sweroside,mangiferin,erythrocentaurin were 0.072-13.39,0.120-4.518,0.060-5.050,0.025-1.518,and 0.031-0.210 μg,respectively.The mean recoveries of five compounds were 97.03%-102.7%(RSD 1.8%-6.2%).There are swertiamarin,gentiopicroside and sweroside in most samples,and mangiferin in half samples.But erythrocentaurin was only detected in a few samples.The contents of five compounds were different in different samples.The contents of swertiamarin in S.mileensis,S.patens,S.yunnanensis and S.delavayi are up to 34.47-118.05 mg· g-1,the contents of gentiopicroside are up to 25.91 mg· g-1 in S.cincta.In S.puncea all contents of swertiamarin,gentiopicroside,sweroside and mangiferin are higher,especially the content of sweroside.There are Xiao-Qingyedans and Da-Qingyedans called in markets,and they can be identified by the contents of swertiamarin,gentiopicroside and sweroside.S.punicea can be identified by the content of sweroside,and the ratio gentiopicroside/total content can be used for identification of S.cincta from other seven Qingyedan species.Conclusion: The method was certified to be accurate and reliable and can be used for identification and quality evaluation of traditional Chinese medicine Qingyedan derived from Swertia species.

关键词(KeyWords)： 青叶胆;HPLC;獐牙菜苦苷;龙胆苦苷;獐牙菜苷;芒果苷;红白金花内酯
Qingyedan;Swertia;HPLC;swertiamarin;gentiopicroside;sweroside;mangiferin;erythrocentaurin

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金-云南省联合基金重点项目(U0832603);; 国家杰出青年科学基金项目(81025023);; 植物化学与西部植物资源持续利用国家重点实验室自主课题项目(P2010-ZZ08);; 中国科学院西部之光“西部博士”启动项目

作者(Authors): 李耀利;尚明英;耿长安;张雪梅;陈纪军;蔡少青;
LI Yao-li1,2,SHANG Ming-ying2,GENG Chang-an1,ZHANG Xue-mei1,CHEN Ji-jun1,CAI Shao-qing2(1.State Key Laboratory of Phytochemistry and Plant Resources in West China,Kunming Institute of Botany, Chinese Academy of Sciences,Kunming 650204,China; 2.State Key Laboratory of Natural and Biomimetic Drugs,School of Pharmaceutical Sciences, Peking University,Beijing 100191,China)

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