不同生长季节下藏药麻花秦艽活性成分含量研究

采用高效液相色谱法测定了野生与栽培藏药麻花秦艽（Gentiana straminea）根中龙胆苦苷、落干酸、獐牙菜苦苷和獐牙菜苷四种环烯醚萜苷类化学成分的含量及其在不同生长季节的变化趋势。结果表明,4种环烯醚萜苷类成分的含量随植物的生长季节而波动,其活性成分含量在野生种与栽培种之间发生了一定的差异。但是,栽培根中的龙胆苦甙已达到药典的标准,可供药用。     

RP-HPLC法测定青海产四种秦艽中獐牙菜苦苷的含量

建立了秦艽中獐牙菜苦苷含量测定的RP-HPLC方法。色谱条件：采用ZorbaxEclipseXDB-C18柱（150mm×4.6mm,5μm）,流动相：甲醇-0.03%磷酸水溶液梯度洗脱（0～25min：15%～23%;25～30min：23%～30%）,流速：1mL/min,柱温：25℃,检测波长240nm,獐牙菜苦苷在0.20～1.8μg范围内成良好的线性关系,r=0.9999,回收率RSD=1.32%。对青海产4种秦艽中獐牙菜苦苷的含量进行了定量分析,测定结果：小秦艽中獐牙菜苦苷的含量最高为：0.62%,麻花秦艽与管花秦艽的含量没有显著差异,分别为：0.43%,0.45%,簧管秦艽的含量最低为：0.32%。     

藏药提宗龙胆和线叶龙胆花中四种苦苷类成分的HPLC测定

应用HPLC分析方法同时测定藏药提宗龙胆和线叶龙胆两种植物花中落干酸、獐牙菜苦苷、龙胆苦苷、獐牙菜苷4种苦苷类成分的含量.采用Econosphere C18色谱柱(250×4.6 mm,5 μm),以甲醇-0.5%乙酸为流动相进行梯度洗脱,流速为1.0 mL/min;检测波长为245 nm.结果表明,除提宗龙胆中未检出獐牙菜苦苷外,其它成分均在两种植物中存在,但含量存在一定的差异.     

栽培藏药材麻花艽中四种苦苷类成分含量的季节性变化

采用高效液相色谱法,测定了栽培藏药材麻花艽根中龙胆苦苷、落干酸、獐牙菜苦苷和獐牙菜苷四种苦苷类成分的含量及其在不同生长期的动态变化。结果表明,四种苦苷类成分随植物的发育节律而波动。龙胆苦苷的含量是评价药材质量的重要指标。龙胆苦苷的含量虽然在7月有个积累的高峰期,但7月为花期,从资源可持续利用的角度考虑,不宜定为采收期,到9月含量又有所增加,故而确定栽培藏药麻花艽根的适宜采收期应在9~10月之间。     

HPLC测定7种龙胆科植物花中龙胆苦苷与獐牙菜苦苷的含量

目的:建立7种龙胆花中龙胆苦苷和獐牙菜苦苷含量测定的HPLC方法。方法:采用微波辅助动态回流法进行提取,色谱条件:Fusion-RP 80 A C18柱(150 mm×4.6 mm,5μm);流动相:甲醇-0.2%磷酸溶液梯度洗脱(0~25 min:15%~30%);流速:1 mL/min;柱温:30℃;检测波长240 nm。结果:7种龙胆花中獐牙菜苦苷和龙胆苦苷的色谱峰与共存组分完全达到基线分离,线性范围分别为0.105~0.945μg(r=0.999 9),0.3~0.7μg(r=0.999 9),平均加样回收率分别为97.8%(RSD=1.02%),98.9%(RSD=1.51%)。结论:所建立的方法测定快速,结果准确可靠。     

滇龙胆中萜类物质积累的动态变化

滇龙胆（Gentiana rigescens）为传统中药材。采用单因素方差分析比较不同生长季节滇龙胆的根部及茎叶部位龙胆苦苷、獐牙菜苦苷及当药苷含量的动态变化。结果表明,在不同生长季节,滇龙胆的根部与茎叶部位3种有效成分的含量具明显差异。龙胆苦苷主要积累于根部;獐牙菜苦苷、当药苷主要积累于茎叶部位。相关性分析表明,龙胆苦苷含量受气候因子的影响,月平均温度与茎叶部位龙胆苦苷含量呈极显著负相关（R=-0.57,P〈0.01）,月降水量与根部、茎叶部位龙胆苦苷含量呈显著（R=-0.48,P〈0.05）或极显著（R=-0.74,P〈0.01）负相关;根中当药苷含量与獐牙菜苦苷含量呈极显著正相关（R=0.62,P〈0.01）,茎叶部位獐牙菜苦苷含量与当药苷含量呈显著正相关（R=0.38,P〈0.05）。研究结果表明,滇龙胆中龙胆苦苷含量受降水量和温度的影响;龙胆苦苷、獐牙菜苦苷及当药苷的含量变动具相关性;9-11月较适宜滇龙胆药材的采收。     

正交法优选调控秦艽次生代谢产物含量变化的条件

为优选可抑制秦艽次生代谢产物含量变化的条件，该文采用3因素4水平正交试验设计方法，共设计16组处理，研究洛伐他汀(MVA途径抑制剂)、膦胺霉素(MEP途径抑制剂)和取样天数对秦艽中马钱苷酸、獐牙菜苷、獐牙菜苦苷、龙胆苦苷4种主要环烯醚萜类化合物含量的影响。结果表明：(1)4种环烯醚萜类化合物含量变化受取样天数影响最大，其次为膦胺霉素浓度，次之为洛伐他汀浓度。(2)以最佳抑制条件处理后，马钱苷酸、獐芽菜苦苷、龙胆苦苷和獐芽菜苷含量分别下降了69%、36%、33%和4%。基于正交法优选的抑制条件，对4种化合物均可抑制。综上所述，可确定调控秦艽次生代谢产物含量变化的最佳抑制条件为膦胺霉素400μmol·L^-1,洛伐他汀50μmol·L^-1,取样天数6 d,该条件为进一步研究MEP和MVA途径在环烯醚萜类化合物代谢合成中的调控机制奠定基础。     

滇龙胆与青叶胆环烯醚萜类物质计量特征分析

龙胆科龙胆属滇龙胆(Gentiana rigescens)与獐牙菜属青叶胆(Swertia mileensis)为我国特有种。采用高效液相色谱法测定其中马钱苷酸、獐牙菜苦苷、龙胆苦苷和当药苷含量,结合多变量分析研究环烯醚萜类成分含量及其构成比例在龙胆科植物种内和种间的差异。研究结果显示,4种环烯醚萜类成分的计量特征呈现种间差异,依据环烯醚萜类成分含量及其构成比例,所有滇龙胆样品可分为4类。马钱苷酸含量与纬度呈极显著负相关(R=-0.348,P0.05),与海拔呈极显著正相关(R=0.307,P0.01);獐牙菜苦苷含量与经度呈极显著负相关(R=-0.592,P0.01),高海拔有利于植株中当药苷构成比例的增加(R=0.245,P0.05);地理因子对植株中龙胆苦苷含量变化影响不显著(P0.05)。     

苦龙胆酯苷的研究进展

苦龙胆酯苷是一种裂环烯醚萜苷类化合物,又称为苦龙苷或龙胆苦酯苷。其分子式为:C27H28O14,分子量为576.52,是已知最苦的裂环烯醚萜苷类化合物。目前已证明可从川东獐牙菜、印度獐牙菜、龙胆草以及辐花肋柱花中提取。辐花肋柱花是最新证明的可提取苦龙胆酯苷的植物。苦龙胆酯苷具有助消化,保肝,抗皮肤肿瘤,抗黑热病等药理活性。在古代印度传统医药以及中药藏药中,苦龙胆酯苷的来源植物是治疗消化系统相关疾病的一味常见的草药,如保肝、抗糖尿病等。在体内药代动力学研究中,兔静脉注射苦龙胆酯苷,在血中有较快的清除率(2.62±0.41 L/h/kg)和广泛的体内组织分布(1.08±0.44 L/kg);采用游离、脂质体和囊泡体形式给仓鼠用药具有明显保护肝肾功能且未发现明显不良反应。     

滇龙胆与青叶胆环烯醚萜类物质计量特征分析?

龙胆科龙胆属滇龙胆( Gentiana rigescens)与獐牙菜属青叶胆( Swertia mileensis)为我国特有种。采用高效液相色谱法测定其中马钱苷酸、獐牙菜苦苷、龙胆苦苷和当药苷含量,结合多变量分析研究环烯醚萜类成分含量及其构成比例在龙胆科植物种内和种间的差异。研究结果显示,4种环烯醚萜类成分的计量特征呈现种间差异,依据环烯醚萜类成分含量及其构成比例,所有滇龙胆样品可分为4类。马钱苷酸含量与纬度呈极显著负相关( R＝－0?348, P<0?05),与海拔呈极显著正相关( R＝0?307, P<0?01);獐牙菜苦苷含量与经度呈极显著负相关( R＝－0?592, P<0?01),高海拔有利于植株中当药苷构成比例的增加( R＝0?245, P<0?05);地理因子对植株中龙胆苦苷含量变化影响不显著( P>0?05)。     

Concentrations of the Active Constituents of the Tibetan Folk Medicine Qinjiao (Gentiana sect. Cruciata) within and between Taxonomic Species across the Qinghai‐Tibetan Plateau

The Tibetan folk medicine Qinjiao is traditionally used to treat various conditions, and its main active constituents comprise four iridoid glycosides, i.e., loganic acid, swertiamarin, gentiopicroside, and sweroside. The traditional crude medicine Qinjiao is derived from the dried roots of three species belonging to Gentiana sect. Cruciata (Gentianaceae) growing in the Qinghai‐Tibetan Plateau (QTP). In this study, we determined by HPLC the contents of the four main active constituents in the dried roots collected from 83 localities at different altitudes across the QTP. The material was classified under the seven taxonomic species G. straminea, G. dahurica, G. crassicaulis, G. waltonii, G. officinalis, G. ihassica, and G. macrophylla. Our results suggested that the four constituents were present in the roots of all seven species for all localities, but their concentrations varied greatly within and between species. The level of gentiopicroside revealed to be the most dominant for all examined localities (2.1–12.4 mg/g), and G. macrophylla Pall . contained the highest concentration of all the four constituents at the species level. Except for loganic acid in G. officinalis, there was no significant correlation between the contents of these constituents and the altitude of the sampling localities. These results suggest that all species of all origins can be used as reliable resource for the crude medicine Qinjiao. However, a few species contain higher concentrations of the main active constituents, irrespective of their origin.     

坚龙胆中的一个新裂环烯醚萜甙

从坚龙胆（Gentiana rigescens）的根中分离得到1个新的裂环烯醚萜甙,命名为坚龙胆甙A（1）,以及8个已知化合物：龙胆苦甙（2）,6＇-O-β-D-吡喃葡萄糖基-龙胆苦甙（3）,马钱子酸（4）,6＇-O-β-D-吡喃葡萄糖基．马钱子酸（5）,獐牙菜甙（6）,2＇-（邻,间-二羟基苯甲酰基）．獐牙菜甙（7）,獐牙菜苦甙（8）,四乙酰开联番木鳖甙（9）。它们的化学结构通过现代波谱解析得以鉴定。化合物3,5和9为首次从坚龙胆中分离得到。     

农林复合系统滇龙胆茎叶化学计量特征研究

以农林复合系统种植的滇龙胆(Gentiana rigescens Franch.ex Hemsl.)为材料,采用高效液相色谱法建立不同栽培系统滇龙胆茎、叶的色谱指纹图谱,并测定其主要活性成分马钱苷酸、獐牙菜苦苷、龙胆苦苷和当药苷含量,研究不同栽培系统滇龙胆茎、叶化学计量特征。采用相关性分析、指纹图谱相似度分析、偏最小二乘判别分析(PLS-DA)、变量投影重要性准则(VIP)等方法进行化学数据分析。结果显示,滇龙胆主要活性成分马钱苷酸含量为(1.85±0.92)mg/g~(7.43±7.64)mg/g,獐牙菜苦苷含量为(1.03±0.17)mg/g~(1.58±0.50)mg/g,龙胆苦苷含量为(15.28±11.34)mg/g~(24.59±7.84)mg/g,当药苷含量为(4.10±1.64)mg/g~(31.67±22.70)mg/g,且叶片中4种活性成分的总含量高于茎;不同栽培系统中,与尼泊尔桤木间作的滇龙胆茎、叶活性成分总含量最高,而与核桃间作的滇龙胆茎、叶活性成分总含量最低。相关性分析显示,植株相同部位和不同部位间的环烯醚萜和裂环烯醚萜含量呈显著(P0.05)或极显著(P0.01)正相关。指纹图谱相似度分析表明,不同栽培系统滇龙胆茎指纹图谱相似度介于0.989~0.992之间、叶指纹图谱相似度为0.988~0.996,相同部位样品化学成分种类相似。PLS-DA分析结果表明,茎和叶片整体化学计量特征具有明显差异;单作及林药间作的样品被区分为不同类群,不同间作模式下滇龙胆茎、叶化学成分具显著差异,叶片高效液相色谱指纹图谱可用于区分不同栽培系统滇龙胆样品。本研究结果可为农林复合系统滇龙胆有效成分含量研究及滇龙胆资源的合理开发利用提供科学依据。     

A New Secoiridoidal Glucosides from Gentiana rigescens (Gentianaceae)

A new acylated secoiridoidal glucoside, named gentiorigenoside A (1 ), was isolated from the root of Gentiana rigescens (Gentianaceae) , together with eight known compounds, gentiopicroside ( 2) , 6′- O-β- D-glucopyranosyl gentiopicroside (3) , loganic acid (4 ) , 6′- O-β- D-glucopyranosyl loganic acid ( 5), sweroside ( 6 ), 2′-( o, m-dihydoxybenzyl) -sweroside (7), swertiamarin (8) and secologanoside (9) . heir structures were elucidated on the basis of detailed spectroscopic analysis. Iridoidal glucosides 3, 5 and 9 were isolated for the first time from the title plant.     

Chemical and Genetic Comparative Analysis of Gentiana crassicaulis and Gentiana macrophylla

Gentiana crassicaulis Duthie ex Burk . and Gentiana macrophylla Pall . are two main sources of Radix Gentianae Macrophyllae (Qinjiao) available in markets, which has a wide range of anti‐inflammatory effects and has been extensively used for fighting rheumatoid arthritis. However, they vary in terms of chemical compositions, pharmacological activities, and biomass. In this study, a combined chemical and genetic (HPLC and DNA barcoding) approach was used to compare these two plants. Four predominant bioactive compounds, namely, gentiopicroside, loganic acid, swertiamarin, and sweroside, were used to assess the chemical variations. Based on chemical variations, 15 samples were clustered into two groups through PCA analyses. DNA barcoding utilizing the variable nuclear ITS2 regions were sequenced, aligned, and compared. Together with 61 sequences collected from GenBank, 76 batches of Qinjiao were clustered in two groups according to species origin. The genetic relationships indicated by the ITS2‐based NJ tree were consistent with the chemical variations. Thus, the chemical profiles determined by HPLC and DNA profiles obtained from ITS2 region could be applied for the quality control of Qinjiao.     

HPLC Determination of Contents of Four Active Constituents in Tibetan Medicine Gentiana straminea ( Gentianaceae) During Different Growing Period

Contents of four iridoid glycosides viz. gentiopicroside, longanic acid, swertiamarin and sweroside of wild and cultivated Gentiana straminea roots gathered in different seasons were analyzed by means of HPLC determination. The results indicated that contents of the four iridoid glycosides varied greatly with different seasons between wild and cultivated Gentiana straminea. And according to the criterion of pharmacopoeia of China, after cultivation content of gentiopicroside in Gentiana str aminea root had accorded with the regulation and can preliminarily replaced the wild species to use as a plant medicine. The mechanism causing the content changes of the four iridoid glycosides may be due to the integrated effects of many factors, such as the ecological factors, the genetic factors.     

Three new secoiridoid glucosides from Eustoma russellianum

Three new secoiridoid glucosides, eustomoside, eustoside and eustomorusside, have been isolated along with three known glucosides, sweroside, swertiamarin and gentiopicroside, as well as one unknown glycoside from Eustoma russellianum.     

Isolation of iridoid and secoiridoid glycosides and comparative study on Radix gentianae and related adulterants by HPLC analysis

HPLC profile guided study led to the isolation of an acylated secoiridoid glycoside, named gentiotrifloroside (1), together with six known compounds, i.e., loganic acid (2), 6-O-beta-d-glucopyranosylgentiopicroside (3), swertiamarin (4), gentiopicroside (5), sweroside (6) and 2 -(o,m-dihydroxybenzyl)-sweroside (7) from Gentiana triflora and Gentiana rigescens. The structure of 1 was deduced from one- and two-dimensional NMR spectroscopic experiments. Compounds 1-7 were used successfully as chemical markers for the comparison of the four species of Gentiana used as Radix gentianae. Additionally, differentiation of Gentiana species mentioned and those used as adulterants was evaluated. The close similarity of chemical composition among the four genuine Gentiana species explain their popular usage as R. gentianae in Chinese medicine. We have also shown that the variation of chemical composition in R. gentianae and related adulterants agree well with their botanical phylogeny.