武当山区野生转家种重楼中4种皂苷含量分析

摘 要 目的：比较武当山区野生转家种重楼不同品种中4种重楼皂苷的含量,为推动本地区重楼药材的规范化种植,优选重楼种质资源提供参考。方法: 采集武当山区种植基地里野生重楼转家种的各个品种重楼药材样本,采用HPLC法测定各个样本中重楼皂苷的含量。结果: 6个重楼品种里只有七叶一枝花（华重楼）和滇重楼中重楼皂苷含量符合药典规定的标准,其他重楼品种中重楼皂苷含量明显低于国家标准。结论:应选择重楼皂苷含量符合中国药典2015年版规定的滇重楼和七叶一枝花推广种植,以达到重楼药材的合理开发利用的目的。     

HPLC测定不同生长年限安徽地产重楼中重楼皂苷的含量

目的:采用HPLC法测定不同生长年限安徽地产重楼中重楼皂苷Ⅰ、Ⅱ、Ⅵ、Ⅶ的含量.方法:采用高效液相色谱法,色谱柱为Agilent SB-C18(4.6mm×250mm,5μm),流动相为乙腈-水(梯度洗脱),检测波长为203nm,流速:1.0mL·min-1,柱温30℃.结果:安徽地产重楼中含有重楼皂苷Ⅰ、Ⅱ、Ⅵ、Ⅶ,但其含量低于药典标准规定,而且不同生长年限中重楼皂苷的含量差异较明显,5年生药材含量最高.结论:研究结果可为重楼资源的开发利用和确保临床用药质量提供依据.     

HPLC测定不同产地滇重楼中的4种重楼皂苷

目的 分析测定不同产地、不同部位滇重楼中重楼皂苷Ⅰ、Ⅱ、Ⅵ、Ⅶ的含量.方法 采用HPLC法,色谱柱为ZorbaxSB-C18(250 mm×4.6 him,5 μm),流动相为乙腈-水(40:60),体积流量1.0 ml·min-1,柱温30℃,检测波长203 nm.结果 4种不同产地滇重楼皂苷的含量差异较大.栽培滇重楼中,4种重楼皂苷的总含鼍高于野生滇重楼,滇重楼不同部位的4种重楼皂苷的总含量为根茎>叶>果实>须根>茎.结论 应积极配合GAP的实施,尽快实现滇重楼人工栽培产业化,保证其质量具有可控性.     

云南漾濞不同地区核桃林下种植重楼的重楼皂苷含量比较

摘 要 目的：对云南漾濞不同地区核桃林下栽种重楼中的4种重楼皂苷含量进行比较。方法: 采集云南漾濞不同地区核桃林下栽种的重楼样品,按中国药典2015年版收载的方法进行含量测定。结果: 云南漾濞不同地区栽种重楼中4种重楼皂苷含量各样品间存在显著差异。样品均为仿野生林下载培,但载培时间长短不一,其中生长年限4~5年的重楼药材中4种重楼皂苷含量明显高于3年生重楼。结论:4种重楼皂苷含量各样品间存在显著差异与重楼生长的生态环境、生长年限和同一物种不同居群间遗传背景等不同有关。     

不同生长年限滇重楼中4种重楼皂苷的含量比较

目的:研究滇重楼中4种重楼皂苷的含量与其生长年限的关系。方法:采用高效液相色谱法测定不同生长年限滇重楼中重楼皂苷Ⅰ、Ⅱ、Ⅵ、Ⅶ的含量。色谱柱为Agilent Zorbax SB-C18(150mm×4.6mm,5μm),流动相为乙腈-水(梯度洗脱),流速为1.0mL·min-1,检测波长为203nm,柱温为35℃。结果:不同生长年限滇重楼中重楼皂苷的含量差异较明显,3年生药材重楼皂苷的含量比较低,4、5年生药材含量明显升高,5年以上药材含量有下降趋势。结论:重楼皂苷含量的变化规律可为确定滇重楼采收年限提供参考依据。     

HPLC法测定14个不同产地滇重楼中薯蓣皂苷元的含量

目的:建立测定滇重楼中薯蓣皂苷元含量的方法,并比较不同产地滇重楼中薯蓣皂苷元的含量。方法:采用高效液相色谱法。色谱柱为Intersil ODS-C18,流动相为乙腈-水(92∶8,V/V),流速为1.0 ml/min,检测波长为203 nm,柱温为35℃,进样量为20μl。结果:薯蓣皂苷元进样量在0.539~13.475μg范围内与其峰面积呈良好的线性关系(r=0.999 9);精密度、稳定性、重复性试验的RSD均≤1.92%;平均加样回收率为98.48%,RSD为1.21%(n=6)。14个不同产地滇重楼中薯蓣皂苷元的含量在3.869 4~21.074 7 mg/g之间。结论:该方法快速、简便,结果准确、可靠,可用于不同产地滇重楼中薯蓣皂苷元的含量测定。薯蓣皂苷元含量受产地、环境因素影响较大,滇重楼品质表现出一定的地域及生境依赖性。     

HPLC法测定不同来源重楼药材中7种甾体皂苷的含量

目的分析不同来源重楼属植物的主要活性成分,比较7种甾体皂苷的含量,为重楼药材的鉴别和品质评价提供参考依据。方法采用HPLC法测定不同来源重楼属15批药材中重楼皂苷Ⅶ、重楼皂苷H、重楼皂苷Ⅵ、重楼皂苷Ⅱ、薯蓣皂苷、重楼皂苷Ⅰ和重楼皂苷Ⅴ的含量。色谱条件为Inertsil ODS-3C18色谱柱(150mm×4.6mm,5μm);流动相为乙腈-水(50∶50);检测波长为203nm;流速为1.0mL·min~(-1);柱温为35℃;进样量为20.0μL。结果 15批样品中,重楼皂苷Ⅶ、重楼皂苷Ⅵ、重楼皂苷Ⅱ和重楼皂苷Ⅰ4种皂苷含量之和均在1.849~20.222mg·g~(-1)之间;样品中7种皂苷含量之和在2.351~37.366mg·g~(-1)之间。精密度、稳定性和重复性实验的RSD值均≤3.0%,7种皂苷的平均加样回收率均在95.92%~103.20%之间(RSD≤3.0%,n=5)。结论因产地或种类不同,重楼皂苷的含量存在一定差异。基于15批重楼样品的HPLC图谱,构建了重楼药材的HPLC指纹图谱,实验结果可用于重楼的真伪鉴别和质量评价。     

浙江七叶一枝花种质资源的化学评价

目的 测定浙江不同采集地野外七叶一枝花中主要活性成分甾体皂苷的含量,为筛选优质种质资源和适宜栽培地提供依据。方法 采用高效液相色谱法,色谱柱为Waters XBridge C₁₈柱(4.6 mm×250 mm,5 μm);流动相为乙腈(A)-水(B),梯度洗脱,柱温：25 ℃,洗脱流速为1.0 mL·min^-1,测定12个不同采集地七叶一枝花中重楼皂苷Ⅰ、Ⅱ、Ⅵ、Ⅶ的含量。结果 来自不同采集地的七叶一枝花种源间的重楼皂苷含量存在显著差异,有一半种源的皂苷含量达不到药典规定的标准,优质种源来自永嘉上桥头村,4种重楼皂苷之和为2.928%,明显高于其他种源。结论 浙南山区较适合七叶一枝花的人工种植。     

20批重楼药材皂苷类成分分析与其TLC鉴别方法探讨

目的：对不同品种、不同产地的重楼药材中的皂苷类成分进行测定,以探讨其标准中TLC鉴别指标成分设定的合理性。方法：按《中国药典》2015年版一部重楼标准项下的TLC法和HPLC法分别检测皂苷类成分的组成和含量。结果：20批重楼药材中只有10批同时检出重楼皂苷Ⅰ、Ⅱ、Ⅵ和Ⅶ 4种皂苷成分,其余10批只检出其中的3种成分。结论：建议修订重楼标准项下相关内容：只要总皂苷含量符合规定,TLC鉴别检出其中两种以上皂苷即可。     

鄂西北地区重楼质量控制与品质评价研究

目的了解鄂西北地区重楼药材的质量情况,建立科学合理的品质评价方法,为道地药材重楼在该地区品种选育与林下仿野生规范化种植提供技术支撑。方法通过DNA条形码技术进行重楼属植物的区分;通过测定重楼药材的水分、总灰分、酸不溶性灰分及8种重楼皂苷的含量对重楼药材进行质量控制与品质评价。结果 DNA条形码序列ITS2能够直观有效的区分重楼属植物;对29批不同产地的重楼药材进行质量综合评价,结果显示茅箭区马家河龙王寨重楼药材质量较好。结论鄂西北地区不同产地品种的重楼质量存在明显差异,不同产地品种的重楼皂苷含量不同,同一产地七叶一枝花和竹溪重楼皂苷含量较高,质量较好,可以作为仿野生种植品种。     

不同生长年限滇重楼HPLC指纹图谱的研究

目的比较不同生长年限滇重楼的高效液相色谱指纹图谱,探索滇重楼的最适宜采收年限。方法采集不同生长年限的滇重楼样品,用高效液相色谱法绘制指纹图谱,对不同生长年限滇重楼进行相似度计算和色谱数据分析。结果不同生长年限的滇重楼质量具有明显差别,其中6年生滇重楼总物质含量较高,各化合物含量变化有所差别。结论所得数据可为滇重楼的采收提供参考。     

蓣知子皂甙IV的结构

从木通科木通属植物白木通[Akebia trifoliata(Thunb.)Koidz.var.australis(Diels)Rehd]种子的乙醇提取物中以硅胶层析等方法得四种三萜皂甙。其中甙IV是新天然产物,命名为蓣知子皂甙IV(yuzhiziosideIV)。根据化学和光谱分析,确定甙IV的结构为3-O-β-D-吡喃木糖基-(1→2)-a-L-吡喃阿拉伯糖齐墩果酸-28-O-β-D-吡喃葡萄糖基-(1→6)-β-D-吡喃葡萄糖酯甙。另外皂甙B(I)、皂甙C(II)和皂甙D(III)为已知物。这些化合物在白木通种子中均是首次得到。     

藤山柳根茎中的三萜成分

目的　研究我国特有的单种属药用植物藤山柳[Clematoclethrascandens (Franch .)Maxim .]根茎的甲醇提取物正丁醇萃取部分的化学成分。方法　将甲醇提取物分散于水中,经石油醚和乙酸乙酯萃取后,再用正丁醇进行萃取,对正丁醇萃取物进行柱色谱分离纯化,用波谱方法鉴定化合物结构。结果　分离鉴定了5个三萜化合物:2α,3β,24-三羟基乌苏-12 ,18-二烯-28-酸-28-O-β-D-吡喃葡糖酯(I) ,niga-ichigoside F₁ (II) ,asiaticacid (III) ,2α,3α,24-三羟基乌苏-12-烯-28-酸(IV) ,2α,3α,19α,24-四羟基乌苏-12-烯-28-酸(V)。结论　以上三萜化合物均为首次从该属植物中获得,其中I是一个新三萜皂苷     

Rapid and simple determination of polyphyllin I,II, VI,and VII in different harvest times of cultivated <Emphasis Type="Italic">Paris polyphylla</Emphasis> Smith var. <Emphasis Type="Italic">yunnanensis</Emphasis> (Franch.) Hand.-Mazz by UPLC-MS/MS and FT-IR

Paris Polyphylla Smith var. yunnanensis (Franch.) Hand.-Mazz (“Dian Chonglou” in Chinese) is a famous herbal medicine in China, which is usually well known for activities of anti-cancer, hemolysis, and cytotoxicity. In this study, Fourier transform infrared (FT-IR) spectroscopy coupled with principal component analysis (PCA) and partial least-squares regression (PLSR) was applied to discriminate samples of P. polyphylla var. yunnanensis harvested in different years and determine the content of polyphyllin I, II, VI, and VII in P. polyphylla var. yunnanensis. Meanwhile, ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was used to study the dynamic changes of P. polyphylla var. yunnanensis harvested in different years (4, 5, 7, 8, 9, 12, and 13 years old). According to the UPLC-MS/MS result, the optimum harvest time of P. polyphylla var. yunnanensis is 8 years, due to the highest yield of four active components. By the PCA model, P. polyphylla var. yunnanensis could be exactly discriminated, except that two 8-year-old samples were misclassified as 9-year-old samples. For the prediction of polyphyllin I, II, VI, and VII, the quantitative results are satisfactory, with a high value for the determination coefficient (R ²) and low values for the root-mean-square error of estimation (RMSEE), root-mean-square error of cross-validation (RMSECV), and root-mean-square error of prediction (RMSEP). In conclusion, FT-IR combined with chemometrics is a promising method to accurately discriminate samples of P. polyphylla var. yunnanensis harvested in different years and determine the content of polyphyllin I, II, VI, and VII in P. polyphylla var. yunnanensis.     

Qualitative and quantitative analysis of steroidal saponins in crude extracts from Paris polyphylla var. yunnanensis and P. polyphylla var. chinensis by high performance liquid chromatography coupled with mass spectrometry

High performance liquid chromatography coupled with electrospray ionization multi-stage tandem mass spectrometry (HPLC–ESI-MSⁿ) and triple quadrupole mass spectrometric detection (HPLC–ESI-MS/MS), respectively, had been employed for the simultaneous identification and quantification of steroidal saponins in the rhizomes of Parispolyphylla var. yunnanensis and P. polyphylla var. chinensis, which are the qualified plants of “Chonglou” in Chinese. The HPLC experiments were performed by means of a reversed-phase C-18 column and a binary mobile phase system consisting of 0.1% aqueous formic acid and acetonitrile under gradient elution conditions. The characteristic fragmentation patterns of diosgenin- and pennogenin-type steroidal saponins were investigated using ESI-MSⁿ in negative ion mode. The MSⁿ data of the [M−H]⁻ ions provided structural information on the sugar sequence of the oligosaccharide chains and the aglycones of steroidal saponins. As a result, ten and seven saponins were determined in P. polyphylla var. yunnanensis and P. polyphylla var. chinensis, respectively, including four unknown compounds. One unknown compound was tentatively identified as diosgenin-3-O-α-l-rhamnopyranosyl(1 → 4) [α-l-rhamnopyranosyl(1 → 2)]-β-d-glucopyranoside and the aglycones of the other three new compounds were reported from Chonglou for the first time. The developed HPLC–ESI-MS/MS method was validated and found to be satisfactorily linear, selective and robust. The limits of detection (LODs) and quantitation (LOQs) ranged, respectively, from 0.5 to 10 ng/mL and 2 to 34 ng/mL depending on six various compounds. The intra- and inter-day precisions of the method were evaluated and were less than 5.0%. Recoveries ranged from 92% to 104% for all compounds. The established quality evaluation method was successfully used for simultaneous quantification of six predominant steroidal saponins in the rhizomes of these two Paris species.     

Preparative separation and purification of steroidal saponins in Paris polyphylla var. yunnanensis by macroporous adsorption resins

Abstract

Context: Saponins are active compounds in natural products. Many researchers have tried to find the method for knowing their concentration in herbs. Some methods, such as solid–liquid extraction and solvent extraction, have been developed. However, the extraction methods of the steroidal saponins from Paris polyphylla Smith var. yunnanensis (Liliaceae) are not fully researched.

Objective: To establish a simple extraction method for the separation of steroidal saponins from the rhizomes of P. polyphylla Smith var. yunnanensis.

Materials and methods: Macroporous adsorption resins were used for the separation of steroidal saponins. To select the most suitable resins, seven kinds of macroporous resins were selected in this study. The static adsorption and desorption tests on macroporous resins were determined. Also, we optimized the temperature and the ethanol concentration in the extraction method by the contents of five kinds of saponins. Then, we compared the extraction method with two other methods.

Results: D101 resin demonstrated the best adsorption and desorption properties for steroidal saponins. Its adsorption data fits best to the Freundlich adsorption model. The contents of steroidal saponins in the product were 4.83-fold increased with recovery yields of 85.47%.

Discussion and conclusion: The process achieved simple and effective enrichment and separation for steroidal saponins. The method provides a scientific basis for large-scale preparation of steroidal saponins from the Rhizoma Paridis and other plants.     

五味子不同炮制品对大鼠肾阳虚的改善作用研究

目的通过比较五味子不同炮制品对肾阳虚大鼠的补益作用,为酒五味子"入补药熟用"提供药理学依据。方法采用甲巯咪唑法造肾阳虚大鼠模型,将成模大鼠随机分为模型组、右归丸(3.7 g/kg)组、生五味子(2.8 g/kg)组、酒五味子(2.8 g/kg)组、醋五味子(2.8 g/kg)组,另设对照组,每组10只。各给药组从第16天开始ig相应药液,给药体积均为10mL/kg,连续30 d。试剂盒测定各组大鼠血清中游离三碘甲状腺原氨酸(FT3)、游离四碘甲状腺原氨酸(FT4)、促甲状腺素(TSH)、一氧化氮(NO)、促甲状腺激素释放激素(TRH)水平;测定血浆中环磷酸腺苷(cAMP)、环磷酸鸟苷(cGMP)水平,计算cAMP/cGMP;观察各组大鼠甲状腺组织病理形态。结果与模型组比较,五味子各炮制品均能不同程度地提高大鼠血清FT3、FT4、NO及下丘脑TRH水平,降低血清TSH水平,两组比较差异具有统计学意义(P0.05、0.01);均可以提高血浆cAMP水平,降低血浆cGMP水平,两组比较差异具有统计学意义(P0.05、0.01)。其中与生五味子、醋五味子组比较,酒五味子作用最优(P0.01)。五味子各炮制品对甲状腺病理形态均有所改善,其中酒五味子、醋五味子较优。结论五味子的补肾作用与改善下丘脑–垂体–甲状腺(HPT)轴功能抑制有关,酒五味子补肾作用最好。     

荔枝叶药材质量标准研究

目的 建立荔枝叶药材的质量标准。方法 建立荔枝叶药材的性状、显微特征、薄层鉴别及含量测定方法;按照中国药典2020年版第四部通则中相关方法,对荔枝叶药材的水分、总灰分、酸不溶性灰分及浸出物等项目进行测定。结果 荔枝叶药材在性状、显微特征和薄层色谱方面均具有专属性特征。药材水分含量为2.66%～6.42%,总灰分含量为2.96%～4.52%,酸不溶性灰分含量为0.17%～0.94%,水溶性浸出物含量为15.38%～19.87%,醇溶性浸出物含量为24.94%～30.33%。没食子酸、原儿茶酸、儿茶素、表儿茶素、芦丁、紫云英苷、槲皮素、山柰酚8种成分分别在0.003 3～0.052 0 mg·mL^–1、0.006 9～0.109 6 mg·mL^–1、0.013 0～0.208 0 mg·mL^–1、0.057 5～0.920 0 mg·mL^–1、0.013 4～0.213 6 mg·mL^–1、0.013 2～0.211 2 mg·mL^–1、0.002 9～0.0...     

青海产小叶黑柴胡中皂苷类成分的含量分析

目的测定青海产小叶黑柴胡中总皂苷、柴胡皂苷a及柴胡皂苷d的含量,为其质量评价和开发利用提供参考。方法采用分光光度法测定小叶黑柴胡中总皂苷的含量;采用高效液相色谱法测定小叶黑柴胡中柴胡皂苷a和柴胡皂苷d的含量。结果小叶黑柴胡中总皂苷含量>2.77%,柴胡皂苷a和柴胡皂苷d的含量分别高于0.54%和0.14%,其中柴胡皂苷a和柴胡皂苷d的含量远高于中国药典中柴胡含量项下的标准。结论本方法简便快速,结果准确、稳定,适用于小叶黑柴胡中皂苷类成分的含量测定。青海产小叶黑柴胡具有较高的药用价值。     

桔梗总皂苷对左氧氟沙星在小鼠体内的药动学及组织分布的影响

目的 采用高效液相色谱（HPLC）法测定小鼠血浆及组织中左氧氟沙星含量,研究桔梗总皂苷对其药动学规律及组织分布的影响。方法 180只昆明小鼠随机分为两组,组Ⅰ为单独ig左氧氟沙星78 mg/kg,组Ⅱ为同时ig桔梗总皂苷65 mg/kg与左氧氟沙星78 mg/kg,采用HPLC法测定给药5、15、30、45 min及1、1.5、2、4、8 h后的血浆及肝、肺、肾组织样品中的左氧氟沙星浓度。结果 血浆中内源性物质对待测物无干扰;血浆及各组织在检测浓度范围内呈良好线性关系（r²>0.999）;方法日内和日间精密度、稳定性及提取回收率均符合生物样品检测标准;组ⅠAUC_0-t为（143.593±16.56）,组ⅡAUC_0-t为（120.339±15.542）,与组Ⅰ比较,组Ⅱ的AUC明显减小,T_1/2Z显著缩短,T_max明显延长,CLz/F显著增加,C_max及Vz/F无显著性差异;肝、肺、肾组织中左氧氟沙星浓度均迅速提高并快速消除,肺组织中消除速度最快,肺中AUC减少最为明显。结论 合用时,桔梗总皂苷使左氧氟沙星在小鼠体内消除速率明显加快,肺组织中尤为明显,故左氧氟沙星在联合含有桔梗成分的中药治疗肺部疾病时,建议分开服用。