高效液相色谱法测定野菊花中绿原酸的含量

目的:建立野菊花中酚酸类主要活性成分绿原酸的含量测定方法。方法:采用RP-HPLC方法,色谱柱为AgilentXDB-C18(250 mm×4.6 mm,5μm),流动相选用乙腈-0.4%磷酸水(13∶87),流速0.8 mL.min-1,检测波长327 nm,柱温为30℃,样品温度20℃。结果:绿原酸在0.088~1.76μg范围内线性关系良好;平均回收率分别为98.8%,RSD小于1.0%;不同产地野菊花中绿原酸的含量均大于1‰。结论:本方法方便、准确、可以用于野菊花药材质量的进一步补充和完善。     

HPLC-DAD-MS/MS对栀子中色素成分的研究

目的鉴定栀子中藏红花素类色素成分。方法色谱条件:Dikma Diamonsil C18色谱柱(4.6 mm×250 mm,5μm);0.15%甲酸水-甲醇梯度洗脱,流速0.6 mL/min,柱温30℃,检测波长440 nm。质谱条件:电喷雾离子源(ESI);负离子模式检测;扫描范围m/z 100～1 400。结合化学成分的紫外-可见吸收光谱特征,与多级质谱(MS/MS)裂解特征分析,对其结构进行确证。结果采用高效液相色谱-二级管阵列检测器(HPLC-DAD)鉴别了栀子中的31个藏红花素类色素成分,并根据MS/MS裂解规律推断了其中的23个化学成分。结论 HPLC-DAD-MS/MS可用于快速筛选和鉴定栀子中的藏红花素类色素成分。     

HPLC-DAD-MS法测定异丙托溴铵原料药中有关物质

目的建立测定异丙托溴铵原料药中有关物质的高效液相色谱–二极管阵列检测器–质谱(HPLC-DAD-MS)法。方法采用HPLC-MS法,Alltima C_(18)色谱柱(150 mm×4.6 mm,5μm);流动相:0.01 mol/L乙酸铵–乙腈,梯度洗脱;体积流量:0.25 m L/min;柱温:35℃,进样量:10μL;检测波长:220 nm。采用离子阱(ESI)正负离子模式进行检测,源温110℃;脱溶剂温度350℃;脱溶剂气体积流量600 L/h;锥孔气体积流量50 L/h。结果异丙托溴铵、托品酸、托品酸乙酯和杂质F在0.01~10.00、0.012~12.000、0.083~8.300、0.05~5.00 mg/m L显示良好的线性关系;异丙托溴铵、托品酸、托品酸乙酯和杂质F的最低检测限分别为1.46、0.57、4.16、5.03 ng;异丙托溴铵、杂质C、托品酸乙酯和杂质F的定量限分别为4.38、1.71、12.48、15.09 ng。结论该方法灵敏度高、选择性高、检测限低,为异丙托溴铵的质量控制提供依据。     

黄连-吴茱萸药对化学成分的HPLC-DAD-MS分析

建立黄连-吴茱萸药对半仿生提取液化学成分的HPLC-DAD-MS分析方法，研究黄连-吴茱萸配伍机制的物质基础。以Hypersil BDS C₁₈为色谱柱，梯度洗脱后，综合分析不同样品的色谱峰保留时间、紫外光谱，一级和二级全扫描质谱图，归属了黄连-吴茱萸(6∶1)半仿生提取液中的17个色谱峰，8个来自吴茱萸，其他9个均来自黄连，确认了4个色谱峰分别为药根碱、羟基吴茱萸碱、巴马汀和小檗碱；推断了3个色谱峰分别为表小檗碱、非洲防己碱和黄连碱，同时测定了小檗碱和巴马汀的含量。黄连配伍吴茱萸后半仿生提取无新物质生成，但小檗碱和巴马汀的溶出量降低。     

Simultaneous Analysis of Thirteen Bioactive Components in Evodia rutaecarpa and Its Varieties by HPLC-DAD-MS

Objective To control the quality of Evodia rutaecarpa better.Methods An HPLC-DAD-MS/MS method was established for the rapid and efficient identification of bioactive constituents and for simultaneous quantitative analysis of four bioactive ingredients including evodiamine,rutaecarpine,dehydroevodiamine,and evodin in E.rutaecarpa,which was applied to evaluating eight samples of E.rutaecarpa and its varieties from different areas.Results Thirteen potentially bioactive constituents including one flavonoid glyc...     

Influence of cultivation conditions, season of collection and extraction method on the content of antileishmanial flavonoids from Kalanchoe pinnata

Ethnopharmacological relevance

Leaves from Kalanchoe pinnata (Lamarck) Persoon (Crassulaceae) are popularly used for healing wounds. Its antileishmanial properties are established in experimental animals, and its active flavonoid components have been identified.

Aim of the study

In this study, we attempted to standardize the extract from K. pinnata leaves by evaluating the influence of season of harvest, sunlight exposure and method of extraction on antileishmanial flavonoids content.

Materials and Methods

HPLC-DAD-MS was used to identify and quantify the active antileishmanial flavonoids in different extracts. ANOVA test for analyses of variance followed by the Tukey test of multiple comparisons were used in the statistical analysis. The antileishmanial potential was assessed by the activation of nitric oxide production by murine macrophage using the Griess method.

Results

We demonstrated that active flavonoids were significantly more abundant when the leaves were collected in the summer, and that aqueous extraction at 50 °C allowed the highest flavonoid extraction. The benefit of sunlight exposure was confirmed in plants cultivated under direct sunlight when compared with those that grown under shade. Under sunny conditions the yield of the most active antileishmanial favonoid quercitrin was increased by 7-fold. All aqueous extracts tested were capable to enhance the macrophage nitric oxide production. However, hot aqueous extract from leaves collected in summer exhibited the higher activity, in agreement with HPLC-DAD-MS analysis tendency. In addition, with the aim of reducing the individual chemical variations of the plant constituents and optimizing the production of the active extract, it was obtained in vitro monoclonal KP specimens that were easily adapted to field conditions and were able to produce antileishmanial flavonoids.

Conclusion

Our study reports the better conditions of cultivation, harvest and extraction protocol for obtaining a K. pinnata extract exhibiting the highest antileishmanial activity. Additionally, we propose the flavonoids quercetin 3-O-α-l-arabinopyranosyl (1 → 2)-α-l-rhamnopyranoside and quercitrin, as satisfactory chemical markers for standardization purposes.     

厚朴及其炮制品HPLC-DAD-MS色谱指纹图谱的研究

目的:厚朴生品与炮制品色谱指纹图谱的建立及应用。方法:应用HPLC-DAD-MS法建立厚朴及其炮制品的色谱指纹图谱,色谱条件为:LC-18分析柱(5μm,250 mm×4.6 mm),柱温30℃,流速1 mL/m in,流动相为水-乙腈,梯度洗脱;测定了10批厚朴生品、10批厚朴炮制品和姜汁样品。结果:建立了厚朴生品HPLC-DAD指纹图谱,10个生品样品有22个共有峰,应用HPLC-MS对其部分主要共有峰进行了初步鉴定;同法得到了厚朴炮制品的指纹图谱,有23个共有峰,与生品比较,峰形相似,炮制品比生品增加了生姜中的有效成分姜酚。结论:在建立的分析条件下获得的色谱指纹图谱可较全面地反映厚朴的化学成分及其炮制前后的差异。     

当归活性成分的血清药物化学研究

用高效液相色谱指纹图谱方法,对中药当归的乙醇提取物及其在灌胃兔子后的血液提取成分进行分析,与二极管阵列检测器和质谱联用仪器提供的色谱,光谱和质谱等多维信息对样品进行分析。通过比较进入血液前后各组分的相对含量变化,可以对血液中测定的各组分的生物活性进行判断和预测。并对当归的药物活性成分进行快速筛选和分析。     

HPLC-DAD-MS研究黄芪的化学成分

目的建立HPLC-DAD-MS快速分离分析黄芪Astragali Radix化学成分的方法。方法色谱柱为Thermo Hypersil-Hypurity C18(150 mm×2.1 mm,5μm)分析柱,柱温40℃,流动相水-甲醇(梯度洗脱),体积流量0.25 mL/min。二极管阵列扫描范围200～370 nm。质谱采用大气压化学电离(APCI+)离子源,扫描范围m/z 100～650。结果通过与已有文献报道的质谱、紫外光谱和保留行为比较可以初步定性5个化合物,分别为calycosin-7-O-β-D-glycoside、ononin、calycosin、formononetin、(3R)-7,2′-dihydroxy-3′,4′-dimethoxyisoflavan。结论因为能够同时提供相对分子质量、紫外光谱和保留行为,所以HPLC-DAD-MS是一种分析中药化学成分的有力方法。