穿心莲药材高效液相色谱指纹图谱研究及其应用

应用反相高效液相色谱(RP-HPLC)法建立穿心莲药材指纹图谱。首先以穿心莲对照药材为对象建立其乙醇提取物特征色谱图,再用5种不同来源穿心莲药材的乙醇提取物验证色谱图的指纹特性。在穿心莲药材乙醇提取物特征色谱图中有共有峰10个,各共有峰的相对保留时间与相对峰面积的相对标准偏差(RSD)均小于4%。对照药材中共有峰峰面积占总峰面积的98.5%以上。该指纹图谱用于清火栀麦胶囊中穿心莲的鉴别专属性良好。     

宁夏枸杞甜菜碱提取物高效液相色谱指纹图谱研究

建立宁夏枸杞甜菜碱提取物高效液相色谱指纹图谱,为鉴别不同来源的宁夏枸杞提供依据。以10批宁夏不同产地的宁夏枸杞主栽品种"宁杞Ⅰ号"样品建立枸杞甜菜碱提取物指纹图谱共有模式,采用"中药色谱指纹图谱相似度评价系统"软件进行数据处理,对15批不同来源的枸杞样品进行了分析。结果表明:8个特征峰构成了宁夏枸杞甜菜碱提取物的色谱指纹图谱,不同产地、不同品种的枸杞样品甜菜碱提取物指纹图谱存在差异;建立的枸杞甜菜碱提取物高效液相色谱(HPLC)指纹图谱对不同产地、不同品种枸杞的鉴别有参考价值。     

用高效液相色谱组方指纹图谱智能预测中药质量的新模式

建立大黄、黄芩、黄连的高效液相色谱(HPLC)组方指纹图谱,确定其融合模型,观察组方融合指纹图谱(CSF)与一清片复方样品指纹图谱的一致性,从而以CSF代替复方整体来智能预测复方制剂质量。用二极管阵列检测器(DAD)同时测定黄芩、大黄、黄连和一清片在268 nm波长下的HPLC指纹图谱,并使用系统指纹定量法进行定性、定量评价。结果 CSF涵盖各单味药主要色谱峰信息,即CSF共有峰(55个)涵盖一清片样品共有峰(50个)的主要指纹图谱信息。15批样品的质量除YQT-S01为5级外,其他质量均为3级及以上。各组合模式CSF质量除CSF-2为6级外,其余均为2级或1级。该文探讨了标准指纹图谱和中药组方融合指纹图谱的相关性,可以与相应的计算机评价软件相结合,通过组方融合指纹图谱所代表复方制剂的整体指纹图谱来实现智能预测中药成方制剂质量的新模式。     

邓老凉茶颗粒的超高效液相色谱质谱联用指纹图谱研究

建立了适用于邓老凉茶颗粒质量控制的超高效液相色谱质谱联用指纹图谱分析方法。样品采用甲醇索氏萃取60 min,萃取液采用超高效液相色谱质谱法进行指纹图谱分析。色谱柱采用Waters ACQUITY HSST3 C18(150 mm×3.0 mm,1.8μm),以0.5%甲酸-乙腈为流动相进行梯度洗脱,流速为0.8 mL/min,柱温35℃。质谱采用负离子ESI模式,选择基峰离子流质量色谱图进行指纹图谱研究。32个共有峰在15 min内得到良好分离,其中15个共有峰通过对照品进行了确证。通过《中药色谱指纹图谱相似度评价系统2004A版》对邓老凉茶颗粒样品进行相似度分析,15个批次样品的相似度均达到0.960以上,表明邓老凉茶颗粒的产品质量稳定性很好。以32个共有峰的相对峰面积进行主成分分析,邓老凉茶颗粒样品之间的细微质量差异得到明显区分。该方法快速、高效、可靠,可有效地用于邓老凉茶颗粒的质量控制。     

养血清脑颗粒的高效液相色谱指纹图谱研究

采用变换波长检测法建立了养血清脑颗粒HPLC指纹图谱,确定了35个共有色谱峰在药材中的归属,并用液相色谱-质谱联用(LC-MS/MS)法鉴定出部分化学成分,同时对制剂中的4种指标性成分进行了定量检测.结果表明,制剂与药材具有良好的色谱峰相关性,4种指标性成分在各自的浓度范围内线性关系良好(r＝0.9997～0.9999...     

朝鲜淫羊藿的色谱指纹图谱及液相色谱/电喷雾串联质谱分析

建立了长白山区朝鲜淫羊藿药材的高效液相色谱指纹图谱的分析方法. 确定了18批朝鲜淫羊藿药材的13个共有峰, 该指纹图谱的精密度、稳定性和重现性的相对标准偏差均低于3.0%. 结合液相色谱/电喷雾串联质谱对特征峰进行了结构确认, 并根据电喷雾串联质谱数据推测了13个特征化合物的结构. 结果表明采用高效液相色谱与质谱联用技术对朝鲜淫羊藿色谱指纹图谱中的特征峰进行结构确认, 使其色谱指纹图谱的特征性更强, 更适合于药材质量的鉴别与评价.     

高效液相色谱指纹图谱法分析南方红豆杉药材的氯仿提取物

建立了南方红豆杉药材氯仿提取物的高效液相色谱(HPLC)指纹图谱分析方法。采用Eurospher 100 C18色谱柱(250 mm×4 mm, 5 μm),以甲醇和水为流动相进行梯度洗脱,流速为1 mL/min,检测波长为232 nm,柱温为30 ℃。以10-脱乙酰巴卡亭III(10-DABIII)为参照物,在相同的色谱条件下测定了10批不同产地的南方红豆杉药材氯仿提取物的指纹图谱,获得了11个共有指纹峰,并利用主成分分析法(PCA)对指纹图谱进行统计分析。结果表明南方红豆杉药材的质量与种植区域有关。该方法稳定、可靠,可用于南方红豆杉药材的质量控制。     

银杏叶提取物的多维指纹图谱研究

利用高效液相色谱/二极管阵列检测/质谱/质谱法,建立了银杏叶提取物的多维指纹图谱,同时建立了总离子流指纹图谱,使同时检测银杏叶中两类有效成分黄酮和内酯类化合物成为可能。采用Zorbax SB-C18色谱柱(250 mm×4.6 mm i.d., 5 μm),流动相为水(含0.5 %甲酸)和乙腈,二元梯度洗脱,流速为1.0 mL/min,350 nm下检测。采用电喷雾离子源,负离子检测模式下采集质谱数据。利用指纹图谱相似度计算软件,以芦丁为内参比峰,考察了不同厂家银杏叶提取物与标准银杏叶提取物EGb 761     

中药指纹图谱共有峰的自动识别

依托时间匹配、光谱信息辅助,利用自编的VB程序,建立并比较了时间匹配和时间匹配参考光谱信息两种识别中药指纹图谱共有峰的方法,并成功地应用于肉苁蓉的高效液相色谱-二极管阵列检测器(HPLC-DAD)法指纹图谱的建立.结果表明,只需要人为设置色谱峰大小下限、保留值相对误差上限和最大吸收波长差值上限三个参数,VB程序编制的指纹分析系统就能快速对所有样本进行共有峰识别和模糊数学处理,实现了无人工干预下的中药指纹图谱共有峰的自动识别,指纹图谱的建立过程客观,并且节省大量时间,其中参考光谱法识别结果更准确.     

银黄口服液的质量控制及其高效液相色谱指纹图谱的研究

利用高效液相色谱方法,建立了复方银黄口服液及其原料药材黄芩和金银花的指纹图谱,测定了银黄口服液中黄芩苷 和绿原酸的含量。以Lichrospher C18色谱柱(250 mm×4.6 mm i.d.,5 μm)为分离柱,以甲醇和0.1%磷酸水溶液为流动 相进行二元梯度洗脱,流速0. 6 mL/min,在254 nm波长下检测。采用夹角余弦和相关系数的方法,计算6批银黄口服液指 纹图谱的相似度均在0.99以上,表明银黄口服液的质量未见显著差异。将银黄口服液指纹图谱中的共有峰与原料药材黄芩 和金银花的指纹图谱中的共有峰相比较,对其归属进行确认,大部分共有峰可以匹配。     

复方板蓝根颗粒化学成分的质谱研究

利用电喷雾多级串联质谱(ESI-MSn)和傅里叶变换离子回旋共振质谱(FT-ICR-MS)技术,对复方板蓝根颗粒经溶剂萃取后的95%(体积分数)醇提和水提部位的化学成分进行了系统研究,鉴定出多种氨基酸成分、糖及其衍生物、有机酸、氨基酸和单糖的梅拉德反应初级产物以及含硫化合物表告依春.该方法灵敏快速,适宜于中药提取物的化学成分分析.     

Fingerprint Chromatogram Analysis of Radix Glehniae by LC Coupled with Hierarchical Clustering Analysis

For the first time a validated liquid chromatography method coupled with hierarchical clustering analysis has been developed for the study of fingerprint chromatograms of extracts from the Radix Glehniae of Glehnia littorali. Liquid chromatography with gradient elution was performed on extracts from 23 plant samples collected from different geographical locations. Ten of twenty-three plant samples were selected using hierarchical clustering analysis and employed to establish the fingerprint. The fingerprint was established and ten chromatographic peaks were selected as characteristic peaks and panaxynol was used as reference standard compounds. The fingerprint chromatograms had a good stability, precision, and reproducibility. This method is a very reliable and useful method for assessment of the quality of Radix Glehniae.     

淫羊藿炮制前后UPLC-PDA-MS 的指纹图谱研究

建立了朝鲜淫羊藿和柔毛淫羊藿药材的超高效液相色谱(UPLC)指纹图谱分析方法, 实验结果表明, 两个品种的指纹图谱差别较大, 朝鲜淫羊藿标出35 个共有峰, 质谱鉴定了26 个峰, 柔毛淫羊藿标出13 个共有峰, 质谱鉴定了9个峰; 研究了朝鲜淫羊藿和柔毛淫羊藿炮制前后UPLC 指纹图谱的变化规律, 指纹图谱发生了五处变化. 该法快速、简便, 重现性好, 从整体上显示淫羊藿药材和炮制品的特征, 为淫羊藿的生品和炮制品的质量控制提供了有效手段.     

Chromatographic Fingerprint Analysis of Varietal Differences among Three Species of Baishouwu and Simultaneous Analysis of Three Bioactive Constituents by Use of LC–DAD

For the first time, a validated high-performance liquid chromatographic method with diode-array detection has been developed for discrimination and quality assessment of three species of Baishouwu. LC with gradient elution was performed on extracts from 23 plant samples collected from different geographic locations. The fingerprint was established and 26 chromatographic peaks were selected as characteristic peaks. Similarities of chromatograms were greater than 0.9 for the same species but much lower than 0.6 for different species. Three bioactive constituents were simultaneously analyzed for further quantitative assessment of the quality of Baishouwu, and amounts of the three analytes in different species of Baishouwu samples were found to be significantly different. This LC–DAD method is thus a very reliable and useful method for assessment of the quality of Baishouwu.     

Chemical fingerprint and quantitative analysis for the quality evaluation of Vitex negundo seeds by reversed‐phase high‐performance liquid chromatography coupled with hierarchical clustering analysis

A simple and efficient method was developed for the chemical fingerprint analysis and simultaneous determination of four phenylnaphthalene‐type lignans in Vitex negundo seeds using high‐performance liquid chromatography with diode array detection. For fingerprint analysis, 13 V. negundo seed samples were collected from different regions in China, and the fingerprint chromatograms were matched by the computer‐aided Similarity Evaluation System for Chromatographic Fingerprint of TCM (Version 2004A). A total of 21 common peaks found in all the chromatograms were used for evaluating the similarity between these samples. Additionally, simultaneous quantification of four major bioactive ingredients was conducted to assess the quality of V. negundo seeds. Our results indicated that the contents of four lignans in V. negundo seeds varied remarkably in herbal samples collected from different regions. Moreover, the hierarchical clustering analysis grouped these 13 samples into three categories, which was consistent with the chemotypes of those chromatograms. The method developed in this study provides a substantial foundation for the establishment of reasonable quality control standards for V. negundo seeds.     

基于遗传算法的色谱指纹峰配对识别方法

指纹峰配对识别是色谱指纹图谱分析中的关键环节之一，本文提出一种基于遗传算法的色谱指纹峰配对识别方法。该法根据对照色谱指纹图谱的峰分布特性初选出若干标定蜂，将其存入一个候选标定蜂库；同时根据这些候选标定峰从待测指纹图谱中选出相应的候选标定峰，也存入候选标定峰库；再用遗传算法从库中选取一组标定峰用于校正待测指纹图谱中各峰的峰位，并自动识别出与对照色谱指纹图谱相对应的各指纹峰。仿真实验及实际分析实验结果均表明，该法识别指纹峰准确可靠，可用于色谱指纹图谱相似度的快速自动计算。     

Fingerprint chromatogram analysis of extracts from the leaves of Tripterygium wilfordii Hook. F. by high performance liquid chromatography

A simple and reliable high performance liquid chromatographic (HPLC) method has been developed and validated for the study of fingerprint chromatograms of extracts from the leaves of Tripterygium wilfordii Hook. F. (TWHF) and for controlling the quality of the herb. HPLC separation of the extracts was performed on a Lichrospher RP-18 column and detected by ultraviolet absorbance at 210 nm. The column temperature was maintained at 35 degrees C. A mobile phase composed of acetonitrile:H2O in the ratio of 39:61 (v/v) was found to be most suitable for this separation at a flow rate of 0.8 mL/min with isocratic elution. Under the chromatographic conditions described, the peak profile of the 10 components collected within 35 min made up the fingerprint of the extracts from leaves of TWHF with universal features. The fingerprint chromatograms had a good stability, precision, and reproducibility. The similarity of the extracts from leaves of TWHF collected in summer and winter was studied with triptolide as a reference peak. The method is suitable for differentiation of extracts from the leaves of TWHF, and can be used as a quality control method for this herb.     

不同产地龙葵药材的高效液相色谱-蒸发光散射检测指纹图谱

采用高效液相色谱-蒸发光散射检测器(HPLC-ELSD)来研究不同产地龙葵药材品质的差异,为龙葵药材质量标准的制定提供参考依据。使用Phenomenex C₁₈色谱柱,流动相为乙腈-20%(v/v)甲醇水溶液(含0.03%(v/v)三乙胺),梯度洗脱,蒸发管温度为40 ℃。利用"中药色谱指纹图谱相似度评价系统"对所采集的色谱图进行相似度比对,选择相似度在0.9~1.0之间的色谱图归一化拟合成龙葵的标准指纹图谱,并利用对照品进行色谱峰指认。研究结果表明,不同产地来源的龙葵药材之间具有显著的差异,依据HPLC-ELSD采集到的非挥发类成分可对中药材进行区分。该方法为龙葵药材的真实性、优良性和稳定性评价提供了一种新的、具有广阔应用前景的技术手段。     

仿刺参中皂苷类成分的高效液相色谱指纹图谱

利用高效液相色谱法建立了仿刺参皂苷类成分的指纹图谱,为仿刺参的质量控制提供了新的方法。采用固相萃取制备供试品溶液,选用Zorbox SB-C18色谱柱(250 mm×4.6 mm, 5 μm),以乙腈-0.1%磷酸水溶液为流动相进行梯度洗脱,检测波长为205 nm,柱温30 ℃。分析了不同产地的10批仿刺参样品,采用国家药典委员会推荐的“中药色谱指纹图谱相似度评价系统(2004 A版)”处理谱图,确定了6个共有峰。计算了10个样本间的指纹图谱相似度,所得相似度计算结果均大于0.97。该方法具有良好的稳定性和重现性,可用于仿刺参的质量控制。