以人血清白蛋白为固定相的分子生物色谱分析几种中药活性成 …

提出用人血清白蛋白分子生物色谱分析中药活性成分的方法,分别比较了当归等４种单味中药的分离结果,考察了提取当归的３种溶剂及提取时间对活性物质提取量的影响,同时建立了当归中阿魏酸与藁本内酯的宣分析方法。     

基于电化学安培法的中药降血糖活性成分筛选

为建立一种简单高效的中药活性成分的提取分离、定性鉴定、含量测定以及降血糖活性研究的新方法,采用绿色无毒的水提酸沉法提取了黄芩的活性成分-黄芩苷,通过薄层色谱法做定性鉴定和紫外分光光度法测定其含量,构建一个高效的电化学葡萄糖传感器,并建立一种基于电化学安培法的降血糖活性研究新方法。 以临床降糖药阿卡波糖论证了电化学安培法检测葡萄糖淀粉酶抑制活性的可靠性和稳定性,并成功应用于黄芩苷的降血糖活性研究。 结果表明,黄芩苷具有较好的葡萄糖淀粉酶抑制活性,其半数抑制浓度(IC₅₀)为0.056 g/L。 本研究为从天然药物中提取分离降血糖活性成分提供了一种新颖的研究方法。     

生物色谱用于当归中活性成分筛选及分离研究

应用分子生物色谱技术研究了筛选和分离当归中的活性成分。以甲基丙烯酸缩水甘油酯为功能单体,通过原位聚合得到了整体柱基质,用二乙胺活化后,以吸附蛋白的方法得到了以牛血清白蛋白(BSA)为固定相的分子生物色谱柱。当归样品经甲醇提取,0.45μm膜过滤后,以BSA分子生物色谱柱为分离柱,以含2%(体积分数)甲醇的Tris-盐酸缓冲溶液为流动相进行分离,用二极管阵列检测器检测。结果表明:活性组分在BSA上的保留机理是疏水作用、静电作用和特定的与空间结构相关的多种因素的共同作用结果。将BSA柱上的保留组分进行了气相色谱-质谱分析,鉴定了8种成分,包括当归中的主要可挥发性成分(藁本内酯)。     

基于微量热法的板蓝根提取方法和活性部位挑选

应用TAM Air微量热系统, 采用微量热法, 考察不同提取方法的板蓝根水煎液及其化学萃取部位在37 ℃时, 对大肠杆菌生长代谢的影响. 结合中医药理论, 分析板蓝根水煎液及其化学萃取部分的药效作用及其差异, 挑选板蓝根药材最佳的提取方法和抑菌最强的活性部位, 并采用试管稀释法求出抑菌活性最强部位的最小抑菌浓度(MIC). 结果表明微量热法可用于常用中药——板蓝根的提取方法和活性部位的挑选, 为进一步研究中药的药性基础提供了一种新的、快速灵敏的技术和方法.     

基于质量标志物的当归抗炎功效近红外快速评价

该文通过筛选当归中对NF-κB具有抑制活性的质量标志物(Q-marker),建立了一种利用近红外光谱技术(NIRS)快速评价当归抗炎功效的方法。采用UPLC/Q-TOF技术结合NF-κB双荧光素酶报告基因对当归中具有NF-κB抑制活性的成分进行筛选,并通过单体、整体及等效性验证确立了关键Q marker。以UPLC分别对多批当归药材中的Q-marker进行含量测定,建立其NIRS定量模型,并进一步将Q-marker的含量与当归的NF-κB抑制活性相关联,构建“量-效”拟合函数。结果表明,绿原酸(X1)、洋川芎内酯I(X3)和Z-藁本内酯(X4)为NF-κB抑制活性的关键Q-marker,其含量波动与整体抗炎活性(Y)的变化相吻合,符合拟合方程：Y=16.13-1484X1+7.981X3+0.112 6X4,且基于NIRS的预测效果良好。该方法通过整合分析实现了基于关键Q-marker的当归药材抗炎功效的快速评价,为中药材品质的快速分析提供了新的研究范式和解决方案。     

红外光谱分析数据特征指纹的可视化表达方法

提出一类红外光谱分析数据特征指纹的可视化表达方法。该方法采用基于贝叶斯准则的多元统计方法对原始数据进行投影变换，再以地维灰度图对变换后数据进行可视化表征，形成基于计算机图像的虚拟化学指纹图谱。将其用于鉴别3种不同产地中药当归样品的结果表明，它能有效提取红外光谱分析数据的特征指纹，实现以虚拟指纹图谱对药材产地的分类鉴别，从而为辨识复杂化学物质体系提供了新的技术手段。     

液相色谱指纹图谱在当归、黄芪及其复方有效成分分析中的应用

将液相色谱指纹图谱与正辛醇-水分配体系模拟中药在人体内的吸收情况相结合分析中药活性成分,分析比较了当归、黄芪单煎液及合煎液在不同靶位酸度下化学成分种类、含量的变化情况,探讨了配伍及酸度变化对中药中各成分的形态分布及在人体中吸收情况的影响.发现不同靶位环境的酸度及不同配伍对中药中的有效物质组分、含量及各组分的吸收都有很大的影响.     

气相色谱-质谱法测定当归粉、当归原药材中的邻苯二甲酸二丁酯

建立了气相色谱-质谱法测定当归粉和当归原药材中邻苯二甲酸二丁酯（DBP）的方法。在国家标准的基础上,通过单因素试验,优化了提取方法和提取溶剂,确定了以正己烷为提取剂从水中液液萃取的方法。方法学试验结果表明,DBP浓度在0．01～1．00μg／mL范围内与色谱峰面积呈良好的线性关关系,线性相关系数r^2=0．9995,检出限为0．05μg／g（全扫描）,不同水平的加标回收率在90％～100％之间,测定结果的相对标准偏差小于5％（n=3）。该方法适合当归粉和当归原药材中DBP的日常检测。     

人参、当归中微量氟的离子色谱分析

近年来,对中药研究的报道大多数侧重于有机成份的分析,而对其中无机成份研究较少。文献报道了126种中药中微量元素的系统分析,但未对人体必需的十四种微量元素之一——氟进行测定。氟是人体必需的微量元素,对牙齿及骨骼的形成和结构,以及钙和磷的代谢均有重要的作用。维持人体内部微量元素平衡的主要来源是食物和药物,为此本文选用人参和当归二种中药,用离子色谱法测定其煎剂中的微量氟,对探讨中药中微量成份的分布与疗效之问的关系有着一定的意义,人参和当归中微量氟的离子色谱分析尚未见文献报道,本文提出了一种简便、快速、准确地测定中药煎剂中微量元素氟的可靠的分析方法。     

FAAS法分析当归、黄芪及当归补血汤中金属元素的形态

采用正辛醇-水分配体系模拟中药水煎液在人体肠胃中的分配情况,应用火焰原子吸收光谱法测定了当归、黄芪及当归补血汤中锌、铁、锰、铜、镁、钙、镍7种金属元素在药材中的总含量,以及水煎液中各金属元素的水溶态、醇溶态含量,探讨了配伍和酸度对水煎液中金属元素含量的影响。实验结果表明,中药中金属元素的形态与中药成分、配伍情况及作用靶位(肠胃)的酸度有关,配伍及酸度对中药中金属元素的溶出率及水煎液中金属元素的形态影响较大。     

Screening and analysis of biologically active compounds inAngelica sinensis by molecular biochromatography

Summary A novel strategy for the screening and analysis of biologically active compounds in traditional Chinese medicine by molecular biochromatography is proposed. Molecular biochromatography with human serum albumin (HSA) immobilized on silica as stationary phase was used to screen and analyse the bioactive compounds in the typical Chinese medicine ofAngelica sinensis (Oliv.) Diels. Ten peaks showed retention on this column, which is based on their affinity for HSA. Ferulic acid and liguistilide were identified as the principal active components, which agrees very well with the results in the literature. A quality control method was also developed based on the simultaneous determination the concentrations of ferulic acid and liguistilide in solutions ofAngelica sinensis (Oliv.) Diels extracted with water and methanol. It was observed that the concentrations of ferulic acid and liguistilide in solution extracted with methanol were 2 and 53 times higher, respectively, than those with water. It was shown that molecular biochromatography is an effective way of analysing and screening biologically active compounds in traditional Chinese medicine.     

Quantification of ligustilides in the roots of Angelica sinensis and related umbelliferous medicinal plants by high-performance liquid chromatography and liquid chromatography-mass spectrometry

A reversed phase high-performance liquid chromatographic method was developed for quantifying E-ligustilide (1) and Z-ligustilide (3) in the roots of Angelica sinensis (Oliv.) Diels with confirmation using UV, atmospheric pressure chemical ionisation (APCI) MS and APCI-MS-MS techniques. Based on the UV spectra of compounds 1, E-butylidenephthalide (2), 3 and Z-butylidenephthalide (4), the absorption at 350 nm was chosen as measuring wavelength in which baseline separation of compounds 1 and 3 could be obtained but avoided the interference of compounds 2 and 4. The identity of compounds 1 and 3 in samples were unambiguously determined by the respective quasi-molecular ions ([M+H]+) in APCI-MS. According to the stability data, acetonitrile was chosen for the preparation of standard solutions in order to minimize the isomerization of compound 3. Compounds 1 and 3 were qualitatively and quantitatively analyzed in seven samples of the roots of Angelica sinensis (Oliv.) Diels, Angelica acutiloba Kitagawa, Angelica acutiloba Kitagawa var. sugiyamae Hikino and the rhizome of Ligusticum chuanxiong Hort. Analysis of an extract from a sample root of Angelica gigas Nakai using LC-MS for the first time could not detect the presence of ligustilide in this herb. The overall analytical procedure is rapid and reproducible which is considered suitable for quantitative analysis of large number of samples.     

Development of pressurized hot water extraction followed by headspace solid-phase microextraction and gas chromatography-mass spectrometry for determination of ligustilides in Ligusticum chuanxiong and Angelica sinensis

In this work, a simple, rapid, solvent-free, and low-cost method was developed for the determination of ligustilides in traditional Chinese medicines (TCMs), which was based on pressurized hot water extraction (PHWE) followed by headspace solid-phase microextraction (HS-SPME) and gas chromatography-mass spectrometry (GC-MS). The two bioactive compounds Z-ligustilide and E-ligustilide in two common TCMs, viz. Ligusticum chuanxiong and Angelica sinensis, were extracted by water at 150 degrees C and 40 bar, followed by concentration with HS-SPME and detection by GC-MS. PHWE and HS-SPME parameters were investigated and method validation (precision and recovery) was studied. It has been shown that the proposed method provides a powerful approach for quantitative analysis of ligustilides in TCMs. The method was applied to determination of ligustildes in the TCMs from different growing areas. The results indicate that PHWE-HS-SPME-GC-MS is a potential tool for TCM quality assessment.     

当归化学成份分析研究——根部非挥发性成份分析

本文报导了甘肃岷县当归根部非挥发性成份的分析结果。共鉴定出37个有机成份,其中15种未见报导。用等离子体发射光谱检出当归中13种微量元素。     

Inhibitory effects of tetramethylpyrazine and ferulic acid on spontaneous movement of rat uterus in situ

Tetramethylpyrazine is one of the alkaloids contained in Ligusticum wallichii Franch. Ferulic acid is a phenolic compound contained in Ligusticum wallichii Franch and Angelica sinensis (Oliv.) Diels. The present study was carried out to examine the effect of tetramethylpyrazine and ferulic acid and the combined effect of both compounds on spontaneous uterine contractions in rats in situ. Tetramethylpyrazine and ferulic acid showed an inhibitory effect on uterine movement when given perorally and intravenously, respectively. The combination of both compounds, at doses individually insufficient to inhibit, synergistically inhibited uterine contraction. It was found that tetramethylpyrazine and ferulic acid inhibited uterine contractions and the inhibitory effect induced by the combination of both was due to the potentiation.     

Improved performance of comprehensive two-dimensional HPLC separation of traditional Chinese medicines by using a silica monolithic column and normalization of peak heights

Performance of comprehensive two-dimensional liquid chromatography system is greatly improved than we reported previously by using a silica monolithic column as for the second dimensional separation. Due to the increase of the elution speed on the second dimensional monolithic column, the first dimensional column efficiency and analysis rate can be greatly improved as comparing with conventionally second dimensional column. The developed system was applied to analysis of methanol extraction of two umbelliferae herbs Ligusticum chuanxiong Hort. and Angelica sinensis (Oliv.) Diels by using CN column as for the first dimensional separation and a silica monolithic ODS column for the second dimensional separation, and the obtained three-dimensional chromatograms were treated by normalization of peak heights with the value of the highest peak or setting a certain value using a software written in-house. It was observed that much more peaks for low-abundant components in TCM extract can clearly be detected here than we reported before, due to the large difference for the amount of components in TCMs' extract. With the above improvements in separation performance and data treatment, totally about 120 components in methanol extraction of Rhizoma chuanxiong and 100 in A. sinensis were separated with UV detection within 130 min. This result meant that both the number of peaks detected increase twice but the analysis time decrease twice if comparing with the previously reported result.     

Antiinflammatory effect of tetramethylpyrazine and ferulic acid.

Tetramethylpyrazine (TMP) is one of the alkaloids contained in Ligusticum wallichii Franch (L. wallichii). Ferulic acid (FA) is a phenolic compound contained in L. wallichii and Angelica sinensis (Oliv.) Diels (A. sinensis). The present study was carried out to examine the antiinflammatory effect and to elucidate the mode of the effect of TMP and FA. Both compounds significantly inhibited the edema induced by carrageenin, the increase of the dye leakage induced by acetic acid and the granuloma formation induced by cotton pellet. And also, TMP and FA inhibited the number of writhes induced by acetic acid. From these results, it is suggested that both compounds have the antiinflammatory effect and the analgesic effect, and both compounds exert an antiinflammatory effect at the early and the late stages of processes in the inflammatory pathology.     

Analysis of Chemical Incredients of Angelica Sinnesis——Analysis of Non-Volatile Constituents of Roots

In the present investigation,the non-volatile constituents of Danguei (Tang-Kuei) roots of Gansu origin [Angelica sinensis (Oliv.) Diels] were analyzed and thirty seven organic constituin s were also identified,fifteen of which have not been recorded in literature before,including a new optically active compound named as Angelicide and an antibiotie,Brereldin A.By plasma emission spectrometry,thirteen minor elements were also detecled     

固定化脂质体色谱研究中药复方的细胞膜通透性成分及其质量控制

提出了固定化脂质体色谱(immobilized liposome chromatography,ILC)研究中药复方的细胞膜通透性成分及其质量控制的方法。以当归补血汤及其组分为例,考察了它们在ILC色谱柱上的分离效果;当归补血汤水提取液和75％乙醇提取液在ILC色谱柱上各有8个保留峰,当归水提取液、甲醇提取液和75％乙醇提取液在ILC色谱柱上分别有3、6、9个保留峰,黄芪水提取液、甲醇提取液和75％乙醇提取液在ILC色谱柱上各有7个保留峰;建立了当归补血汤及其组分中阿魏酸和蒿本内酯的定量分析方法,当归补血汤水提取液和75％乙醇提取液中阿魏酸的含量分别为0．0743％和0．0688％,蒿本内酯的含量分别为0．0472％和0．457％,当归水提取液和75％乙醇提取液中阿魏酸的含量分别为0．0694％和0．0691％,蒿本内酯的含量分别为0．0781％和0．455％。     

Hypothesis of active components in volatile oil from a Chinese herb formulation, 'Shao-Fu-Zhu-Yu decoction', using GC-MS and chemometrics

Traditional Chinese medicine (TCM) with few or no side effects has increasingly attracted attention all over the world. However, the bioactive components and the therapeutic mechanisms are usually not understood because of the complex chemical compositions of these medicines. In this paper, GC-MS coupled with a chemometric method was developed for analysis of active components in volatile oil from a Chinese herb formulation, "Shao-Fu-Zhu-Yu Decoction". The volatile oils, obtained by hydrodistillation from "Shao-Fu-Zhu-Yu Decoction" (SFZYD) and its constituent herbs with abundant volatile oil (Angelica sinensis, Ligusticum chuanxiong, Cinnamomum cassia, Foeniculum vulgare, Zingiber officinale), were chemically analyzed using GC-MS and bioassayed using oxytocin-induced uterine contraction assay in vitro. Then, a mathematic model relating the chemical compositions and their activities in inhibiting mice uterine contraction was established for hypothesis of the bioactive compounds based on chemometrics. As a result, nine compounds which might contribute to the inhibition of oxytocin-induced uterine contraction were selected, and the activities of some of them were further confirmed by our experiments and/or the literature. The data suggest that the developed method is helpful for screening bioactive components from complex mixtures, such as the extracts of TCM.