基于指纹图谱和化学计量法评价不同产地番石榴叶的质量

番石榴叶为桃金娘科植物番石榴(Psidium guajava Linn.)的干燥叶,主要分布于广东、广西、福建、海南、四川及云南等地。本研究通过高效液相色谱法(HPLC)指纹图谱对不同产地的番石榴叶进行质量评价。采用相似度评价、聚类分析、主成分分析和正交偏最小二乘法-判别分析对指纹图谱数据进行统计分析。结果表明31批番石榴叶样品确定了14个共有峰,其相似度处于0.720～0.928,其中相似度小于0.8的番石榴叶样品主要产自广西、广东,相似度大于0.9的番石榴叶样品主要产自四川、云南;聚类分析及主成分分析提示番石榴叶样品可根据产地大致分为两类,包括沿海地区(广西、广东、福建、海南)和西南地区(四川、云南),不同产地番石榴叶药材质量存在差异;正交偏最小二乘法-判别分析结果表明9、4、12、11、13、3、10号峰是引起番石榴叶产地质量差异的主要成分,并鉴定出9号峰为鞣花酸,11号峰为异槲皮苷,13号峰为番石榴苷,12号峰为瑞诺苷,10号峰为金丝桃苷。定量分析鞣花酸、异槲皮苷、番石榴苷、瑞诺苷、金丝桃苷发现西南地区的5种成分总量高于沿海地区,具有显著性差异。本研究建立的番石榴叶HPLC指纹图谱结合化学计量法的方法简便准确,可为番石榴叶质量控制和品质评价提供参考依据。     

基于指纹图谱结合化学模式识别法评价不同产地睡莲花药材质量CSCD

建立睡莲花HPLC指纹图谱,并对5种成分进行含量测定,结合化学模式识别法,综合分析各产区睡莲花药材的成分差异,为睡莲花药材的进一步开发利用奠定基础。采用Waters C_(18)色谱柱(250 mm×4.6 mm, 5μm),以乙腈(A)-0.2%磷酸水溶液(B)为流动相,梯度洗脱,流速1.0 mL/min,柱温30℃,检测波长273 nm、350 nm。在273 nm下标定23个共有峰,指认了5种化学成分,350 nm标定17个共有峰,指认了4种化学成分。对15批样品聚类分析,在273 nm下分为4类,350 nm下分为3类。正交偏最小二乘法判别分析(OPLS-DA)共找到273 nm的10个可能影响睡莲花药材质量的差异性标志物。15批样品相似度为0.972~0.999,表明不同批次睡莲花样品存在差异。该方法可有效评价不同产地睡莲花药材的质量差异,为其质量控制提供参考。     

太子参红外指纹图谱的双指标序列分析

本文采用傅里叶变换红外光谱仪测定了四大种植主产区(安徽、江苏、福建、贵州)的栽培品及河南的野生品,共13个批次太子参甲醇提取物,获得不同产地太子参栽培品的FTIR指纹图谱,并对指纹图谱进行了相似度的计算,同时结合(共有峰率,变异峰率)双指标序列分析方法,对不同产地太子参的FTIR光谱进行了研究。结果表明,该方法所建立的太子参FTIR指纹图谱能够区分栽培品和野生品的差异,双指标序列分析法能够为产区间和产区内太子参品质差异提供参考,两种分析方法结合可为该药材的质量分析、评价与控制提供依据。     

马鞭草药材UPLC指纹图谱建立及指标性成分的测定CSCD

建立马鞭草药材指纹图谱,并对4种有效成分的含量进行测定,为马鞭草药材质量控制提供参考。采用超高效液相色谱(UPLC)法建立马鞭草药材指纹图谱,并利用高分辨质谱对共有峰进行指认,以15批马鞭草药材指纹图谱共有峰的相对峰面积为变量,进行化学计量学分析,并对15批马鞭草药材有效成分戟叶马鞭草苷、马鞭草苷、毛蕊花糖苷和异毛蕊花糖苷的含量进行测定。结果显示,马鞭草药材指纹图谱有16个共有峰,并指认出其中6个成分,聚类分析(HCA)和主成分分析(PCA)将15批药材分为3类,正交偏最小二乘法-判别式分析(OPLS-DA)共筛选出7个VIP>1.0的差异性成分。含量测定结果显示,不同产区的马鞭草药材戟叶马鞭草苷、马鞭草苷、毛蕊花糖苷和异毛蕊花糖苷的含量存在较大的差异。该方法能有效分析不同产地马鞭草药材的质量差异,为不同产地马鞭草药材质量的评价提供参考。     

天然植物黄蜀葵提取物对变形链球菌生长及黏附抑制作用的实验研究

目的通过研究黄蜀葵花总黄酮对变形链球菌生长及黏附的影响,为临床龋病预防提供实验基础。方法采用二倍稀释法观察不同浓度的黄蜀葵花总黄酮对变形链球菌生长的影响,测定该药物的最小抑菌浓度（MIC）;以低于MIC的5个浓度梯度配置含药的TPY液体培养基,接种变形链球菌,厌氧培养24h,计算黄蜀葵花总黄酮对变形链球菌的黏附抑制率。结果一定浓度的黄蜀葵花总黄酮能够抑制变形链球菌的生长,MIC为2．5g／L;变形链球菌的黏附率随培养基中黄蜀葵花总黄酮浓度的升高而下降,且抑菌作用呈现明显的浓度依赖性。结论天然植物黄蜀葵提取物黄蜀葵花总黄酮对变形链球菌的生长和黏附都有一定的抑制作用。     

狼毒大戟二萜类成分HPLC指纹图谱及聚类分析

为建立狼毒大戟药材的特征图谱,科学评价其质量,本文采用高效液相色谱法,乙腈-水梯度洗脱,测定了11批不同来源的狼毒大戟样品二萜类化合物,通过相似度分析建立了狼毒大戟提取物的高效液相指纹图谱,并对11批药材进行系统聚类分析。本文采用乙腈-水为流动相,梯度洗脱,检测波长:230 nm,流速:1.0mL/min,进样量:10μL,柱温:30℃。相似度评价结果显示11批不同产地的狼毒大戟相似度均高于0.9,并对11批样品建立共有模式,确定了23个特征共有峰,聚类分析结果将狼毒大戟样品分为2类,与药材产地相关。本文所建立狼毒大戟的指纹图谱方法简便、可靠,可作为狼毒大戟药材的基源鉴定参考方法。     

利用红外光谱和高效液相色谱比较分析不同产地红根草丹参酮ⅡA含量

红根草是广西道地药材,丹参酮ⅡA是红根草的主要活性作用成分之一,因此,筛选出丹参酮ⅡA含量高的优良种质,对解决红根草资源紧缺问题及提高其药材质量均有重要作用。本研究利用傅里叶红外光谱技术对不同产地红根草化学成分的红外吸收特征峰进行了测定,利用HPLC技术不同产地红根草中丹参酮ⅡA含量,同时,借助化学计量学方法比较分析了不同产地样品的红外光谱吸收峰与丹参酮ⅡA相关性。结果表明,不同产地红根草化学成分组成比较相似,但化学成分的含量却有较大差异。多元对数相关分析技术显示,1 734 cm~(-1)、1 632 cm~(-1)、1 548 cm~(-1)、1 515 cm~(-1)、830 cm~(-1)及789 cm~(-1)等处附近的吸收强度与实际测定不同产地红根草丹参酮ⅡA含量均有密切关系。两种方法均一致显示,丹参酮ⅡA含量以江西南丰类型最高,其次是广西钟山类型,其中以江西奉新类型含量最低。     

基于HPLC指纹图谱结合一测多评法的葛花质量控制研究

建立葛花药材的指纹图谱,并同时建立其中10个主要黄酮类成分含量的一测多评分析方法(QAMS),用于不同产地葛花药材整体质量评价。采用高效液相色谱法(HPLC)对不同产地葛花中主要化学成分进行分析,确定最佳色谱条件,建立指纹图谱;在此基础上,以葛花苷为参照峰,建立QAMS测定葛花10个黄酮类成分含量;并与外标法(ESM)测定结果比较,验证QAMS的准确性。建立了葛花药材指纹图谱,20批葛花和10批粉葛花药材的相似度均大于0.85,标定共有峰分别为18和27个。以葛花苷为参照峰,各个成分校正因子的重复性、耐用性均良好;应用一测多评法测定10个成分的含量,与外标法测定结果比较,含量无明显差异(P>0.05),相对偏差<3.0%。不同产地来源的葛花药材中10个成分含量均存在较大差异,说明应提高葛花药材的质量控制标准。所建立的指纹图谱结合一测多评分析方法简便、可行,可客观、有效的用于不同产地葛花药材质量评价,可以为葛花质量标准修订、优质葛花种质资源的选育和育种提供参考。     

基于3种香茶菜属药用植物高效液相色谱的主成分分析

对香茶菜(Isodon amethystoides)、显脉香茶菜(I.nervosa)和大萼香茶菜(I.macrocalyx)不同产地和器官(根、茎、叶)共21个样品进行高效液相色谱分析,以出峰时间-峰面积为指标,以样品为对象进行主成分分析,比较不同样品间的差异程度。结果发现,(1)香茶菜、大萼香茶菜、显脉香茶菜在高效液相色谱上虽然有一定的差异,但这种差异并不明显;(2)基于HPLC显示的香茶菜不同种群间差异比三个种间的差异更为明显,说明不同产地对香茶菜属植物样品的植化组成有很大的影响,不同种在植化上的相似性使它们在一定程度上可以作为替代药材;(3)研究反映出基于高效液相色谱的PCA在反映不同样品植物化学组成差异程度上具有应用价值。     

安徽东至中药材粗榧高效液相指纹图谱和化学成分分析

建立安徽东至粗榧药材高效液相指纹图谱,比较不同产地、不同干燥方法、不同药用部位粗榧药材的质量.以Kromasil C18为分析柱,用甲醇-乙腈(1:1)和0.1%醋酸水溶液梯度洗脱进行色谱分离,利用有效部位HPLC图谱辅助对共有峰进行指认,标示出13个共有峰,使用计算机辅助相似度评价软件进行数据处理.通过对不同产地、不同干燥方法、不同药用部位粗榧药材指纹图谱的分析,发现不同来源粗榧药材化学组成相似,其相对比例具有差异.通过HPLC-UV-MS/MS联用分析以及分离纯化鉴定两个主要化学成分的结构,它们分别为芹菜素-5-O-α-L-吡喃鼠李糖基-(1→4)-β-D-吡喃葡萄糖苷,芹菜素-5-O-α-L-吡喃鼠李糖基-(1→4)-6"-O-乙酰基-β-D-吡喃葡萄糖苷,干燥过程可能使它们发生转化.指纹图谱用于粗榧药材的质量评价切实可行,方法重现性好.     

Metabolite identification strategy of non-targeted metabolomics and its application for the identification of components in Chinese multicomponent medicine Abelmoschus manihot L.

Identification of multicomponent in traditional Chinese medicine (TCM) is complex and time-consuming. The inspection of the full-scan mass chromatograms was usually performed manually, which is labor-intensive. It is difficult to distinguish low response signals from complex chemical background. Furthermore, this process is typically based on earlier knowledge of the chemical composition of TCM, and those molecules that have not been characterized earlier were thus ignored. In this paper, a strategy using UPLC-MS combined with pattern recognition analysis was developed to simplify and quicken the identification of multicomponent in Abelmoschus manihot (L.) Medik. First, complex signals obtained by UPLC-MS were processed using automated data mining algorithm and further processed with multivariate chemometric methods. Multicomponent in Abelmoschus manihot L. can be clearly displayed in S- and VIP-plot. Using this method, 320 peaks which present in Abelmoschus manihot L. were detected. In the next step, accurate mass spectra of the characteristic markers acquired by QTOF MS were used to estimate their elemental formulae and enable structure identification. By searching in METLIN database, 41 components were tentatively identified in Abelmoschus manihot L. Our results showed that UPLC-MS based-pattern recognition analysis approach can be used to quickly identify TCM multicomponent and for standardization of herbal preparations.     

Separation of botulinum-positive and -negative fish samples by means of a pattern recognition method applied to headspace gas chromatograms.

A gas chromatographic headspace technique was used to analyze the gas produced during putrefaction of pond-raised, degutted trout, incubated in evacuated plastic pouches. The following samples were analyzed; 10 samples which, due to natural contamination with Clostridium botulinum, were toxic when injected into mice, 10 samples which were nontoxic when injected, and 9 samples inoculated with one strain of C. botulinum type E. The gas chromatograms showed the presence of 118 compounds in most samples. Quantitative differences among most chromatograms could be observed, but no compound was unique to any of the three groups. By means of a specific pattern recognition method, all negative samples were shown to fall into one group and were distinctly separated from the toxic samples. No differences could be observed between the two groups of inoculated and naturally contaminated trout samples. The results suggest that headspace analysis combined with pattern recognition analysis might prove to be a valuable method for screening studies of foods containing living cells of C. botulinum.     

Separation of botulinum-positive and -negative fish samples by means of a pattern recognition method applied to headspace gas chromatograms.

A gas chromatographic headspace technique was used to analyze the gas produced during putrefaction of pond-raised, degutted trout, incubated in evacuated plastic pouches. The following samples were analyzed; 10 samples which, due to natural contamination with Clostridium botulinum, were toxic when injected into mice, 10 samples which were nontoxic when injected, and 9 samples inoculated with one strain of C. botulinum type E. The gas chromatograms showed the presence of 118 compounds in most samples. Quantitative differences among most chromatograms could be observed, but no compound was unique to any of the three groups. By means of a specific pattern recognition method, all negative samples were shown to fall into one group and were distinctly separated from the toxic samples. No differences could be observed between the two groups of inoculated and naturally contaminated trout samples. The results suggest that headspace analysis combined with pattern recognition analysis might prove to be a valuable method for screening studies of foods containing living cells of C. botulinum.     

Comparative Investigation for Raw and Processed Products of Euodiae Fructus Based on High-Performance Liquid Chromatography Fingerprints and Chemical Pattern Recognition

As a traditional Chinese medicine, Euodiae Fructus is widely used due to its analgesic, anti-inflammatory, and antihypertensive effects. However, Euodiae Fructus has also been documented to be toxic, and the toxic effects can be reduced by processing. To distinguish Euodiae Fructus from its processes products and study the changes of raw and processed products before and after processing, we evaluated four auxiliary material processing methods including vinegar, Zingiberis Rhizoma, Coptidis Rhizoma, and Glycyrrhizae Radix et Rhizoma. The raw Euodiae Fructus and four processed Euodiae Fructus samples were analyzed and compared based on the high-performance liquid chromatography (HPLC) fingerprints combined with chemometrics, including principal component analysis (PCA), partial least squares-discriminant analysis (PLS-DA), and principal component analysis-class (PCA-Class). A total of 27 common peaks were obtained by fingerprint analysis. The fingerprint similarity of raw and processed samples was between 0.86–0.999. We also determined the contents of the main active ingredients - Evodiamine and Rutaecarpine. PCA and PLS-DA analyses were used to distinguish between the raw and processed samples of Euodiae Fructus, and 14 chemical markers were screened out. Four kinds of processed products were further analyzed and the results showed that they could be successfully distinguished under the established models, and 12 chemical markers were labeled. PCA-Class results revealed that the classification models constructed in this study had adequate discrimination ability. The method combined with HPLC fingerprinting and multi-component chemical pattern recognition technology could be used to differentiate raw and processed Euodiae Fructus with adequate predictive power. Our findings confirmed the rationality of the pharmacopoeial method and provided a reference for the quality control of the Glycyrrhizae Radix et Rhizoma processed Euodiae Fructus.     

芦荟的HPLC指纹图谱建立、化学模式识别分析及其含量测定

采用Phenomenex C₁₈色谱柱(250 mm×4.6 mm,5μm),以甲醇-乙腈-0.3%磷酸水为流动相进行梯度洗脱,建立芦荟的指纹图谱。运用化学模式识别方法对不同产地芦荟药材质量控制方法进行评价。结果表明:12批芦荟HPLC指纹图谱共标定23个共有峰,并通过对照品指认其中6个成分;除了广西的3批药材之外,其他药材相似度都在0.93以上;聚类分析和主成分分析将12批芦荟分为3类;利用正交偏最小二乘判别法筛选出芦荟药材差异的5个色谱峰,并以相同HPLC法对其进行含量测定。本文将HPLC指纹图谱与化学模式识别相结合的方法简便准确,为芦荟药材的质量控制和品质评价提供依据。     

Analytical pyrolysis-pattern recognition for the characterisation of leafy spurge (Euphorbia esula L.) biotypes

Curie-point pyrolysis-gas chromatography-pattern recognition was used to elucidate chemical variations within leafy spurge (Euphorbia spp.). Hierarchical cluster analysis (HCA) readily identified two major clusters corresponding to E. esula and E. cyparissias. The E. esula cluster further separated into three distinct subclusters. Results from principal components analysis (PCA) and fuzzy c-varieties (FCVPC) pattern recognition were similar, verifying the presence of three biotypes among the E. esula samples studied. It is suggested that analytical pyrolysis in combination with pattern recognition may predict the behaviour of biocontrol agents introduced into fields to control leafy spurge.     

Identification of the potential active components of Abelmoschus manihot in rat blood and kidney tissue by microdialysis combined with ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry

In this paper, microdialysis combining with ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS) was applied to simultaneously identify components in blood and kidney dialysis after oral administration of Abelmoschus manihot extract. Microdialysis probe was implanted in the jugular vein and the kidney medulla, respectively; microdialysis samples were collected continuously, transferred to microtubes and analyzed by UPLC-Q-TOF/MS. The components in microdialysis samples were separated by an UPLC HSS T3 column and eluted with acetonitrile and water (containing 0.1% formic acid) at a flow rate of 0.4 mL/min. The results showed that unbound constituents in blood circulation of the rat include hyperoside, isoquercitrin, quercetin monoglucuronide, quercetin-3'-O-glucoside, quercetin, myricetin, and hibifolin while unbound constituents in kidney are hyperoside, isoquercitrin, quercetin monoglucuronide, which might be the potential active components in vivo. The developed method was simple and reliable, and could be adopted to rapidly screen and identify potential active components contributing to pharmacological effects of TCM and to better clarify its action mechanism.     

Corolla Is a Novel Protein That Contributes to the Architecture of the Synaptonemal Complex of Drosophila

In most organisms the synaptonemal complex (SC) connects paired homologs along their entire length during much of meiotic prophase. To better understand the structure of the SC, we aim to identify its components and to determine how each of these components contributes to SC function. Here, we report the identification of a novel SC component in Drosophila melanogaster female oocytes, which we have named Corolla. Using structured illumination microscopy, we demonstrate that Corolla is a component of the central region of the SC. Consistent with its localization, we show by yeast two-hybrid analysis that Corolla strongly interacts with Cona, a central element protein, demonstrating the first direct interaction between two inner-synaptonemal complex proteins in Drosophila. These observations help provide a more complete model of SC structure and function in Drosophila females.     

Fourier transform magnitudes are unique pattern recognition templates

Fourier transform magnitudes are commonly used in the generation of templates in pattern recognition applications. We report on recent advances in Fourier phase retrieval which are relevant to pattern recognition. We emphasise in particular that the intrinsic form of a finite, positive image is, in general, uniquely related to the magnitude of its Fourier transform. We state conditions under which the Fourier phase can be reconstructed from samples of the Fourier magnitude, and describe a method of achieving this. Computational examples of restoration of Fourier phase (and hence, by Fourier transformation, the intrinsic form of the image) from samples of the Fourier magnitude are also presented.