桑叶HILIC特征指纹图谱研究

应用亲水色谱法(HILIC)建立了桑叶药材的指纹图谱,并对10批桑叶样品进行分析,为桑叶药材的真伪鉴别及质量控制提供了新方法。采用HILIC XBridgeTMAmide色谱柱,以乙腈-水(含0.2%甲酸、20mmol/L甲酸铵、20%甲醇)为流动相,梯度洗脱,流速为0.8 mL/min,柱温为25℃,检测波长为322 nm,进样量为20μL。该方法具有良好的精密度、重现性和稳定性,检测的10批桑叶指纹图谱有17个共有峰,4个特征峰,采用ESI-TOF/MS对4个特征峰进行了指认,结合相似度分析可以用于不同产区桑叶药材的鉴别。桑叶HILIC指纹图谱有望成为桑叶药材真伪鉴别及质量控制的有力工具。     

超高效液相色谱指纹图谱结合化学计量学优选掌叶大黄饮片炮制工艺

优选掌叶大黄饮片的炮制工艺.建立基于UPLC(超高效液相色谱)的掌叶大黄饮片主要指标成分指纹图谱测定方法,结合相似度评价、聚类分析、主成分分析和方差分析等化学计量学方法,对不同润制时间、不同干燥方式、不同烘干温度及时间掌叶大黄饮片的指纹图谱信息进行分析.不同润制时间掌叶大黄饮片的指纹图谱之间存在明显差异;聚类分析可将不...     

气相色谱-质谱指纹图谱在鉴别人体气味中的应用

利用顶空固相微萃取-气相色谱/质谱法(SPME-GC/MS)分析了人体腋窝气味。以标准品十七烷进行定位,通过谱图分析和相对保留时间计算,确定了八强峰。根据八强峰和指纹图谱分区相似度对指纹图谱进行分析比较,建立了区别不同个体的新方法。该方法能较全面地反映人体气味的特征峰和整体信息,初步探讨了用仪器鉴别不同个体的可能性。     

利用样本成分耗散物的非线性化学指纹图谱原理及相似度计算与评价

在恒温恒压条件下,以丙酮和样本中底物作为主要耗散物的不同成分的样本对非线性化学反应机理产生不同影响,从而引起反应体系电位-时间曲线形状不同变化为特征的B-Z化学振荡体系为例,就非线性化学指纹图谱原理进行了详细研究和讨论,并提出了计算非线性化学指纹图谱系统相似度的通用方法.利用系统相似度和欧氏距离、相关系数及夹角余弦对不同生产批次古汉养生精和18种其他样本的非线性化学指纹图谱的相似度进行了计算与分析.结果表明,相关系数和夹角余弦都不能用来作为评价非线性化指纹图谱相似度的指标.利用欧氏距离公式计算指纹图谱的非参数型相似度时,能正确反映指纹图谱的特征差异,但用其计算参数型相似度时,则有时不能正确反映样本非线性化学指纹图谱特征差异的相对程度.系统相似度能最真实反映样本指纹图谱之间差异程度,是4种相似度计算方法中最好的,可用于非线性化学指纹图谱相似度计算与评价.成功提出了一种经济、简便、易行和有效的鉴别样本真伪与评价其质量的科学方法.     

气相色谱-质谱指纹图谱在鉴别贵州茅台酒中的应用

应用气相色谱-质谱(GC-MS)建立了贵州茅台酒的指纹图谱,确证了贵州茅台酒中35种特征组分,并采用浙江大学的中药指纹图谱相似度计算软件对样品图谱之间的相似度进行了评价和鉴别。方法的精密度及重复性良好。研究考察了38个批次贵州茅台酒、5种由贵州茅台酒股份有限公司生产的酱香型系列白酒以及12种由其他厂家生产的白酒的指纹图谱与贵州茅台酒指纹图谱模板的相似度。结果表明,通过酒的特征组分比较和基于“夹角余弦法”的指纹图谱相似度分析,可以区分贵州茅台酒和其他不同酒精度、不同香型的白酒。所建立的方法为贵州茅台酒的真伪鉴定提供了技术储备。     

阿胶、龟甲胶中脂溶性成分的高效液相色谱指纹图谱

采用高效液相色谱法研究并建立东阿阿胶、龟甲胶脂溶性成分的指纹图谱,为药用动物胶的质量控制提供了有效的方法。采用液-液-液三相静态萃取方法制备样品,以水-乙腈为流动相进行二元梯度洗脱,检测波长为205 nm,柱温25 ℃,分析时间为60 min。采集20批样品的色谱图并对其进行相似度和聚类分析。分别标定了阿胶、龟甲胶的共有峰,其相似度分析及聚类分析结果显示两种胶间存在着明显的差异。该方法稳定可靠,可以有效地区别不同种属的药用动物胶,为动物胶剂的鉴别及质量控制提供了依据。     

非线性化学指纹图谱技术用于菖蒲的鉴别及二元混合物中不同菖蒲含量的测定

采用非线性化学指纹图谱技术获得了石菖蒲、水菖蒲和九节菖蒲的特征指纹图谱,直观差异明显;将石菖蒲分别与水菖蒲和九节菖蒲进行二元掺杂,利用指纹图谱定量信息和各菖蒲含量拟合得到线性回归方程,相关系数0.9990.用该方程预测了二元混合物中不同菖蒲的含量,相对误差≤4.17%,RSD≤2.73%.非线性化学指纹图谱技术具有良好的重现性和特征性,同种菖蒲指纹图谱间系统相似度≥0.9989,不同种菖蒲指纹图谱间相似度≤0.9123.此外,该技术摒弃传统方法复杂的前处理过程,可高效、快速进行鉴别,从整体性角度对样本进行定性鉴别与定量分析.     

新疆紫草的毛细管电泳指纹图谱研究

建立了基于毛细管电泳技术的新疆紫草指纹图谱的质量控制方法.优化后的电泳条件:分离柱为50μm×40 cm未涂层毛细管柱,运行缓冲液为pH 8.0、100 mmol/L硼酸盐缓冲液(含25 mmol/L SDS及20%(V/V)无水乙醇),进样量0.5 psi×5 s,分离电压25 kV,检测波长214 nm.应用此条件,35 min内可实现新疆紫草有效成分的高效分离.在方法学验证的基础上,建立了新疆紫草指纹图谱.以新疆紫草对照药材指纹图谱为对照图谱,通过特征指纹峰、SFˊ相似度评价、聚类分析对不同购买地的新疆紫草进行质量评价和鉴别.此研究结果与其它中药鉴定方法对照结果一致.本方法准确、可靠、用时短,且具有良好的重现性,为新疆紫草的质量控制与评价提供了一种新的快速有效的鉴别方法.     

基于矿物元素指纹图谱鉴定陈皮产地的方法研究

采用电感耦合等离子体质谱法测定了新会、广西、湖南3地42批陈皮样品中51种矿物元素的含量,运用方差分析和偏最小二乘法-判别分析（PLS-DA）研究了不同产地陈皮中矿物元素的差别,以筛选得到的9种元素为溯源指标,将不同数量级矿物元素含量进行分类和系数缩放后,构建了陈皮中矿物元素的指纹图谱,并基于各产地矿物元素含量的平均值构建了新会、广西、湖南产地陈皮中矿物元素的标准指纹图谱。通过采用SPSS20.0计算18批新会陈皮矿物元素指纹图谱与其标准指纹图谱的相似度,确定新会陈皮矿物元素指纹图谱的相似度阈值为0.941,基于此阈值建立了新会陈皮的鉴别模型。采用24批样品对所建立的鉴别模型进行准确性验证,正确率为91.6%。该方法操作简单直观,满足实验分析要求,可为陈皮的产地溯源提供参考。     

铁皮石斛的裂解气相色谱指纹图谱及其系统聚类分析

采用裂解气相色谱/质谱法(Py-GC/MS)测定了10种不同产地的铁皮石斛并结合系统聚类分析法比较了这些铁皮石斛的指纹图谱,采用释放气体分析法考察了裂解温度对指纹图谱的影响。结果表明,0.4 mg样品在450 ℃下可瞬间裂解,10种样品的指纹图谱具有相似性,且重现性好;采用系统聚类分析能区别不同产地的样品。本法快速、简便、准确,不失为药材质量控制的良好方法。     

An approach to comparative analysis of chromatographic fingerprints for assuring the quality of botanical drugs

The present study was focused on developing the chemometric methods for analysis of the chromatographic fingerprint to control the quality of botanical drugs, which has gained attention in Asia and other countries. We developed a novel approach to generate a set of fingerprint features, called Fisher components (FCs) that were extracted from the chromatographic fingerprint. The method greatly reduces the dimensionality of the fingerprint vector, and the resulting FCs still retain most discriminatory information of the original fingerprint. Choosing an example of relevance to contemporary botanical drugs, we applied the FCs to a set of Shenmai injection samples. We successfully identified the manufacturers of the samples using two classifiers, linear discriminant analysis (LDA) and k-Nearest Neighbor (k-NN) based on the FCs. We also applied a similarity assessment together with the visual analysis using the FCs to exam the products from different manufacturers. We found that the lot-to-lot consistency of products can be accurately determined using the FCs. Finally, we demonstrated that the application of chemometric methods for chromatographic fingerprinting offers reliability to detect suspected fraud samples. In summary, we demonstrated that the presented approaches could be useful to determine the identity, consistency, and authenticity of Shenmai injection through chromatographic fingerprinting. The methods are equally applicable to other botanical drugs.     

Quality control method for commercially available wild Jujube leaf tea based on HPLC characteristic fingerprint analysis of flavonoid compounds

Flavonoids are the main active components of natural medicinal plants with many physiological functions. In this study, an HPLC fingerprinting method based on the distribution and relative amount of 11 bioactive flavonoids was established for the quality evaluation of commercially available wild Jujube leaf tea (JLT) from China. Separation of the crude flavonoid extract was achieved on a column filled with C₁₈ material with a high carbon content. The flavonoids in wild JLT were identified based on UV spectroscopy and accurate mass measurements by TOF‐MS. Twenty‐one batches of practical samples collected from different habitats were analyzed by using the developed HPLC method to construct the HPLC characteristic fingerprint of wild JLT. Then, combined with clustering and similarity analyses, the HPLC characteristic fingerprint was used for the authentication and quality evaluation of commercial wild JLT. Results indicated that the proposed HPLC characteristic fingerprint reflected the inherent characteristics of wild JLT collected from different regions. Authenticity identification and quality control of commercially available wild Jujube tea were achieved based on the HPLC characteristic fingerprint analysis. This new approach to bioactive component profiling provided a promising reference method for the quality evaluation of commercial wild flower and plant tea.     

Chemical fingerprint analysis for quality assessment and control of Bansha herbal tea using paper spray mass spectrometry

Chemical fingerprinting methodology is an approach for quality assessment and control of herbal medicines and related products based on the holistic chemical profile obtained by various analytical techniques. This study demonstrates the first application of paper spray mass spectrometry (PS-MS) as a chemical fingerprinting methodology for tracing the origins, establishing the authenticity, and assessing the overall quality of a famous herbal product, Bansha herbal tea (BHT). A negative ion PS-MS spectrum yielded the best chemical profiling information and was most appropriate for fingerprint analysis of BHT. In addition to the identification of active ingredients, various compounds present in BHT were simultaneously detected without any sample pretreatment and chromatographic separation, providing valuable information for the quality assessment and control of this herbal product. According to the principal component analysis of the PS-MS fingerprints, two sources of commercially available BHT products made by different manufacturers were easily differentiated. Qualified and expired products from the two manufacturers were also successfully distinguished, and the consistency of the quality between the manufacturers was assessed. Our experimental data demonstrated that the PS-MS chemical fingerprinting is a simple, rapid, and robust methodology for pharmaceutical analysis, with promising prospects for quality assessment and control of herbal medicines and related products with high-throughput.     

硅烷化GC-MS指纹图谱在卷烟品牌分析中的应用

采用硅烷化衍生化法结合气相色谱-质谱(GC-MS)法对卷烟烟丝中的主要化学成分进行检测,获得了21个卷烟样品的烟丝硅烷化GC-MS指纹图谱数据,并应用聚类分析和主成分分析法对烟丝硅烷化GC-MS指纹图谱数据进行综合评价。结果表明,该方法可用于不同品牌卷烟的比较和区分,硅烷化成分的含量分布特征能反映不同品牌卷烟的特性,可为卷烟品牌的风格表征、品质维护和真伪鉴别提供参考。     

Chemical Fingerprint Analysis for Quality Control of Herba Ephedrae Based on HPLC-DAD Combined with Chemometrics Methods

A method based on high performance liquid chromatography with photodiode array detector (HPLC-DAD) was developed for chemical fingerprinting analysis of Herba Ephedrae. The index of chromatographic fingerprint's information content was utilized to optimize the fingerprint detection conditions, which reduced the time of analysis and increased the veracity of analysis greatly. Then, the similarity analysis of fingerprints was used in quality consistency evaluation of Herba Ephedrae samples. Moreover, hierarchical clustering analysis (HCA) was applied to classify the samples according to their sources and varieties. In addition, the overlapped chromatographic peaks were resolved with the help of heuristic evolving latent projection (HELP) method in order to gain the better quantitative evaluation. The results indicated that the samples could be successfully grouped in accordance with their varieties and sources. Furthermore, five marker constituents were firstly screened out to be the main chemical markers, which importantly contribute to the classification of Herba Ephedrae samples. This investigation shows that the developed methodology can be generalized to the research of quality control of herbal medicines.     

Ecosystem discrimination and fingerprinting of Romanian propolis by hierarchical fuzzy clustering and image analysis of TLC patterns

The fingerprinting capacity of thin layer chromatography (TLC) and image analysis in the case of propolis samples collected in different area in Romania has been investigated. Fuzzy divisive hierarchical clustering approach was used as a powerful tool of samples discrimination and fingerprinting according to the geographical origin and local flora. The fuzzy partition and patterns obtained by membership degrees plot were in a very good agreement with floral origin and geographic location of Romanian propolis samples, and clearly illustrate the fuzziness concerning their similarities and difference. The results obtained strongly support that TLC via image analysis can be successfully employed in the fingerprinting methodologies if they are combined with appropriate fuzzy clustering method. The method developed in this paper might be also extended in the authenticity and origin control of fruits, herbs or derived products.     

Development of the Fingerprints for the Quality Evaluation of Scutellariae Radix by HPLC-DAD and LC-MS-MS

A high-performance liquid chromatography coupled with photodiode array detection and electrospray ionization tandem mass spectrometry (HPLC-DAD-ESI-MS²) method was firstly developed for a chemical fingerprint analysis of Scutellariae Radix and rapid identification of major compounds in the fingerprints. The experimental data for chromatography were used for hierarchical clustering analysis and similarity calculation, and those for UV and MS spectra were applied for identifying characteristic peaks. Twenty samples including S. baicalensis Georgi., S. viscidula Bge. and S. amoena C. H. Wright were classified into three groups. By comparing the UV and MS spectra data with those of the authentic standards and literature, 20 main peaks in the fingerprints were identified for the first time. The developed fingerprint assay was specific and could be readily utilized for comprehensive evaluation of Scutellariae Radix.     

Fingerprint Analysis of Euonymus alatus (Thuhb) siebold by LC–DAD and LC–ESI–MS

A liquid chromatography coupled with photodiode array detection and electrospray ionization tandem mass spectrometry method was first developed for a chemical fingerprint analysis of Euonymus alatus (Thuhb) siebold (EAS) and rapid identification of major compounds in the fingerprints. Fingerprint profiles were found to be consistent for the herbs acquired from different locations, but the relative abundance of peaks was varied. Twelve peaks were chosen as the common peaks. Quercetin and rutin were detected by comparing the retention times, MS and UV spectra with the standards. The relative retention time and relative peak area of the 12 peaks in the fingerprint were calculated by setting the quercetin as the reference peak. The experimental data were used for similarity calculation and hierarchical clustering analysis. By comparing the UV and MS spectra data with those of the authentic standards and literature, five main peaks in the fingerprints were identified. Finally, five medicinal portions of the herb (leaf, fruit, stem, pterygium and root) were also analyzed by this method. It was found that there were similar chemical components in different parts of this herb but the contents were very different. The developed fingerprint assay was specific and could be readily utilized for comprehensive evaluation of EAS, as well as to distinguish different medicinal portions.     

Anatomy of fingerprint search calculations on structurally diverse sets of active compounds

Similarity searching using molecular fingerprints is a widely used approach for the identification of novel hits. A fingerprint search involves many pairwise comparisons of bit string representations of known active molecules with those precomputed for database compounds. Bit string overlap, as evaluated by various similarity metrics, is used as a measure of molecular similarity. Results of a number of studies focusing on fingerprints suggest that it is difficult, if not impossible, to develop generally applicable search parameters and strategies, irrespective of the compound classes under investigation. Rather, more or less, each individual search problem requires an adjustment of calculation conditions. Thus, there is a need for diagnostic tools to analyze fingerprint-based similarity searching. We report an analysis of fingerprint search calculations on different sets of structurally diverse active compounds. Calculations on five biological activity classes were carried out with two fingerprints in two compound source databases, and the results were analyzed in histograms. Tanimoto coefficient (Tc) value ranges where active compounds were detected were compared to the distribution of Tc values in the database. The analysis revealed that compound class-specific effects strongly influenced the outcome of these fingerprint calculations. Among the five diverse compound sets studied, very different search results were obtained. The analysis described here can be applied to determine Tc intervals where scaffold hopping occurs. It can also be used to benchmark fingerprint calculations or estimate their probability of success.     

多元曲线分辨-交替最小二乘法应用于延胡索药材指纹图谱中重叠峰分辨及延胡索乙素的含量测定

采用光谱相关色谱法对延胡索药材指纹图谱中的有效组分延胡索乙素峰进行确认;利用多元曲线分辨 交替最小二乘法（MCR-ALS）对延胡索乙素重叠峰簇进行解析,得到延胡索乙素的真实色谱峰,其光谱还原率大于0.999,并对其进行准确定量,该方法可作为指纹图谱的辅助方法,为中药的真伪鉴定和质量客观评价提供可靠依据;最后讨论了药材的炮制对延胡索乙素含量的影响。