利用样本成分耗散物的非线性化学指纹图谱的检测方法、条件、特点和应用

分析了适用于测定样本非线性化学指纹图谱的热力学体系和动力学模型,结果表明,在远离平衡的封闭体系和无耗散物补充的敞开体系中,阻尼非线性化学反应对样本化学成分的群集表征和整体含量分析具有重要作用．以丙酮和葡萄糖等为耗散物的阻尼B—Z振荡非线性化学反应为例,研究了反应物种及其浓度、电极类型、温度、搅拌速率、样本种类及用量等影响非线性化学指纹图谱的因素,并对指纹图谱中样本化学成分的定量信息、指纹图谱的特点和应用等进行了研究．成功提出了用于鉴别和评价样本的非线性化学指纹图谱的检测条件和方法．     

Principle of nonlinear chemical fingerprint by using dissipative components in samples as well as calculation and evaluation of similarity

Under the conditions of constant temperature and pressure,different influences of samples with different chemical components on the mechanism of nonlinear chemical reaction will cause different changes of the potential-time relationship curve of the nonlinear chemical reaction system.Using it as the character,and using the B-Z nonlinear chemical system to use acetone and substrates in samples as main dissipative substances qua an example,the principle of nonlinear chemical fingerprint has been researched and discussed in detail.At the same time,the general method for calculating the system similarity about nonlinear chemical fingerprint was also put forward,and similarities of nonlinear chemistry fingerprints of different batches of Guhan Yangshengjing and 18 sorts of other samples were calculated by Euclidean distance,correlation coefficient,included angle cosine and system similarity,at the same time,the various similarities were analyzed.The results showed that,both of correlation coefficient and included angle cosine are unable to be used as the criterion for quantitatively evaluating the similarity of nonlinear chemistry fingerprint;as non-parametric similarity,Euclidean distance can accurately reflect the feature differences in the fingerprints,but as parametric similarity,sometimes,Euclidean distance can not accurately reflect the relative extent of characteristic difference in the nonlinear chemical fingerprints;system similarity can most truthfully reflect the characteristic difference in the nonlinear chemical fingerprints,and is the best evaluating method among the four ones.Therefore,system similarity can be used to quantitatively calculate the similar extent between the nonlinear chemical fingerprints.An economical,simple and convenient,easy pushing and effective method for identifying and evaluating complicated samples has successfully been put forward.     

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.     

A multi-evaluating strategy for raw and processed Veratrum nigrum L.: Fingerprinting combined with quantitative analysis based on multivariate chemometric methods

Veratrum nigrum L. (VN) is a well-known herbal medicine and rich in chemical components with multiple pharmacological activities including antihypertensive, anticancer, and antifungal effects. In the current experiment, the quality of VN from different habitats was evaluated based on combinative method of fingerprint, multi-component quantification and chemical pattern recognition. Fifteen batches of VN were collected, and intrinsic chemical composition were identified using ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry, which is a method for analyzing the similarity between samples, coupled with fingerprint of traditional Chinese medicine. The fingerprint similarity model show that 22 common peaks were selected covering 15 batches of and the similarity > 0.963. The total of 22 joint components were tentatively identified by comparison with standard substances or literature. A ultra-high performance liquid chromatography coupled with triple quadrupole mass spectrometry method for simultaneous determination of 8 compounds was established to evaluate the contents of raw and processed Veratrum nigrum L. Multivariate analysis was then applied to compare different batches of herbs based on ultra-high performance liquid chromatography coupled with triple quadrupole mass spectrometry data. All raw and processed samples were classified by partial least squares discriminant analysis based on the 8 analyzed compounds. The findings suggested that veratramine and polydatin with a variable importance for the project (VIP) > 1 were identified as significant constituents, the presence of which can be used to differentiate between raw and processed Veratrum nigrum L. samples. These results indicate that processing methods show important effects on the composition of Veratrum nigrum L..     

Discrimination of Alismatis Rhizoma based on chromatographic fingerprint,multiple components quantification,and pattern recognition analysis

A simple, sensible, and reliable HPLC–DAD method was first developed for fingerprint analysis of Alismatis Rhizoma, and then applied to analyze 85 samples from three main cultivated areas. In all, 40 common fingerprint peaks were designated, and six of which were definitely identified. Then, the combinatory analysis using similarity evaluation, principal component analysis, and orthogonal partial least square discriminant analysis revealed clear chemical consistency between samples from Fujian and Jiangxi provinces and substantial differences between those from Fujian/Jiangxi and Sichuan provinces. Furthermore, six components were dug out as potential chemical markers for distinguishing Alismatis Rhizoma from different areas, among which five were qualified for quantitative analysis. In conclusion, the combination of chemical fingerprint, multiple components quantification, and pattern recognition analysis was rather powerful and useful in discriminating Alismatis Rhizoma from different regions, which was a benefit for quality control.     

Integrated evaluation of HPLC and UV fingerprints for the quality control of Danshen tablet by systematic quantified fingerprint method combined with antioxidant activity

Danshen tablet, which consists of Salviae Miltiorrhizae Radix et Rhizoma, Notoginseng Radix et Rhizoma and Borneolum syntheticum , has been widely used in the therapy of cardiovascular disease. The aim of this study was to develop comprehensive evaluation methods for the quality control of Danshen tablet. First, five‐wavelength fusion fingerprint was established to avoid one‐sidedness of a single wavelength. Then, the ultraviolet spectrum fingerprint was applied to reflect the information of unsaturated bond and conjugated system of chemical substances in Danshen tablet. The similarity analyses of these two fingerprints were performed by systematic quantified fingerprint method in terms of qualitative and quantitative aspects. After that, the evaluation results of high‐performance liquid chromatography and ultraviolet fingerprints were integrated by the mean algorithm, which could reduce the error caused by single method. The integrated evaluation results showed that 30 batches of samples were classified into seven grades. Finally, the fingerprint–efficacy relationship was established using an on‐line antioxidant system and partial least‐squares model to explore the connection between chemical components and antioxidant activities. The methods established in this paper were found suitable for the analysis of Danshen tablet.     

Different fingerprinting strategies to differentiate Porana sinensis and plants of Erycibe by high‐performance liquid chromatography with diode array detection,ultra high performance liquid chromatography with tandem quadrupole mass spectrometry,and chemometrics

Plants of Erycibe are widely used in traditional Chinese medicine for the treatment of joint pain and rheumatoid arthritis. With the reduction of Erycibe resources in the wild, Porana sinensis has been widely used as a substitute. However, it is important to understand the chemical distinctions between the two kinds of plants and identify their individual chemical markers. In this study, multiwavelength chromatographic fingerprint and precursor ion fingerprint techniques were used in conjunction with chemometric tools to fingerprint and thus differentiate between plant samples. The similar results obtained from different fingerprints prove the reliability of the two fingerprints. Results obtained from principal component analysis and orthogonal projection to latent structures discriminant analysis identified similarities between the chemical components of P. sinensis and plants of Erycibe. However, concentrations of 4‐caffeoylquinic acid, 3,5‐dicaffeoylquinic acid, 3,4‐dicaffeoylquinic acid, and 4,5‐dicaffeoylquinic acid were higher in P. sinensis than in plants of Erycibe, suggesting that P. sinensis may be more effective in medical treatments of some diseases than Erycibe.     

气相色谱-质谱指纹图谱法研究普洱茶醇溶物挥发性成分

以普洱茶7542为参考样本, 乙醇为溶剂, 超声提取制备普洱茶醇溶物, 建立了普洱茶7542醇溶物气相色谱-质谱(GC-MS)指纹图谱, 同时对其指纹图谱进行相似度计算和主成分分析.试验结果表明, 普洱茶醇溶物的最佳提取方法:采用50 mL 95%乙醇超声提取30 min, 顶空进样最佳条件为振荡箱温度110℃, 振荡时间20 min.通过对9个不同年限、不同批次的普洱茶7542系列样品醇溶物挥发性成分GC-MS指纹图谱进行相似度分析, 测定样品醇溶物挥发性成分图谱与对照图谱之间的相似度在0.706~0.906之间, 说明不同年限、不同批次的9个普洱茶7542醇溶物挥发性成分和参照样本相比发生了改变, 但变化较小, 但其共有成分峰面积百分含量存在差异.通过对9个不同年限、不同批次普洱茶7542醇溶物挥发性成分共有成分进行主成分分析, 提取了2个成分, 即2-羟甲基-2-甲基-吡咯烷-1-甲醛和N-丁基苯磺酰胺, 其特征值大于1, 累计方差贡献率达90.2%, 能较好的代表 9个不同年限、不同批次普洱茶7542的全部信息, 可以用主成分分析来反映样品的全部信息.因此可通过测定9个不同年限、不同批次普洱茶7542醇溶物中2-羟甲基-2-甲基-吡咯烷-1-甲醛和N-丁基苯磺酰胺的含量差异评价普洱茶7542的质量.     

Chemical test methods for determining components of liquids

The reported data on determining substances by chemical test methods are summarized. The reagents used; methods of their immobilization on supports; and principles, devices, and methods for determining components are considered. The main analytical characteristics are discussed.     

Study on Absorption of Radix Astragali in Rabbit Plasma by Alternative Moving Window Factor Analysis and LC–DAD–MS

In the present paper, study on the multiple absorbed bioactive components of Radix Astragali in rabbit was performed by alternative moving window factor analysis (AMWFA) and LC–DAD–MS. AMWFA was used to confirm the common components both in its abstract fingerprint and plasma fingerprint. This method could be utilized not only to determine the number of common components in different systems but also to identify their corresponding spectra. As can be seen from the results, the method determines the common components in different chromatographic fingerprints more efficient and reliable, especially for none or only weak selective information of some chromatographic peak clusters. With the aid of AMWFA both its abstract fingerprint and plasma fingerprint, one could find that 25 chemical components in RA were absorbed into the rabbit body after oral solution of RA administration.     

Comparison of the volatile constituents of Artemisia capillaris from different locations by gas chromatography-mass spectrometry and projection method

The volatile chemical constituents of Artemisia capillaries (an important traditional Chinese medicine) were determined by gas chromatography-mass spectrometry (GC-MS) and sub-window factor analysis (SFA). Seventy-five components were separated and 43 of them were qualitatively and quantitatively determined, which represented about 89.03% of the total content. This profile was then used to identify and assess the consistency of the herb by using an orthogonal projection method. Four different sources of A. capillaries were analyzed and compared with each other. Among the components determined, there were 51 components coexisting in all samples although the relative peak areas of a few showed variations. It is the first time to apply orthogonal projection method to the comparison of different samples, and it reduces the burden of qualitative analysis as well as the subjectivity. The results showed a fair consistency in their GC-MS fingerprint. A. capillaris was distinguished from Artemisia sacrorum L., a possible substitute in traditional Chinese medicine by comparing the fingerprints with each other.     

Quality assessment of cortex cinnamomi by HPLC chemical fingerprint, principle component analysis and cluster analysis

HPLC fingerprint analysis, principle component analysis (PCA), and cluster analysis were introduced for quality assessment of Cortex cinnamomi (CC). The fingerprint of CC was developed and validated by analyzing 30 samples of CC from different species and geographic locations. Seventeen chromatographic peaks were selected as characteristic peaks and their relative peak areas (RPA) were calculated for quantitative expression of the HPLC fingerprints. The correlation coefficients of similarity in chromatograms were higher than 0.95 for the same species while much lower than 0.6 for different species. Besides, two principal components (PCs) have been extracted by PCA. PC1 separated Cinnamomum cassia from other species, capturing 56.75% of variance while PC2 contributed for their further separation, capturing 19.08% variance. The scores of the samples showed that the samples could be clustered reasonably into different groups corresponding to different species and different regions. The scores and loading plots together revealed different chemical properties of each group clearly. The cluster analysis confirmed the results of PCA analysis. Therefore, HPLC fingerprint in combination with chemometric techniques provide a very flexible and reliable method for quality assessment of traditional Chinese medicines.     

Quality Evaluation of Atractylodis Macrocephalae Rhizoma Based on Combinative Method of HPLC Fingerprint,Quantitative Analysis of Multi-Components and Chemical Pattern Recognition Analysis

A method for the quality evaluation of Atractylodis Macrocephalae Rhizoma (AMR) based on high-performance liquid chromatography (HPLC) fingerprint, HPLC quantification, and chemical pattern recognition analysis was developed and validated. The fingerprint similarity of the 27 batches of AMR samples was 0.887–0.999, which indicates there was very limited variance between the batches. The 27 batches of samples were divided into two categories according to cluster analysis (CA) and principal component analysis (PCA). A total of six differential components of AMR were identified in the partial least-squares discriminant analysis (PLS-DA), among which atractylenolide I, II, III, and atractylone counted 0.003–0.045%, 0.006–0.023%, 0.001–0.058%, and 0.307–1.175%, respectively. The results indicate that the quality evaluation method could be used for quality control and authentication of AMR.     

Establishment of GC-MS fingerprint of fresh Houttuynia cordata

Fresh Houttuynia cordata THUNB. is a Chinese materia medica generally used in Chinese medicine therapy. It possesses the actions of clearing heat, eliminating toxins, reducing swelling, discharging pus and relieving stagnation. However, dry H. cordata has traditionally been used in clinical application instead of the fresh counterpart. In this paper, the chemical profiles of H. cordata were established using fingerprinting techniques. A modified GC-MS method was developed in the comparison of fingerprints among fresh and dry herbs of H. cordata. It was shown that the varieties, as well as relative levels of chemical components, in the fresh herb were more abundant than in the dry counterpart. Fingerprinting profiles were found to be consistent for fresh herbs acquired from various production areas, but the relative abundance of peaks were varied. Besides, the chemical components among different medicinal portions of fresh herbs were found to be inconsistent. The developed fingerprint can be successfully applied to distinguish between fresh and dry herbs, as well as determining differentiation among different medicinal portions.     

Toxicological classification of urine samples using pattern recognition techniques and capillary electrophoresis

In toxicology, hazardous substances detected in organisms may often lead to different pathological conditions depending on the type of exposure and level of dosage; hence, further analysis on this can suggest the best cure. Urine profiling may serve the purpose because samples typically contain hundreds of compounds representing an effective metabolic fingerprint. This paper proposes a pattern recognition procedure for determining the type of cadmium dosage, acute or chronic, administrated to laboratory rats, where urinary profiles are detected using capillary electrophoresis. The procedure is based on the composition of a sample data matrix consisting of areas of common peaks, with appropriate pre-processing aimed at reducing the lack of reproducibility and enhancing the potential contribution of low-level metabolites in discrimination. The matrix is then used for pattern recognition including principal components analysis, cluster analysis, discriminant analysis and support vector machines. Attention is particularly focussed on the last of these techniques, because of its novelty and some attractive features such as its suitability to work with datasets that are small and/or have low samples/variable ratios. The type of cadmium administration is detected as a relevant feature that contributes to the structure of the sample matrix, and samples are classified according to the class membership, with discriminant analysis and support vector machines performing complementarily on a training and on a test set.     

Establishment of HPLC-DAD-MS fingerprint of fresh Houttuynia cordata

A HPLC-DAD-MS fingerprint method of fresh Houttuynia cordata THUNB. was developed basing on the consistent chromatographic features among 11 batches of authentic samples. Major chemical components including phenolic compounds, flavones and alkaloids were simultaneously analyzed. Eleven common peaks in the fingerprint were chosen and identified by comparing their UV and ESI-MS data with the authentic compounds. The unique properties of this HPLC-DAD-MS fingerprint were successfully applied to analyze and differentiate samples from different geographical origins, processing methods and various medicinal parts of H. cordata. The results showed that these variations will give rise to differences in identities and/or abundance of chemical compounds, indicating that a comprehensive quality evaluation of those major ingredients in H. cordata is critical to assess and represent its overall quality.     

化学指纹图谱的相似性测度及其评价方法

提出化学指纹图谱相似性测度概念,并以峰数弹性、峰比例同态性和峰面积同 态性为评价指标,用于多角度评价化学指纹图谱相似性测试优劣,根据这些评价指 标,用计算机仿真方法研究比较了6种相似性测度,结果表明夹角余弦测试用于度 量指纹图谱间谱峰比例的波动较适宜,峰匹配测度可较灵敏地检测小峰个数波动, 而欧氏距离测试则具有较好的综合评价能力。最后,采用实测的参麦注射液色谱分 析谱图。研究考察了上述计算机仿真实验结果,证明仿真结果符合实际情况,这表 明本研究方法可用于评价化学指纹图谱相似性测度。     

Statistical mixture design optimization of extraction media and mobile phase compositions for the characterization of green tea

The influence of different solvents on the extraction medium and the RP-HPLC mobile phase composition were investigated by statistical mixture designs to optimize solvent proportions to prepare the fingerprint of a medicinal herbal extract. For modeling, the number of peaks was used as a measure of fingerprint information. The optimum compositions of solvent to extract chemical substances from green tea and for mobile phase chromatographic analysis were ethyl acetate/ ethanol/dichloromethane (20:5:75 v/v/v) and MeOH/ACN/water (7.5:57.5:35 v/v/v), respectively. This system results in 26 peaks in the chromatographic fingerprint. These results show that an incorrect choice of modifiers for mobile phase composition and solvent extraction hampers the detection of a maximum number of peaks and produces a poor chromatographic fingerprint.     

Fingerprint developing of coffee flavor by gas chromatography-mass spectrometry and combined chemometrics methods

In this paper, chromatographic fingerprint was firstly used for quality control of tobacco flavors. Based on gas chromatography-mass spectrometry (GC-MS) and combined chemometrics methods, a simple, reliable and reproducible method for developing chromatographic fingerprint of coffee flavor, one of tobacco flavors, was described. Six coffee flavor samples obtained from different locations were used to establish the fingerprint. The qualitative and quantitative analysis of coffee flavor sample from Shenzhen was completed with the help of subwindow factor analysis (SFA). Fifty-two components of 68 separated constituents in coffee flavor sample from Shenzhen, accounting for 88.42% of the total content, were identified and quantified. Then, spectral correlative chromatography (SCC) was used to extract the common peaks from other five studied coffee flavor samples. Thirty-eight components were found to exist in all six samples. Finally, the method validation of fingerprint analysis was performed based on the relative retention time and the relative peak area of common peaks, sample stability and similarity analysis. The similarities of six coffee flavor samples were more than 0.9104 and showed that samples from different locations were consistent to some extent. The developed chromatographic fingerprint was successfully used to differentiate coffee flavor from coco flavor and some little difference sample prepared with coffee flavor and coco flavor by both similarity comparison and principal component projection analysis. The developed method can be used for quality control of coffee flavor.