菌陈挥发油GC-MS指纹图谱分析

建立了菌陈挥发油GC-MS指纹图谱，将正交投影算法用于不同样本中共有峰的提取，提高了共有峰的识别能力，降低了由于保留时间的漂移、质谱检索匹配度不高以及色谱峰重叠而引起的共有峰识别的难度．采用共有峰率、变异率及相似度等几个指标对4个不同产地的菌陈挥发油成分进行了评价，从共性、差异和整体性等多个方面全面表征菌陈挥发油的化学模式特征．结果表明，不同产地的菌陈挥发油成分大致相同，该指纹图谱可用于菌陈中挥发油成分的质量控制．     

茵陈挥发油GC-MS指纹图谱分析

建立了茵陈挥发油GC-MS指纹图谱,将正交投影算法用于不同样本中共有峰的提取,提高了共有峰的识别能力,降低了由于保留时间的漂移、质谱检索匹配度不高以及色谱峰重叠而引起的共有峰识别的难度.采用共有峰率、变异率及相似度等几个指标对4个不同产地的茵陈挥发油成分进行了评价,从共性、差异和整体性等多个方面全面表征茵陈挥发油的化学模式特征.结果表明,不同产地的茵陈挥发油成分大致相同,该指纹图谱可用于茵陈中挥发油成分的质量控制.     

Comparative analysis of the volatile components in cut tobacco from different locations with gas chromatography-mass spectrometry (GC-MS) and combined chemometric methods

A combined approach of subwindow factor analysis and orthogonal projection resolution was used to analyze the volatile components of cut tobacco samples from different sources. After extracted with simultaneous distillation and extraction method, the volatile components in cut tobacco from five different locations were detected by GC-MS. Then, the qualitative and quantitative analysis of the volatile components of cut tobacco from Changde area was completed with the help of subwindow factor analysis resolving two-dimensional original data into pure mass spectra and chromatograms. One hundred and two volatile components among 138 separated peaks were identified and quantified, accounting for about 88.90% of the total content. Finally, orthogonal projection method was used to extract the common peaks from different locations. Among the identified components, there were 74 components coexisting in five studied samples although the relative content of each component showed difference to some extent. The results showed a fair consistency in their GC-MS fingerprints. It was the first time to apply orthogonal projection method to compare different cut tobacco samples, and it reduced the burden of qualitative analysis as well as the subjectivity. The obtained results proved the combined approach powerful for the analysis of complex cut tobacco samples. The developed method can be used to compare the sameness and differences of cut tobacco from different sources and for quality control of cigarette production and materials.     

Liquid exclusion adsorption chromatography, a new technique for isocratic separation of nonionic surfactants. III. Two-dimensional separation of fatty alcohol ethoxylates

In this paper, a novel procedure for qualitative and quantitative analysis of the two-dimensional data obtained from GC-MS is investigated to determine chemical components of essential oils in Cortex Cinnamomi from four different producing areas. A new method named iterative optimization procedure (IOP) specially used to resolve embedded peaks is also developed. With the help of IOP and other chemometric techniques, such as heuristic evolving latent projections, evolving factor analysis, sub-window factor analysis and orthogonal projection resolution, and etc., the detection of the purity of chromatographic peaks can be first addressed, and then the overlapping peaks are resolved into the pure chromatogram and mass spectrum of each component. The similarity searches in the MS database are finally conducted to qualitatively determine the chemical components. The results obtained showed that the accuracy of qualitative and quantitative analysis could be greatly enhanced by chemometric resolution methods. The chemometric resolution techniques upon the two-dimensional data can be quite promising tools for the analysis of the complex samples like traditional Chinese medicine.     

HPLC Fingerprint with Multi-components Analysis for Quality Consistency Evaluation of Traditional Chinese Medicine Si-Mo-Tang Oral Liquid Preparation

Si-Mo-Tang(SMT) oral liquid preparation, a traditional Chinese medicine, was prepared from four crude herbal drugs, Fructus Aurantii Submaturus, Radix Aucklandiae, Semen Arecae and Radix Linderae Aggregatae. A combinative method using HPLC fingerprint and quantitative analysis was developed and validated for quality consistency evaluation of SMT. Individual HPLC chromatograms were evaluated against the mean chromatogram generated via a similarity evaluation computer program. Data from chromatographic fingerprints were also processed with principal component analysis(PCA) and hierarchical cluster analysis(HCA). Additionally, six components (naringin, isonaringin, hesperidin, neohesperidin, norisoboldine and potassium sorbate) in SMT were simultaneously determined to interpret the quality consistency. For fingerprint analysis, 20 peaks were selected as the characteristic peaks to evaluate the similarities of 26 SMT collected from different manufacturers. Among the 20 characteristic peaks, 10 peaks were assigned to be naringin, hesperidin, neohesperidin, isonaringin, neoeriocitrin, tangeretin, nobiletin, norisoboldine, 5-(ethoxymethyl)furan-2-carbaldehyde and potassium sorbate, respectively. The results of similarity analysis, PCA and HCA, indicate that the samples from different manufacturers were consistent with each other in composition. The results from the quantitative data show that the contents of six compounds were significantly different in SMT oral liquid preparations from different manufacturers. The combinative method of chromatographic fingerprint with quantitative analysis developed here offered an efficient way for the quality consistency evaluation of the traditional Chinese medicine SMT.     

Determination of the volatile chemical constituents of Notoptergium incium by gas chromatography-mass spectrometry and iterative or non-iterative chemometrics resolution methods

The qualitative and quantitative determination of the chemical constitutes in traditional Chinese medicine (TCM) is an important task, which builds the foundation of the theory of pharmacological activity. The hyphenated chromatography instruments combined with the related chemometric methods provide powerful tools for the resolution of such complex systems. The familiar chemometrics methods can be roughly divided into two different kinds, the iterative one such as orthogonal projection approach (OPA) and non-iterative one representing by evolving window orthogonal projection (EWOP). One can use different kinds of methods according to overlapping condition, and then the measured data matrix can be resolved into pure concentration profiles and mass spectra of the chemical components with relative high efficiency and acceptable accuracy. One kind of TCM, named Notoptergium incium (NI) was analyzed by gas chromatography-mass spectrometry (GC-MS) and resolved by above chemometric approach. Experiment results show the efficiency and convenience of the proposed approach. 65 of the 98 separated constituents in essential oil, accounting for 92.13%, were identified by mass spectroscopy (MS).     

Quantitative analysis combined with chromatographic fingerprint and antioxidant activities for the comprehensive evaluation of Compound Danshen Tablets

The composition of traditional Chinese medicine is extremely complex, so it is difficult to ensure quality consistency. We took Compound Danshen Tablets as the object of the study, by using high‐performance liquid chromatography to establish multiwavelength fusion fingerprints. Characteristic fingerprints of 30 batches of samples were generated at four wavelengths and evaluated by systematic quantified fingerprint method. An on‐line antioxidant determination method was used for the determination of the antioxidant components in Compound Danshen Tablets. The fingerprint analysis of the marker compounds can reflect the content of the marker compounds, which were determined by using the external standard method. This study elucidated that multiwavelength fusion fingerprint profiles and multiple markers compound analysis in conjunction with the assay of antioxidant activity offered a reliable and efficient approach to quantitatively evaluate the quality consistency of the traditional Chinese medicine and herbal preparations.     

GC-MS与交互移动窗口因子分析法(AMWFA)用于3种陈皮挥发油成分的比较分析

将GC-MS法与新近提出的用于二维数据比较和解析的交互移动窗口因子分析法(AMWFA)结合， 对3种不同原植物来源的陈皮挥发油成分的共性和差异性进行分析, 并对重叠峰进行解析, 总共分辨出138个峰, 定性出78个峰, 其中共有组分44个.     

Separation and identification of volatile constituents in Artemisia argyi flowers by GC-MS with SPME and steam distillation

Artemisia argyi leaf is a widely used traditional Chinese medicine (TCM). In this work, for the first time, the separation and identification of volatile constituents in Artemisia argyi flowers is performed. Gas chromatography-mass spectrometry (GC-MS) with solid-phase microextraction (SPME) is developed for the fast analysis of volatile constituents in the flowers. Several headspace SPME parameters, including fiber coating, extraction temperature, and extraction time, are optimized. Forty-nine compounds in the flowers are re-identified by SPME-GC-MS. At the same time, in order to compare with the SPME, steam distillation is used for analysis of the volatile constituents in the flowers, and forty-seven are detected. The total fifty-three compounds in the flowers, which mainly include cylcofenchene, alpha-pinene, alpha-myrcene, D-limonene, caryophyllene, and germacrene D, are identified by the two methods. Compared to the volatile components in Artemisia argyi leaves, the main components (including the two active compounds of borneol and borneol acetate) are also found in Artemisia argyi flowers. These results show that Artemisia argyi flowers as well as leaves might be used as TCM.     

Analytical comparison of different parts of Radix Angelicae Sinensis by gas chromatography coupled with mass spectrometry

The three parts of Radix Angelicae Sinensis (Danggui) were reported to have different therapeutic effects. In order to investigate their chemical compositions of different parts of Danggui, gas chromatography-mass spectrometry (GC-MS) combined with chemometrics data analysis was applied to provide a more detailed study. Subwindow factor analysis (SFA) and a modified augmented evolving window orthogonal projection (AEWOP) method were used to resolve the batch GC-MS data sets from the Danggui samples. Then, t-test, Wilcoxon rank sum test, and principal component analysis (PCA) were applied to carry out the comparison job. The results indicate which components are found to have significant differences among the three parts. These findings may be helpful for further research of the pharmacological activities of Danggui.     

中药注射剂荧光光谱法的快速鉴别和热稳定性研究

红外光谱指纹图谱技术应用中药注射剂的快速鉴别中，某些试样因难于成膜造成制样困难，为此，直接或仅经水稀释后测定了12种21批次的常用中药注射剂的荧光光谱。结果表明：由于中药注射剂常含荧光性物质，但因不同注射剂所含具体荧光性物质不同，同种注射剂又因厂家的具体配方的差异或制备工艺条件的波动，均会使其特征的荧光激发、发射光谱不同或其强度呈现差异；从而可充分利用荧光分析的高灵敏度，使其作为中药整体红外指纹识别的一种辅助手段，根据注射剂的荧光图谱的差异达到快速鉴别、认定和控制配方、工艺的目的。比较而言，荧光法表现出来的差异更为一目了然，易于判断。此外，荧光光谱法还可用于中药注射液的热稳定性的研究。     

Assessment of quality consistency in traditional Chinese medicine using multi‐wavelength fusion profiling by integrated quantitative fingerprint method: Niuhuang Jiedu pill as an example

Under the wave of the revival of traditional Chinese medicine, there is a quite imperative duty to study an integrated and comprehensive method of fingerprint data processing and analysis on the quality consistency of traditional Chinese medicine. So, we proposed six parameters from two aspects (qualitative and quantitative), three levels (biased to strong peaks, biased to weak peaks, no obvious bias), to comprehensively evaluate the similarity of the two fingerprints. On this basis, another five parameters were proposed to evaluate the integrated effects (consistency, volatility, and similarity). This method was applied to 22 batches of Niuhuang Jiedu pill samples. Next, a practical and convenient multi‐wavelength fusion method was designed to provide more information, and the generated fusion profilings were used for subsequent evaluation. The characteristics of the parameters were confirmed by correlation analysis. The results of both hierarchical clustering analysis and principal component analysis for raw data and standardized data were consistent with integrated quantitative fingerprint method results. At the same time, this method gave a reasonable explanation for abnormal and dissimilar samples. This work illustrated that the proposed method was particularly suitable for similarity analysis of fingerprints and capable of ensuring the quality consistency in traditional Chinese medicine.     

不同产地莪术挥发油的有效成分

用HP6890/5973气相色谱-质谱联用仪分析了四川、福建、浙江产的莪术挥发油的有效成分，结果表明不同产地莪术挥发油的成分差异较大，提示在选择莪术油为原粒制备制剂时要规定产地。     

Capillary electrophoresis fingerprinting coupled with chemometrics to evaluate the quality consistency and predict the antioxidant activity of Sanhuang tablet as part of its quality control

A capillary electrophoresis fingerprint was constructed for Sanhuang tablet, a Chinese traditional patent medicine, that was commonly used in clinical practice, where the isosceles trapezoid method was first applied for the optimization of background electrolyte solution, and the resolution index was performed to assess the experimental conditions; furthermore, a novel linear quantitative fingerprint method was established for accurate qualitative and quantitative discrimination of the test samples from diverse commercial brands. The fingerprint analysis coupled with quantitative determination of two components was employed to elucidate that the quality consistency of the products was relatively good within one manufactory, but poor among different companies for the 30 batches of samples. In addition, the fingerprint–efficacy relationship between chemical components and antioxidant activity in vitro was investigated using partial least squares analysis, and the calibration and prediction of the antioxidant activity of the selected samples via fingerprint data were presented with the desired results. This work illustrates that the proposed fingerprint analysis based on linear quantitative fingerprint method can be applied for the quality evaluation of traditional Chinese medicine and herbal preparations as part of their quality control, and the constructed mathematical model is particularly suitable for depicting the fingerprint–efficacy relationship.     

Resolution of the essential constituents of Ramulus cinnamomi by an evolving chemometric approach

The analysis of complex mixtures, such as those of traditional Chinese medicine (TCM), is difficult by conventional methods of analysis. Chemometric methods provide a new way to solve such problems. Subwindow factor analysis (SFA) paired with the evolving window orthogonal projection (EWOP) method, has been used as a new evolving approach to the resolution of volatile components of Ramulus cinnamomi (RC). Compared with conventional chromatographic analysis, the chromatographic separation conditions necessary are greatly mitigated in our approach, yet the accuracy of qualitative and quantitative results is improved, because the measured data matrix has been resolved into chromatograms and mass spectra of the chemical components. Our method is, moreover, friendly to use and easy to program. Experimental results show the efficiency and convenience of the proposed approach. Forty-seven of the seventy-eight separated constituents in an essential oil, accounting for 89.55% of the sample, were identified by mass spectroscopy (MS).     

Quantification and semiquantification of multiple representative components for the holistic quality control of Allii Macrostemonis Bulbus by ultra high performance liquid chromatography with quadrupole time‐of‐flight tandem mass spectrometry

Allii Macrostemonis Bulbus (named Xiebai in China) is a folk medicine with medicinal values for the treatment of thoracic obstruction and cardialgia, and a food additive as well. However, there is even no quantitative standard for Allii Macrostemonis Bulbus recorded in the current edition of the Chinese Pharmacopeia. Hence, simultaneous assay of multiple components is urgent. In this study, chemometric methods were firstly applied to discover the components with significant fluctuation among multiple Allii Macrostemonis Bulbus samples based on optimized fingerprints. Meanwhile, the major components and main absorbed components in rats were all selected as its representative components. Subsequently, a sensitive method was established for the simultaneous determination of 54 components (15 components for quantification and 39 components for semiquantification) by ultra high performance liquid chromatography coupled with quadrupole time‐of‐flight tandem mass spectrometry. Moreover, the validated method was successfully applied to evaluate the quality of multiple samples on the market. It became known that multiple Allii Macrostemonis Bulbus samples varied significantly and showed poor consistency. This work illustrated that the proposed approach could improve the quality of Allii Macrostemonis Bulbus, and it also provided a feasible method for quality evaluation of other traditional Chinese medicines.     

综合线性定量指纹法评价退热解毒注射液融合指纹图谱研究

该研究引入综合线性定性相似度Sl、综合线性定量相似度Pl%和指纹变异系数α3个参数建立了1种新颖的综合线性指纹图谱评价方法,从定性定量的角度全面评价了中药的整体质量。方法采用高效液相二极管阵列(HPLC/DAD)采集了退热解毒注射液样品在210、254、265、330、360 nm波长下的指纹图谱并进行数据融合,同时定量分析了绿原酸、连翘酯苷A、丹皮酚和柴胡皂苷D 4个指标成分。通过综合线性指纹法鉴定样品质量。结果显示,10批注射液样品被分成5个等级,10批中间体样品被分成4个等级。此外,4个指标成分的定量分析结果与融合指纹图谱的定量相似度Pl%具有高度相关性,表明综合线性定量指纹法具有代替标准物质用于样品定量分析的潜力。该方法为中药整体质量控制提供了一种新思路。     

Offline Two-Dimensional RP/RPLC Method to Separate Components in Dracaena cochinchinensis (Lour.) S.C.Chen Xylem Containing Resin

An offline two-dimensional RP/RPLC system was developed for the separation of components in Dracaena cochinchinensis (Lour.) S.C.Chen xylem containing a resin that is an important raw material in Chinese traditional medicine. In this separation method, a CN-column was used in the first dimension and a C₁₈-column in the second dimension. Fractions collected from the CN-column were resolved into many additional components, which indicated that the two-dimensional RP/RPLC system based on the CN- and C₁₈-columns is orthogonal. The method provides higher peak capacity and better resolution. It is suitable for the analysis of complex samples such as those found in traditional Chinese medicines.     

Metabolomic comparison between natural Huaier and artificial cultured Huaier

Vanderbylia robiniophila (Murrill) B.K. (Huaier) is a kind of higher fungal fruiting body that is parasitic on the trunk of Sophora japonica and Robinia pseudoacacia L.. As a traditional Chinese medicine with a history of more than 1,600 years, Huaier has attracted wide attention for its excellent anticancer activity. A systematic study on the metabolome differences between natural Huaier and artificial cultured Huaier was conducted using liquid chromatography–mass spectrometry in this study. Principal component analysis and orthogonal projection on latent structure-discriminant analysis results showed that cultured Huaier evidently separated and individually separated from natural Huaier, indicating metabolome differences between natural and cultured Huaier. Hierarchical clustering analysis was further performed to cluster the differential metabolites and samples based on their metabolic similarity. The higher contents of amino acids, alkaloids and terpenoids in natural Huaier make it an excellent choice as a traditional Chinese medicine for anticancer or nutritional supplementation. The results of the Bel-7,402 and A549 cell cytotoxicity tests showed that the anticancer activity of natural Huaier was better than that of cultured Huaier. This may be due to the difference in chemical composition, which makes the anticancer activities of natural and cultured Huaier different.     

Identification and quantification of 13 components in Angelica sinensis (Danggui) by gas chromatography-mass spectrometry coupled with pressurized liquid extraction

Angelica sinensis (Danggui in Chinese), a well-known traditional Chinese medicine, is also used as a health food product for women's care in Europe and America. Therefore, the demand for Danggui is enormous throughout the world. Due to the shortage of Angelica sinensis, Angelica acutiloba and Angelica gigas are commonly used as the substitutes of Danggui in the market of southeast Asia. However, the three common Angelica roots showed variation in their genetic and chemical composition. Up to date, it is thought that ferulic acid, ligustilide and other phthalides such as butylidenephthalide are the biologically active components of Danggui. In this paper, the contents of 13 compounds including ferulic acid, Z-ligustilide, E-ligustilide, Z-butylidenephthalide, E-butylidenephthalide, 3-butylphthalide, 3-butylidene-4-hydroxyphthalide, senkyunolide A, 6,7-epoxyligustilide, senkyunolide F, senkyunolide H, senkyunolide I, and 6,7-dihydroxyligustilide were determined or estimated by using gas chromatography-mass spectrometry (GC-MS) coupled with pressurized liquid extraction (PLE). The results showed that GC-MS coupled with PLE offered a simple, rapid and high sensitive method to analysis of components in Angelica root. And the contents of investigated compounds in Angelica sinensis, Angelica acutiloba and Angelica gigas, which are used as Danggui in China, Japan and Korea, respectively, were highly variant. It is thought that interaction of multiple chemical compounds contributes to the therapeutic effects of Chinese medicines. However, the overall clinical efficacy of these different Danggui has not been determined. Therefore, comparison of chemical components and pharmacological activities of different Angelica root is helpful to elucidate the mechanism of therapeutic effects of Danggui.