木香挥发性化学成分的气相色谱-质谱分析

采用水蒸气蒸馏法从木香中提取挥发油,用气相色谱-质谱联用仪对木香挥发油进行化学成分的分析。利用化学计量学方法对重叠峰进行分辨,对各个色谱峰定性,并用色谱峰总体积积分和归一化法获得各化合物的相对含量。共鉴定了60种化合物,占挥发油总成分的97%以上。其中主要有土木香内酯、巴西菊内酯、(Z)6,(Z)9-十五二烯-1-醇等。     

Quality evaluation of raw and processed Crataegi Fructus by color measurement and fingerprint analysis

Crataegi Fructus and its processed products have been used as a traditional medicine for a long time, and numerous active components are responsible for their curative effects. However, a comprehensive and fast method for the quality control of its processed products is still lacking. In this study, two analytical methods based on color measurements and fingerprint analysis are established. In the color measurements, the color values of the peel and flesh of Crataegi Fructus were evaluated spectrophotometrically. Based on the results, a color reference range was established using percentiles, and the standard color difference value was established using the median color values. Then, the color values of Crataegi Fructus and its processed products were analyzed using Bayes linear discriminant analysis and mathematical functions were built in order to predict the degree of processing. Moreover, high‐performance liquid chromatography fingerprint analysis was performed on a Hibar C₁₈ column, and a high‐performance liquid chromatography fingerprint pattern was obtained, from which nine peaks were identified. Chemometric methods were successfully applied to differentiate raw and processed Crataegi Fructus.     

Quality assessment of raw and processed Arctium lappa L. through multicomponent quantification,chromatographic fingerprint,and related chemometric analysis

In traditional Chinese medicine, raw and processed herbs are used to treat different diseases. Suitable quality assessment methods are crucial for the discrimination between raw and processed herbs. The dried fruit of Arctium lappa L. and their processed products are widely used in traditional Chinese medicine, yet their therapeutic effects are different. In this study, a novel strategy using high‐performance liquid chromatography and diode array detection coupled with multivariate statistical analysis to rapidly explore raw and processed Arctium lappa L. was proposed and validated. Four main components in a total of 30 batches of raw and processed Fructus Arctii samples were analyzed, and ten characteristic peaks were identified in the fingerprint common pattern. Furthermore, similarity evaluation, principal component analysis, and hierachical cluster analysis were applied to demonstrate the distinction. The results suggested that the relative amounts of the chemical components of raw and processed Fructus Arctii samples are different. This new method has been successfully applied to detect the raw and processed Fructus Arctii in marketed herbal medicinal products.     

Establishing the chromatographic fingerprint of traditional Chinese medicine standard decoction based on quality by design approach: A case study of Licorice

A novel analytical quality by design approach for developing a chromatographic fingerprint was established for analyzing complex traditional Chinese medicine, using a licorice standard decoction as an example. Considering the characteristics of integrity and ambiguity, the resolution of eight common peaks, total peak number, capacity factor distributions, and peak purity were selected as potential critical method attributes for assessing the quality of the chromatographic fingerprint. A central composite design was used to evaluate the relationship between critical method attributes and critical method parameters, including column temperature, wavelength, flow rate, formic‐acid concentration, and gradient parameters. A standard probability method was employed to calculate the design space of the fingerprint analysis parameters and evaluate the robustness of the methodology. The optimized high‐performance liquid chromatography fingerprint conditions were acetonitrile and 0.1% formic acid water gradient elution (0‐5 min, 5–19% A; 5–10 min, 19% A; 10–50 min, 19–42% A; 50–54 min, 42–100% A; 54–60 min, 100% A), column temperature 25±5°C, detection wavelength 265 nm. The design space of fingerprint analytical method based on the analytical quality by design approach not only met the requirements of the fingerprint analysis, but also improved the robustness and applicability of the fingerprint method.     

Monitoring quality consistency of Liuwei Dihuang Pill by integrating the ultraviolet spectroscopic fingerprint,a multi‐wavelength fusion fingerprint method,and antioxidant activities

Liuwei Dihuang Pill, a classical traditional Chinese medicine, has been widely used to treat kidney yin deficiency in China for hundreds of years. Little attention, however, has been paid to quality control methods for this formulation. In the present study, we aimed to establish a comprehensive and practical quality evaluation system for Liuwei Dihuang Pill. Thus, ultraviolet spectroscopic fingerprints and multi‐wavelength fusion fingerprints have been proposed for quality consistency evaluation of the popular patent. The similarity analysis of Liuwei Dihuang Pill samples was evaluated by systematic quantitative fingerprint method from qualitative and quantitative perspectives. The results showed that 26 batches of samples were classified into five grades for fusion fingerprints and three grades for ultraviolet spectroscopic fingerprints. Moreover, the fingerprint–efficacy correlation was also established and confirmed through the partial least squares model to visualize the antioxidant activity of Liuwei Dihuang Pill in vitro. In conclusion, these results indicated that integrating ultraviolet spectroscopic fingerprints and multi‐wavelength fusion fingerprints coupled with antioxidant activities provide a rapid and effective approach to monitor the quality consistency of Liuwei Dihuang Pill.     

Quality assessment of Herba Leonuri based on the analysis of multiple components using normal‐ and reversed‐phase chromatographic methods

A novel, improved, and comprehensive method for quality evaluation and discrimination of Herba Leonuri has been developed and validated based on normal‐ and reversed‐phase chromatographic methods. To identify Herba Leonuri , normal‐ and reversed‐phase high‐performance thin‐layer chromatography fingerprints were obtained by comparing the colors and R _f values of the bands, and reversed‐phase high‐performance liquid chromatography fingerprints were obtained by using an Agilent Poroshell 120 SB‐C18 within 28 min. By similarity analysis and hierarchical clustering analysis, we show that there are similar chromatographic patterns in Herba Leonuri samples, but significant differences in counterfeits and variants. To quantify the bio‐active components of Herba Leonuri , reversed‐phase high‐performance liquid chromatography was performed to analyze syringate, leonurine, quercetin‐3‐O‐robiniaglycoside, hyperoside, rutin, isoquercitrin, wogonin, and genkwanin simultaneously by single standard to determine multi‐components method with rutin as internal standard. Meanwhile, normal‐phase high‐performance liquid chromatography was performed by using an Agilent ZORBAX HILIC Plus within 6 min to determine trigonelline and stachydrine using trigonelline as internal standard. Innovatively, among these compounds, bio‐active components of quercetin‐3‐O‐robiniaglycoside and trigonelline were first determined in Herba Leonuri . In general, the method integrating multi‐chromatographic analyses offered an efficient way for the standardization and identification of Herba Leonuri .     

High‐performance liquid chromatography–based fingerprint analysis with chemical pattern recognition for evaluation of Mahonia bealei (Fort.) Carr

A simple and efficient high‐performance liquid chromatography method combined with chemical pattern recognition was established for quality evaluation of Mahonia bealei (Fort.) Carr. A common pattern of 30 characteristic peaks was applied for similarity analysis, hierarchical cluster analysis, principal component analysis, and partial least squares discriminant analysis in the 37 batches of M. bealei (Fort.) Carr. to discriminate wild M. bealei (Fort.) Carr., cultivated M. bealei (Fort.) Carr., and its substitutes. The results showed that partial least squares discriminant analysis was the most effective method for discrimination. Eight characteristics peaks with higher variable importance in projection values were selected for pattern recognition model. A permutation test and 26 batches of testing set samples were performed to validate the model that was successfully established. All of the training and testing set samples were correctly classified into three clusters (wild M. bealei (Fort.) Carr., cultivated M. bealei (Fort.) Carr., and its substitutes) based on the selected chemical markers. Moreover, 26 batches of unknown samples were used to predict the accuracy of the established model with a discrimination accuracy of 100%. The obtained results indicated that the method showed great potential application for accurate evaluation and prediction of the quality of M. bealei (Fort.) Carr.     

Total detection of Tianma Toutong tablets for quality consistency by a five‐wavelength fusion fingerprint and chemometrics

A fingerprint method was developed and combined with chemometrics for quality evaluation of Tianma Toutong tablets, which are herbal medicine tablets used to treat migraine. Samples were analyzed by high‐performance liquid chromatography, where five single‐wavelength profiles (203, 232, 254, 280 and 310 nm) were fused to generate a five‐wavelength fusion fingerprint and were also used for the quantitative analysis of seven chemical markers (gastrodin, caffeic acid, hesperidin, isoimperatorin, chlorogenic acid, ferulic acid and imperatorin). A systematic quantitative fingerprint method and principal component analysis were used to analyze the generated data. Samples could be well distinguished from different manufacturers by analyzing the chromatographic data sets. In addition, the partial least squares model can serve as an antioxidant activity evaluation of Tianma Toutong tablets, as well as a reference for the selection of active constituents to analyze the spectrum–activity relationship. In summary, the integrated use of the fingerprint and chemometric analysis provides a reliable method for the identification of markers and the quality control of Tianma Toutong tablets.     

Quality assessment of crude and processed Arecae semen based on colorimeter and HPLC combined with chemometrics methods

Raw Arecae Semen, the seed of Areca catechu L., as well as Arecae Semen Tostum and Arecae semen carbonisata are traditionally processed by stir‐baking for subsequent use in a variety of clinical applications. These three Arecae semen types, important Chinese herbal drugs, have been used in China and other Asian countries for thousands of years. In this study, the sensory technologies of a colorimeter and sensitive validated high‐performance liquid chromatography with diode array detection were employed to discriminate raw Arecae semen and its processed drugs. The color parameters of the samples were determined by a colorimeter instrument CR‐410. Moreover, the fingerprints of the four alkaloids of arecaidine, guvacine, arecoline and guvacoline were surveyed by high‐performance liquid chromatography. Subsequently, Student's t test, the analysis of variance, fingerprint similarity analysis, hierarchical cluster analysis, principal component analysis, factor analysis and Pearson's correlation test were performed for final data analysis. The results obtained demonstrated a significant color change characteristic for components in raw Arecae semen and its processed drugs. Crude and processed Arecae semen could be determined based on colorimetry and high‐performance liquid chromatography with a diode array detector coupled with chemometrics methods for a comprehensive quality evaluation.     

Quantitative analysis of multicomponents by single marker combined with HPLC fingerprint qualitative analyses for comprehensive evaluation of Aurantii Fructus

The present study aimed to develop a strategy involving quantitative analysis of multicomponents by single marker in combination with high‐performance liquid chromatography fingerprint qualitative analysis for performing the quality control of Aurantii Fructus. The content of 12 components (eriocitrin, neoeriocitrin, narirutin, naringin, hesperidin, neohesperidin, meranzin, poncirin, naringenin, nobiletin, tangeretin, and auraptene) in samples was determined using reliable relative correction factors that were obtained using naringin as an internal reference standard. The new method demonstrated good applicability, and no significant differences were observed between the external standard method and the new method as determined by calculating standard method difference. Qualitative evaluation of samples was conducted using similarity analysis, hierarchical cluster analysis, and quality fluctuation analysis. Chromatographic fingerprint data were divided into three groups by similarity and hierarchical cluster analyses, and seven components may have a more significant impact on the quality of Aurantii Fructus in quality fluctuation analysis. Overall, the study suggests that the qualitative and quantitative analyses of multicomponents using quantitative analysis of multicomponents by single marker combined with chromatographic fingerprinting can be considered good quality criteria for performing quality control and providing technical support for the further pharmacological and pharmaceutical research of Aurantii Fructus.     

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.     

Qualitative and quantitative analysis of Alpiniae Oxyphyllae Fructus by high-performance liquid chromatography coupled to Fourier transform-ion cyclotron resonance mass spectrometry

Alpiniae Oxyphyllae Fructus, as a homology of medicine and food, has been widely used in China for thousands of years. However, the existing qualitative and quantitative methods are difficult to evaluate the quality of Alpiniae Oxyphyllae Fructus samples from multiple sources. In this paper, an high-performance liquid chromatography fingerprint was established for assessing the quality of Alpiniae Oxyphyllae Fructus from different areas. Then, high-performance liquid chromatography was coupled to Fourier transform-ion cyclotron resonance mass spectrometry for characterization of the chemical compositions in Alpiniae Oxyphyllae Fructus. In fingerprint analysis, 54 common peaks were confirmed and six chromatographic peaks of them were identified. The similarity of 14 samples from different areas was between 0.990 and 1.000. Moreover, a total of 30 chemical components were characterized by high-performance liquid chromatography coupled to Fourier transform-ion cyclotron resonance mass spectrometry method, six compounds of which were decisively identified. Finally, the content of nootkatone was determined by high-performance liquid chromatography. In conclusion, the methods used in this study are efficient for qualitative and quantitative analysis of Alpiniae Oxyphyllae Fructus. Also, these methods can be used to control the quality of other traditional Chinese medicines.     

穿山甲色谱指纹谱的建立及其在基源鉴别和质量等级区分方面的应用

中药穿山甲为鳞鲤科动物穿山甲的鳞甲,常用于治疗乳汁不通、痈肿疮毒、风湿痹痛等症,饮片来源上经常存在基源不清、等级不明的问题。针对以上问题,目前已发展出了多种分析方法,包括薄层色谱、质谱和DNA检测,但由于特异性低、设备成本高、步骤繁琐等不足,以上方法均不适合作为穿山甲质量监测的常规手段,而色谱指纹谱技术恰好可以弥补以上不足。该文旨在建立穿山甲的色谱指纹谱,并探讨其在等级及基源鉴别方面的可行性。穿山甲粉末经1 mol/L HCl加热水解得供试液,色谱柱采用Waters Symmetry 300 C18,以0.1%(v/v)三氟乙酸/水、0.1%(v/v)三氟乙酸/乙腈为流动相,280 nm为检测波长,采用梯度洗脱获得穿山甲指纹谱。经考察,该方法的精密度、日内日间重复性及样品稳定性均表现良好(RSD<5%)。以12批中华穿山甲一等品的平均指纹谱为对照指纹谱,同时确认17个共有峰,继而以共有峰绝对峰面积为原始数据,计算样品指纹谱与对照指纹谱的相似度。结果表明,中华穿山甲与其他动物来源穿山甲的相似度不高于0.776,体现了相似度评价在基源鉴别上的有效性;但不同等级穿山甲之间交叉严重,表明...     

Qualitative analysis of Psoraleae Fructus by HPLC‐DAD/TOF‐MS fingerprint and quantitative analysis of multiple components by single marker

A variety of bioactive substances may account for the recognized efficacy and wide clinical application of Psoraleae Fructus in China. A high‐performance liquid chromatography–diode array detector (HPLC‐DAD) fingerprint method was developed to present the comprehensive phytochemical profile of the crude drug. Thirteen major compounds were separated and identified by HPLC coupled with time‐of‐flight mass spectrometry (HPLC/TOF‐MS), namely psoralenoside (PO), isopsoralenoside (IPO), psoralen (PS), isopsoralen (IPS), neobavaisoflavone (NBF), bavachin (BC), corylin (CN), bavachromene (BCM), psoralidin (PD), isobavachalcone (IBC), bacachinin (BCN), corylifol A (CA) and bakuchiol (BK). Then quantitative analysis of multiple components by single marker (QAMS) was applied in content determination of PO, IPO, PS, IPS, BC, IBC, BCN, CA and BK, with NBF as the internal standard. The calculation results indicated no significant difference from the traditional external standard method (p > 0.05, RSD < 2.62%), suggesting that QAMS is a reliable and convenient method for content determination of multiple chemical compositions, especially when there is a shortage of reference substances. In conclusion, simultaneous qualitative and quantitative analysis of Psoraleae Fructus may be fulfilled through the newly proposed method of QAMS combined with HPLC‐DAD/TOF‐MS fingerprint.     

Quality assessment of Fructus Ligustri Lucidi by the simultaneous determination of six compounds and chemometric analysis

A comprehensive strategy was designed for the quality assessment of Fructus Ligustri Lucidi, a well‐known and commonly used herbal medicine in clinical practice in China. First, a simple and stable method of high‐performance liquid chromatography was developed for the simultaneous quantitative analysis of six compounds, namely, salidroside, nuzhenide, specnuezhenide, oleanic acid, ursolic acid, and acetyl oleanic acid in Fructus Ligustri Lucidi. The separation of analytes was conducted on a C₁₈ column (200 mm × 4.6 mm, 5 μm) at 30°C, and the wavelength of UV detector was set at 210 nm. In quantitative analysis, all of the calibration curves showed good linear regression (R² > 0.9994) within the tested ranges, and the mean recoveries of three different concentrations ranged from 95.21–102.34%. The described method was applied to determine 11 batches of samples collected from different stores in China. Then multiple chemometrics analysis including hierarchical cluster analysis and principal component analysis were performed to classify samples and search significant compounds. Three notable compounds, specnuezhenide, oleanic acid, and acetyl oleanic acid, were discovered for better quality control compared with those stated in the China pharmacopeia. The results demonstrated that this strategy could be readily utilized for the comprehensive quality control of Fructus Ligustri Lucidi.     

Quantitative fingerprinting based on the limited‐ratio quantified fingerprint method for an overall quality consistency assessment and antioxidant activity determination of Lianqiao Baidu pills using HPLC with a diode array detector combined with chemometric methods

Lianqiao Baidu pills are widely used herbal medicinal preparation that were analyzed to develop a quality consistency technique. The characteristic fingerprints of 28 batches of Lianqiao Baidu pill samples were established at five wavelengths and simultaneously assessed by using a limited‐ratio quantified fingerprint method using 15 marker compounds. The principal component analysis and fingerprinting results were compared, and the qualitative classification of the samples by principal component analysis agreed with their quantitative evaluation by the limited‐ratio quantified fingerprint method. Furthermore, the antioxidant activities of the samples were surveyed and determined using a 2,2‐diphenyl‐1‐picrylhydrazyl radical‐scavenging approach. A relationship between the common peaks in the fingerprints and the antioxidant activities was established using a partial least squares model. The relationship can be used both to determine the antioxidant activities of the Lianqiao Baidu pill preparations in vitro and as a reference for the selection of active constituents for sample quality classification. The classification results for the samples based on principal component analysis agreed with the quantitative evaluation by the limited‐ratio quantified fingerprint method, which demonstrated that the method can be applied to the holistic quality control of traditional Chinese medicine and herbal preparations.     

中药质量评价关键问题与分析方法探讨

该文针对我国目前中药质量评价体系现状,总结了中药质量评价的关键科学问题,并围绕"发现"与"控制"有效成分的中药质量评价方法进行探讨.笔者结合多年中药质量分析研究工作经验,分析了中药"谱-效"关系研究、全时段等基线多波长融合指纹图谱、生物活性测定法3种质量评价与检测方法的研究内容、手段、适用范围等,为丰富、完善中药质量控...     

Evaluation of the quality consistency of powdered poppy capsule extractive by an averagely linear‐quantified fingerprint method in combination with antioxidant activities and two compounds analyses

A novel averagely linear‐quantified fingerprint method was proposed and successfully applied to monitor the quality consistency of alkaloids in powdered poppy capsule extractive. Averagely linear‐quantified fingerprint method provided accurate qualitative and quantitative similarities for chromatographic fingerprints of Chinese herbal medicines. The stability and operability of the averagely linear‐quantified fingerprint method were verified by the parameter r . The average linear qualitative similarity (improved based on conventional qualitative “Similarity”) was used as a qualitative criterion in the averagely linear‐quantified fingerprint method, and the average linear quantitative similarity was introduced as a quantitative one. was able to identify the difference in the content of all the chemical components. In addition, was found to be highly correlated to the contents of two alkaloid compounds (morphine and codeine). A simple flow injection analysis was developed for the determination of antioxidant capacity in Chinese Herbal Medicines, which was based on the scavenging of 2,2‐diphenyl‐1‐picrylhydrazyl radical by antioxidants. The fingerprint–efficacy relationship linking chromatographic fingerprints and antioxidant activities was investigated utilizing orthogonal projection to latent structures method, which provided important pharmacodynamic information for Chinese herbal medicines quality control. In summary, quantitative fingerprinting based on averagely linear‐quantified fingerprint method can be applied for monitoring the quality consistency of Chinese herbal medicines, and the constructed orthogonal projection to latent structures model is particularly suitable for investigating the fingerprint–efficacy relationship.     

色谱指纹图谱与中药质量控制

本文综述了近年来色谱指纹图谱研究中多种色谱技术、相关化学计量学方法的发展及其在中药质量控制中的应用,并对中药质量稳定性和有效性的全面评价方法和可行性进行了初步探讨。提出以色谱指纹图谱为核心,依托现代色谱技术、化学计量学和系统生物学方法,建立中药化学成分信息与药效(毒性)信息相关关系的中药质量控制研究策略,深层次揭示中药的内在作用机理,期望真正实现中药质量的可控、安全和有效。