色谱技术在中药有效成分辨识中的应用进展

中药效应成分辨识是中药质量控制研究的关键,本文从化学特性、生物靶标和整体作用三个层面综述了2000年以来色谱技术在中药效应成分辨识研究中的应用进展,并提出未来发展方向.     

Measurement uncertainty for total ash and acid-insoluble ash determination of Chinese materia medica

The total ash and acid-insoluble ash contents of Chinese materia medica (CMM) have been widely used as one of the indices to illustrate the quality as well as purity of herbal medicines. Such an approach has also been adopted by many official organizations in writing monographs for pharmacopoeia worldwide. In the present study, a full validation on the methodology was conducted with the inclusion of various commonly encountered matrices in herbs, namely, leaf, flower, fruit, stem and root. The practice ensures method suitability for a wider scope of matrix variation in CMM. Moreover, in order to comply with the technical requirement of International Organization of Standardization 17025 quality assurance system, the concept of measurement uncertainty has also been incorporated in the present study. Measurement uncertainties for total ash and acid-insoluble ash contents have been estimated.     

中药质量控制技术发展展望

本文从中药产业需求、现代化需求、技术需求、机遇与挑战等方面概括了中药质量控制技术发展的背景;讨论了中药质量控制技术对于提高中药药效和安全性、推动产业发展和推进中药国际化的意义;综述了中药质量控制技术的现状,分析了在过程控制、安全性控制、标准品和对照品制备、指纹图谱技术等方面的不足;提出了中药质量控制技术应重点发展以分离和表征技术为主的中药质量控制关键技术、中药安全性控制技术、中药质量控制标准体系、中药质量控制原创性技术和中药标准品、对照品生产技术,制定技术标准,建立具有中药特色的过程控制和产品质量控制标准。     

发光分析与中药质量控制

回顾了近年来发光(荧光，室温磷光)分析技术在中草药质量控制中的应用，总结其特点并展望了可能的应用。     

Chemometrics and modernization of traditional Chinese medicine

Development of chromatographic fingerprinting and its related chemometric methods in the research of quality control of traditional Chinese medicines(TCMs) are discussed. The quality control methods for guarantying the authentication and stability of products and semi-products of TCMs are firstly assessed. The technique based on chromatographic fingerprinting is essentially a kind of high-through put and integral tools to explore the complexity of herbal medicines. In order to further control the comprehensive quality of TCMs,confirmation and identification of their important chemical components are necessary. Some new strategies are proposed to trace the chemical changes of chromatographic fingerprints both in product processing and/or after their administration by modern chromatographic techniques and chemometrics. Combined with systems biology and bioinformatics,it seems possible for one to reveal the working mechanism of TCMs and to further control their intrinsic quality comprehensively.     

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。     

基于定量指纹图谱技术的中药质量控制

定量指纹图谱技术是中药指纹图谱技术与多指标成分定量分析相结合的中药质量控制模式。定量指纹图谱技术的发展包括定量组分的制备、过程控制的指纹图谱技术和产品含量测定3个主要部分。本文以丹参为例,通过水提、醇沉、过膜、大孔树脂分离和工业色谱分离5个工艺流程制备了丹参定量组分,对各个工艺步骤以指纹图谱技术考察其稳定性和重复性。对丹参定量组分中的原儿茶醛、迷迭香酸、丹酚酸B 3个成分进行含量测定。3个成分的含量总和大于50%。定量组分的制备以现有的活性成分为目标,经过去粗存精的工艺过程,其质量标准得到了有效的提高。     

色谱及相关技术在中药质量评价中的应用

对中药物质基础的研究方法——化学物质组学,以及中药质量评价的关键技术——指纹图谱技术做了介绍,并在此基础上综述了中药化学信息获取的关键技术——色谱及相关技术在中药质量评价中的应用。论述的结果表明,以化学物质组学为指导,结合色谱及相关技术获取指纹图谱信息及多指标成分定量信息的研究模式,对中药质量评价体系的建立具有指导意义。     

Qualitative and quantitative analysis in quality control of traditional Chinese medicines

Separation techniques with high efficiency and sensitive detection have been widely used for quality control of traditional Chinese medicines (TCMs). High-performance liquid chromatography, gas chromatography, and capillary electrophoresis are commonly used to separate various components in TCMs. Ultraviolet detection, fluorescence detection, evaporative light-scattering detection, mass spectrometry and nuclear magnetic resonance can be applied to separation techniques for qualitative and quantitative analysis of TCMs. The development of quality control for TCMs based on quantitative and qualitative analysis from 2000 to 2007 are reviewed; the fingerprint technique is also discussed due to its broad application in the quality control of TCMs. Prospects for further research based on our primary results are also discussed.     

中药指纹图谱学体系在中药创制中的作用

从整体论、系统论和复杂科学的角度论述了中药指纹图谱学体系的构成背景和核心任务。中药指纹图谱学体系具有体系科学性质,是从系统性和整体性角度研究中药(包括植物药)的物质基础和作用机制以及药代动力学规律和相关制剂技术的崭新学科。该体系以中药指纹图谱信息学为核心和桥梁,包括中药指纹图谱测试学、中药指纹图谱质控学、中药指纹图谱谱效学、中药指纹药物动力学、中药指纹药剂学和中药生物指纹图谱学。该体系的研究需采用复杂科学和体系科学的理论和方法,突破线性思维和还原分析,强调宏观和系统的有机综合,应用数学科学原理和方法来开辟中药创制的新模式。中药指纹图谱学的理论体系和方法是解析中药的主导技术和实现中药现代化的核心力量,该体系理论和技术的成熟及完善可为现代中药创制提供强有力的理论和技术支撑。中药指纹图谱学体系的最终目的是为人类和有益生物研制出疗效显著、安全、可控的现代化中药。     

Recent analytical approaches in quality control of traditional Chinese medicines—A review

Traditional Chinese medicines (TCMs) are gaining more and more attention all over the world, due to their specific theory and long historical clinical practice. But the uncontrollable quality is a bottleneck for its modernization and globalization. This paper reviewed the recent analytical methods in the quality control of TCMs, including screening strategies of bioactive markers from TCMs through biochromatographic methods, the traditional chromatographic methods, DNA methods, as well as the spectroscopic methods, including FT-IR, NIR and NMR. The comprehensive methods, such as fingerprint and multi-component quantification are emphasized; hyphenated techniques, like HPLC-MS, GC-MS, CE-MS, LC-NMR, chemometric methods, and combination of chemical and biological methods, such as biofingerprint, metabolic fingerprint are now more and more widely used in TCMs. In a few word, the analysis and quality control of TCMs are moving towards an integrative and comprehensive direction, in order to better address the inherent holistic nature of TCMs.     

Chromatographic fingerprinting and related chemometric techniques for quality control of traditional Chinese medicines

Development of chromatographic fingerprint (CF) and related chemometric methods and their applications to quality control of traditional Chinese medicines (TCMs) were discussed. CF is essentially a kind of quality control method for TCMs (or Chinese herbal medicines). Also, it is a quality‐relevant‐data high‐throughput and integral tool to explore chemically the complexity of TCMs. With the help of chemometrics, some difficulties in evaluation and analysis of CFs, such as calculation of information content, peak alignment, pattern analysis, deconvolution of overlapping peaks, etc. could be well solved. To further explore TCMs synergic quality, intensive study of CF coupled with chemometrics will create the possibility to achieve the aim to reveal the working mechanisms of TCMs and to further control and strengthen TCMs' intrinsic quality in a comprehensive manner.     

中药指纹图谱研究的某些进展与展望

在中药现代化的过程中,中药的质量控制标准直接影响到中药的国际化问题。近年来,国内很多学者对中药指纹图谱用于质量控制的问题进行了深入的研究,取得了很多成果。本文着重就近几年来中药指纹图谱的某些进展进行了综述,内容包括气相色谱指纹图谱、液相色谱指纹图谱以及其他指纹图谱。最后就指纹图谱研究的发展提出了几点看法。共引用文献161篇。     

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.     

灰色系统理论在中药色谱指纹图谱模式识别中的应用研究

建立了基于灰色系统理论的中药色谱指纹图谱模式识别模型。运用基于范数与信息熵赋权法的灰色关联分析,求出各待比较图谱特征变量数据序列与理想图谱特征变量数据序列之间的灰色关联度,并依据模糊匹配中的“最大匹配度”原则进行判断,从而达到品种识别和质量评价的目的。该模型在56批次不同品种化橘红药材样品的高效液相色谱分析中取得较满意的结果: 熵权与范数的灰色关联分析均能准确识别出毛橘红与光橘红两个品种,克服了传统相似度或灰色关联在化橘红色谱指纹图谱分析中的误判问题;对药材中化学成分的种类与含量十分接近的毛橘红不同栽培品种的识别率超过92.85%。此外,该模型计算量较少,整个模式识别过程通过计算机编程实现,操作简单。实验结果显示,灰色系统理论在中药色谱指纹图谱模式识别中有良好的适应性。     

Primary study on the application of Serum Pharmacology in Chinese traditional medicine

In the paper, two main methods, which are Serum Pharmacology and Traditional Pharmacology, were adopted to study Chinese traditional medicine, such as Ginkgo biloba extract (GBE), ginsenosides (GS) and compound GG (GBE + GS), pharmacology in vitro. The results showed that there were evident difference between the results of Serum Pharmacology and that of Traditional Pharmacology. There was no significant difference between the drug effect of crude GS on nitric oxide (NO) production in ECV304 and that of crude GBE, and the drug effect of GG was superior to that of GS and GBE, respectively. But, compared with GBE serum, the GS serum up-regulation of NO production in ECV304 increased significantly, and the GG serum up-regulation of the NO production in ECV304 was inferior to that of GS serum and GBE serum significantly. The results suggested that Serum Pharmacological study should be adopted in the pharmacological investigation on the Chinese traditional medicine and the drug screening of the Chinese traditional medicine.     

Determination of strychnine and brucine in traditional Chinese medicine preparations by capillary zone electrophoresis with micelle to solvent stacking

A novel micelle to solvent stacking on-line sample preconcentration technique in capillary zone electrophoresis（MSS-CZE） has been developed to determine the strychnine and brucine in traditional Chinese medicine preparations.The optimal running buffer was 30 mmol/L H₃PO₄ containing 20%acetonitrile at pH 4.0.The sample matrix was 8 mmol/L H₃PO₄ containing 5 mmol/L sodium dodecyl sulfate（SDS） at pH 3.0.The established MSS-CZE method afforded more than 50-fold improvements in concentration sensitivity compared with typical CZE-UV analysis.The calibration curve was linear in the range from 0.2 to 15.0μg/ mL for both strychnine and brucine,with correlation coefficients of 0.9984 and 0.9976,respectively.The limits of detection（5/ N = 3:1） for strychnine and brucine were 0.02 and 0.05μg/mL,respectively.The MSS-CZE method has been successfully applied to the analysis of strychnine and brucine in Chinese medicinal preparations.     

Direct analysis in real time mass spectrometry and multivariate data analysis: A novel approach to rapid identification of analytical markers for quality control of traditional Chinese medicine preparation

The paper presents a novel strategy to identify analytical markers of traditional Chinese medicine preparation (TCMP) rapidly via direct analysis in real time mass spectrometry (DART-MS). A commonly used TCMP, Danshen injection, was employed as a model. The optimal analysis conditions were achieved by measuring the contribution of various experimental parameters to the mass spectra. Salvianolic acids and saccharides were simultaneously determined within a single 1-min DART-MS run. Furthermore, spectra of Danshen injections supplied by five manufacturers were processed with principal component analysis (PCA). Obvious clustering was observed in the PCA score plot, and candidate markers were recognized from the contribution plots of PCA. The suitability of potential markers was then confirmed by contrasting with the results of traditional analysis methods. Using this strategy, fructose, glucose, sucrose, protocatechuic aldehyde and salvianolic acid A were rapidly identified as the markers of Danshen injections. The combination of DART-MS with PCA provides a reliable approach to the identification of analytical markers for quality control of TCMP.     

中药真菌毒素质量控制概况、限量标准制定及有关问题的思考

建立超高效液相色谱-串联质谱法同时测定地龙药材中黄曲霉毒素B₁、B₂、G₁、G₂的含量,并对阳性结果进行确证. 样品经70%甲醇提取、免疫亲和柱净化、超高效液相色谱柱分离、三重四极杆质谱检测,并采用离子峰度比进行阳性结果确证. 结果表明,4种黄曲霉毒素的线性关系良好（r>0.999 5）,回收率在91.2%~95.6%范围内. 黄曲霉毒素B₁、B₂、G₁、G₂的检出限分别为0.10、0.038、0.11、0.038 μg/kg. 方法专属性好,灵敏度高,准确性好,可以进行最终阳性检出的判定. 15批地龙药材中6批黄曲霉毒素确证阳性检出, 证明地龙药材较易被黄曲霉毒素污染.

     

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.