中药质量控制的研究策略和色谱技术

综述了色谱技术作为现代高效分离分析技术在中药药效物质基础研究和中药质量控制中的重要作用。提出中药药效物质组学研究策略,以实现中药药效物质基础与作用机理研究的一体化,进而从深层次揭示和全面控制中药的内在质量。论述结果表明,现代色谱及其联用技术是中药质量研究的重要技术平台之一。     

中药活性成分研究是中药现代化的重要组成部分

中药现代化是一个逐步发展、不断完善和充实的长期的系统工程。中医药在我国民族生存和繁衍的几千年历程中起着至关重要的作用,具有自己的特色和优势,是中华民族的宝贵财富,既要继承,又要创新和发展。中药现代化可以有多种模式,只要有利于人民健康,有利于传承和发展中医药,不管采取何种模式均应得到鼓励和欢迎。中药的活性成分研究是中药现代化的重要组成部分：一方面,一些活性成分能直接或作为先导化合物间接发展成药物;另一方面,活性成分又为中药复方现代化提供物质基础。这体现在活性成分既在现代化中药的制造和生产中起到标准化作用,同时活性成分也是中药复方专利最重要的核心组成部分。     

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。     

高效液相色谱-线性指纹定量法评价人参归脾丸的三波长指纹图谱

以样品指纹向量与对照指纹向量的多元化学组分间存在线性函数关系为基础,引入相关系数r、斜率a、截距b及线性定性相似度S_L、线性定量相似度M_L等概念,建立线性指纹定量法(linearly quantified fingerprint method, LQFM)模型以定性、定量控制中药质量。用高效液相色谱-二极管阵列检测法(HPLC-DAD)同时测定了10批人参归脾丸(Renshen Guipi Wan, RSGPW)分别在203、228和326 nm下的高效液相色谱指纹图谱,运用LQFM理论模型,鉴定RSGPW质量。LQFM能显著区别不同来源的产品,同一厂家产品质量控制一致。鉴定结果与已知的系统指纹定量法评价结果对比,LQFM对中药多元化学成分的复杂系统能实现客观的定性、定量评价。多波长指纹图谱能最大限度地兼顾中药各成分的紫外吸收特征,对中药质量控制具有更全面和可靠的特点。     

Understanding the traditional aspect of Chinese medicine in order to achieve meaningful quality control of Chinese materia medica

Although sophisticated and technologically advanced, current quality control methods for Chinese medicines (syn. Chinese materia medica or CMM) lack comprehensiveness and practicability. They are more suited for analyzing single-chemical drugs or specific, known chemical components that have already been isolated. While these methods can fully satisfy the modern scientific requirements for identity, purity and quality in the assessment of chemical drugs, they are not suitable for handling the complex chemical nature of traditional CMM whose multifunctional components along with their inherent holistic activities are frequently unknown and thus are not adequately analyzed by these methods. In order to assess properly and meaningfully the identity and quality of complex CMM (also known as Chinese herbs and Chinese herbal medicines), additional measures that can retain the traditional aspect of CMM need to be included. This requires a basic understanding of traditional Chinese medicine (TCM).     

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。     

质谱技术在中药研究中的应用进展

中药是中华传统医学的瑰宝,在我国已广泛应用于疾病预防和治疗中,但其存在化学成分复杂、作用机制不明确等问题,制约了中药现代化及国际化的发展进程。质谱技术具有高灵敏度、高选择性、高稳定性及高通量的特点,特别适合于中药复杂成分及其代谢物的定性定量分析。该文综述了近年来质谱技术在中药成分鉴定及质量控制、中药代谢组学及中药药代动力学研究方面的典型应用,并对存在的问题及改进方法进行了展望。     

Global detection and identification of components from crude and processed traditional Chinese medicine by liquid chromatography connected with hybrid ion trap and time‐of‐flight‐mass spectrometry

We herein present a chemical profiling method to efficiently process the information acquired by ultra fast liquid chromatography (UFLC)‐electrospray ionization source in combination with hybrid ion trap and high‐resolution time‐of‐flight mass spectrometry (UFLC‐(ESI)‐IT‐TOF/MS), facilitating the structural determination of serial components contained in crude or processed traditional Chinese medicine (TCM). Under the optimized UFLC and IT‐TOF‐MSⁿ conditions, over 39 compounds were separated and detected in crude or processed Fructus corni within 25 min. The components were identified by comparing the mass spectra and retention time with reference compounds, or tentatively assigned by elucidating low‐energy collision‐induced dissociation (CID) fragment ions and matching empirical molecular formula with that of the published compounds. Several factors in the processing procedure were examined. The experimental results demonstrate that the chemical reactions that occurred in the processing procedure can be used to elucidate the processed mechanism of F. corni, which is regularly affected by the processing conditions. This study provides a novel approach and methodology to identify the complicated components from various complex mixtures such as crude TCM, processed TCM, and biological samples. It can be used as a valid analytical method for further understanding the processing mechanism of TCM, along with the intrinsic quality control of TCM and its processed product.     

基因工程中的液相色谱

 对液相色谱用于基因工程生产蛋白质分子构象、变性蛋白的复性、蛋白折叠中间体等方面的研究进展,以及液相色谱在基因工程样品的质量 检验、制备方面的新进展分别加以评述。全文引用了42篇文献。     

Modified Jiu Wei Qiang Huo decoction improves dysfunctional metabolomics in influenza A pneumonia‐infected mice

In order to study the effective mechanism of a traditional Chinese medicine (TCM), modified Jiu Wei Qiang Huo decoction (MJWQH), against H1N1‐induced pneumonia in mice, we chose a holistic approach. A reverse‐phase liquid chromatography with quadruple time‐of‐flight mass spectrometry (LC‐Q‐TOF‐MS) was developed to determine metabolomic biomarkers in mouse serum for the MJWQH effects. Thirteen biomarkers of H1N1‐induced pneumonia in mice serum were identified, which comprised l ‐valine, lauroylcarnitine, palmitoyl‐l ‐carnitine, l ‐ornithine, uric acid, taurine, O‐succinyl‐l ‐homoserine, l ‐leucine, l ‐phenylalanine, PGF2α, 20‐ethyl‐PGE2, arachidonic acid, and glycerophospho‐N‐arachidonoyl ethanolamine. Among them, metabolites of amino acids, fatty acids and arachidonic acid had the most relevant changes in mice with H1N1‐induced pneumonia. MJWQH effectively improved weight loss, lung index, biomarkers and inflammatory mediators such as prostaglandin E2 and phospholipase A2 in the infected mice. Importantly, MJWQH reversed the elevated biomarkers to the control levels from the infection, which provided a systematic view and a theoretical basis for its prevention or treatment. The results suggest that the protective effect of MJWQH against H1N1‐induced pneumonia is possibly through regulation of pathways for amino acid, fatty acid and arachidonic acid metabolism. They also suggest that the LC‐MS‐based metabolomic strategy is a powerful tool for elucidation of the mechanisms of TCM. Copyright © 2013 John Wiley & Sons, Ltd.     

色谱在药物-机体复杂巨系统研究中的应用进展

色谱是一门以分离分析为主,旨在追求复杂事物纯而净的分析化学的重要分支学科。其经过百余年的发展,理论与技术日臻完善,集科学、技术与艺术于一体。近年来,色谱及其与质谱、核磁共振波谱、原子发射光谱等联用技术极大推动了环境、食品、石油化工、生物医药等领域中所涉及复杂体系的研究进展。作为我国传统文化的核心代表,中医药为中国乃至世界人民的健康服务逾千年,从古至今历经上千年临床考验,疗效经久不衰。近年来,中国政府强调继承与创新,加大推进中医药的现代化与国际化。然而中药自身的多成分协同起效复杂性及其与机体时刻新陈代谢变化的复杂性往往相互作用,由此形成了药物-机体复杂巨系统。该复杂巨系统的分析研究是中医药现代化进程的关键瓶颈。色谱的优势在于复杂成分的分离与分析,此恰能为上述复杂巨系统提供技术支撑,色谱及其联用技术已成为推动中医药分子化、数字化、信息化乃至现代化的主流技术。该文综述了色谱及其联用技术在中药复杂体系、复杂生命过程及药物-机体复杂巨系统中的应用进展,介绍了笔者研究团队对中医药现代化的认识、研究思路和研究工作,最后笔者结合对于百年色谱与千年中医药文化之现代化交织的感悟,对色谱技术在此领域的前景做出了展望。     

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。     

高分辨率电喷雾飞行时间质谱测定丹参脂溶性组分

丹参 [1] (Salvia miltiorrhiza Bage)为唇形科鼠尾草属多年生草本植物 ,临床上用于治疗妇科病、冠心病、缺血性中风及动脉粥样硬化等症 .近年来 ,飞行时间质谱 (TOF- MS)的分辨率优于 1 0 4 [2 ] ,尤其是再配合 ESI/MAL DI/APCI及新型脉冲电喷雾 [3 ]等离子源 ,使之成为当今最     

阐明中药科学内涵 推进中药现代化与创新药物研究进程

众所周知,中药对中华民族的繁衍昌盛发挥了不可磨灭的功绩。但是,作为经验医学,中药具有与生俱来的不足,诸如：缺乏采用循证医学原则提供的现代临床试验数据和结果;作用机理和作用物质尚不清楚;缺乏严格、科学的质量监控标准和方法。因此,难以保证达到作为药物所必须具备的“安全、有效、稳定、可控”的质量要求。这个问题如果不能妥善解决,不仅难于保证国人对疾病防治的需求,更难走向国际医药主流社会。通过多年的思考与实践,为揭示中药的科学内涵,推动中药现代化和创新药物研究,我们提出了一些新的研究思路和方法。     

高效液相色谱-四极杆-飞行时间质谱快速筛查鉴别食品中非法添加的62种中药材北大核心CSCD

建立了高效液相色谱-四极杆-飞行时间质谱快速筛查鉴别食品中非法添加的62种中药材的方法。依据卫生部关于进一步规范保健食品原料管理的通知(卫法监发[2002]51号)中可用于保健食品的物品名单,确定食品中可能非法添加的62种中药材原料清单,再从62种中药材中筛选其特征组分,获得不同中药材对应的特征组分清单。62种对照药材经甲醇超声提取后,于Thermo Accucore aQ色谱柱(150 mm×2.1 mm,2.6μm)上分离,在电喷雾正负离子扫描模式下,分别以0.1%(v/v)甲酸水溶液-乙腈和水-乙腈为流动相梯度洗脱,进行一级质谱和二级质谱全扫描检测,采用Library View软件建立不同中药材对应的特征组分的一级精确质量数据库和二级碎片质谱库。样品同法处理后上样分析,采用Peak View软件将样品高分辨数据与自建数据库中的质谱图、精确分子离子质量数、碎片离子质量数、保留时间等相关参数进行快速筛查鉴别分析。该工作通过建立“中药材-特征组分”对应清单,构建了食品中易非法添加的62种中药材中共388种特征组分的高分辨质谱库,每种中药材包括5~10种特征组分,通过对实际食品样品配制酒、代用茶及饮料进行筛查分析,1批次配制酒样品与淫羊藿中药材的7种特征组分匹配一致,推断该配制酒样品中加入了淫羊藿中药材。该法可实现无标准品情况下中药材的定性筛查,具有高通量、准确、简便、快捷的特点,解决了食品中非法添加中药材难以识别和确证的难题,可以实现食品中非法添加中药材的快速筛查鉴别分析。     

Comprehensive chemical profiling and quantification of Shexiang Xintongning tablets by integrating liquid chromatography-mass spectrometry and gas chromatography-mass spectrometry

Shexiang Xintongning tablet (SXXTN) is a traditional Chinese medicine (TCM) preparation for the treatment of coronary heart disease (CHD) angina pectoris. However, due to the complexity of the compounds in SXXTN, the active chemical components responsible for the therapeutic effect are still ambiguous. The purpose of our study was to characterize the chemical profile of SXXTN and quantify the representative chemicals. The high-performance liquid chromatography coupled with time-of-flight mass spectrometry (HPLC-QTOF MS) method and gas chromatograph coupled with mass spectrometry (GC–MS) method were utilized to identify the chemical constituents of SXXTN. A total of 140 compounds including alkaloids, ginsenosides, organic acids, esters, triterpenes, phthalides and amino acid were identified in accordance with their retention times, accurate masses and characteristic MS/MS fragment patterns. Forty-four volatile components were characterized by GC–MS through NIST database matching. In the further research of quantitative analysis, 40 non-volatile compounds and 17 volatile compounds were determined and successfully applied for detecting in 7 batches of SXXTN samples by high performance liquid chromatography coupled with triple-quadrupole tandem mass spectrometry (HPLC-QQQ MS) and gas chromatograph coupled with triple-quadrupole tandem mass spectrometry (GC-QQQ MS) in multiple reaction monitoring (MRM) mode, respectively. The quantitative methods were verified in linearity, precision, repeatability stability and recovery. The above results indicated that the established method was practical and reliable for synthetical quality evaluation of SXXTN. In addition, our study might supplement the chemical evidence for disclosing the material basis of its therapeutic effects.     

二维色谱技术及其在中药领域中的应用

二维色谱是近年发展起来的多维色谱分离技术,因其高分辨率、高峰容量、高灵敏度等优点,在复杂样品的分离分析中发挥了巨大作用．中药是一个复杂的化学体系,包含多种类化学成分,因此,分析和阐明中药中的化学成分是中药现代研究的一个关键问题．二维色谱技术在中药等复杂体系的应用中显示了重要的应用价值,也具有广阔的发展前景．本文对二维色谱的分类、主要部件以及在中药领域中的应用进行综述,重点介绍二维气相色谱和二维液相色谱在中药研究中的应用．     

大孔树脂精制6种含黄连复方水提液体系的吸附动力学比较研究

研究含黄连的不同复方水提液体系经大孔吸附树脂精制后,黄连中主要指标性成分生物碱的变化,阐明不同复方组合-配伍对药方中组成药味指标性成分的影响,为大孔树脂精制技术在中药复方中的应用提供实验依据.选用黄连用量和复方总用量均相同,组方药味不同的复方1～6.通过考察5种树脂AB-8、S-8、X-5、NKA-II、NKA-9对黄连中盐酸小檗碱的静态吸附率,确定AB-8树脂作为复方1～6吸附动力学比较研究的树脂;通过比较复方1～6水提液体系在相同树脂环境下的静态吸附过程,考察6种不同复方组成药味对复方中指标性成分分离纯化的影响.实验结果表明,由于体系本身溶液环境的差异,复方1～6各指标性成分达到吸附平衡的时问均有差异.AB～8树脂对复方1～6水提液体系的静态吸附在开始阶段吸附量的增加比较明显,在40min后,由于吸附竞争点饱和的原因,吸附量的增加均比较平稳;黄连与不同药材配伍时,虽然配伍药材所含的化合物类型差异较大,但黄连中生物碱类成分均可以在树脂上被选择性吸附,可知在复方水提液复杂体系中可以选择性地分离纯化生物碱类有效成分.