Chinese Journal of Natural Medicines 2013, 11(6): 0596−0607

Chinese Journal of Natural Medicines

Quality control of traditional Chinese medicines: a review SONG Xin-Yue 1, 3, LI Ying-Dong 2, SHI Yan-Ping 1, JIN Ling 2 *, CHEN Juan 1* 1

Key Laboratory of Chemistry of Northwestern Plant Resources and Key Laboratory for Natural Medicine of Gansu Province, Lanzhou Institute of Chemical Physics, Chinese Academy of Sciences, Lanzhou 730000, China; 2 Gansu College of Traditional Chinese Medicine, Lanzhou 730000, China; 3 University of Chinese Academy of Sciences, Beijing 100039, China Available online 20 Nov. 2013

[ABSTRACT] Traditional Chinese medicines (TCMs) are in great demand all over the world, especially in the developing world, for primary health care due to their superior merits such as low cost, minimal side effects, better cultural acceptability, and compatibility with humans. However, Chinese medicines consist of several herbs which may contain tens, hundreds, or even thousands of constituents. How these constituents interact with each other, and what the special active ones are, may be the biggest bottleneck for the modernization and globalization of TCMs. Valid methods to evaluate the quality of TCMs are therefore essential and should be promoted and be developed further through advanced separation and chromatography techniques. This paper reviews the strategies used to control the quality of TCMs in a progressive perspective, from selecting single or several ingredients as the evaluation marker, to using different kinds of chromatography fingerprint methods. In summary, the analysis and quality control of TCMs are developing in a more effective and comprehensive manner to better address the inherent holistic nature of TCMs. [KEY WORDS] Traditional Chinese Medicines; Quality control; Bioactive ingredients; Chromatography fingerprint

[CLC Number] R914

1

[Document code] A

[Article ID] 1672-3651(2013)06-0596-12

Introduction

Traditional Chinese Medicines (TCMs) have attracted interest and acceptance in many countries with the merits of few side effects, affordability, and local availability [1]. In addition, their long historical clinical practice and reliable therapeutic efficacy make them excellent sources for discovering natural bioactive compounds [2]. However, many people, especially who prefer to use Western medicines, hold an opposite opinion due to indistinct drug effects and the poor quality control of TCMs. To globalize their use, integrative and convincing quality control is urgently needed. In the former years, researchers often choose one or more chemical markers to assess the quality of TCMs. However, the clinical application of a particular plant medicine is the synergistic

[Received on] 28-Mar.-2013 [Research funding] This project was supported by the National Key Technology Research and Development Program of China (No. 2011BAI05B02) and the National Natural Science Foundation of China (No. 21105106). [*Corresponding author] CHEN Juan: Prof., Tel: 86-931-4968208, Fax: 86-931-4968094, E-mail: [email protected]; JIN Ling: Prof., E-mail: [email protected]. These authors have no any conflict of interest to declare. Published by Elsevier B.V. All rights reserved

effect of multiple chemical compositions [3]. Therefore, the simple quantitative analysis of one or several chemical markers in plant medicines can not represent their quality well [4]. Generally, samples with a similar chromatographic fingerprint would have similar properties. Consequently, chromatographic fingerprinting has the potential to establish the identity, authenticity, and lot-to-lot consistency of herbal medicines [5]. Thus, multiple patterns of chromatographic fingerprinting have developed, such as multiple chromatographic fingerprints, and bio- and meta-fingerprints. Based on the comprehensive description of all the characteristics of TCMs, chromatographic fingerprinting is considered an effective method to control their quality [4]. In this review, the development and application of effective control methods in the past twenty years, with an emphasis on the application of novel techniques, are discussed. The development of fingerprint techniques and the potential problems in further research should also be considered.

2

A specific Component as an Evaluation Marker

Evaluation markers are characteristic constituents responsible for the biological, or even the therapeutic effects of Chinese plant medicines. Since the therapeutic or bioactive components of many plant medicines have not been fully elucidated, characteristic, principal, synergistic, correlative, toxic, and general components may be selected [6]. Selecting

SONG Xin-Yue, et al. /Chinese Journal of Natural Medicines 2013, 11(6): 596−607

a single specific component as an evaluation marker was employed for evaluating the quality and authenticity of herbal medicines, identifying the single plant or plant medicine preparations and assessing the quantitative plant composition of a plant product [7]. Among 1203 raw materials of TCMs, recorded in the Chinese Pharmacopoeia (2010), the quality of 529 herbs is evaluated based on the use of a single evaluation marker [8]. Zhao et al [9] designed a simple, sensitive, and accurate liquid chromatography combined with photodiode-array detection (LC-PDA) method to determine dehydroevodiamine in Euodia rutaecarpa (A.Juss.) Benth. A four-factor-three-level orthogonal array design was adopted to optimize the extraction conditions, thus leading to an excellent linear behavior and low LOD. Sharma [10] applied a novel and rapid high-performance thin-layer chromatography method (HPTLC) to determine pyrethroid in several commercial emulsifiable concentrate formulations of cypermethrin, α-cypermethrin and λ-cyhalothrin. In 2004, our group [11] developed a LC-PDA to determine daphnetin in extracts of Daphne tangutica Maxim. and its medicinal preparation, which was demonstrated to be a powerful technique. These satisfactory results proved that the determination of a single chemical marker to control the quality of TCMs is a sensitive and fast method.

3

Multiple Components as Evaluation Markers

Despite possessing simple characteristics, a single evaluation marker cannot provide sufficient and convincing information for TCMs which contain several hundred chemical components. Considering the synergistic effects of multiple components on the effectiveness or therapeutic function of TCMs, more chemical markers or active ingredients should be determined by feasible separation and detection methods to assess their bioactivities and possible side effects. Wang et al [2] used LC-mass spectrometry (LC-MS) to simultaneously quantify four active schisandra lignans from a traditional Chinese medicine Schisanda chinensis (Turcz.) Baill. Since Si-Wu-Tang is one of the most popular TCM formulae for woman’s health, an effective and sensitive method to control its quality is necessary. Wang et al [12] applied a high performance liquid chromatography-MS (HPLC-MS) to simultaneously determine nine identified active ingredients in the raw herbs and products of Si-Wu-Tang. The obtained results demonstrated this method to be a sensitive and rapid quantification approach. Bulbus Fritillariae is the most commonly used antitussive traditional Chinese medicinal herb in which isosteroidal alkaloids are the main bioactive ingredients. As the contents and structure types of these bioactive alkaloids vary in different species, quality control of these active principles is urgent to ensure its effectiveness in clinical practice. Lin et al [13] used HPLC-evaporative light scattering detection method (HPLC-ELSD) to quantify six isosteroidal alkaloids as evaluation markers, and this simple and selective assay was readily used as a suitable quality control method. More ex-

amples [14-22] clarified that the separation and determination of multiple components is an effective and feasible method to control the quality of TCMs. However, some certified standards of evaluation markers are difficult to be separated, expensive, or have poor stability, which makes their supply and use inaccessible. To solve the above problems, Wang et al [23] proposed a novel method named quantitative analysis of multi-components by single-marker (QAMS), which applied the single accessible evaluation marker to simultaneously detect multiple inaccessible ingredients due to active ingredients in plants having inherent functional and proportional relationships. The relative retention values between the target compounds and the single marker were used for qualitative analysis, while their relative correction factors (RCFs) were applied to quantitative analysis. In the Chinese Pharmacopeia (2010), this was used to determine the contents of five alkaloids, such as berberine, palmatine, coptisine, epiberberine and jatrorrhizine [24] . Based on this method, Wang et al [25] established a quantitative method to determine ginsenosides Rg1, Rb1, Rd, Re, and notoginsenoside R1, for the purpose of the quality control of Panax notoginseng (Burkill) F.X.Chen ex. C.Y.Wu & K.M.Feng. The durability of the method was evaluated with five different HPLC instruments, five various C18 chromatographic columns, and four detective wavelengths. The results illustrated that this method had relatively good durability and suitability among various apparatus and chromatographic conditions. Subsequently, the novel QAMS method was applied to determine the content of five saponins in forty-three P. notoginseng samples, and compared them with traditional external standard methods in order to evaluate the accuracy of the QAMS method. There was no significant difference between the contents of the five ingredients calculated by the two methods, which suggested that this method was highly accurate, and provided a reliable basis for application in the quality control of TCMs. This method has been applied to determine multiple components in other plants, including Schisandra chinensis, Angelica dahurica var. formosana (H.Boissieu) Yen, Salvia miltiorrhiza Bunge, and Paeonia lactiflora Pall. [26-29] 3.1 The ways to select bioactive markers The markers for quality control should be strongly correlated to the safety of TCMs, and be responsible for their therapeutic or harmful effects. Consequently, how to determine these active ingredients efficiently is an important and valuable task. Recently, several ways have been developed to achieve this goal. 3.1.1 Pharmacological screening The traditional method for screening bioactive chemical markers is to isolate the compounds first, and then screen their activities in vitro or in vivo through animal, organ, tissue, receptor, and/or enzyme models. However, these methods have proved to be time-consuming, laborious, and inappropriate for clarifying the synergistic action of TCMs. There-

SONG Xin-Yue, et al. /Chinese Journal of Natural Medicines 2013, 11(6): 596−607

fore, some more useful ways have been developed. 3.1.2 Colon carcinoma cell line (Caco-2) scheme As is known, active ingredients should possess perfect permeability and be absorbed from the gastrointestine to ensure they are largely used by the body. Based on this, a new method called the Caco-2 scheme was developed, where Caco-2 cells are cultured as a confluent monolayer on the porous membrane of an inner well situated within an outer well. Transport studies are performed by placing the studied compounds in the apical side, and then do qualitative and quantitative analysis in the basolateral side. The bioactivity of Oldenlandia diffusa (Willd.) Roxb. was investigated by the production of post-absorption samples using Caco-2 intestinal epithelial monolayers combined with HPLC-MS [30]. 3.1.3 Liposome equilibrium dialysis (LED) Qi et al [3] suggested that the screening and analysis of multiple bioactive compounds could be achieved by characterizing the interaction between plants and biological systems with LED. Active ingredients in TCMs would combine with liposomes, and these combined active compounds would not penetrate a dialysis membrane, while free compounds would. Therefore, multiple interactive components would be simultaneously predicted by comparing the chromatograms of the TCM extract before and after interaction with liposome membranes. Based on an LED system, several constituents, including phenolic acids, diterpenoid quinones, and saponins which have interacted with liposomes in Fufang Danshen prescription were analyzed and identified by HPLC-MS [3]. The results proved this technique was a powerful tool to study active-membrane interactions for screening active compounds. 3.1.4 Interaction with proteins Most compounds in TCMs could bind to plasma proteins reversibly. In addition, the degree of binding has significant effects on the pharmacokinetic and pharmacodynamic properties of some active ingredients, i.e. genistein and daidzein with estrogen receptors alpha and beta [31]; pyridostigmine bromide and N, N’-diethyl-3-toluamide with bovine brain tubulin [32]; ingenol-3-angelate with protein kinase C [33]. The Li group [34] used a microdialysis-HPLC system to study the interaction of the potential active ingredients in Danggui Buxue decoction with bovine serum albumin. By comparing the binding degrees with bovine serum albumin, nine compounds were found to possess potential activities. The current study reports that curcumin, as the main pharmacologically active ingredient of turmeric, and diacetylcurcumin can bind to human serum albumin and bovine serum albumin. The apparent binding constants, and the number of substantive binding sites, were evaluated by the fluorescence quenching method [35]. Compared to the traditional pharmacological screening, some superiorities of this method in the screening of bioactive compounds from complex natural products are noticeable. It avoids the most time-consuming step: purifying each compound. More importantly, it combines high separation performance and the biological identi-

fication [36]. 3.1.5 Interaction with target cells and molecules Modern pharmacological studies have revealed that active ingredients can bind with some receptors, channels, and enzymes on target-cell membranes and (or) inside cells. When a TCM extract is incubated together with target cells, the potential bioactive candidates in the TCMs should selectively combine with the cells. Those cell-combining components can be deposited together with target cells when centrifuged. After denaturalization under suitable conditions, the bound components dissociate from receptors and the former are then detectable by effective methods [5]. Using the proposed approach, the potential bioactive components of Danggui Buxue decoction, a commonly used TCM for anemia, and its constituent plants, Radix Angelica Sinensis and Radix Astragli, were investigated. Six compounds in the extract of Danggui Buxue decoction were detected since the components could selectively combine with endothelial cells, among which two contributed by Radix Angelica Sinensis and four by Radix Astragli. The results indicated that the proposed approach could be applied to predict the bioactive candidates in TCMs [37]. In view of the two key steps of drugs action, absorption by intestinal epithelium cells and interaction with target cells, this rapid and reliable method could be utilized to predict the bioactive constituents in TCMs. More importantly, it agreed with the characteristics of TCMs as being multi-component, with multi-target sites and multi-channel actions [38]. In addition, Zhang et al [39] applied cell extract combined with gas chromatography-MS (GC-MS) to screen and analyze bioactive compounds in Elemene emulsion injection. The results showed that this novel developed method could screen the potential bioactive components in TCMs by interacting with the target cells quickly. Although these methods were proved to be effective and sensitive to screen active ingredients, they are unavoidably confronted with some limitations. Firstly, since only one type of target is immobilized in the solid phase, the active ingredients absorbed by other targets are lost. Secondly, the short lifetime of these stationary phase requires special materials or gels be used. Thirdly, the separation ability is weaker than that of HPLC, leading to some overlapped peaks occurring in the chromatograms. Finally, due to the limitation of the mobile phase, these methods can not connect to MS directly, which brings some difficulties to on-line detection [6]. 3.1.6 TLC-bioautography The TLC-bioautographic method combines chromatographic separation and in situ activity determination, giving quick access to information concerning both the activity and localization of active ingredients in complex plant matrices [39]. It is usually used to screen for the activities of antimicrobial and antibacterial compounds, and antioxidant or acetylcholinesterase inhibitors by absorbing the active ingredients onto the surface of chromatographic plates, and then placing these plates directly in contact with a medium which was inoculated

SONG Xin-Yue, et al. /Chinese Journal of Natural Medicines 2013, 11(6): 596−607

with bacterial or fungal cultures or spraying certain chemicals on plates. Since the active ingredients inhibit microbial growth, oxidation capacity, or enzymatic activity, different colors will appear by virtue of suitable indicators. Then powerful detection techniques and software can be used to record and analyze the spot information for quantitative analysis. With its distinctive advantages of simplicity, low cost, high sensitivity, and specificity, this method has received wide application to determine the active ingredients in many herbs like Dysoxylum grande Hiern, Eugenia jambolana Lam., Bauhinia purpurea L., Angelica archangelica L., and Perilla frutescens var. acuta (Odash.) Kudô [40-44]. 3.1.7 High-throughput screening (HTS) Nowadays, high-throughput screening (HTS), combined with data analysis method, has become a valuable way to screen for the active ingredients. The Liu group [6] introduced a strategy to produce high-quality fractionated libraries. An off-line, two-dimensional LC was used to separate mediumand low-polar extracts of Chinese medicinal plant formulas, while HPLC-diode array detctor (DAD) and MS analysis were used to obtain ultra-violet (UV) and MS spectra of the library components. The detected components were characterized by retention, molecular weight, and UV absorbance assisted by WiseProcessor, a customer-developed software to automatically process analytical data. Recently, highly effective separation methods have been applied to enhance the applicability of HTS. Ultra-performance liquid chromatography (UPLC) was evaluated as an efficient screening approach to facilitate the development of a method for drug candidates. By virtue of its simplicity, high efficiency, and quick separation, UPLC serves as a feasible and productive alternative to conventional multi-parametric chromatographic screening approaches for many compounds in the early stages of drug development [45]. Beside effective separation methods, sensitive detection techniques have also been developed, such as MS coupled to a low-temperature plasma probe ion source. Without sample preparation or pretreatment, the active ingredients of eleven types of commercial pharmaceuticals were directly desorbed/ionized, and then detected by a linear ion trap mass spectrometry. The sensitive analysis demonstrated that a low-temperature plasma probe ion source combined with MS was an effective HTS method to screen pharmaceuticals, and could be potentially applied to on-line quality control in the pharmaceutical industry [46]. At present, HTS programs for drug discovery depend mainly on compound libraries from combinational chemistry. Unfortunately, the time-consuming manual process involved in the isolation of active ingredients from the highly complex plant extracts made a considerable deterioration to the quality of plant drugs. So the miniaturized and integrated microchip has been shown to be a substantial improvement in HTS, e.g., Herbochip®, DNA chip, protein chip, cell chip [47]. 3.1.7 Outliers discriminated by robust principal component analysis (RPCA)

In a well-controlled experiment, outliers discriminated by RPCA represent the compounds in samples which are of particular quality and distinguishable from the remainder. Therefore, the chemical constituents in a medicinal plant which causing a discrimination between the outliers and the majority of samples could be considered as possible analytical markers for quality control. Based on this strategy, a novel approach for rapidly exploring characteristic analytical markers was proposed for the quality control of extract granules of Radix Salviae Miltiorrhizae. In this study, a number of samples were analyzed through UPLC-MS. RPCA was performed on three groups of samples, the raw material, the in-house prepared aqueous extract of Radix Salviae Miltiorrhizae, and a commercial sample, in order to determine the variation of special constituents between the raw material and the final products. Then RPCA was performed on the commercial products to explore the applicability of the identified characteristic markers. Candidate markers were chosen by RPCA and their molecular formulae were determined by high resolution electrospray ionization-MS (HRESI-MS) analysis. The suitability of the identified markers was then evaluated by determining the relationship between the quantities of the markers with their antioxidant activities biologically, and further confirmed in a variety of samples. Based on this theory, five active ingredients were found [48]. So the powerful software is beneficial for the exploration of characteristic analytical markers for quality control. However, some disadvantages of this method may hinder its wide application. In some cases, outliers are attributed to experimental errors instead of active ingredients with unique characteristics [48]. In addition, to obtain persuasive results, significant amounts of data and scientific data analysis are needed, which requires a relatively long time and much effort. 3.2 Separation and detection techniques of evaluation markers Separation techniques with high efficiency and sensitive detection methods have been widely used for the quality control of TCMs. HPLC, GC, capillary electrophoresis (CE), and TLC are commonly used separation methods, while UV, fluorescence detector (FD), ELSD, MS, and nuclear magnetic resonance (NMR) are widely used as detection techniques to analyze TCMs qualitatively and quantitatively. 3.2.1 TLC and relative techniques TLC has been widely applied to the qualitative identification and semi-quantitative analysis due to its simplicity, low cost, high sample capacity, rapid availability of results, and good selectivity. Although chromatography techniques including HPLC, GC, and CE possess higher separation ability and selectivity, TLC has its unique advantages. Firstly, the disposable property helps TLC avoid cross-over contamination. Particularly, all substances adsorbed onto the stationary phase of column chromatography will decrease the life of column and result in poor peak shape. Secondly, due to the easy operation of TLC, a relatively short time is needed to

SONG Xin-Yue, et al. /Chinese Journal of Natural Medicines 2013, 11(6): 596−607

train operators to acquire the desire results. Thirdly, visible light, as well as sensitive visualization reagents, can be easily used to identify and characterize almost all of the compounds. Recently, the development of TLC has focused primarily on miniaturization, high-throughput, speed acceleration, and the combination with advanced detectors, such as MS. Following these trends, many novel TLC-related techniques have been developed, such as ultra-TLC (UTLC), HPTLC, multi-step TLC, and micro-TLC (MTLC). As for UTLC, the layers have a monolithic structure based on a silica gel matrix, rather than granular adsorbents. Therefore, UTLC will increase the sensitivity, shorten analysis time drastically, and reduce the quantity of consumables per analysis. However, UTLC will only fully perform its powerful functions when appropriate and efficient equipment is available, which needs further development [49]. In order to improve selectivity, special developing chambers like automatic multiple development have been developed [50]. Over-pressured layer chromatography (OPLC) is an instrumental planer liquid chromatographic technique which could eliminate extra clean-up steps before chromatographic separation, and ensure high sample throughput and low operating costs [51]. In addition, various detectors, such as densitometers, video, photo, and MS make it possible for TLC to achieve more accurate qualitative and quantitative results. Vermaak et al [50] used HPTLC to authenticate Hoodia gordonii raw material and its products. Through active research, they found that the presence of the bioactive marker-P57 was a guarantee for the quality of H. gordonii. HPTLC analysis of several raw material samples collected from different locations and weight-loss products was carried out on silica gel plates and developed in a suitable mobile phase, followed by Liebermann-Burchard (L-B) reagent. 3.2.2 GC and relative techniques Chinese herbs consist of many volatile and low boiling point active compounds, where GC performs an important function. Compared with liquid, gas chrmoatograhy has some remarkable advantages, such as low viscosity, large diffusion coefficient which contributes to low flow resistance and rapid mass transfer velocity. Therefore it is advantageous for the analysis of compounds with low molecular mass. Unavoidably, GC has its own drawbacks, including limited analytes and stationary phases. The introduction of advanced sample pretreatment, such as micro-extraction techniques, is an important developing trend towards enlarging the application of GC, decreasing impurity contamination, and enhancing the selectivity. Solid phase micro-extraction (SPME) is a solvent-free method for the isolation and concentration of volatile compounds in the headspace. Deng et al. applied a novel, solvent-free, and low-cost headspace-SPME (HS-SPME) to determine the ligustilides in Ligusticum chuanxiong and Angelica sinensis. The two bioactive compounds Z-ligustilide and E-ligustilide were first extracted by pressurized hot water, and then concentrated with HS-SPME, and detected with

GC-MS. The high extraction efficiency proved HS-SPME to be a powerful approach for the quantitative analysis of ligustilides in TCMs [52]. This method was also applied to qualify and quantify the contents of eucalyptol, camphor, and borneol in chrysanthemum flowers [53]. 3.2.3 HPLC and related techniques LC-DAD/MS is a powerful way to identify ambiguous compounds by comparison with standards. Ding and colleagues [54] have determined the alkaloids in Corydalis yanhusuo W. T. Wang using LC-MS/MS and LC-DAD. In addition, Lee et al [55] used LC-MS to rapidly determine the aristolochic acid content in plant medicines. LC-NMR has distinct superiority to unambiguously identify the structures of compounds. However, this method has some disadvantages, such as relatively low sensitivity and expensive cost. HPLC is widely applied to assess the chemical composition of TCMs with the advantages of simplicity, stability, and durability. Furthermore, HPLC can be equipped with multiple detectors such as UV detector, ELSD, FD, and so on. Fan et al [56] chose ten major active components in Carthamus tinctorius L. to carry out a qualitative and quantitative evaluation based on HPLC-DAD. The RSD of peak area for each of the ten bioactive compounds was calculated, respectively to validate the precision of the method. A comprehensive, two-dimensional HPLC-MS method offers more effective information to identify the unknown compounds. Qi et al [57] applied HPLC-MS to analyze seven isoflavonoids and six saponins in Radix Astragali and its preparations. Compared with conventional HPLC-DAD-ELSD, this new method provided lower LOD and LOQ with an excellent linearity response. Based on the contents of thirteen bioactive ingredients, they analyzed ten commercial crude drug samples and ten prepared samples. The conclusion was drawn that different origins, sources, cultural influences, and harvest time would result in different qualities. Recently, the advent of charged aerosol detection (CAD) has attached much interest. Its principle involves the charging of aerosol particles via corona discharge with subsequent electrometer-based measurement [58]. As a quality sensitive type detector, the response obtained with CAD depends mainly on the quality of the sample and does not depend significantly on the individual analyte properties. Therefore it can detect all of the non-volatile compounds and many of the semi-volatile compounds. Additionally, it has high sensitivity, a wide dynamic response range of up to 4 orders of magnitude, good reproducibility, easy of use, and reliable operation [59]. However, the main disadvantage of this detector is the lack of spectral information, so it cannot be applied to identify a certain peak, or to perform peak purity analysis [60]. Overall, owing to its superiorities, it can serve as a vigorous complementarity to other LC detectors, like UV and MS. Bai et al [61] established a HPLC-CAD method for the simultaneous determination of seven saponins in Radix et Rhizoma Notoginseng. Meanwhile, the LODs and LOQs of three detectors, UV, ELSD,

SONG Xin-Yue, et al. /Chinese Journal of Natural Medicines 2013, 11(6): 596−607

and CAD, were compared. The results showed that the CAD method exhibited a lower LOD and LOQ than the other two detectors. Consequently, it was expected to be a sensitive and universal method for the analysis of TCMs containing weakly UV-absorbing compounds. High-speed counter-current chromatography (HSCCC) is a liquid-liquid partition chromatographic technique. Compared with the other methods, it offers lower solvent consumption, simpler separation conditions, larger loading, and no sample adsorption. Furthermore, the emergence of 2D-HSCCC provides significantly higher resolving power and peak capacity. However, with the development of countercurrent chromatography, there are two main factors restricting its wide application. Firstly, there is no mature and systematic theory to guide a suitable choice of solvent systems. Therefore, many experiments need to be carried out to choose a satisfactory solvent combination. Moreover, the separation mechanism of this technique requires in-depth theoretical investigation to be suitable for industrial application [62]. Turbulent flow chromatography (TFC) was introduced by Quinn and Takarewski in the late 1990s as a fast technique for the direct injection of complicated matrices, like biological fluids [63]. Nowadays, TFC acts as an effective, high-throughput, sample preparation technique which makes use of high flow rates in 0.5 or 1.0 mm internal diameter columns packed with particles in the size range of 30–60 µm [64]. Owing to the suitable flow rate and the special columns, turbulence will form. Under turbulent flow conditions, small molecules diffuse more extensively than macromolecules so that they are driven into the pores of the sorbent while the larger molecules and matrix constituents are flushed to waste without diffusing into the pores, so the small molecules are separated from macromolecules [63]. The main bottleneck of this novel technique is the relatively poor chromatographic separation capacity due to the turbulent flow columns with lower theoretical plate numbers than analytical columns. For this reason, two column methods are preferred in most cases. In the two-column approach, the analytes are first separated from the macromolecules and retained on the turbulent flow column. They are then eluted to a second analytical column for analysis [65-67]. The outstanding advantage of TFC is the direct injection and elution of the analyte, which removes time-consuming off-line steps, like evaporation, reconstitution, and preparation. Therefore, it is more efficient, automated, and requires far less time than the off-line counterparts [62, 68]. 3.2.4 Capillary electrophoresis and related techniques Capillary electrophoresis (CE) is becoming more and more popular for the separation and quantification of mixtures of natural compounds. Compared with other analytical methods, it has superiority, including high peak capacity, low sample and reagent consumption, high speed analysis and efficiency, excellent mass sensitivity, and cost-effectiveness. Therefore, CE has been widely used in the identification and separation of medicinal plant components, such as alkaloids,

polyphenols, carbohydrates, lipid, and terpenes, accompanied with capillary zone electrophoresis (CZE), micellar electrokinetic chromatography (MEKC), non-aqueous CE (NACE), capillary electrochromatography (CEC), and pressurized capillary electrochromatography (p-CEC). However, the small injection volume and a short optical path length in the most commonly used UV detector result in the poor sensitivity of CE. Consequently, two main methods have been developed to overcome this drawback. One is to use more sensitive detection methods, such as laser-induced fluorescence, electrochemical, MS, fluorescence, and chemiluminescence detectors, the other is pre-concentration of the test samples, such as stacking, sweeping, and field-amplified sample injection (FASI) [2]. Recently a novel hybrid technique, known as p-CEC coupled with a micro-HPLC pump, was proposed to increase the speed of separation and avoid bubble formation. Lu et al. [69] separated and determined the structurally-related anthraquinones in rhubarb (Rheum sp.) by p-CEC. Additionally, a monolithic column was designed through in situ polymerization which offered a unique pore structure with high permeability and favorable mass transfer characteristics. The anthraquinone compounds in rhubarb were formerly determined by TLC and HPLC. However, neither of the methods was entirely adequate because of either poor resolution or consuming-time analysis.

4

Chromatographic Fingerprint Method

Although selecting several active ingredients as evaluation markers is an effective analytical approach, the matrix of chemical constituents in TCMs is complex, which makes their separation and screening extremely difficult. Since different plants may contain the same compounds, the former method would fail to confirm the identity of a specific plant, let alone determine its quality accurately. Therefore it is essential to develop a more demanding and feasible way. Fingerprint analysis is considered as a more effective method, since it emphasizes more of the characteristics of TCM plant materials. Recently, chromatographic fingerprinting has gained great attention and been widely accepted by WHO, German Commission E, the British Herbal Medicine Association, the Indian Drug Manufacturers’ Association, and some other official or non-official organizations [70-74]. In addition, compulsory fingerprint analysis was also proposed by the Chinese State Food and Drug Administration (SFDA) to control the quality of Chinese medicine injections in 2004 [6]. The development of fingerprint methods focuses on two key issues. One is how to acquire more effective and reliable information, while the other is how to evaluate the similarity with chemometric methods. Generally, chromatographic fingerprinting is made by analyzing a number of samples to determine the common and stable compounds. In this review, more attention is paid to the development of fingerprint analysis with chromatographic methods.

SONG Xin-Yue, et al. /Chinese Journal of Natural Medicines 2013, 11(6): 596−607

4.1 Fingerprint analysis combined with multiple chromatographic separation and detection methods The development of multiple chromatographic separation and detection methods has provided great progress in fingerprint analysis. HPLC, CE, GC, etc. perform an important function in the determination and separation of active ingredients. Sun et al [75] employed CE to develop the fingerprint of Flos Carthami, while HPLC was applied for the fingerprint analysis of Psoralea corylifolia L. [76]. Ten common peaks were identified by MS data. The fingerprint profile was then used to identify and assess the differences among the plant materials grown in various areas of China. A chromatographic fingerprint of coffee flavor was developed with GC-MS and chemometric methods. Then the developed chromatographic fingerprint was successfully used to differentiate coffee flavor from coco flavor by both similarity comparison and principal reserved [77]. 4.2 Multiple-patterns of chromatographic fingerprinting Undoubtedly, chromatographic fingerprinting has become one of the most powerful methods to ensure the efficacy and safety of plant medicines. However, most of reported chromatographic fingerprints are carried out as a single chromatogram, which sometimes may be inadequate for multi-herb botanical products such as TCMs. Fan et al. [78] developed with multiple-chromatographic fingerprints which consist of more than one chromatographic fingerprint to represent the whole characteristics of the chemical constitutions of complex medicines. They adopted two extraction methods to create different HPLC chromatograms, then used the retention time, UV absorbance, and MS spectra of [M – H]– ions to make the qualitative and quantitative analysis. A date-level information fusion method was employed to acquire a fuses vector which represented the integrated chemical characteristics of binary fingerprints. Consequently, the lot-to-lot consistency and fraudulent materials were determined, either by using similarity measures or by a chemometrics approach. Undoubtedly, multiple-chromatographic fingerprints will give more detailed and accurate information about plant materials. At the same time, complicated data processing will be a challenging issue. 4.3 Fingerprint with multi-ingredients quantitative analysis As is known, fingerprint analysis can offer holistic and integrated characteristic information of traditional Chinese medicines. However, one main shortcoming is that it only shows the results of the similarity calculated on the relative value using a pre-selected marker compound as the reference standard. Therefore minor differences between very similar chromatograms might not be distinguishable [79]. Multiple-ingredient analysis would address this problem. Yang et al [80] developed a novel method combining HPLC fingerprint and multiple-ingredient quantitative analysis for the quality evaluation of Shuang-huang-lian oral liquid formulation. For fingerprint analysis, forty-five peaks were selected as the common peaks to evaluate the similarities

among several different Shuang-huang-lian oral liquid preparations collected from different manufacturers. In addition, simultaneous quantification of eleven markers, including chlorogenic acid, caffeic acid, rutin, forsythiaside, scutellarin, baicallin, forsythin, luteoloside, apigenin, baicalein, and wogonin was performed. Statistical analysis of the obtained data demonstrated that the developed method had achieved the desired linearity, precision, and accuracy. 4.4 Bio-fingerprint and meta-fingerprint The group of Zou and Li [34] proposed a novel bio-fingerprint to screen and analyze multiple bioactive compounds by comparing the difference between the fingerprint chromatograms of the extract of TCMs before and after interaction with biological systems, such as DNA, protein, cells, etc. According to the different sampling methods, the bio-fingerprint analyses were classified into three types, including microlysis, equilibrium dialysis, and cell or target molecule extraction. Based on these novel methods, Angelica sinensis (Oliv.) Diels, Cordyceps sinensis , combined formulations of Danggui Buxue decoction, Shaofu Zhuyu decoction, etc. were analyzed by the different cell extraction methods [81-83] . Similarly, meta-fingerprint analysis is a sensitive way to determine the bioactive ingredients through comparing chemical profiles of TCMs and their metabolite profiles. It is of great importance to clarify the effects of the active constituents on pharmacological systems, in order to guarantee the clinical efficacy of plant medicines. Through comparing the chemical and metabolic fingerprint of Danshen injection, a polyphenolic acid was found to be the bioactive ingredient [83] . Therefore, based on this study, the indexes for fingerprinting could be optimized to improve the quality control efficiency. However, this method may be laborious and time-consuming. Tan et al [84] developed a feasible method to evaluate the quality of Uncaria sinensis Havil. from different sources by primary and secondary meta-chromatography fingerprints. Firstly, two primary metabolic fingerprints were developed using a GC-MS method, and four secondary metabolic fingerprints were developed using HPLC-MS. Then, principal component analysis (PCA) was used to discriminate the samples since the score plot showed a clear batch-to-batch differentiation between the samples. Thus, the combination of primary and secondary meta-fingerprinting is a suitable method to assess the quality of TCMs comprehensively and powerfully. However, the metabolites may not be reproducible due to the collection time or to the variation between individual plants, which needs further scientific experiment design and data analysis [82]. 4.5 Statistic analysis for chromatographic fingerprinting Chromatographic fingerprinting can provide comprehensive and integrated information of plant materials. Therefore, powerful statistical analysis should be applied to acquire the useful inferences successfully [85]. Li et al [86] used LC-MS, combined with hierarchical cluster analysis, to characterize the distribution of steroidal alkaloids in Fritillaria species

SONG Xin-Yue, et al. /Chinese Journal of Natural Medicines 2013, 11(6): 596−607

and related compound formulas. Due to the strong identification ability of LC-MS, twenty-seven steroidal alkaloids in seventeen Fritillaria species and twelve Bulbus Fritillariae-containing compound formulas, produced a perfect separation. Using this method, it was possible to set up a chromatographic fingerprint of Fritillariae species, and then use hierarchical cluster analysis of the Statistical Product and Service Solutions (SPSS) statistical package to classify sixty-two specimens of seventeen examined Fritillaria species into three groups. Thus, statistical analysis performs an essential role in acquiring meaningful information from a large volume of data by virtue of powerful professional software.

5 Holistic Analysis of Multiple Constituents from Each Component Plant in a Single Run Although chromatography fingerprinting can offer integrated and adequate information, it is a time-consuming process, and typically needs special software and analytical methods. Recently, aimed at attaining more effective quality control of TCM preparations, this group has proposed a new strategy, named as holistic analysis of multiple constituents from each component plant in a single run. According to the theory and medicinal practice of TCM, all of the medicinal plants in a composition are responsible for the therapeutic effects. Therefore, certain bioactive constituents from each medicinal plant in the composition are selected as evaluation markers, and then their quantitative information is obtained in a single run with an appropriate analytical method (the strategy is outlined in Figure 1) [87]. This strategy was successfully applied for assessing the quality of the Chinese medicine Qin-Bao-Hong antitussive tablet, the Yiqing granule, and the Wuji pill. It was proven to be a simple, reasonable, feasible, and comprehensive technique for the quality control of TCM preparation [87-89]. Qin-Bao-Hong antitussive tablet, a Chinese medicinal preparation, contains three Chinese plants, including Scutellaria baicalensis Georgi roots, Syringa amurensis Rupr. bark, and Rhododendron dauricum L. leaves. According to the proposed strategy, baicalin and baicalein from S. baicalensis, syringin from S. amurensis, and hyperoside, quercetin, and farrerol from R. dauricum were selected as the evaluation markers, and their quantitative information was obtained in a single run with HPLC-PDA. The method was then successfully used for the quality control of Qin-Bao-Hong antitussive tablet from different production batches. In 2010, a simple, accurate, and reliable UPLC method with PDA was developed for the simultaneous quantification of twelve evaluation markers in Yiqing granule, namely, berberine, palmatine, and jatrorrhizine from Rhizoma Coptidis, aloe emodin, rhein, emodin, chrysophanol, and physcion from Radix et Rhizoma Rhei, baicalin, baicalein, wogonoside, and wogonin from Radix Scutellariae. The developed method was applied to evaluate the quality of the products from different manufac

Fig. 1 Holistic analysis of multiple constituents from each component plant in a single run [87]

Fig. 2 Chromatograms obtained from a mixed standard solution (A) and from Yiqing granule sample solution (B). The peaks marked 1-12 were baicalin, wogonoside, baicalein, aloe emodin, wogonin, rhein, jatrorrhizine, palmatine, berberine, emodin, chrysophanol, and physcion, respectively [89]

turers. As shown in Fig. 2, the twelve components were well-separated owing to the high separation efficiency of UPLC. Likewise, the holistic analysis of twelve constituentsfrom different manufacturers in a single run by UPLC was applied to a quality control study.

6 Significant Issues in Further Research on the Quality Control of TCMs 6.1

Tedious statistical analysis Quality control of TCMs involves troublesome statistical analysis, such as the process of HTC to select bioactive ingredients, and the similarity analysis of fingerprint chromatography. For the similarity analysis of fingerprint chromatography, the shift and repeatability of the chromatographic peaks increases the challenge to identify the compounds. Although many methods have been developed to solve this problem, how to obtain enough useful information

SONG Xin-Yue, et al. /Chinese Journal of Natural Medicines 2013, 11(6): 596−607

from chromatographic fingerprinting, and then to use that information to arrive at a powerful conclusion for the quality control, are also challenging. Fortunately, computer-based software such as Computer Aided Software Engineering (CASE) and SPSS allow for peak identification and matching, and background and retention time correction, which perform a vital function in developing fingerprints. 6.2 Absence of powerful theory support Unquestionably, TCMs are beneficial to our health, but how the active ingredients interact with each other to perform the curative efficacy is difficult to elucidate. The interaction between curative efficacy and the structures of compounds should be elucidated clearly to assist in the wide acceptance of TCMs. Finally, the relationship between chromatographic fingerprinting and the safety and efficacy of TCMs is another unsolved question. Consequently, the strategies and theories for the quality control of TCMs need to be under continuous development.

Abbreviations SFDA, Chinese State Food and Drug Administration; SPSS, Statistical Product and Service Solutions; CASE, Computer Aided Software Engineering; EM, Evaluation Marker; CF, Chromatographic Fingerprint.

[10]

[11]

[12]

[13]

[14]

[15]

[16]

[17]

References [1]

[2]

[3]

[4]

[5]

[6]

[7] [8]

[9]

Li P, Qi LW, Liu EH, et al. Analysis of Chinese herbal medicines with holistic approaches and integrated evaluation models [J]. Trends Anal Chem, 2008, 27 (1): 66-77. Wang BL, Hu JP, Tan W, et al. Simultaneous quantification of four active Schisandra lignans from a traditional Chinese medicine Schisandra chinensis (Wuweizi) in rat plasma using liquid chromatography/mass spectrometry [J]. J Chromatogr B, 2008, 865 (1-2): 114-120. Jiang Y, David B, Tui PF, et al. Recent analytical approaches in quality control of traditional Chinese medicines - A review [J]. Anal Chim Acta, 2010, 657 (1): 9-18. Wen XD, Qi LW, Chen J, et al. Analysis of interaction property of bioactive components in Danggui Buxue Decoction (DBD) with protein by microdialysis coupled with HPLC-DAD-MS [J]. J Chromatogr B, 2007, 852 (1-2): 598-604. Razmovski-Naumovski V, Tongkao-on W, Kimble B, et al. Multiple chromatographic and chemometric methods for quality standardisation of Chinese herbal medicines [J]. World Sci Technol, 2010, 12 (1): 99-106. Li SL, Han QB, Qiao CF, et al. Chemical markers for the quality control of herbal medicines: an overview [J]. Chin Med, 2008, 3: 7. Liang YZ, Xie PS, Chan K. Quality control of herbal medicines [J]. J Chromatogr B, 2004, 812 (1-2): 53-70. Liu L, Li YF, Cheng YY. A method for the production and characterization of fractionated libraries from Chinese herbal formulas [J]. J Chromatogr B, 2008, 862 (1-2): 196-204. Zhao Y, Zhou X, Chen HG, et al. Determination of dehydroevodiamine in Evodia rutaecarpa (Juss.) Benth. by high performance liquid chromatography and classification of the sam-

[18]

[19]

[20]

[21]

[22]

[23]

[24]

[25]

ples by using hierarchical clustering analysis [J]. Fitoterapia, 2009, 80 (7): 415-420. Sharma KK. Determination of active ingredient in synthetic pyrethroid formulations by high-performance thin-layer chromatography/densitometry [J]. J AOAC Int, 2002, 85 (6): 1420-1424. Chen J, Liu X, Shi YP. Determination of daphnetin in Daphne tangutica and its medicinal preparation by liquid chromatography [J]. Anal Chim Acta, 2004, 523 (1): 29-33. Wang ZJ, Wo SK, Wang L, et al. Simultaneous quantification of active components in the herbs and products of Si-Wu-Tang by high performance liquid chromatography-mass spectrometry [J]. J Pharm Biomed Anal, 2009, 50 (2): 232-244. Lin G, Li P, Li SL, et al. Chromatographic analysis of Fritillaria isosteroidal alkaloids, the active ingredients of Beimu, the antitussive traditional Chinese medicinal herb [J]. J Chromatogr A, 2001, 935 (1-2): 321-338. Zhao HY, Jiang, JG. Application of chromatography technology in the separation of active components from nature derived drugs [J]. Mini-Rev Med Chem, 2010, 10 (13): 1223-1234. Li HX, Ding MY, Lv K, et al. Determination of the active ingredients in Chuanxiong by HPLC, HPLC-MS and EI-MS [J]. J Lip Chromatogr Related Technol, 2001, 24 (13): 2017-2031. Guo YH, Luo X, Yu MY, et al. Active ingredients and efficacies of Ganoderma lucidum cultivated on non-medicinal parts of Chinese medicinal herbs [J]. Acta Microbial Sin, 2011, 51 (6): 764-768. Cao YH, Lou CG, Fang YZ, et al, Determination of active ingredients of Rhododendron dauricum L. by capillary electrophoresis with electrochemical detection [J]. J Chromatogr A, 2002, 943 (1): 153-157. Cui YY, Feng SY, Zhao G, et al. HPLC analysis of the active ingredients of Forsythia suspensa [J]. Acta Pharm Sin, 1992, 27 (8): 603-608. Cao YH, Zhang X, Fang YH, et al. Determination of active ingredients of Apocynum venetum by capillary electrophoresis with electrochemical detection [J]. Mikrochim Acta, 2001, 137 (1-2): 57-62. Tang YP, Zhu M, Yu S, et al. Identification and comparative quantification of bio-active phthalides in essential oils from Si-Wu-Tang, Fo-Shou-San, Radix Angelica and Rhizoma Chuanxiong [J]. Molecules, 2010, 15 (1): 341-351. Peng YY, Liu FH, Ye JN. Determination of phenolic acids and flavones in Lonicera japonica Thumb, by capillary electrophoresis with electrochemical detection [J]. Electroanalysis, 2005, 17 (4): 356-362. Ohtaka N, Nakai Y, Yamamoto M. Separation and isolation methods for analysis of the active principles of Sho-saiko-to (SST) oriental medicine [J]. J Chromatogr B, 2004, 812 (1-2): 135-148. Wang ZM, Gao HM, Wang WG. Multi-components quantitation by one marker new method for quality evaluation of Chinese herbal medicine [J]. Chin J Chin Mat Med, 2006, 31 (23): 1925-1928. National Pharmacopoeia Committee. Chinese Pharmacopoeia [M]. Chinese Medical Science and Technology Publishing Company, 2010: 285. Wang CQ, Jia XH, Chen J, et al. Systematic study on QAMS method for quality control of Panax notoginseng [J]. Chin J Chin Mat Med, 2012, 37 (22): 3438-3445.

SONG Xin-Yue, et al. /Chinese Journal of Natural Medicines 2013, 11(6): 596−607 [26] He FC, Li SX, Zhao ZQ, et al. Simultaneous quantitative analysis of four lignanoids in Schisandra chinensis by quantitative analysis of multi-components by single marker [J]. Chin J Chin Mat Med, 2012, 47 (7): 930-933. [27] Yang F, Wan L, Hu YC, et al. Simultaneous determination of three coumarins in Angelica dahurica var. formosana by QAMS [J]. Chin J Chin Mat Med, 2012, 37 (7): 956-960. [28] Li Q, Liu W, Luo ZL, et al. Simultaneous determination of four tanshinones in Salvia miltiorrhiza by QAMS [J]. Chin J Chin Mat Med, 2012, 37 (6): 824-828. [29] Huang SJ, Yang QW, Shi YH, et al. Simultaneous assay of paeoniflorin and albiflorin in Paeoniae Radix Alba by QAMS [J]. Chin J Chin Mat Med, 2011, 36 (6): 780-783. [30] Ganbold M, Barker J, Ma R, et al. Cytotoxicity and bioavailability studies on a decoction of Oldenlandia diffusa and its fractions separated by HPLC [J]. J Ethnopharmacol, 2010, 131 (2): 396-403. [31] Casanova M, You L, Gaido KW, et al. Developmental effects of dietary phytoestrogens in Sprague-Dawley rats and interactions of genistein and daidzein with rat estrogen receptors alpha and beta in vitro [J]. Toxicol Sci 1999, 51 (2): 236-244. [32] Prasad V, Scotch R, Chaudhuri AR, et al. Interactions of bovine brain tubulin with pyridostigmine bromide and N, N’-diethyl-m-toluamide [J]. Neurochem Res, 2000, 25 (1): 19-25. [33] Kedei N, Lundberg DJ, Toth A, et al. Characterization of the interaction of ingenol 3-angelate with protein kinase C [J]. Cancer Res, 2004, 64 (9): 3243-3255. [34] Zhou P, Li P, Wei YH. Interactions of Fufang Danshen Prescription with liposome biomembrane [J]. Chin J Nat Med, 2008, 6 (4): 278-282. [35] Mohammadi F, Bordbar AK, Divsalar A, et al. Analysis of binding interaction of curcumin and diacetylcurcumin with human and bovine serum albumin using fluorescence and circular dichroism spectroscopy [J]. Protein J, 2009, 28 (3-4): 189-196. [36] Su XY, Kong L, Li X, et al. Screening and analysis of bioactive compounds with biofingerprinting chromatogram analysis of traditional Chinese medicines targeting DNA by microdialysis/HPLC [J]. J Chromatogr A, 2005, 1076 (1-2): 118-126. [37] Li SL, Li P, Sheng LH, et al. Live cell extraction and HPLC-MS analysis for predicting bioactive components of traditional Chinese medicines [J]. J Pharm Biomed Anal, 2006, 41 (2): 576-581. [38] Zhang X, Qi LW, Yi L, et al. Screening and identification of potential bioactive components in a combined prescription of Danggui Buxue decoction using cell extraction coupled with high performance liquid chromatography [J]. Biomed Chromatogr, 2008, 22 (2): 157-163. [39] Zhang H, Hu CX, Liu CP, et al. Screening and analysis of bioactive compounds in traditional Chinese medicines using cell extract and gas chromatography-mass spectrometry [J]. J Pharm Biomed Anal, 2007, 43 (1): 151-157. [40] Wah LK, Abas F, Cordell GA, et al. Steroids from Dysoxylum grande (Meliaceae) leaves [J]. Steroids, 2013, 78 (2): 210-219. [41] Bag A, Bhattacharyya SK, Pal NK, et al. In vitro antibacterial potential of Eugenia jambolana seed extracts against multidrug-resistant human bacterial pathogens [J]. Microbiol Res, 2012, 167 (6): 352-357. [42] Annegowda HV, Mordi MN, Ramanathan S, et al. Effect of extraction techniques on phenolic content, antioxidant and an-

[43]

[44]

[45]

[46]

[47]

[48]

[49]

[50]

[51]

[52]

[53]

[54]

[55]

[56]

[57]

timicrobial activity of Bauhinia purpurea: HPTLC determination of antioxidants [J]. Food Anal Meth, 2012, 5 (2): 226-233. Wszelaki N, Paradowska K, Jamroz MK, et al. Bioactivityguided fractionation for the butyrylcholinesterase inhibitory activity of furanocoumarins from Angelica archangelica L. roots and fruits [J]. J Agric Food Chem, 2011, 59 (17): 9186-9193. Gu LH, Wu T, Wang ZT. TLC bioautography-guided isolation of antioxidants from fruit of Perilla frutescens var. acuta [J]. LWT Food Sci Technol, 2009, 42 (1): 131-136. Al-Sayah MA, Rizos P, Antonucci V, et al. High throughput screening of active pharmaceutical ingredients by UPLC [J]. J Sep Sci, 2008, 31 (12): 2167-2172. Liu YY, Lin ZQ, Zhang SC, et al. Rapid screening of active ingredients in drugs by mass spectrometry with low- temperature plasma probe [J]. Anal Bioanal Chem, 2009, 395 (3): 591-599. Naoghare PK, Song JM. Chip-based high throughput screening of herbal medicines [J]. Comb Chem High T Scr, 2010, 13 (10): 923-931. Song JZ, Li SL, Zhou Y, et al. A novel approach to rapidly explore analytical markers for quality control of Radix Salviae Miltiorrhizae extract granules by robust principal component analysis with ultra-high performance liquid chromatographyultraviolet-quadrupole time-of-flight mass spectrometry [J]. J Pharm Biomed Anal, 2010, 53 (3): 279-286. Hauck HE, Bund O, Fischer W, et al. Ultra-thin layer chromatography (UTLC)-a new dimension in thin-layer chromatography [J]. J Planar Chromatogr–Mod TLC, 2001, 14 (4): 234-236. Vermaak I, Hamman JH, Viljoen AM. High performance thin layer chromatography as a method to authenticate Hoodia gordonii raw material and products [J]. S Afr J Bot, 2010, 76 (1): 119-124. Papp E, Bagocsi B, H-Otta K, et al. The role of over-pressuredlayer chromatography among chromatographic methods for the determination of the aflatoxin content of fish [J]. J Planar Chromatogr–Mod TLC, 1999, 12 (5): 383-387. Deng CH, Ji J, Wang XC, et al. Development of pressurized hot water extraction followed by headspace-solid-phase microextraction and gas chromatography-mass spectrometry for determination of ligustilides in Ligusticum chuanxiong and Angelica sinensis [J]. J Sep Sci, 2005, 28 (11): 1237-1243. Dong L, Wang JY, Deng CH, et al. Gas chromatography-mass spectrometry following pressurized hot water extraction and solid-phase microextraction for quantification of eucalyptol, camphor, and borneol in chrysanthemum flowers [J]. J Sep Sic, 2007, 30 (1): 86-89. Ding B, Zhou TT, Fan GR, et al. Qualitative and quantitative determination of ten alkaloids in traditional Chinese medicine Corydalis yanhusuo WT Wang by LC-MS/MS and LC-DAD [J]. J Pharm Biomed Anal, 2007, 45 (2): 219-226. Lee MC, Tsao CH, Lou SC, et al. Analysis of aristolochic acids in herbal medicines by LC/UV and LC/MS [J]. J Sep Sci, 2003, 26 (9-10): 818-822. Fan L, Zhao HY, Xu M, et al. Qualitative evaluation and quantitative determination of ten major active components in Carthamus tinctorius L. by high-performance liquid chromatography coupled with diode array detector [J]. J Chromatogr A, 2009, 1216 (11): 2063-2070. Qi LW, Li P, Ren MT, et al. Application of high-performance liquid chromatography-electrospray ionization time-of-flight

SONG Xin-Yue, et al. /Chinese Journal of Natural Medicines 2013, 11(6): 596−607

[58]

[59]

[60]

[61]

[62]

[63]

[64]

[65]

[66]

[67]

[68]

[69]

[70] [71] [72] [73] [74]

mass spectrometry for analysis and quality control of Radix Astragali and its preparations [J]. J Chromatogr A, 2009, 1216 (11): 2087-2097. Vehovec T, Obreza A. Review of operating principle and applications of the charged aerosol detector [J]. J Chromatogr A, 2010, 1217 (10): 1549-1556. Gamache PH, Mccarthy RS, Freeto SM, et al. HPLC analysis of nonvolatile analytes using charged aerosol detection [J]. LCGC North Am, 2005, 23 (2): 150. Vervoort N, Daemen D, Torok G. Performance evaluation of evaporative light scattering detection and charged aerosol detection in reversed phase liquid chromatography [J]. J Chromatogr A, 2008, 1189 (1-2): 92-100. Bai CC, Han SY, Chai YY, et al. Sensitive determination of saponins in Radix et Rhizoma Notoginseng by charged aerosol detector coupled with HPLC [J]. J Liq Chromatogr Relat Technol, 2009, 32 (2): 242-260. Di DL, Zheng YY, Chen YF, et al. Advance in application of high-speed countercurrent chromatography in separation and purification of flavonoids [J]. Chin J Anal Chem, 2011, 39 (2): 269-275. Zimmer D, Pickard V, Czembor W, et al. Comparison of turbulent-flow chromatography with automated solid-phase extraction in 96-well plates and liquid-liquid extraction used as plasma sample preparation techniques for liquid chromatography-tandem mass spectrometry [J]. J Chromatogr A, 1999, 854 (1-2): 23-35. Stolker AAM, Peters RJB, Zuiderent R, et al. Fully automated screening of veterinary drugs in milk by turbulent flow chromatography and tandem mass spectrometry [J]. Anal Bioanal Chem, 2010, 397 (7): 2841-2849. Moeller BC, Stanley SD. The development and validation of a turbulent flow chromatography-tandem mass spectrometry method for the endogenous steroid profiling of equine serum [J]. J Chromatogr B, 2012, 905: 1-9. Xu Y, Willson KJ, Musson DG. Strategies on efficient method development of on-line extraction assays for determination of MK-0974 in human plasma and urine using turbulent-flow chromatography and tandem mass spectrometry [J]. J Chromatogr B, 2008, 863: 64-73. Ynddal L, Hansen SH. On-line turbulent flow chromatography high-performance liquid chromatography-mass spectrometry for fast sample preparation and quantitation [J]. J Chromatogr A, 2003, 1020 (1): 59-67. Ceglarek U, Lembcke J, Fiedler GM, et al. Rapid simultaneous quantification of immunosuppressants in transplant patients by turbulent flow chromatography combined with tandem mass spectrometry [J]. Clin Chim Acta, 2004, 346 (2): 181-190. Lu HX, Wang JB, Wang XC, et al. Rapid separation and determination of structurally related anthraquinones in rhubarb by pressurized capillary electrochromatography [J]. J Pharm Biomed Anal, 2007, 43 (1): 352-357. WHO. General Guidelines for Methodologies on Research and Evaluation of Traditional Medicine (2000). FDA. Guidance for Industry—Botanical Drug Products (Draft Guidance) (2000). EMEA. Final Proposals for Revision of the Note for Guidance on Quality of Herbal Remedies (1999). Forward. British Herbal Medicine Association (1996). Indian Drug Manufacturers’ Association (1998).

[75] SunY, Guo T, Sui Y, et al. Fingerprint analysis of Flos Carthami by capillary electrophoresis [J]. J Chromatogr B, 2003, 792 (2): 147-152. [76] Zhao LH, Huang CY, Shan Z, et al. Fingerprint analysis of Psoralea corylifolia L. by HPLC and LC-MS [J]. J Chromatogr B, 2005, 821 (1): 67-74. [77] Huang LF, Wu MJ, Zhong KJ, et al. Fingerprint developing of coffee flavor by gas chromatography-mass spectrometry and combined chemometrics methods [J]. Anal Chim Acta, 2007, 588 (2): 216-223. [78] Fan XH, Cheng YY, Ye ZL, et al. Multiple chromatographic fingerprinting and its application to the quality control of herbal medicines [J]. Anal Chim Acta, 2006, 555 (2): 217-224. [79] Wang BL, Hu JP, Tan W, et al. Simultaneous quantification of four active schisandra lignans from a traditional Chinese medicine Schisandra chinensis in rat plasma using liquid chromatography/mass spectrometry [J]. J Chromatogr B, 2008, 865 (1-2): 114-120. [80] Yang DZ, An TQ, Jiang XL, et al. Development of a novel method combining HPLC fingerprint and multi-ingredients quantitative analysis for quality evaluation of traditional Chinese medicine preparation [J]. Talanta, 2011, 85 (2): 885-890. [81] Chio YH, Sertic S, Kim HK, et al. Classification of Ilex species based on metabolomic fingerprinting using nuclear magnetic resonance and multivariate data analysis [J]. J Agric Food Chem, 2005, 53 (4): 1237-1245. [82] Choi YH, Kim HK, Hazekamp A, et al. Metabolomic differentiation of Cannabis sativa cultivars using 1H NMR spectroscopy and principal component analysis [J]. J Nat Prad, 2004, 67 (6): 953-957. [83] Zhang JL, Cui M, He Y, et al. Chemical fingerprint and metabolic fingerprint analysis of Danshen injection by HPLC-UV and HPLC-MS methods [J]. J Pharm Biomed Anal, 2005, 36 (5): 1029-1035. [84] Tan SN, Yong JWH, Teo CC, et al. Determination of metabolites in Uncaria sinensis by HPLC and GC-MS after green solvent microwave-assisted extraction [J]. Talanta, 2001, 83 (3): 891-898. [85] Sun MJ, Bai L, Liu DQ. A generic approach for the determination of trace hydrazine in drug substances using in situ derivatizaion-headspace GC-MS [J]. J Pharm Biomed Anal, 2009, 49 (2): 529-533. [86] Li HJ, Jiang Y, Li P. Characterizing distribution of steroidal alkaloids in Fritillaria spp. and related compound formulas by liquid chromatography-mass spectrometry combined with hierarchial cluster analysis [J]. J Chromatogr A, 2009, 1216 (11): 2142-2149. [87] Chen J, Sun JN, Song XY, et al. Holistic analysis of seven constituents from three medicinal herbs composing Wuji pills in a single run by ultra performance liquid chromatography: application to quality control study [J]. Anal Meth, 2012, 4 (9): 2989-2995. [88] Chen J, Wang GY, Shi YP. Method development and validation for simultaneous HPLC analysis of six active components of the Chinese medicine Qin-Bao-Hong antitussive tablet [J]. Acta Chromatogr, 2009, 21 (2): 341-354. [89] Chen J, Yang Y, Shi YP. Simultaneous quantification of twelve active components in Yiqing granule by ultra-performance liquid chromatography: application to quality control study [J]. Biomed Chromatogr, 2011, 25 (9): 1045-1053.

SONG Xin-Yue, et al. /Chinese Journal of Natural Medicines 2013, 11(6): 596−607

中药质量控制方法的进展 宋昕玥 1, 3, 李应东 2, 师彦平 1, 晋

玲 2 *, 陈

娟 1*

1

西北特色植物资源化学重点实验室和甘肃省天然药物重点实验室,中国科学院兰州化学物理研究所,兰州 730000;

2

甘肃中医学院,兰州 730000;

3

中国科学院大学,北京 100039

【摘 要】 中药在世界特别是发展中国家需求量大,凭借其优越的特点例如价廉、副作用少,更好的文化可接受性和人体 适应性,被广泛应用于医疗保健中。然而,复方中药由多种中药材组成,而每一种中药材中就含有几百甚至几千种化学成分。 这些化学成分是如何相互作用的, 以及哪些又是其发挥药效的活性成分等问题是限制中药现代化和国际化的主要瓶颈。因此, 建立有效、合理的方法评价和控制中药质量是必需的.随着分离和色谱技术的发展,质控方法也在不断完善。该综述从发展的角 度概述了中药质量控制方法,从单指标成分到多指标成分分析,再到新兴的指纹图谱技术。总之,中药的分析和质量控制方法 正朝着更有效、更全面的方向发展,以更好的反映中药的内在整体性质。 【关键词】 中药; 质量控制; 活性成分; 指纹图谱技术

【基金项目】

“十二五”国家科技支撑计划项目(No. 2011BAI05B02)和国家自然科学基金(No. 21105106)资助

Quality control of traditional Chinese medicines: a review.

Traditional Chinese medicines (TCMs) are in great demand all over the world, especially in the developing world, for primary health care due to their ...
937KB Sizes 0 Downloads 0 Views