Trends in Analytical Chemistry 61 (2014) 207214

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Trends in Analytical Chemistry

j o u r n a l h o m e p a g e : w w w. e l s e v i e r. c o m / l o c a t e / t r a c

Metabolomics and traditional Chinese medicine

Chunxiu Hu, Guowang Xu *
Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 457 Zhongshan Road,
Dalian 116023, China

A R T I C L E

I N F O

Keywords:
Analytical characterization
Bioactive component
Biochemistry
Chinese herbal medicine
Chromatographic ngerprinting
Holistic bioactivity
Metabolomics
Multi-component mixture
TCM syndrome
Traditional Chinese medicine

A B S T R A C T

Traditional Chinese medicine (TCM) has attracted increasing interest throughout the world because of
its potential as complementary therapy of choice and a source for discovery of new drugs. However, the
analytical characterization of TCM is still in its infancy due to its chemical nature of multi-component
mixtures that often possess their own inherent holistic bioactivities. Metabolomics incorporates stateof-the-art approaches enabling systemic molecular characterization in complex samples and provides
the option for an integrated view of the biochemistry in TCM. In this review, we present current
metabolomics technologies in research on TCM related to chemical composition, bioactive components
and ecacy. We also highlight the potential role of metabolomics technologies in evidence-based studies
of TCM syndromes.
2014 Elsevier B.V. All rights reserved.

Contents
1.
2.
3.

4.

5.

Introduction ........................................................................................................................................................................................................................................................
Advances in metabolomics technologies ..................................................................................................................................................................................................
Applications of metabolomics in systems-based TCM .........................................................................................................................................................................
Chemical characterization of CHM ................................................................................................................................................................................................
3.1.
Bioactive component screening of CHM .....................................................................................................................................................................................
3.2.
Ecacy evaluation of CHM ..............................................................................................................................................................................................................
3.3.
Evidence-based studies in TCM syndromes ............................................................................................................................................................................................
TCM pattern diagnosis .......................................................................................................................................................................................................................
4.1.
TCM pattern intervention .................................................................................................................................................................................................................
4.2.
Future perspectives ..........................................................................................................................................................................................................................................
Acknowledgments ............................................................................................................................................................................................................................................
References ............................................................................................................................................................................................................................................................

Abbreviations: APCI, Atmospheric pressure chemical ionization; APPI, Atmospheric pressure photoionization; CE, Capillary electrophoresis; CHM, Chinese herbal
medicine; DART-MS, Direct analysis in real time mass spectrometry; EI, Electron ionization; ESI, Electrospray ionization; FAB, Fast atom bombardment; FT-ICR, Fourier
transform ion cyclotron resonance; GC, Gas chromatography; GC GC, Comprehensive two-dimensional gas chromatography; GK, Goto-Kakizaki; HILIC, Hydrophilic
interaction chromatography; IT, Ion trap; LC, Liquid chromatography; LC LC, Comprehensive two-dimensional liquid chromatography; MALDI, Matrix-assisted laser
desorption/ionization; MS, Mass spectrometry; NMR, Nuclear magnetic resonance; OPLS-DA, Orthogonal partial least squares discriminant analysis; PCA, Principal component analysis; Q, Quadrupole; QQQ, Triple quadrupole; TCM, traditional
Chinese medicine; TOF, Time-of-ight; UPLC, Ultra-performance liquid chromatography; WM, Western medicine.
* Corresponding author. Tel.: +86 411 84379530; Fax: +86 411 84379559.
E-mail address: xugw@dicp.ac.cn (G. Xu).
http://dx.doi.org/10.1016/j.trac.2014.06.007
0165-9936/ 2014 Elsevier B.V. All rights reserved.

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1. Introduction
Interest in study of traditional Chinese medicines (TCMs), or so
called Chinese herbal medicines (CHMs), recently became widespread due to their potential as a source of new drug discovery and
complementary therapy of choice, although the history of TCM can
be dated back to thousands of years ago. TCM holds a holistic health
care philosophy (i.e., looking at the behavior of the system as a
whole). It regards disease as energetic imbalance and adheres to
the concept of cure instead of treatment. The components of CHM
are very complex; CHMs usually consist of hundreds or even thousands of different compounds. The therapeutic effects of CHM can
usually be attributed to the combination of multiple elements

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C. Hu, G. Xu/Trends in Analytical Chemistry 61 (2014) 207214

(i.e., multi-component mixture) of the herb or the formula acting

together.
Traditionally, a TCM is administered as an individually prepared water decoction containing multiple herb materials in certain
proportions. Notably, the nal item that is taken orally by a patient
is an extract of herbs that is inuenced by not only the botanical
origin of its herbal components but also the procedure that is used
to treat the herbal material [1]. Any minor changes in these two
aspects will greatly affect the therapeutic ecacy of the products.
Such complexity in TCM administration brings a great challenge in
establishing suitable analytical approaches for the development of
modernization and standardization of TCMs and their natural products. Besides, TCM possesses two unique features multidimensional pharmacology {i.e., synergetic effects [2], and combinatorial
effects and opposing principles [3]} and personalized or individual treatment. Such features of TCM determine that the traditional
analysis based on major components is not sucient to provide information on bioactivity of herbal products. Analytical characterization of TCM is still in its infancy due to the chemical nature
of multi-component mixtures that often possess inherent holistic
bioactivities.
To meet this challenge, metabolomics which aims at comprehensive characterization of the total metabolome in a biological
system and dynamic metabolomics responses to alterations of inner
and/or external factors [4] is in great demand. As an emerging eld
of science in the post-genomics era, metabolomics has been broadly
applied in many elds {e.g., toxicological survey [5], functional genomics [6,7], clinical diagnostics [8], plant and microbes [911], and
nutritional biochemistry [12,13]}. The most commonly used two technologies in metabolomics applications are nuclear magnetic resonance (NMR) [14] and mass spectrometry (MS) [15]. Although its
sensitivity is less than that of MS, NMR is still popular in many applications due to its rapidity, high throughput, easy operation, being
non-destructive of samples, and quantitation of metabolites from
different groups. However, in most cases, MS is preferred because
of its advantages of unparalleled sensitivity, super resolution and
structural specicity. In practical applications, chromatographic separation technologies [e.g., gas chromatography (GC), liquid chromatography (LC) and capillary electrophoresis (CE)] are often coupled
to MS depending on the properties of the analytes being measured.
As the data matrix generated in metabolomics studies is usually
very large, data processing and data analysis are crucial for their
nal outcome. Many available software packages {e.g., JDAMP [16],
MetAlign [17], MSFACTs [18], Mzmine [19], and XCMS [20]} were
developed for processing raw data from metabolomics. On this basis,
multivariate (e.g., PCA, PLS-DA, PLS, and OPLS) and univariate (e.g.,
T-test and ANOVA) analyses were carried out for classication and
differential metabolite discovery. In combination with advanced bioinformatics tools, many studies demonstrated that metabolomics
incorporating state-of-the-art analytical approaches enables systemic molecular characterization of complex samples [21,22] and
provides an integrated biochemical view of TCM.
In this review, we summarize metabolomics methods based on
GC-MS and LC-MS and applied in research on TCM. We highlight
the subjects related to chemical composition, bioactive components and the ecacy of CHM, and metabolomics applications in
evidence-based studies of TCM syndromes.
2. Advances in metabolomics technologies
As early as the 1970s, the embryonic form of metabolomics (at
that time called metabolic prole) was coming into being. A typical
example is that GC-MS became feasible to measure qualitatively and
quantitatively metabolic proles of body uids and tissue extracts. Concurrently, rapid improvement was achieved in NMR spectroscopy due to the increasing strengths of magnetic eld and magic

angle spinning, which remarkably improved the sensitivity and made

NMR a leading analytical tool to detect metabolites of many different classes in biological samples.
The initiation of study on MS-based metabolomics in a real sense
was carried out by van der Greef et al. using eld desorption and
fast atom bombardment (FAB)-MS for proling complex, nonvolatile metabolites in body uids to study gender difference [23].
They pioneered the utilization of pattern-recognition techniques to
MS-based datasets. Meanwhile, the NMR-based metabonomics approach was pioneered in the study of vertebrate blood and plasma
by Nicholson et al. [24].
Although there were some sporadic applications, the limitation in early analytical technologies (e.g., low sensitivity, low specicity and low throughput) made it a big challenge to qualify and
to quantify complex metabolites in biological samples. Analytical
techniques with super sensitivity and selectivity, high specicity,
good reliability, and easy automation are required to detect and to
identify metabolites in complex biological systems. The revolution in soft-ionization technologies for MS included matrixassisted laser desorption/ionization (MALDI) [25] and electrospray
ionization (ESI) [26] as well as atmospheric pressure chemical ionization (APCI) [27]. Metabolomics has largely been driven by these
technologies, as they possess better resolution and sensitivity, and
higher specicity and mass accuracy than earlier technologies. These
MS instruments are still undergoing rapid advances in terms of sensitivity, resolution, mass accuracy and dynamic range.
As complexity increases in the composition of metabolome the
whole set of endogenous and exogenous metabolites with small molecular weight in an individual organism modern technologies that
allow for qualitative and quantitative measurement of a vast number
of metabolites in complex biological systems are required for
metabolomics studies. Modern metabolomics platforms usually comprise approaches using chromatography-based technologies such
as GC and LC, including high-performance LC (HPLC), ultraperformance LC (UPLC), hydrophilic interaction chromatography
(HILIC)) and CE coupled to MS or MS/MS. Notably, frequently used
ion sources are, for GC-related techniques, electron ionization (EI),
and, for LC- and CE-based MS systems, ESI. Occasionally, APCI and
atmospheric pressure photoionization (APPI) are coupled to LC for
detection of non-polar compounds. Frequently used mass analyzers in current metabolomics include quadrupole (Q), ion trap (IT),
time-of-ight (TOF), Fourier transform ion cyclotron resonance (FTICR), orbital trap and a combination of them [e.g., triple quadrupole (QQQ), Q-TOF, IT-TOF, IT-FT-ICR].
Besides, NMR is also frequently applied in modern metabolomics
studies. In recent decades, many studies using NMR for metabolomics
were carried out by Nicholson and his group. The related
metabolomics studies generated many valuable ndings, ranging
from early diagnosis to disease treatment, drug toxicity, gene function, microbial, physiological and epidemiological aspects [6,2831].
3. Applications of metabolomics in systems-based TCM
Differing from Western medicine (WM) that is based on the
concept of one size ts all (i.e., 1 disease 1 target 1 drug), TCM
follows the principle of multiple components multiple target interactions and has been developed into a holistic healthcare system
[32]. Although attractive worldwide as a rich source of leading molecules for complementary and alternative medicine, TCM suffers from
inextricable obstacles in modern CHM research [33]. In CHM, most
of the active components are not well dened. The constituents and
their related concentrations in each herbal medicine can vary greatly
{e.g., depending on species [34], plant-growth conditions, and processing procedure [35]}. Metabolomics provides a valuable chance
for studying TCM in terms of characterizing the chemical components, dening the multiple active components and evaluating the

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209

Fig. 1. Workow for mass spectrometry-based metabolomics in research on systems-based traditional Chinese medicine.

ecacy. Fig. 1 shows the workow of MS-based metabolomics in

systems-based TCM.
3.1. Chemical characterization of CHM
The targets in TCM research, no matter what the crude materials, formulas or body uids from different TCM syndromes, are very
complex samples. Metabolomics, incorporating advanced analytical instruments, is capable of simultaneously determining and
identifying thousands of metabolites in complex samples.
Chromatography-MS-based technologies are the mainstream for
analysis of endogenous and exogenous metabolites in complex
samples.
Volatile oils are one of the main pharmacologically active components of many herb plants. GC- or comprehensive two-dimensional
GC (GC GC)-MS-based technologies are preferred for their analysis. GC-MS in selected ion mode and GC GC-TOF/MS were applied
for qualitative and quantitative analysis of volatile compounds of
Cyperus rotundus [36]. The yield and the quality of three extraction methods [i.e., hydrodistillation, pressurized liquid extraction
(PLE) and supercritical uid extraction (SFE)] were compared by the
chemical constituents of different extracts. It was found that PLE
had the highest extraction eciency, whereas SFE had the best selectivity for extraction of two compounds (-cyperone and
-cyperone). In another study, Qiu et al. characterized and quantied chemical compositions of volatile oils in radix ginseng at different growth ages using GC GC-TOF/MS metabolomics combined
with an unsupervised pattern-recognition method [37]. Chemical
compositions of samples were clearly discriminated among the three
growth ages and a large number of compounds were found to contribute to the differentiation. It was found that volatile compounds of -cadinol, -bisabolol, thujopsene, and n-hexadecanoic
acid showed positive correlation with the growth of the age of panax
ginseng. These studies demonstrate that coupled GC with a
chemometric method can be an ideal tool for characterization of
certain specic bioactive components of a CHM.
LC is a separation technology that is most commonly used in CHM
research due to its universal adaptability to most compounds without
need to take the volatility or the stability of the sample into consideration. Monolithic column LC, capillary LC and UPLC were developed to enhance the separation eciency. In a newly published
review on untargeted metabolomics to determine the composition of herbal products, Commisso et al. summarized LC-MSbased untargeted metabolomics approaches in comprehensive
analysis of complex plant tissues, unraveling environment-dependent
plant cell composition and analysis of natural remedies [38]. Wang

et al. combined the technologies of MALDI-MS with LC-MS to semiquantitate and to identify the alkaloid in a Chinese herbal medicine, Fuzi, the processed lateral roots of Aconitum carmichaeli Debx
(A. carmichaeli) [39]. The contents of alkaloids were found signicantly different in 14 batches of Fuzi analyzed, and decreased with
the length of the processing period. In another study, a UPLC-QTOF/
MS-based metabolic proling approach with pattern-recognition
analysis successfully discriminated Fuzi from its different preparations by comprehensive chemical composition analysis of Fuzi and
its processed products [40]. Besides, metabolomics analytical technology by direct analysis in real time mass spectrometry (DARTMS) in conjunction with multivariate analysis was a novel, rapid
method for monitoring batch-to-batch reproducibility of 12 tested
batches of Danshen alkaline precipitations [41]. Articial variations (e.g., starting material change and process fault) were successfully identied under this method. In other applications, the
metabolomics technologies of DART-MS and HPLC-DAD-ESI/MS were
applied for monitoring processing procedures of herbs [4244].
The studies discussed above demonstrate that MS-based
metabolomics technologies can be applied for characterization of
chemical components of CHM from different growth conditions,
growth ages, batches and processing procedures.
3.2. Bioactive component screening of CHM
A particular feature of TCM is holistic interactions (i.e., the synergetic interactions of the bioactive multi-components of the herb
or the formula). As bioactive components in CHM generally have
low abundance, how to rapidly screen the bioactive components at
a systematic level is key in modernizing TCM.
Traditionally, procedures to nd out bioactive components are:
(1) to extract the effective compound groups from the complex
mixture of CHMs;
(2) to purify the compounds from the extractions; and,
(3) to perform animal pharmacological experiments on the
bioactivity of the puried compounds singly.
Such a screening method for single target constituents of CHMs
is not only inecient, but also does not match the holistic concept
of multi-targets and systems of TCM theory [45]. Metabolomics, as
has been described, can provide an optimal platform for achieving this aim by incorporating state-of-the-art technologies.
A GC-MS metabolomics approach (Fig. 2) has been developed
for screening bioactive components from zedoary turmeric oil
and glucose injection using extracts from living cells [46]. Elemene

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Fig. 2. Flow scheme for screening bioactive components in traditional Chinese medicines based on cell extract and GC-MS [46]. [Reproduced by permission of Elsevier (License
Number: 3406820710544)].

emulsion injection, a representative TCM with known bioactive components, was used to validate the method by interacting with the
murine ascites hepatocarcinoma cell strain with high metastatic potential. It was concluded that curzerene and -elemene were two
major bioactive compounds in zedoary turmeric oil and glucose injection. Metabolomics utilizing HPLC-TOF/MS screened the potential bioactive constituents in a combined prescription of Danggui
Buxue decoction using cell extraction [47]. Validation of the method
was achieved using HL-7702 cells, RAW 264.7 cells and Caco-2 cells
that were incubated together with the extract of TCM. The results
demonstrated that 9, 7 and 13 potential bioactive compounds were
found in these three cell extractions, respectively.
LC-MS-based metabolomics has also been widely applied in
screening bioactive components in CHM. An LC-ESI-MS(n) system was
used for systematic screening and characterization of the major
bioactive compounds of Poria cocos and their metabolites in rats
[48]. A total of 34 compounds were detected and identied from
Poria cocos extract. In vivo, seven compounds were observed in rat
urine while one was detected in rat plasma.
Another study employed a UPCL-Q-TOF/MS analytical method
and macrophage cell extraction to screen and to analyze the potential bioactive components from Reduning Injection [49]. Chlorogenic acid, 3,4-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid and
4,5-dicaffeoylquinic acid were identied as potentially bioactive components from TCM injection. In addition, LC/DAD-ESI/MS was used
in screening bioactive components of Baihe Zhimu Tang [50], HPLCQ-TOF/MS and human umbilical vein endothelial cell were used for
screening the potentially bioactive components from a Shaofu Zhuyu
decoction [51], and HPLC-ESI-MS/MS was used for screening and
analyzing the multiple absorbed bioactive compounds and metabolites of Jitai tablets [52]. These studies demonstrated that LCMS-based metabolomics can be a valuable tool in screening and
identifying potentially bioactive components accountable for therapeutic effects of TCM, further promoting the development of TCM
and facilitating modern drug discovery.
3.3. Ecacy evaluation of CHM
TCM is a complex and interacted system, which is frequently used
in the form of formula (the combination of several different herbs).
The strategy of TCM is to utilize a system therapy to support the
self-healing capabilities of the body to regain its resilience from an

imbalanced state [53]. Such system therapy possesses multicomponent and multi-target characteristics in the treatment of diseases as each herbal drug comprises multiple bioactive constituents
that have concerted actions [54]. To date, TCM has been an empirical healthcare system with a long documented history. Such a traditional medical system can provide only a certain type of evidence
to support its alleged healthcare benets. By introducing systembased metabolomics into TCM research, we expect that there is a
better chance of evaluating the ecacy of TCM.
Many applications using metabolomics have been reported to
be revealing the ecacy of CHMs in the treatment of diseases. Two
approaches to LC-MS-based metabolomics combined with measuring biochemical parameters were used to study the biological
effects of Panax ginseng upon type 2 diabetes [55]. Panax ginseng
roots harvested after being grown for 36 years were selected to
treat Goto-Kakizaki (GK) rats with spontaneous type 2 diabetes for
a nine-week period. Plasma lipid proling of all studied GK rats and
ginsenoside proling of the root extracts of Panax ginseng with different growth ages were obtained on LC-IT-TOF/MS and UPLC-QTOF/
MS systems, respectively. Compared to the untreated controls,
patients treated with 4- and 6-year-old ginseng showed a signicantly benecial effect on glucose disposal and on regulation of
triacylglycerol lipid species, while 5-year-old ginseng signicantly
improved the level of high-density lipoprotein cholesterol. Besides,
6-year-old ginseng signicantly decreased the level of very low
density lipoprotein cholesterol and improved plasma-glycated hemoglobin in type 2 diabetic GK rats. Such results demonstrated that
ginseng roots have growth age-dependent therapeutic effects on two
major issues in diabetic GK rats hyperlipidemia and hyperglycemia. Ginseng roots 4-year-old contain bioactive natural components that may be valuable in drug discovery and dietary
supplements. After combining the results of the treatment effects
of ginseng with the proles of ginseng extracts, it can be argued
that the age-dependent therapeutic effects be correlated with the
variation in the contents of several ginsenosides in ginseng roots
at different growth ages. Fig. 3 shows the therapeutic effects of
ginseng observed [55] and the correlated ginsenoside biological
effects reported in literature [5659].
In another case, UPLC-MS-based metabolomics was applied with
a statistical technique to evaluate the therapeutic effects of
Epimedium brevicornum Maxim, a herbal medicine used for strengthening bones and toning kidney, on rats with kidney-deciency

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211

Fig. 3. Observed ginseng therapeutic effects and the correlated ginsenoside biological effects [55]. [Reproduced by permission of the Royal Society of Chemistry
(http://pubs.rsc.org/en/content/articlelanding/2011/mb/c1mb05254c)].

syndromes [60]. The chemical prole of Epimedium brevicornum

Maxim and plasma metabolic proles of rats from three different
treatments (pre- and post-hydrocortisone intervention and treatment with Epimedium brevicornum Maxim) were obtained by UPLCMS. PCA was used for comparison of the proles. Four compounds
that were regarded as the active constituents of the herb were found
in the plasma of kidney-decient rats, and two of them were also
observed in urine.
In other examples, a metabolomics study based on UPLC-QTOF/
MS was used to evaluate the therapeutic effect and possible action
mechanism of a TCM preparation Xindi soft capsule on a rat model
with acute blood stasis [61]. Urinary metabolomics study of myocardial ischemia and treatment effects of compound Danshen Tablets
in rats was conducted on a UPLC-QTOF/MS system [62]. The details
of these examples and several selected metabolomics applications in the study of the therapeutic effects of TCM are summarized in Table 1 [6369].
All of these studies illustrate that MS-based metabolomics is very
powerful in exploring the therapeutic effects of TCM and provides
insights into the potential action mechanism of TCM.
4. Evidence-based studies in TCM syndromes
Therapy with syndrome differentiation is the essence of a TCM
cure. As the physiopathological changes of a disease usually engender a corresponding syndrome in the body, the differences of
disease symptoms are sure to be reected by the metabolome in
the body system. As an approach of studying the metabolome in a

holistic context, metabolomics allowing a systemic study of all sorts

of small molecular metabolites matches well with the holistic nature
of TCM treatment, so it has potential as a useful tool for studying
TCM syndromes [45,70].
4.1. TCM pattern diagnosis
Taking the application of metabolomics on diagnosis and treatment of psoriasis patients with blood stasis syndrome in TCM as
an example, an HPLC-MS metabolomics method in combination with
OPLS-DA and molecular docking analysis was applied to investigate the biological characterization of the patient urine and the therapeutic metabolomics mechanism of the optimized Yinxieling
formula that was taken orally by patients after 0, 4, 8, and 12 weeks
[71]. A total of 60 subjects (i.e., 41 patients versus 19 healthy volunteers) participated in this study. Three potential biomarkers involved in glucose metabolism, eight involved in fat metabolism and
four involved in amino-acid and nucleotide metabolism were identied from LC-MS metabolomics data. The analyzed results suggest
that the deciency of bile acids and the consequent cytokine release
and endotoxin translocation might play a role in the pathogenesis
of psoriasis [71].
Wei et al. used a GC-MS urine metabolomics approach with PCA
and CM diagnosis to identify diagnostic biomarkers for prediabetic sub-types [72]. Three pre-diabetic groups including QiYin deciency and Qi-Yin deciency with dampness (sub-type
A) and Qi-Yin deciency with stagnation (sub-type B) were distinctly separated. The majority of metabolites (i.e., sugars and amino

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Table 1
Selected metabolomics applications in evaluating the therapeutic effects of traditional Chinese medicine (TCM)
Metabolomics technique

Samples

TCM

Disease

GC-MS

Rat plasma

Compound Danshen dripping

pills

Myocardial infarction

UPLC-QTOF-MS

Rat urine

Acute blood stasis

UPLC-QTOF/MS

Rat plasma

A TCM preparation Xindi soft

capsule
Compound Danshen tablets

UPLC-QTOF/MS and
UPLC-SQ-MS

Human serum

Berberine

Type 2 diabetes

LC-MS

Rat urine

Rehmanniae glutinosa

Kidney yin deciency

UPLC-QTOF/MS

Rat urine and serum

Yunnan Baiyao

Rheumatoid arthritis

LC-IT-TOF/MS

Rat urine

Tongxinluo

Endothelial dysfunction

LC-FTMS

Mouse plasma
and liver

A multi-components
preparation of eight natural
ingredients

Early stage of obesity

HILIC/RPLC-MS

Rat urine

Ginsenoside Rg3

Liver tumor

acids) were at higher levels in urine of sub-type B as compared to

sub-type A, indicating more disorders in carbohydrate metabolism and renal function in sub-type B versus sub-type A. These studies
demonstrated that metabolomics technology combined with multivariate statistical analysis is helpful in identifying potential diagnostic biomarkers for diseases, enhancing our understanding of the
biological response to TCM syndrome and the action mechanism
of CHM.
4.2. TCM pattern intervention
A good example of metabolomics applications in TCM syndrome is on the study of pyretic pulmonary syndrome model interfered by Radix Scutellariae using a UPLC-Q-TOF/MS-based
approach and PCA method to analyze the urinary metabolome of
Wistar rats treated differently [73]. Radix Scutellariae is a wellknown medicinal herb in China and can alleviate heat syndrome.
Six specic biomarkers were determined that can represent Streptococcus pneumonia-induced pyretic pulmonary syndrome in rats.
The results of statistical analysis showed that the Chinese herb could
substantially make the disease-related increased biomarkers come
back to normal levels.
More specically, a UPLC-MS-based metabolomics strategy was
successfully employed to characterize the kidney-yang deciency
(LYD) syndrome induced by high dose of hydrocortisone and the
ecacy of Rhizoma Drynariae [74]. PCA of urine metabolomics data
of rats from hydrocortisone-induced group and the pre-dose group
showed that there are distinct metabolic patterns between them.
The time-dependent recovery effect of Rhizoma Drynariae on KYD
syndrome rats was observed from day 3 to day 15 after therapy. The
metabolites that were considered to contribute to the differentiation between the two groups were found to be involved in energy

Myocardial ischemia

Findings

Ref.

Compound Danshen dripping pills show

therapeutic ecacy in the regulation of energy
metabolism, glycolysis, and lipid metabolism
damaged by myocardial ischemia.
The TCM preparation showed a tendency to
recover acute blood stasis to healthy status.
Compound Danshen Tablets exhibited
protective effects on myocardial ischemia by
reversing potential biomarkers close to normal
levels, especially for the four metabolites in
the pathway of purine metabolism.
Berberine treated type 2 diabetes by downregulating the high levels of free fatty acids
and mediating glucose and lipid metabolism in
patient serum.
5-hydroxymethyl-2-furfural was the main
chemical marker of Rehmanniae glutinosa in
the treatment of type 2 diabetes.
Yunnan Baiyao exhibit therapeutic effect on
rheumatoid arthritis by suppressing some AAderived proinammatory mediator excretion
from osteoblasts.
Tongxinluo exerted protective effect on
endothelial dysfunction by adjusting pathways
of fatty acid oxidation, proteolysis of collagen,
adenine, and phenylalanine and porphyrin
metabolites close to their normal state.
The multi-component preparation produced
anti-atherogenic effects by regulating lipid
metabolism in mice in the early stage of
obesity.
Rg3 in a beta-cyclodextrin-based formulation
induced considerable change in the urine
metabolic pattern in cancer rats.

[63]

[61]
[62]

[64]

[65]

[66]

[67]

[68]

[69]

metabolism, amino acid metabolism and gut microora. Such ndings provide valuable information for illustrating the therapeutic
mechanism of Rhizoma Drynariae.
Moreover, Kuang et al. studied Cold and Hot syndromes (i.e., four
properties, such as cold-cool and hot-warm) in TCM using a UPLCQTOF/MS-based metabolomics strategy in combination with bioinformatics tools [75]. Some 56 male Sprague-Dawley rats (n = 8 per
group) received the extracts of six herbs for 30 days, respectively.
One group without treatment was used as the controls. Unsupervised PCA and supervised orthogonal partial least squares discriminant analysis (OPLS-DA) multivariate data analyses of the global urine
proles of rats on the 30th day from each group were performed to
evaluate sample clustering according to the group information, and
obtain discrimination between different groups as well as nd the
signicantly differential metabolites that might be potential
biomarkers. The Cold and Hot medication-treated rats were distinguished clearly by the differences in the metabolic proles, proving
the existence of the four properties of TCM.
Taken together, the ndings of these studies show that MSbased metabolomics strategy is very promising in studying the
essence of TCM syndrome. For CHM and TCM studies, a better way
[53,76,77] is utilizing the conception of systems biology to combine
the chemical ngerprint, biological response and biological
biomarkers together to control quality, to dene active components, to shed light on active mechanisms, and to evaluate pharmaceutical effects (Fig. 4).
5. Future perspectives
Obviously, the applications of MS-based metabolomics (e.g., GCMS or LC-MS) have made a signicant impact on TCM research
in characterizing herb chemical compositions, screening and

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213

Fig. 4. Core of Herbal Medicine research on the basis of metabolomics.

identifying potential bioactive components and evaluating therapeutic effects of CHM and its products. Tremendous advances in
recent hybrid analytical techniques with unparalleled sensitivity, resolution, reliability and specicity will further extend metabolomics
applications in TCM research domains, such as sample processing,
batch-to-batch variation, quality control, safety, toxicity, stability etc.
Growing evidence from recent publications indicates that, by combining advanced bioinformatics tools, MS-based metabolomics has
the opportunity to examine the synergetic effects of CHM using
animal experiments, preclinical or clinical trials. However, many challenges exist in metabolomics applications to study TCM due to the
adverse factors including: (1) the bioactive multi-components of
CHM are not well dened; (2) the action mechanisms of TCM remain
unclear; (3) quality control of TCM lacks unied criteria, (4) safety
of TCM is mainly empirical; (5) the multi-component interactions
of TCM are obscure; and, (6) the scientic basis of symptom classication is ambiguous. Accordingly, crucial core technologies (e.g.,
hybrid MS instruments equipped with two or more mass analyzers used for detection, comprehensive bidimensional or multidimensional chromatographic techniques used for separation) are
required for analytical characterization of TCM in relation to these
aspects.
Acknowledgments
The authors gratefully acknowledge support from the China International Science and Technology Cooperation Program
(2009DFA41250) funded by the Ministry of Science and Technology of the Peoples Republic of China, and the Creative Research
Group Project (No. 21321064) of the National Natural Science Foundation of China.
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