RESUMO
BACKGROUND: Lycium barbarum polysaccharide (LBP) is an active ingredient extracted from Lycium barbarum that inhibits neuroinflammation, and Lycium barbarum glycopeptide (LbGp) is a glycoprotein with immunological activity that was purified and isolated from LBP. Previous studies have shown that LbGp can regulate the immune microenvironment, but its specific mechanism of action remains unclear. AIMS: In this study, we aimed to explore the mechanism of action of LbGp in the treatment of spinal cord injury through metabolomics and molecular experiments. METHODS: SD male rats were randomly assigned to three experimental groups, and after establishing the spinal cord hemisection model, LbGp was administered orally. Spinal cord tissue was sampled on the seventh day after surgery for molecular and metabolomic experiments. In vitro, LbGp was administered to mimic the inflammatory microenvironment by activating microglia, and its mechanism of action in suppressing neuroinflammation was further elaborated using metabolomics and molecular biology techniques such as western blotting and q-PCR. RESULTS: In vivo and in vitro experiments found that LbGp can improve the inflammatory microenvironment by inhibiting the NF-kB and pyroptosis pathways. Furthermore, LbGp induced the secretion of docosahexaenoic acid (DHA) by microglia, and DHA inhibited neuroinflammation through the MAPK/NF-κB and pyroptosis pathways. CONCLUSIONS: In summary, we hypothesize that LbGp improves the inflammatory microenvironment by regulating the secretion of DHA by microglia and thereby inhibiting the MAPK/NF-κB and pyroptosis pathways and promoting nerve repair and motor function recovery. This study provides a new direction for the treatment of spinal cord injury and elucidates the potential mechanism of action of LbGp.
Assuntos
Medicamentos de Ervas Chinesas , Lycium , Traumatismos da Medula Espinal , Animais , Masculino , Ratos , Ácidos Docosa-Hexaenoicos/metabolismo , Medicamentos de Ervas Chinesas/farmacologia , Glicopeptídeos , Lycium/química , Lycium/metabolismo , Doenças Neuroinflamatórias , NF-kappa B/metabolismo , Piroptose , Traumatismos da Medula Espinal/complicações , Traumatismos da Medula Espinal/tratamento farmacológicoRESUMO
Aim: The aim of this study was to elucidate the potential mechanism of Rg1 in alleviating hepatic ischemia-reperfusion (HIRI) through the mitophagy pathway. Methods: The HIRI rat models were established and divided into 4 groups: the sham group, sham+Rg1 group, ischemia/perfusion (I/R) group and I/R+Rg1 group. Then the activities of aspartate transaminase (AST) and alanine aminotransferase (ALT) were detected by automatic serum analyzer. Meanwhile, cell apoptosis and changes in liver tissues were checked by TUNEL assay and histopathological analysis, respectively. The relative protein levels were detected by western blotting. Subsequently, cell counting Kit-8 assay and cytometric analysis were used to investigate cell viability and apoptosis of liver cells. Finally, the time points of the strongest mitochondrial autophagy were explored and the mitochondrial morphology was observed by the mitochondrial transmembrane potential (MMP) in vivo and in vitro. Results: The mitophagy aggravated hepatocyte damage during liver I/R in vivo. In addition, Rg1 alleviated liver damage after liver I/R, maintained the stability of MMP and inhibited mitochondrial autophagy and signaling pathways during liver I/R in vivo. Furthermore, Rg1 could effectively increase cell viability, inhibit cell apoptosis and stabilize MMP after OGD/R injury in vitro Moreover, Rg1 exerted its protective effect on HIRI by regulating the PINK1/Parkin signaling pathway and the mitochondrial autophagy. Conclusion: Rg1 could further improve its mechanism of alleviating HIRI in apoptosis and autophagy, 2 types of regulated programmed cell death via the mitochondrial pathway.
Assuntos
Traumatismo por Reperfusão , Ratos , Animais , Ratos Sprague-Dawley , Traumatismo por Reperfusão/tratamento farmacológico , Traumatismo por Reperfusão/metabolismo , Traumatismo por Reperfusão/patologia , Isquemia/metabolismo , Isquemia/patologia , Fígado/metabolismo , Apoptose , Autofagia , ReperfusãoRESUMO
To systematically evaluate the efficacy and safety of traditional Chinese medicine injections combined with antiviral wes-tern medicine in the treatment of influenza by Bayesian network Meta-analysis. Four Chinese databases(CNKI, Wanfang, VIP, CBM) and three English databases(PubMed, EMbase, Cochrane Library) were retrieved by computer for randomized controlled trials(RCT) about the comparison between the combined administration of traditional Chinese medicine injections with antiviral western medicine(or with placebo) and the single administration of antiviral western medicine in the treatment of influenza, or the comparison between two different traditional Chinese medicine injections combined with antiviral western medicine in the treatment of influenza. The retrieval time was from the establishment of the databases to July 2020. Two researchers independently screen out the literatures and extracted the data according to the inclusion criteria and exclusion criteria. The included studies were evaluated by the Cochrane 5.1 bias risk assessment tool, and data analysis was conducted by ADDIS 1.16.8 and Stata 14.0. A total of 49 literatures about 5 061 cases were included finally, involving six traditional Chinese medicine injections, namely Reduning Injection, Xiyanping Injection, Tanreqing Injection, Yanhuning Injection, Qingkailing Injection and Shuanghuanglian Injection. The results of network Meta-analysis,(1)in terms of total effective rate, 6 traditional Chinese medicine injections combined with antiviral western medicine was better than antiviral western medicine alone. In terms of probability sorting, Shuanghuanglian Injection combined with antiviral western medicine ranked first.(2)There was no significant statistical difference in adverse reaction rate among all groups, but Reduning Injection ranked first in the probability sorting, due to the least side effects.(3)In terms of antipyretic time, Yanhuning Injection, Xiyanping Injection and Tanreqing Injection combined with antiviral western medicine was better than antiviral western medicine alone and Xiyanping Injection ranked first in the probability sorting.(4)In terms of cough relief time, Qingkailing Injection and Tanreqing Injection combined with antiviral western medicine was better than antiviral western medicine alone and Qingkailing Injection was superior to Yanhuning Injection. Qingkailing Injection combined with antiviral western medicine ranked first in the probability sorting.(5)In terms of the length of hospital stay, Xiyanping Injection, Tanreqing Injection and Reduning Injection combined with antiviral western medicine was superior to antiviral western medicine alone, and Reduning Injection ranked first in the probability sorting due to the shortest hospital stay. Because of the small quantity and low quality of included studies, the results of this study shall be regarded cautiously and comprehensively. In the future, multi-center, large-sample and clinical randomized controlled trials shall be conducted to verify the results.
Assuntos
Medicamentos de Ervas Chinesas , Influenza Humana , Antivirais , Teorema de Bayes , Humanos , Influenza Humana/tratamento farmacológico , Medicina Tradicional Chinesa , Metanálise em RedeRESUMO
Hepatic ischemia-reperfusion (IR) injury remains the major cause of liver damage post-liver surgery or transplantation. Diminishing oxidative stress and inflammatory responses is a powerful channel to reduce the rate of morbidity and mortality. Gastrodin (GSTD), a bioactive compound extracted from the traditional Chinese herbal agent with a long history of clinical application in nervous system diseases, is suggested to possess anti-oxidative effects on liver diseases, such as nonalcoholic fatty liver disease. However, the therapeutic potential of GSTD in liver IR injury remains unclear. In this paper, we performed surgery to set up the 70% hepatic IR injury models in mice after a three-day pretreatment of GSTD. We found the administration of GSTD reduced liver damage, which correlated with lower histological Suzuki's score, lower serum alanine transaminase (AST) and alanine transaminase (ALT) levels, less oxidative stress, and cell apoptosis in a dose-responsive manner, as compared to the parallel control. Meanwhile, we observed a great induction of heme oxygenase-1 (HO-1) and an activation of the p38 mitogen-activated protein kinases/nuclear factor erythroid 2-related factor 2 (p38MAPK/Nrf2) pathway in response to the GSTD pretreatment, while the protective effects upon GSTD diminished in mice with HO-1 heterozygous mutation. In addition, GSTD inhibited IR induced toll-like receptor (TLR) 4, but not TLR2 in a HO-1 dependent manner, leading to a down-regulation of cytokines, such as interleukin (IL)-6 and TNF-[Formula: see text]. Collectively, our findings revealed GSTD attenuated liver IR injury via activation of the HO-1 pathway, providing a novel therapeutic strategy to minimize the IR induced oxidative stress in the process of liver transplantation.
Assuntos
Antioxidantes , Álcoois Benzílicos/administração & dosagem , Álcoois Benzílicos/farmacologia , Glucosídeos/administração & dosagem , Glucosídeos/farmacologia , Fígado , Fator 2 Relacionado a NF-E2/metabolismo , Fitoterapia , Traumatismo por Reperfusão/prevenção & controle , Transdução de Sinais/efeitos dos fármacos , Animais , Citocinas/metabolismo , Modelos Animais de Doenças , Regulação para Baixo/efeitos dos fármacos , Medicamentos de Ervas Chinesas , Heme Oxigenase-1/metabolismo , Camundongos Endogâmicos C57BL , Estresse Oxidativo/efeitos dos fármacos , Cuidados Pré-Operatórios , Receptor 4 Toll-Like/metabolismo , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismoRESUMO
Nuclear factor erythroid-related factor 2 (Nrf2) has been implicated in several detoxifying and antioxidant defense processes. Nrf2-mediated heme oxygenase-1 (HO-1) expression was demonstrated to play a key role against oxidative stress. Gastrodin (GSTD) is a well-known active compound isolated from the roots of Rhizoma gastrodiae, a plant used in ancient Chinese traditional medicine. The aim of this work was to investigate whether GSTD could alleviate H2O2-induced oxidative stress in mouse liver sinusoidal endothelial cells (LSECs). In LSECs exposed to 1 mM H2O2, treatment with GSTD (1, 10, or 50 µM) resulted in higher cell viability than the untreated control. Treated cells maintained a higher Bcl2/Bax ratio and suppressed caspase-9 expression compared with untreated cells, reducing cell apoptosis. GSTD was protective for H2O2-induced oxidative injury by reducing the generation of intracellular reactive oxygen species and malondialdehyde. HO-1 and Nrf2 expressions were synergistically upregulated by GSTD. Inhibition of HO-1 by 10 µM zinc protoporphyrin resulted in less protective effects on cell viability and malondialdehyde reduction by GSTD treatment in H2O2-exposed LSECs. Additionally, phosphorylated p38 in LSECs exposed to H2O2 was elevated by GSTD. Inhibition of p38 phosphorylation by SB203580 did not induce Nrf2 and HO-1 expression after 1 or 10 µM GSTD treatment and the protective effect on cell viability and malondialdehyde reduction in H2O2-exposed LSECs was reduced. The data conclusively demonstrated that GSTD-induced HO-1 and Nrf2 expression is involved in protection of LSECs from H2O2-induced oxidative injury, which may be regulated by p38 phosphorylation.
Assuntos
Álcoois Benzílicos/farmacologia , Células Endoteliais/efeitos dos fármacos , Glucosídeos/farmacologia , Heme Oxigenase-1/metabolismo , Peróxido de Hidrogênio/farmacologia , Fator 2 Relacionado a NF-E2/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , Animais , Apoptose/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Fígado/citologia , Fígado/efeitos dos fármacos , Malondialdeído/metabolismo , Camundongos , Modelos Teóricos , Estresse Oxidativo/fisiologia , Fosforilação/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Regulação para Cima/efeitos dos fármacos , Proteínas Quinases p38 Ativadas por Mitógeno/sangueRESUMO
Nuclear factor erythroid-related factor 2 (Nrf2) has been implicated in several detoxifying and antioxidant defense processes. Nrf2-mediated heme oxygenase-1 (HO-1) expression was demonstrated to play a key role against oxidative stress. Gastrodin (GSTD) is a well-known active compound isolated from the roots of Rhizoma gastrodiae, a plant used in ancient Chinese traditional medicine. The aim of this work was to investigate whether GSTD could alleviate H2O2-induced oxidative stress in mouse liver sinusoidal endothelial cells (LSECs). In LSECs exposed to 1 mM H2O2, treatment with GSTD (1, 10, or 50 µM) resulted in higher cell viability than the untreated control. Treated cells maintained a higher Bcl2/Bax ratio and suppressed caspase-9 expression compared with untreated cells, reducing cell apoptosis. GSTD was protective for H2O2-induced oxidative injury by reducing the generation of intracellular reactive oxygen species and malondialdehyde. HO-1 and Nrf2 expressions were synergistically upregulated by GSTD. Inhibition of HO-1 by 10 µM zinc protoporphyrin resulted in less protective effects on cell viability and malondialdehyde reduction by GSTD treatment in H2O2-exposed LSECs. Additionally, phosphorylated p38 in LSECs exposed to H2O2 was elevated by GSTD. Inhibition of p38 phosphorylation by SB203580 did not induce Nrf2 and HO-1 expression after 1 or 10 µM GSTD treatment and the protective effect on cell viability and malondialdehyde reduction in H2O2-exposed LSECs was reduced. The data conclusively demonstrated that GSTD-induced HO-1 and Nrf2 expression is involved in protection of LSECs from H2O2-induced oxidative injury, which may be regulated by p38 phosphorylation.
Assuntos
Animais , Coelhos , Álcoois Benzílicos/farmacologia , Células Endoteliais/efeitos dos fármacos , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , Heme Oxigenase-1/metabolismo , Glucosídeos/farmacologia , Peróxido de Hidrogênio/farmacologia , Regulação para Cima/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Apoptose/efeitos dos fármacos , Fígado/citologia , Fígado/efeitos dos fármacos , Malondialdeído/metabolismo , Modelos TeóricosRESUMO
Belamcanda chinensis has been extensively used as antibechic, expectorant and anti-inflammatory agent in traditional medicine. Irisflorentin is one of the major active ingredients. However, little is known about the metabolism of irisflorentin so far. In this work, rat liver microsomes (RLMs) were used to investigate the metabolism of this compound for the first time. Seven metabolites were detected. Five of them were identified as 6,7-dihydroxy-5,3',4',5'-tetramethoxy isoflavone (M1), irigenin (M2), 5,7,4'-trihydroxy-6,3',5'-trimethoxy isoflavone (M3), 6,7,4'-trihydroxy-5,3',5'-trimethoxy isoflavone (M4) and 6,7,5'-trihydroxy-5,3',4'-trimethoxy isoflavone (M5) by means of NMR and/or HPLC-ESI-MS. The structures of M6 and M7 were not elucidated because they produced no MS signals. The predominant metabolite M1 was noted to be a new compound. Interestingly, it was found to possess anticancer activity much higher than the parent compound. The enzymatic kinetic parameters of M1 revealed a sigmoidal profile, with Vmax = 12.02 µm/mg protein/min, Km = 37.24 µm, CLint = 0.32 µL/mg protein/min and h = 1.48, indicating the positive cooperation. For the first time in this work, a new metabolite of irisflorentin was found to demonstrate a much higher biological activity than its parent compound, suggesting a new avenue for the development of drugs from B. chinensis, which was also applicable for other herbal plants. Copyright © 2016 John Wiley & Sons, Ltd.
Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Isoflavonas/metabolismo , Microssomos Hepáticos/metabolismo , Espectrometria de Massas em Tandem/métodos , Animais , Técnicas In Vitro , Masculino , Ratos , Ratos Sprague-DawleyRESUMO
A chemometric strategy has been developed to discover component difference and similarity between two chromatograms (correlation) by using comprehensive two-dimensional (2D) gas chromatography with time-of-flight mass spectrometry (GC×GC-TOFMS). It allows for rapid determination of the presence or absence of analytes of interest in both pure and overlapping peak clusters, and then locates elution windows of target components. First, representative elution windows of analytes are extracted from the 2D GC×GC map to characterize the spectral space and further construct an orthogonal projection matrix for analysis. Next, multi-component spectral correlative chromatography (MSCC) is employed to scan the whole or pre-selected GC×GC-TOFMS data range to obtain component features. An auto-correlative projection curve is proposed to assess the projection residual from MSCC by defining a new evaluation index as reference, based on fixed-size moving window evolving factor analysis. In principle, the method can also be utilized to locate specific compounds whose known spectra are available. It is not restricted by data with high homoscedastic and heteroscedastic noise. Simulated GC-MS data and an extremely complicated herbal product mixture comprising 9 herbs demonstrates that the two-dimensional correlative distribution graph is effective for chemical interpretation between GC×GC-TOFMS data. It allows discovery of information buried in this type of highly complex dataset, especially for rapid and effective data comparison, where specific molecular identity might otherwise be hidden.
Assuntos
Cromatografia Gasosa-Espectrometria de Massas/métodos , Modelos Teóricos , Preparações de Plantas/análiseRESUMO
Alternative moving window factor analysis (AMWFA) has shown the powerfulness for comprehensive comparison and individual identification of chemical components among different but related mixture systems. However, quantification of these components can only be attained after extraction of all spectra of pure components in samples with least square technique. In this study, a novel two-step iterative constraint method (TICM) is developed for independent quantification of the interested target analytes. The pure chromatographic profiles of the components can be mined out from mixtures with high complexity using a two-step iterative operation and stepwise purification of the targets from interferers. Some effective constraints of chromatographic profiles, such as non-negative and single-peaked properties, as well as zero-concentration outside of elution windows of components, are employed to further improve the efficiency of the method. One of the strong advantages of TICM is simplification of complex mixtures to several sub-systems for processing easily with the help of AMWFA, as well as bi-linear property of data sets obtained from coupled chromatographic instruments. It meets the urgent requirements and challenges of qualitative and quantitative analysis of complicated systems with multi-component in the investigation of herbal medicines (HMs), metabonomics and systems biology. From the results of simulated LC-DAD data, GC-MS data of volatile chemical components in three kinds of ginseng with different growth conditions, and four different medicinal parts of the same herb, good performance of the proposed method is achieved.
Assuntos
Misturas Complexas/análise , Análise Fatorial , Cromatografia Gasosa-Espectrometria de Massas , Cromatografia , Medicina Herbária , Metabolismo , Panax/química , Biologia de SistemasRESUMO
Quality control of herbal medicines (HMs) is a big big headache because of the high complexity and unknown mechanism on disease treatment. In this work, mass spectral profiling, a new tool for data processing is proposed to help a lot in solving this problem as gas chromatography-mass spectroscopy (GC-MS) is used to detect both the active and non-active ingredients buried in HMs. The main idea of mass spectral profiling is employment of target m/z points of GC-MS data on the extraction of chromatographic profiles of pure and/or mixed compositions concerned. Further, the absolute or relative abundance at these m/z points can be utilized for results interpretation. With the help of this tool, the qualitative and quantitative information of chemical components within complicated HMs will be mined out effectively. It can then be recommended as reference indices to assess the importance of target compositions in HMs, such as efficacy evaluation on disease treatment of the active constituents. Mass spectral profiling with less data points significantly improves the possibility to get the rich information with no strong requirements of data preprocessing procedures, like alignment of shift of retention times among different chromatographic profiles. It is powerful for quality control of HMs coupled with pattern recognition techniques on high-throughput data sets. In this study, a commonly used herbal medicine, Houttuynia cordata Thunb and its finished injection products, were used to deliver the strategies. Absolutely, the working principles can be extended to the investigation of metabonomics with gas chromatography-time-of-flight-mass spectrometry (GC-MS-TOF). The good performance of mass spectral profiling shows that it can be a promising tool in the future studies of complex mixture systems.
Assuntos
Medicamentos de Ervas Chinesas/química , Cromatografia Gasosa-Espectrometria de Massas/métodos , Medicina Herbária , Controle de Qualidade , HouttuyniaRESUMO
The volatile components between stems and roots and also among five Clematis species from China were studied and analyzed by gas chromatography-mass spectrometry (GC-MS) combined with alternative moving window factor analysis (AMWFA), a new chemometric resolution method. Identification of the compounds was also assisted by comparison of temperature-programmed retention indices (PTRIs) on HP-5MS with authentic samples included in our own laboratory database under construction. A total of 153 different compounds accounting for 86.6-96.5% were identified and significant qualitative and quantitative differences were observed among the samples. The major volatile components in different essential oils from Clematis species were n-hexadecanoic acid and (Z,Z)-9,12-octadecadienoic acid. Our work further demonstrated chemometric resolution techniques upon the two-dimensional data and PTRIs can provide a complementary and convenient method for fast and accurate analysis of complex essential oils.
Assuntos
Clematis/química , Medicamentos de Ervas Chinesas/análise , Óleos de Plantas/análise , Cromatografia Gasosa-Espectrometria de Massas/métodos , Ácido Linoleico/análise , Óleos Voláteis/análise , Ácido Palmítico/análiseRESUMO
It is a crucial issue to determine the origins of herbal medicinal materials and identify the quality grades and fakes of their final products collected from different pharmaceutical corporations. Pattern recognition technique may assist the manufacturers to achieve this purpose and effectively control the quality of their products. In this work, a widely used method in chemometrics, orthogonal projection (OP) technique, was applied to discrimination analysis and identification of fingerprints of the herbal medicine houttuynia cordata Thunb. (HCT) and its final injection products. The advantages of the OP technique are clearly shown after comparing with the conventional methods such as principal component analysis (PCA), Mahalanobis distance (MD), and similarity comparison method (SCM). Three different sources of medicinal material HCT and its final injection products from six different manufacturers were studied under 'sixfold', 'threefold' and 'threefold-bis' cross-validation procedures. The good performance of the proposed method in determination and identification of unknown samples shows it could be a powerful tool for quality control in herbal medicine production and other related research fields.
Assuntos
Medicina Herbária , Plantas Medicinais , Técnicas de Química Analítica/métodos , Cromatografia Gasosa/métodos , Análise por Conglomerados , Houttuynia/metabolismo , Modelos Químicos , Modelos Estatísticos , Fitoterapia , Análise de Componente Principal , Controle de QualidadeRESUMO
In this investigation, a novel chemometric method is developed for the analysis of five possible relationships of components or spectral features between two correlative but different hyphenated chromatographic systems. It is very helpful for comparison study of components present in different complex systems in both chemistry and systems biology. The proposed method, named alternative moving window factor analysis (AMWFA), could be utilized to determine the number of common components between different samples and then to identify their corresponding spectra half-automatically. AMWFA can alternatively be employed to mind for the selective information hiding in anyone of the two compared data X and Y, and to self-verify the resolution results by changing the extracted target matrices in analysis. From the results of comparison of simulated hyphenated chromatographic data, volatile chemical components in drug pair rhizoma ligustici chuanxiong-radix paeoniae rubra (RLC-RPR) and its single herbal medicines, and analysis of Angelica oral solution and its plasma sample after oral intake to rabbit, powerful ability of the proposed method is shown.
Assuntos
Cromatografia Gasosa-Espectrometria de Massas/métodos , Angelica/química , Animais , Análise Fatorial , Extratos Vegetais/administração & dosagem , CoelhosRESUMO
A "chromatographic fingerprint" of a herbal medicine is essentially its chromatographic spectrum: a characteristic representation of its chemical components, some of which are pharmacologically active. Since a wide variety of factors, such as the geographical location, the harvest season, and the part used can influence the chemical constituents (and therefore the pharmacological activity) of any particular herbal medicine and its products, these fingerprints provide a way to compare and contrast the compositions of different variants of the same herbal medicine. In particular, it is possible to ascertain whether particular components present in one herbal fingerprint are also present in another fingerprint. In this work we use a novel method-modified window target-testing factor analysis (MWTTFA), based on the use of target factor analysis (TFA), fixed-size moving window evolving factor analysis (FSMWEFA) and a Gaussian shape correction to the chromatographic profiles-to achieve this end. To demostrate the strategy, the fingerprints of samples from garlics produced in different geographical locations were compared, as well as the fingerprints of samples taken from above-ground and below-ground parts of Houttuynia cordata Thunb. The results from these comparisons clearly show that four chemical components present in Hunan common edible garlic are absent in Xingping base garlic, while seven components are present in Xingping base garlic but absent in Hunan common edible garlic. Also, eleven components are present in the sample from the above-ground part of Houttuynia cordata Thunb but not in the sample from the below-ground part, while seven components are present in the sample from the below-ground part of Houttuynia cordata Thunb that are not present in the sample from the above-ground part. These interesting conclusions should be very useful for future pharmacological and clinical research into these herbal medicines, and the novel MWTTFA technique can also be used for quality control purposes.