Traditional Chinese Medicine Extracts Research Articles

An approach to optimize the batch mixing process for improving the quality consistency of the products made from traditional Chinese medicines.

The efficacy of traditional Chinese medicine (TCM) is based on the combined effects of its constituents. Variation in chemical composition between batches of TCM has always been the deterring factor in achieving consistency in efficacy. The batch mixing process can significantly reduce the batch-to-batch quality variation in TCM extracts by mixing them in a well-designed proportion. However, reducing the quality variation without sacrificing too much of the production efficiency is one of the challenges. Accordingly, an innovative and practical batch mixing method aimed at providing acceptable efficiency for industrial production of TCM products is proposed in this work, which uses a minimum number of batches of extracts to meet the content limits. The important factors affecting the utilization ratio of the extracts (URE) were studied by simulations. The results have shown that URE was affected by the correlation between the contents of constituents, and URE decreased with the increase in the number of targets and the relative standard deviations of the contents. URE could be increased by increasing the number of storage tanks. The results have provided a reference for designing the batch mixing process. The proposed method has possible application value in reducing the quality variation in TCM and providing acceptable production efficiency simultaneously.

Study on directional separation of picroside Ⅱ from extract of traditional Chinese medicine by molecularly imprinted technology

Picroside II, separated from Chinese herbal medicine, is an active compound with neroprotective activity. Molecularly imprinted polymers (MIPs) have high affinity toward template molecules synthesized by molecularly imprinted technology for its specific combined sites, which can overcome the shortcomings of traditional separation methods, such as complex operation and low efficiency. In this paper, MIPs were prepared by precipitation polymerization with picroside II as the template molecule, 1-vinylimidazole (1-Vinyl) as functional monomer, ethylene glycol dimethacrylate (EDMA) as cross-linker. The morphology of MIPs was characterized by scanning electronmicroscope (SEM) and its static adsorption capacity was measured by the scatchard equation. The results showed that picroside II MIPs have spherical shape, and most of them are uniform in size. Furthermore, the maximum binding capacity (Q(max)) of MIPs is 3.02 mg x g(-1), higher than that of non-imprinted polymers (NIPs). This result indicated that picroside II MIPs with good morphology and high targeted affinity toward the template molecules can be prepared by precipitation polymerization, which can be used to separate picroside II and its analogies from extract of Chinese herbal medicine. In addition, this method has the advantages of good environment and simple operation, which might offer a novel method for the efficient separation of picroside II in the traditional herbal medicines.

Efficient optimization of ultra‐high‐ performance supercritical fluid chromatographic separation of Rosa sericea by response surface methodology

An approach for rapid optimization of ultra-high-performance supercritical fluid chromatographic (UHPSFC) gradient by response surface methodology was developed for fast separation of complex crude extracts of the leaves of Rosa sericea. The optimization was performed with Box-Behnken designs and the multicriteria response variables were described using Derringer's desirability. Based on factorial design experiments, five factors were selected for Box-Behnken designs to optimize the UHPSFC conditions, which led to 46 experiments being performed within 8 h. An evaporative light-scattering detector (ELSD) was used, and quantitative analysis of main components in R. sericea samples was employed to evaluate the statistical significance of the parameters on UHPSFC-ELSD analytes response. The results indicated that the optimized UHPSFC-ELSD method is very sensitive with LODs and LOQs below 1.19 and 4.55 μg/mL, respectively. The overall intra- and interday variations were less than 3.91 and 6.41%, respectively. The recovery of the method ranged from 95.66 to 104.22%, with RSD < 5.91%. This newly developed UHPSFC-ELSD method was demonstrated to be fast and sensitive in analyzing complex herbal extracts of Traditional Chinese Medicines.

Study on registration administration of extracts of traditional Chinese medicine and its preparation

This paper studied the status of registration administration of extracts of traditional Chinese medicine (TCM) and its preparation, discussed some factors which affect the quality of extracts of TCM and its preparation, and puted forward some suggestions to strengthen the registration administration of extracts of TCM and its preparation such as clear positioning and legal basis, improving and enhancing quality standards, implementing of record management and so on.

Constituents from Commonly used Traditional Chinese Medicines

The isolation and structure elucidation of chemical constituents from traditional Chinese medicines (TCMs) are still important and attractive, especially for finding lead compounds with a potential use of new drug development. TCMs are used for treatment of diseases from ancient times up to today (thousands of years), and are clinically effective. However, studies have only been carried out for about 20 years, and the active components are seldom identified. Because the chemical components of TCMs and pharmacological responses of a drug are very complex, it is extremely difficult to identify an active compound from a TCM to represent its clinical efficacy. This situation is actually like mechanisms of many diseases, which are not known yet. As a part of a program to assess the chemical and biological diversity of traditional Chinese medicines, we focused on minor components, particularly on those from water extracts of TCMs based on their traditional use of decoctions. Our efforts have led to the characterization of a great numbers of previously unknown constituents, such as homosecoiridoids (1–5) from the flower buds of Lonicera japonica [1], indole alkaloids (6–9) from the root of Isatis indigotica [2,3], and conformational isomers of aconitane diterpenoids (10–15) from the lateral root of Aconitum carmichaelii [4]. In this presentation recent works on these commonly used TCMs will be outlined. Acknowledgements: Financial support from the National Natural Sciences Foundation of China (No. 30825044) is acknowledged.

Rapid screening of bioactive compounds from natural products by integrating 5-channel parallel chromatography coupled with on-line mass spectrometry and microplate based assays

A high throughput method was developed for rapid screening and identification of bioactive compounds from traditional Chinese medicine, marine products and other natural products. The system, integrated with five-channel chromatographic separation and dual UV–MS detection, is compatible with in vitro 96-well microplate based bioassays. The stability and applicability of the proposed method was validated by testing radical scavenging capability of a mixture of seven known compounds (rutin, dihydroquercetin, salvianolic acid A, salvianolic acid B, glycyrrhizic acid, rubescensin A and tangeretin). Moreover, the proposed method was successfully applied to the crude extracts of traditional Chinese medicine and a marine sponge from which 12 bioactive compounds were screened and characterized based on their anti-oxidative or anti-tumor activities. In particular, two diterpenoid derivatives, agelasine B and (−)-agelasine D, were identified for the first time as anti-tumor compounds from the sponge Agelas mauritiana, showing a considerable activity toward MCF-7 cells (IC50 values of 7.84±0.65 and 10.48±0.84μM, respectively). Our findings suggested that the integrated system of 5-channel parallel chromatography coupled with on-line mass spectrometry and microplate based assays can be a versatile and high efficient approach for the discovery of active compounds from natural products.

Hollow Fiber Liposome Microscreening and Its Application in Screening and Fishing Membrane Permeation Ingredient of Coumarins and Lignans in Traditional Chinese Medicine

Abstract A novel method was proposed to screen and analyze the membrane permeable ingredient group by hollow fiber liposome microscreening (HFLMS) coupled with high performance liquid chromatography (HPLC). In the HFLMS coupled with HPLC, the hollow fiber filled with liposome was used as a screening device, and the membrane permeable ingredient group were screened, fished and isolated from traditional Chinese medicine (TCM) extracts in phosphate buffer (pH 7.4). In this work, the surface nature of the hollow fiber with liposome was characterized, the nonspecific binding between activity enter of hollow fiber and the target compound was discussed, and the reproducibility of HFLMS coupled with HPLC was studied. Using this method, the coumarins and lignans compound groups that could permeate through the liposome membrane were screened and fished from TCM extracts, respectively. Some structures of the lignans and coumarins combined with liposomes in TCMs were identified by the comparison to the retention times of the reference substances.

Impact of Lycium Barbarum Polysaccharide and Danshensu on vascular endothelial growth factor in the process of retinal neovascularization of rabbit.

To discuss the impact of Lycium Barbarum Polysaccharide (LBP) and Danshensu purified from Traditional Chinese Medicine (TCM) on vascular endothelial growth factor (VEGF) of rabbits with retinal neovascularization. Forty rabbits were divided into normal control group, model control group, LBP group and Danshensu group. Animals in the normal control group were fed in the normal oxygen environment. Animals in the other three groups were put into the environment with 70% oxygen for 5 days in order to build the model of oxygen-induced vascular proliferation retinopathy. And then different TCM extract was injected into the abdominal cavities of these annimals. After 7 days, the VEGF content of in the serum of rabbit was measured by double antibody sandwich method. DATA ANALYSIS INDICATED THAT VEGF CONTENT WAS AS FOLLOWS: Danshensu group was lower than model control group (12.92±3.84ng/L vs 19.32±4.15ng/L, P<0.05); LBP group and normal control group were lower than model control group (12.92±3.84ng/L, 9.26±1.61ng/L vs 19.32±4.15ng/L, P<0.01); total blood viscosity, plasma viscosity, cholesterol content, fibrinogen content and triacylglycerol content after peritoneal injection of LBP and Danshensu were obviously lower than before injection. TCM extract-LBP and Danshensu can prominently reduce the content of VEGF in the process of vascular proliferative retinopathy of rabbit; can prevent the occurrence of retinal microvascular disease by improving partial oxygen-deficient environment or affecting all kinds of new growth factor.

Cell-based high-throughput proliferation and cytotoxicity assays for screening traditional Chinese herbal medicines

A simple, reliable, high-throughput screening method was developed and used to assess the pharmaceutical effects of extracts of traditional Chinese herbal medicines (TCHMs). This method is based on 3-dimensional (3-D) cultures of mouse embryonic stem (mES) and human colon cancer and breast cancer cells expressing enhanced green fluorescent protein (EGFP) in polyethylene terephthalate (PET) fibrous scaffolds on modified 384-well plates with online monitoring of culture fluorescence for dynamic responses of cells to drugs present in culture media. Cell responses to deoxycholic acid and the extracts of 3 TCHMs (Ganoderma lucidum spores, Ginkgo biloba, and Epimedium brevicornum) at various concentrations were investigated for their effects on proliferation and cytotoxicity. The screening results, i.e., the growth responses of cancer cells to those drugs, were consistent with what have been reported in the literature, confirming the reliability of the new screening approach. Different from previous screening methods for both TCHMs and western medicines that used animal models or 2-D cell-based assays with single cell lines, this 3-D cell-based screening method employs both cancer and normal cells and thereby provides a way for quick, direct evaluation of the anticancer effects of TCHMs. This method also offers assessment on the side effects of TCHMs.

Hollow fiber cell fishing with high performance liquid chromatography for screening bioactive compounds from traditional Chinese medicines

A novel hollow fiber cell fishing method with high performance liquid chromatography was proposed and used to screen, isolate, and analyze bioactive compounds from Traditional Chinese Medicines (TCMs). The active compounds that interact with the living cells acceptor inside the hollow fiber lumen were screened and isolated from the TCM extracts in phosphate buffer solution (pH 7.4). Subsequently, the active compounds bound to the cells were desorbed with methanol, and were analyzed using HPLC. HFCF with HPLC was introduced for the screening and analysis of lignans in Schisandra chinensis (Turcz) Baill and coumarins in Fructus Cnidii and Fructus Psoraleae. The surface properties of the hollow fibers filled with living cells were characterized. The nonspecific binding between the active centers of the hollow fibers and the bioactive compounds were investigated. The cell survival rates were determined before and after the screening. The repeatability of the method was tested. Some structures of the lignans and coumarins screened from TCMs were identified by the comparison to the retention times of the reference substances. HFCF-HPLC is a simple, fast, effective, and reliable method for the screening and analysis of bioactive compounds, and it can be extended to screen other bioactive compounds from TCMs.

Simultaneous Screening and Identifying Four Categories of Particular Flavonoids in the Leaves of Murraya exotica L. by HPLC-DAD-ESI-MS-MS

Polymethoxylated flavonoids (PMFs), the particular flavonoid subclass in which all or almost all hydroxyls are capped by methylation, have high oral bioavailability and various activities. A sensitive high-performance liquid chromatography-diode array detection-electrospray ionization tandem mass spectrometry (HPLC-DAD-ESI-MS-MS) method was established to screen and identify the PMFs in the leaves of Murraya exotica. Eight PMF standards, including two polymethoxylated flavones, two polymethoxylated flavanones, two polymethoxylated chalcones and two PMF glycosides, were first to be analyzed in positive mode by collision-induced dissociation MS-MS. On the basis of the ESI-MS, characterizations were deduced, and in the results of the extracted ion chromatogram MS-MS experiment, 26 PMFs, including 18 flavones, five flavanones or chalcones and three PMF glycosides, were screened out from the complex extract of the leaves of M. exotica. Among them, 24 PMFs were hydroxylated polymethoxyflavonoids, whereas the rest were all permethoxylated PMFs. This was the first systematic report on the presence of PMFs in the leaves of M. exotica. The results indicated that the established analytical method could be adopted as a rapid, effective technique for the structural characterization of PMFs from the complex extracts of traditional Chinese medicines.

A strategy of EIC-MS coupled with diagnostic product ions analysis for efficient discovery of new hydroxylated polymethoxyflavonoid glycosides from the leaves of Murraya paniculata L. using HPLC-DAD-MS/MS

Hydroxylated polymethoxyflavonoid glycosides (OH-PMFGs) are a rare kind of highly methoxylated flavone glycosides, which demonstrate many health-promoting bioactivities. Here we aim to identify OH-PMFGs from the leaves of Murraya paniculata using EIC (extracted ion chromatogram)-MS coupled with diagnostic product ions (DPIs) analytical method. Respective DPIs for glycosides of polymethoxyflavones, polymethoxyflavanones and polymethoxychalcones were obtained from the fragmentation pathways of eleven PMF (polymethoxylated flavonoid) standards. A sensitive HPLC-DAD-ESI-MS/MS was established for simultaneous qualitative and quantitative determination of the main OH-PMFGs in the extract. 54 OH-PMFGs including 49 flavone glycosides and 5 flavanone or chalcone glycosides were screened and identified from the leaves of M. paniculata. Meanwhile, the contents of three main OH-PMFGs in the extract were determined by HPLC-UV. It was the first systematic report of the presence of rare OH-PMFGs in the plants of the genus Murraya. The results indicated that the methodology developed could be employed as an effective technique for structural characterization of OH-PMFGs in complex extracts of TCMs (traditional Chinese medicines).

Intervention of Traditional Chinese Medicine Extract in Hepatic Fibrosis Through the TGFβ/Smad Signal Pathway

Intervention of Traditional Chinese Medicine Extract in Hepatic Fibrosis Through the TGFβ/Smad Signal Pathway

A Simulation Study for Effect of Particle Size on On-line Measurement Sampling System of Traditional Chinese Medicine Extraction Process

As the objective of simulation study,the samples of gardenia fructus extract were collected every 5 min while boiling for 60 min.Each sample was filtrated by different sizes of sieve meshes,and then scanned by the near infrared spectra.Using the HPLC determined value for reference,quantitative calibration models of geniposide were established by the partial least square method.Based on on-line near infrared spectral technology,the effects of by-pass preprocess system and filter mesh on the accuracy of calibration model were further studied.By the analysis of paired t test,the filtrated sample was compared with original extract,which illustrated that the filtrated sample and the original extract weren' t significant different,so the sample after filtration could be representative of the original gardenia fructus extract for the data analysis.In addition,after the sample was filtrated by a sieve with meshes size of 0.049 mm,quantitative prediction model,which was built by raw spectrum pretreatment,could reach the ideal prediction result( Root Mean Square Error of Cross Validation( RMSECV) was 0.1962,Root Mean Square Error of Prediction( RMSEP) was 0.1867,Bias was 0.0256).It was recommended that proper size of sieve mesh could be selected by the simulation study,and then the feasibility of sample pretreatment system could be verified on-line,finally the stable preprocess system could be established.These steps could improve the performance of prediction models dramatically.

Idiopathic pulmonary fibrosis: The current status of its epidemiology, diagnosis, and treatment in China.

Idiopathic pulmonary fibrosis (IPF) is a type of intractable and rare disease, and its epidemiology in China is still unclear. The diagnosis and treatment of IPF has received considerable attention and two editions of guidelines on IPF diagnosis and treatment have been published by the Chinese Society of Respiratory Diseases. Treatment of IPF with Traditional Chinese Medicine (TCM) has been widely investigated in China and several types of TCM extracts are reported to be effective in animal models. One effective treatment is lung transplantation; this treatment has been successfully performed in China, yielding satisfactory long-term survival.

Rapid Screening of Lipase Inhibitors from Fructus aurantii Using Lipase-immobilized Magnetic Beads Coupled with High-performance Liquid Chromatography-Mass Spectrometry

A new method for the rapidly screening of lipase inhibitors from extracts of traditional Chinese medicine using lipase-immobilized magnetic beads coupled with high-performance liquid chromatography-mass spectrometry(LC-MS) was proposed.After the lipase-immobilized magnetic beads were prepared,they were incubated with total flavonoid of Fructus aurantii to fish the ligands of lipase.Then,the eluent of magnetic beads' screening was analyzed by LC-MS.Four compounds,as naringin,hesperidin,neohesperidin and poncirin,were identified as the potential lipase inhibitors.Furthermore,the in vitro inhibitory activities against lipase of four compounds were tested.The results showed that hesperidin,neohesperidin and poncirin exhibit lipase inhibitory activities,while poncirin was firstly reported to be a lipase inhibitor.Our research showed that active compounds could be rapidly screened from extracts of traditional Chinese medicine via lipase-immobilized magnetic beads coupled with LC-MS.

Insulin-secretagogue activity of eleven plant extracts and twelve pure compounds isolated from Aralia taibaiensis

AimsTo investigate the insulinogenic activities of the eleven saponins enriched traditional Chinese medicine (TCM) extracts. Main methodsRadioimmunoassay and trypan blue exclusion assay were used to investigate the insulinogenic activity and cytotoxic effects respectively. Key findingsThe total saponin extract of Aralia taibaiensis (sAT) exhibited highest insulinogenic activity and no cytotoxicity was recorded. Twelve pure compounds from sAT stimulated insulin secretion from a mouse insulinoma βTC3 cells in a concentration-dependent manner. TA35 outperformed the other compounds which suggested that the active insulinogenic ingredient of sAT was probably TA35. In addition, both sAT and TA35 markedly potentiated glucose-induced insulin secretion. SignificanceOur study is the first to show that sAT dramatically stimulated insulin secretion and its antidiabetic activity may be related to its high saponin content. These findings suggested that sAT and the compound TA35 isolated from sAT may provide novel therapeutic tools for the treatment of non-insulin dependent diabetes mellitus (NIDDM).

Antioxidant and anti-inflammatory activities of selected Chinese medicinal plants and their relation with antioxidant content

BackgroundThe main aim of this study is to evaluate the antioxidant and anti-inflammatory properties of forty four traditional Chinese medicinal herbal extracts and to examine these activities in relation to their antioxidant content.MethodsThe antioxidant activities were investigated using DPPH radical scavenging method and yeast model. The anti-inflammatory properties of the herbal extracts were evaluated by measuring their ability to inhibit the production of nitric oxide and TNF-α in RAW 264.7 macrophages activated by LPS and IFN- γ, respectively. The cytotoxic effects of the herbal extracts were determined by Alomar Blue assay by measuring cell viability. In order to understand the variation of antioxidant activities of herbal extracts with their antioxidant contents, the total phenolics, total flavonoids and trace metal (Mg, Mn, Cu, Zn, Se and Mo) quantities were estimated and a correlation analysis was carried out.ResultsResults of this study show that significant levels of phenolics, flavonoids and trace metal contents were found in Ligustrum lucidum, Paeonia suffuticosa, Salvia miltiorrhiza, Sanguisorba officinalis, Spatholobus suberectus, Tussilago farfara and Uncaria rhyncophylla, which correlated well with their antioxidant and anti-inflammatory activities. Some of the plants displayed high antioxidant and anti-inflammatory activities but contained low levels of phenolics and flavonoids. Interestingly, these plants contained significant levels of trace metals (such as Zn, Mg and Se) which are likely to be responsible for their activities.ConclusionsThe results indicate that the phenolics, flavonoids and trace metals play an important role in the antioxidant activities of medicinal plants. Many of the plants studied here have been identified as potential sources of new antioxidant compounds.

Cytoprotective properties of traditional Chinese medicinal herbal extracts in hydrogen peroxide challenged human U373 astroglia cells

Age is the leading risk factor for many of the most prevalent and devastating diseases including neurodegenerative diseases. A number of herbal medicines have been used for centuries to ameliorate the deleterious effects of ageing-related diseases and increase longevity. Oxidative stress is believed to play a role in normal ageing as well as in neurodegenerative processes. Since many of the constituents of herbal extracts are known antioxidants, it is believed that restoring oxidative balance may be one of the underlying mechanisms by which medicinal herbs can protect against ageing and cognitive decline. Based on the premise that astrocytes are key modulators in the progression of oxidative stress associated neurodegenerative diseases, 13 herbal extracts purported to possess anti-ageing properties were tested for their ability to protect U373 human astrocytes from hydrogen peroxide induced cell death. To determine the contribution of antioxidant activity to the cytoprotective ability of extracts, total phenol content and radical scavenging capacities of extracts were examined. Polygonum multiflorum, amongst others, was identified as possessing potent antioxidant and cytoprotective properties. Not surprisingly, total phenol content of extracts was strongly correlated with antioxidant capacity. Interestingly, when total phenol content and radical scavenging capacities of extracts were compared to the cytoprotective properties of extracts, only moderately strong correlations were observed. This finding suggests the involvement of multiple protective mechanisms in the beneficial effects of these medicinal herbs.

Study on reference extracts for quality assessment on traditional Chinese medicines based on akebiae saponin

On the basis of analysis and summary of current application of reference extracts of traditional Chinese medicine, reference extracts for quality assessment of traditional Chinese medicines are divided into three classes by purpose: 1. herbal or volatile oil reference extracts for TLC identification of original herbal medicines or patent traditional Chinese medicines containing original herbal medicines. 2. component reference extracts for identification of certain components or positioning of chromatographic peaks. 3. reference extracts marked with component content to be determined for quantitative analysis on samples. For the third kind of reference extracts, akebiae saponin was taken as an example for preparation thought and methods in such aspects as preparation process, component identification and content analysis.