An ultra-robust fingerprinting method for quality assessment of traditional Chinese medicine using multiple reaction monitoring mass spectrometry / 药物分析学报

Chromatographic fingerprinting has been perceived as an essential tool for assessing quality and chemical equivalence of traditional Chinese medicine.However,this pattern-oriented approach still has some weak points in terms of chemical coverage and robustness.In this work,we proposed a multiple reaction monitoring(MRM)-based fingerprinting method in which approximately 100 constituents were simultaneously detected for quality assessment.The derivative MRM approach was employed to rapidly design MRM transitions independent of chemical standards,based on which the large-scale finger-printing method was efficiently established.This approach was exemplified on QiShenYiQi Pill(QSYQ),a traditional Chinese medicine-derived drug product,and its robustness was systematically evaluated by four indices:clustering analysis by principal component analysis,similarity analysis by the congruence coefficient,the number of separated peaks,and the peak area proportion of separated peaks.Compared with conventional ultraviolet-based fingerprints,the MRM fingerprints provided not only better discriminatory capacity for the tested normal/abnormal QSYQ samples,but also higher robustness under different chromatographic conditions(i.e.,flow rate,apparent pH,column temperature,and column).The result also showed for such large-scale fingerprints including a large number of peaks,the angle cosine measure after min-max normalization was more suitable for setting a decision criterion than the unnormalized algorithm.This proof-of-concept application gives evidence that combining MRM tech-nique with proper similarity analysis metrices can provide a highly sensitive,robust and comprehensive analytical approach for quality assessment of traditional Chinese medicine.

Topical administration of traditional Chinese medicine and the methods for the promotion of drug penetration / 中国药科大学学报

@#Monomer component extracted from the herb is the main effective component of traditional Chinese medicine(TCM). Topical administrations of monomer component of TCM has attracted more and more attention due to the convenience of administration and the concentration enrichment in lesion. The main task for the studies of topical drug delivery system is to design the methods which can promote the penetration of the drugs. Currently, the main methods used to improve the penetration of monomer component of TCM includes the synthesis of different dosage forms and the application of physical and chemical techniques to facilitate the penetration of the drugs. This review summarizes the progress in different indications and mechanisms of diverse monomer components of TCM, different dosage forms, and physicochemical techniques used to facilitate drug penetration.

Experimental Study of Optimized Compatibility Component of Water Extractives of Liqi Huoxue Medicinals by Uniform Designed Method / 世界科学技术-中医药现代化

This experiment was designed to search and identify the active principle as well as the optimal proportion of water-soluble extractives of traditional Chinese medicine (water-soluble extractives) Liqi Huoxue medicinals com-patibility (Qixue Bingzhi Fang-CWQB) in the prevention and treatment of atherosclerosis (As) by optimal uniform design method. The water-soluble extractives of CWQB were divided into 6 sections (A, B, C, D, E, F) through macroporous resin. The effect intensity and step of every component were compared through its effect on blood fat level, platelet aggregation, inflammatory factors, vascular endothelial growth factor (VEGF) and so on among hyper-lipoidemia rat models. The pharmacological experimental results and statistical analysis showed that CWQB water-soluble extractives of component D (mainly is paeoniflorin, accounted for 49.12%), component E (mainly is total flavonoids, accounted for 30.0%) compatibility had better effects on decreasing blood fat and triglyceride (TG). Com-pared with the model group, there was significant difference (P < 0.01). It also had inhibiting effect on endothelin (ET) and maximum platelet aggregation rate (P < 0.01). The component F (mainly is total acids, accounted for 32.7%) had inhibiting effect on serum IL-6 and IL-8 (P< 0.01). It was concluded that different compatibility of wa-ter-soluble extractives of CWQB can be applied to different targets or steps of the body. The active principle extrac-tives include main component of paeoniflorin, flavonoids and total acids. The best proportion is about 1:1:1.

Building methodology for discovering and developing Chinese medicine based on network biology / 中国中药杂志

In this article, we introduced the concept and technology of network biology and network pharmacology, and proposed the possible strategy for building the methodology for discovering and developing Chinese medicine based on network biology. The concept of network pharmacology of Chinese medicine and its potential application in the modernization of traditional Chinese medicine was also discussed.

Prediction of multi-target of Aconiti Lateralis Radix Praeparata and its network pharmacology / 中国中药杂志

<p><b>OBJECTIVE</b>To predict multi-targets by multi-compounds found in Aconiti Lateralis Radix Praeparata and construct the corresponding multi-compound-multi-target network.</p><p><b>METHOD</b>Based on drug-target relationships of FDA approved drugs, a model for predicting targets was established by random forest algorithm. This model was then applied to predict the targets of Aconiti Lateralis Radix Praeparata and construct the multi-compound-multi-target network.</p><p><b>RESULT</b>The predicted targets of 22 compounds of Aconiti Lateralis Radix Praeparata are validated by literature. Each compound in the established network was correlated with 16. 3 targets on average, while each target was correlated with 4. 77 compounds on average, which reflects the "multi-compound and multi-target" characteristic of Chinese medicine.</p><p><b>CONCLUSION</b>The proposed approach can be used to find potential targets of Chinese medicine.</p>

Network pharmacology study on major active compounds of Fufang Danshen formula / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the correlations between multi-compounds of Fufang Danshen formula and their multi targets and multi diseases.</p><p><b>METHOD</b>Literature knowledge of nine major active compounds from Fufang Danshen formula, including tanshinone II(A), salvianolic acid B, protocatechuic aldehyde, danshensu, cryptotanshinone, notoginsenoside R1, ginsenoside Rg1, ginsenoside Rb1 and borneol were collected from PubMed. Combined with cardiovascular related diseases and genes from OMIM database, the corresponding multi-compound-multi- target-multi-disease network was constructed and visualized by Cytoscape software.</p><p><b>RESULT</b>AND CONCLUSION: Network analysis showed that the 9 compounds could modulate 42 cardiovascular associated genes (e. g. PPARG, ACE, KCNJ11, KCNQ1, ABCC8, et al), which related to 30 cardiovascular associated diseases including non-insulin-dependent diabetes mellitus, hyperinsulinemic hypoglycemia, hypertension, and coronary heart disease. These results suggested new potential indications of Fufang Danshen formula.</p>

Symptom-based traditional Chinese medicine slices relationship network and its network pharmacology study / 中国中药杂志

<p><b>OBJECTIVE</b>To discover and interpret the correlations between traditional Chinese medicine (TCM) slices and their properties such as function, symptom and channel tropism by constructing the relationship network using network pharmacology approaches.</p><p><b>METHOD</b>TCM slices related information was extracted from Chinese Pharmacopeia (2010 edition, volume I) by text mining, and was used to construct the TCM slices-symptom relationship network. The corresponding network analysis was also performed.</p><p><b>RESULT</b>Three thousands and sixteen pair of TCM slice-symptom correlation associated with 646 TCM slices was discovered, and the constructed network unfolded the complex relationships between TCM slices. Further network analysis results indicated that the un-annotated function and channel tropism of TCM slice can be revealed by proposed symptom-based network.</p><p><b>CONCLUSION</b>Network pharmacology approaches can be applied in TCM research to discover and interpret the relationships between TCM slices and their properties.</p>

Discovering active components from traditional Chinese medicine by component-knockout approach / 中国中药杂志

<p><b>OBJECTIVE</b>A component-knockout approach was established to discover active components from traditional Chinese medicine.</p><p><b>METHOD</b>According to the principle of gene knockout technique, an experiment workflow for component-knockout method was developed, which is distinct from the bio-guided screening method. The differences of therapeutic efficacies between different combinations of individual components were analyzed by some statistical methods including Analysis of Variance (ANOVA), whilst a set of criterias were established to assess active components. By comparing the difference of drug efficacy between the original formulae and the mixture being knockout certain component, the active components can be identified.</p><p><b>RESULT</b>The presented component-knockout method was applied to discover the active components of Shenmai formulae for the synergistic effects on the cyclophosphamide chemotherapy for S180 tumor-bearing mice. The results indicated that panoxadiol, a type of ginsenosides, were the effective components of Shenmai formulae.</p><p><b>CONCLUSION</b>A new method for identifying effective components from Chinese medicinal formulae was developed and successfully applied to discover the active components of Shenmai Formulae, which possesses the synergistic actions towards chemotherapy process.</p>

Analysis of myocardial proteomic alteration after Qishenyiqi formula treatment in acute infarcted rat hearts / 中国中药杂志

<p><b>OBJECTIVE</b>Use proteomic technique to research the effects of Qishenyiqi formula in acute myocardial infarcted rats, and further explore its protein-level mechanism.</p><p><b>METHOD</b>The rat hearts of sham, myocardial infarcted and Qishenyiqi formula treated groups were collected. Analyzing and comparing the differences of protein expressions in each groups with two-dimensional electrophoresis, computer-assisted image analysis and matrix-assisted laser desorption/ionization mass spectrometry.</p><p><b>RESULT</b>Compared with myocardial infarcted group, there were 14 protein expressions changed in Qishenyiqi formula treated group. The result showed that 9 protein expressions were up-regulated, and 5 ones were down-regulated significantly. These proteins are associated with energy metabolism, mitochondrial function, oxidative stress, cytoskeleton and others.</p><p><b>CONCLUSION</b>Myocardial protective effects of Qishenyiqi formula in myocardial infarcted procedure might be closely related to the recovery of energy supply, the reduction of oxidative stress, as well as the promotion of cell recovery.</p>

Chemical constituents from Corydalis yanhusuo / 中国中药杂志

<p><b>OBJECTIVE</b>To study the chemical constituents from the 60% ethanol extract of Corydalis yanhusuo.</p><p><b>METHOD</b>The chemical constituents were separated and purified by various chromatographic techniques, and their structures were elucidated by chemical evidence and spectroscopic analysis.</p><p><b>RESULT</b>Nine alkaloids were obtained and determined as 7-formyldidehydroglaucine (1), nantenine (2), (+)-O-methylbulbocapnine (3), d-corydaline (4), tetrahydrocoptisine (5), 8-oxocoptisine (6), palmatine (7), tetrahydropalmatine (8), and dehydrocorydaline (9), respectively.</p><p><b>CONCLUSION</b>Compound 1 was obtained from natural products for the first time, and its NMR data were firstly reported. Compound 3 was reported from Papaveraceae for the first time, and compound 6 was firstly reported from Corydalis yanhusuo.</p>

Identification of major constituents in the traditional Chinese medicine "QI-SHEN-YI-QI" dropping pill by high-performance liquid chromatography coupled with diode array detection-electrospray ionization tandem mass spectrometry.

An HPLC-DAD-ESI-MS(n) method was developed for simultaneous analysis of major chemical constituents in "QI-SHEN-YI-QI" dropping pill, a traditional Chinese medicine (TCM) widely used for treating cardiovascular diseases. The chromatographic separation was performed on an intertsil ODS-3 C(18) column (4.6 mm x 250 mm, 5 microm), whilst water with 0.05% acetic acid and acetonitrile were used as mobile phase. On the basis of the characteristic UV absorption profile, the information of molecular weight, and structure provided by ESI-MS(n), 31 constituents derived from Astragalus membranaceus, Radix Salviae Miltiorrhizae, and Panax notoginseng, were detected and 20 of them were identified in this study. The proposed method contributes to the quality control of "QI-SHEN-YI-QI" dropping pill.

Effects of water extractives of a qi and blood regulating prescription on early atherosclerosis of apolipoprotein E-deficient mice / 中西医结合学报

OBJECTIVE: To investigate whether the water extractives of regulating qi and blood prescription (WQBP) had effects on early atherosclerosis of apolipoprotein E-deficient mice (ApoE-mice) at the age of 19 weeks or not, and to explore the possible mechanisms. METHODS: Forty ApoE-mice, six weeks of age, were given high-fat diet and randomly divided into four groups: high-dose WQBP-treated group (360 mg/kg), low-dose WQBP-treated group (72 mg/kg), simvastatin-treated group (25 mg/kg) and untreated group, with ten mice in each group. Meanwhile, ten C57BL/6 mice of same genetic background were allocated to normal control group. Mice in the high- and low-dose WQBP-treated groups and simvastatin-treated group were administered with corresponding drugs from the 15 to 19 weeks. Mice in the untreated and normal control groups were administered with isovolumic water. Sacrificed at 19 weeks, the level of blood-lipid, the plaque construction, plaque integral, and the contents of plaque macrophages and vessel smooth muscle cells of the mice were analyzed by immunohistochemical method and a computer picture processing system. RESULTS: Compared to the untreated group, high-dose WQBP group could obviously decrease the level of low-density lipoprotein cholesterol (LDL-C). Simvastatin group could decrease the levels of LDL-C and total cholesterol (TC) (P<0.01). In high-dose WQBP-treated group and simvastatin-treated group, the thickness of fiber cap and the quantities of vessel smooth muscle cells increased (P<0.05), the quantities of plaque macrophages and the ratio of lipid and plaque reduced (P<0.01). CONCLUSION: WQBP and simvastatin can interfere in early atherosclerosis of ApoE-mice, attenuate and stabilize plaque in some extent. The mechanisms may include adjusting blood lipid, decreasing macrophage number and increasing the quantities of vessel smooth muscle cells.

Predicting Caco-2 permeability using support vector machine and chemistry development kit.

PURPOSE: To predict Caco-2 permeability is a valuable target for pharmaceutical research. Most of the Caco-2 prediction models are based on commercial or special software which limited their practical value. This study represents the relationship between Caco-2 permeability and molecular descriptors totally based on open source software. METHODS: The Caco-2 prediction model was constructed based on descriptors generated by open source software Chemistry Development Kit (CDK) and a support vector machine (SVM) method. Number of H-bond donors and three molecular surface area descriptors constructed the prediction model. RESULTS: The correlation coefficients (r) of the experimental and predicted Caco-2 apparent permeability for the training set and the test set were 0.88 and 0.85, respectively. CONCLUSION: The results suggest that the SVM method is effective for predicting Caco-2 permeability. Membrane permeability of compounds is determined by number of H-bond donors and molecular surface area properties.

LC/MS fingerprinting of Shenmai injection: a novel approach to quality control of herbal medicines.

Chromatographic fingerprinting has been recommended as a potential and reliable strategy for the quality control of herbal medicines. Although varieties of chromatographic techniques, particularly HPLC, have been widely employed, hyphenated chromatographic approach has not been sufficiently exploited in chromatographic fingerprinting. In this work, LC/MS fingerprinting of Shenmai injection was developed. Thirty ginsenosides as well as seven ophioponins were selected to construct the LC/MS fingerprint using selective ion monitoring (SIM) mode, while previous HPLC fingerprint [H.J. Zhang, Y.J. Wu, Y.Y. Cheng, J. Pharm. Biomed. Anal. 31 (2003) 175-183] only represents the ginsenosides. Subsequently, the proposed LC/MS fingerprints were applied to identifying the product manufacturers. All the samples were accurately classified based on their LC/MS fingerprints in conjunction with principal components analysis (PCA). This study would be potentially helpful to improve the quality control ability of fingerprinting-based strategy for complex herbal medicines.

Fractal fingerprinting of chromatographic profiles based on wavelet analysis and its application to characterize the quality grade of medicinal herbs.

Extracting chemical fingerprints is an important step for representing and interpreting chromatographic data. In this paper, the chromatographic profile is decomposed into components at different resolution levels using wavelet analysis, then the fractal dimensions of these components are computed as the chemical fingerprints. The chromatographic fingerprint is characterized by the vector composed of these chemical fingerprints, which can represent the chemical patterns of different categories of complex samples. Computer simulations reveal that the fractal fingerprints are more stable than the original chromatographic profile data with respect to variations of peak retention time. To demonstrate the validity of this method, the evaluation of the quality of the medicinal herb Angelica sinensis (Oliv.) diels is investigated. Principal component analysis of the fractal fingerprints indicates that samples belonging to the same quality grade are clustered together, while those belonging to different quality grades are separated. Using these fractal fingerprints taken from the chromatographic scans as inputs for an artificial neural network (ANN). The quality grades of two sets of the herbs were verified by cross-validation, indicating that 96.7% of the herbs are correctly identified with respect to their quality grades evaluated by experienced experts, and 100.0% of the herbs are correctly identified with respect to their quality grades determined by pharmacodynamical evaluation.

Analysis of 'SHENMAI' injection by HPLC/MS/MS.

An HPLC/MS/MS method was developed for the analysis of 'SHENMAI' injection, composed of red ginseng and ophiopogon. The constituents of 'SHENMAI' were found to be similar with those of ginseng and 39 ginsenosides were detected. By the studies of MS and MS/MS spectra and the comparison with literature data, most of these ginsenosides were identified. Based on this study, suggestions were put forward to improve the quality control system of 'SHENMAI' injection.

Studies on integrated analysis of serum proteomics and urinary metabonomics of breast cancer patients / 中华检验医学杂志

Objective To study on the diagnosis methodology of breast cancer using integrated analysis of proteomics and metabonomics approach.Methods The serum samples from the 14 breast cancer patients and 7 healthy people were analyzed by two-dimensional electrophoresis in order, to find differential expressed proteins. On the other hand, an HPLC-based urinary metabonomics profiling was employed to pick out chemical patterns between breast cancer patients and healthy people.Results Glutathione S-transferase M5 was found highly expressed in the breast cancer patients′serum.The concentration pattern of S-Adenosylmethionine, Orotidine, 1-Methyladenosine and N2-Methylguanosine in patients′ urine were peculiarly different from that in the healthy people.Conclusion There are peculiar chemical pattern differentiae between the body fluid of breast cancer patients and the body fluid of healthy people. Synthetic study of cancer proteomics and metabonomics can contribute not only to prognosis of cancer but also to the pathogenesis of cancer.

Determination of ginsenoside in ethanol extracting process of red ginseng using near-infrared spectroscopy / 中草药

Objective Near-infrared (NIR) spectroscopy was used as a fast analytical technique in the ethanol reflux-extraction process of red ginseng. Methods The NIR spectra of the extracting solution of red ginseng were obtained and the reference measurements of the active constituent in the extracting solution were performed by the colorimetric method. Firstly, the interference information in the spectra was detected by orthogonal signal correction (OSC) method. Then a calibration model between NIR spectra and reference measurements was established by partial least square regression. Results The results showed that the predictive accuracy of NIR calibration model used for the determination of ginsenoside in ethanol extracting process of red ginseng was good. Conclusion NIR Spectroscopy could be applied to the fast analysis for ethanol extracting processes of red ginseng.

Optimization for vacuum belt drying process of Radix Salvia Miltiorrhiza extract / 中草药

Objective To select the optimum conditions of the vacuum belt drying process of Radix Salvia Miltiorrhiza(RSM) extract.Methods The process was studied by using orthogonal test design and grading method for multi-index on the parameters of the water content of dried product and drying rate of RSM extract,the average quantity of vapour during unit time span,as the index.Results The optimum process determined by the grading method was listed as follows: water content of the extract before drying was 40%,the feeding speed was 1.5 mL/s,the belt speed was 5 cm/min.Conclusion This technology can increase the average quantity of vapour during unit time span and the drying product has high quality with lower water content and desirable drying rate.

Fast quality assessment of Radix Ginseng Rubra by near-infrared spectroscopy / 中草药

Objective Near-infrared (NIR) reflectance spectroscopy was used to develop a fast quality assessment method by simultaneous material identification and moisture quantification of Radix Ginseng Rubra (RGR). Methods The sample was identified by the comparison of its spectrum with a standard NIR spectral library. Similarity measurement was used as the discriminating parameter. The moisture content of sample was quantified by a partial least square (PLS) calibration model, correlative spectrum calculation was used for wavelength selection and multiplicative signal correction (MSC) was applied for pretreatment in the calibration model. Primary reference data were obtained using the traditional loss on drying (LOD) method. Results The NIR library can distinguish RGR from the counterfeit successfully. The optimized eight-factor PLS calibration model of NIR spectra has a high correlation coefficient (R=(0.999 7).) Conclusion The proposed method is rapid, accurate and can be used routinely in the traditional Chinese materia medica manufacturers for quality control of raw materials.