Strategy for analysis and screening of bioactive compounds in traditional Chinese medicines

Traditional Chinese medicines (TCMs), due to their long time clinic test and reliable therapeutic efficacy, are attracting increased global attention served as excellent pools of bioactive compounds for the discovery of new drugs. However, hundreds or even thousands of components are usually contained in traditional Chinese medicines and only a few compounds are responsible for the pharmaceutical and/or toxic effects. The large numbers of other components in traditional Chinese medicines make the screening and analysis of the bioactive components extremely difficult. By the way, the combination effect of bioactive components on the pharmacological activity makes it very difficult to clear the therapeutic mechanism of TCMs. Therefore, some strategies have to design for screening of bioactive compounds in traditional Chinese medicines, which further leads to disclose the therapeutic mechanism of TCMs in molecular level. The review will summarize the present state of the art of screening strategy for active compounds in traditional Chinese medicines, and the chromatography methods for screening and analysis of bioactive compounds in traditional Chinese medicines will be emphasized. (C) 2004 Elsevier B.V. All rights reserved.

The use of CE-electrochemiluminescence with ionic liquid for the determination of bioactive constituents in Chinese traditional medicine

CE/tris(2,2-bipyridyl) ruthenium(ll) (Ru(bpy)(3)(2+)) electrochemiluminescence (ECL), CEECL, with an ionic liquid (IL) detection system was established for the determination of bioactive constituents in Chinese traditional medicine opium poppy which contain large amounts of coexistent substances. A minimal sample pretreatment which involves a one-step extraction approach avoids both sample loss and environmental pollution. As the nearby hydroxyl groups in some alkaloid such as morphine may react with borate to form complexes and IL, as a high-conductivity additive in running buffer, could cause an enhanced field-amplified effect of electrokinetic injection. Running buffer containing 25 mM borax-8 mM 1-ethyl-3-methylimidazolium tetrafluoroborate (EMImBF(4)) IL (pH 9.18) was used which resulted in significant changes in separation selectivity and obvious enhancement in ECL intensities for those alkaloids with similar structures. Sensitive detection could be achieved when the distance between the Pt working electrode and the outlet of separation capillary was set at 150 mu m and the stainless steel cannula was fixed approximately 1 cm away from the outlet of the capillary. Quantitative analysis of four alkaloids was achieved at a detection voltage of 1.2 V and a separation voltage of 15 kV in less than 7 min.

Facile separation and determination of Aconitine alkaloids in traditional Chinese medicines by CE with tris(2,2 '-bipyridyl) ruthenium(II)-based electrochemi-luminescence detection

A facile CE method coupled with tris(2,2'-bipyridyl) ruthenium(ll)-based electrochem iluminescence [Ru(bpy)(3)(2+)] detection was developed for simultaneous determination of Aconitum alkaloids, i.e., hypaconitine (HA), aconitine (AC), and mesaconitine (MA) in baseline separation. The optimal separation of these Aconitum alkaloids was achieved in a fused-silica capillary column (50 cm x 25 mu m id) with 30 mM phosphate solution (pH 8.40) as running buffer at 12 kV applied voltage. The three alkaloids can be determined within 10 min by a single run. The calibration curves showed a linear range from 2.0 x 10(-7) to 2.0 x 10(-5) M for HA, 3.4 x 10(-7) to 1.7 x 10(-5) M for AC, and 3.8 x 10(-7) to 1.9 x 10(-5) M for MA. The RSDs; for all analytes were below 3.01%. Good linear relationships were found with correlation coefficients for all analytes exceeding 0.993. The detection limits were 2.0 x 10(-8) M for HA, 1.7 x 10(-7) M for AC, and 1.9 x 10(-7) M for MA under optimal conditions. This method was successfully applied to determine the three alkaloids in Aconitum plants.

Molecular authentication of the traditional Tibetan medicinal plant Swertia mussotii

Swertia mussotii is an important species in Tibetan folk medicine. However, it is quite expensive and frequently adulterated, so reliable methods for authentication of putative specimens and preparations of the species are needed to protect consumers and to support conservation measures. We show here that the chloroplast (cp) DNA rpl16 intron has limited utility for differentiating S. mussotii from closely related species, since the cpDNA rpl16 sequences are identical in S. mussotii and two other species of Swertia. However, the rDNA internal transcribed spacer (ITS) sequences differ significantly between S. mussotii and all of 13 tested potential adulterants. Thus, the ITS region provides a robust molecular marker for differentiating the medicinal S. mussotii from related adulterants. Therefore, a pair of allele-specific diagnostic primers based on the divergent ITS region was designed to distinguish S. mussotii from the other species. Authentication by allele-specific diagnostic PCR using these primers is convenient, effective and both simpler and less time-consuming than sequencing the ITS region.

各向同性与各向异性介质中地震波旅行时正演模拟与反演成像

By seismic tomography, interesting results have been achieved not only in the research of the geosphere with a large scale but also in the exploration of resources and projects with a small scale since 80'. Compared with traditional inversion methods, seismic tomography can offer more and detailed information about subsurface and has been being paid attention by more and more geophysicists. Since inversion based on forward modeling, we have studied and improved the methods to calculate seismic traveltimes and raypaths in isotropic and anisotropic media, and applied the improved forward methods to traveltime tomography. There are three main kinds of methods to calculate seismic traveltime field and its ray path distribution, which are ray-tracing theory, eikonal equation by the finite-difference and minimum traveltime tree algorithm. In ray tracing, five methods are introduced in the paper, including analytic ray tracing, ray shooting, ray bending, grid ray tracing and rectangle grid ray perturbation with three points. Finite-difference solution of eikonal equation is very efficient in calculation of seismic first-break, but is awkward in calculation of reflection traveltimes. We have put forward a idea to calculate traveltimes of reflected waves using a combining way of eikonal equation method and other one in order to improve its capability of dealing with reflection waves. The minimum traveltime tree algorithm has been studied with emphases. Three improved algorithms are put forward on the basis of basic algorithm of the minimum traveltime tree. The first improved algorithm is called raypath tracing backward minimum traveltime algorithm, in which not only wavelets from the current source but also wavelets from upper source points are all calculated. The algorithm can obviously improve the speed of calculating traveltimes and raypaths in layered or blocked homogeneous media and keep good accuracy. The second improved algorithm is raypath key point minimum traveltime algorithm in which traveltimes and raypaths are calculated with a view of key points of raypaths (key points of raypths mean the pivotal points which determine raypaths). The raypath key point method is developed on the basis of the first improved algorithm, and has better applicability. For example, it is very efficient even for inhomogeneous media. Another improved algorithm, double grid minimum traveltime tree algorithm, bases upon raypath key point scheme, in which a model is divided with two kinds of grids so that the unnecessary calculation can be left out. Violent undulation of curved interface often results in the phenomenon that there are no reflection points on some parts of interfaces where there should be. One efficacious scheme that curved interfaces are divided into segments, and these segments are treated respectively is presented to solve the problem. In addition, the approximation to interfaces with discrete grids leads to large errors in calculation of traveltimes and raypaths. Noting the point, we have thought a new method to remove the negative effect of mesh and to improve calculation accuracy by correcting the traveltimes with a little of additional calculation, and obtained better results.

改造型盆地油气成藏机理与油气预测－以银根－额济纳旗盆地为例

Reformed basin is a basin that underwent multiple immense reformation after the sedimentary stage, the major geologic elements of the petroleum system in the prototyped basin are destroyed to a certain extent, and their petroleum system has been reconstructed. This type of basin is frequently found in the course of exploration both home and abroad. In China, especially in the western and southern part of China, the basins in which oil explorations have been conducted are mostly reformed basins. The reformed basins from Paleozoic, Late Mesozoic to Cenozoic are widely distributed in West and South China. They are, and moreover, will be a challenge for oil and gas exploration. The conventional investigation and exploration techniques used in the slightly reconstructed basin just don't work well when facing the reformed basin. Therefore, the study on the reformed basin, especially the study on the pool-forming mechanism and reservoir prediction becomes a focus and one of difficulties for the geologists overseas and domestic. Yingen-Ejinaqi Basin is a typical case of the Late Mesozoic and Cenozoic reformed basins in China. It locates in West China and is a exploration frontier with difficulties and no break through is made for years. A comprehensive research on it will be of significance for oil and gas exploration in similar basins of China. The late research for reformed basin in China now is mainly concentrated on basin classification, formation mechanism, geologic features, and survey technique, distribution regularity of oil accumulation and its dominating factors, assessment of oil exploration prospect and target zones, etc. On the other hand, the study on the pool-forming mechanism and reservoir prediction seems insufficient in systematization, and the research is deficient in methodology and combination of qualitative and quantitative studies, as well as the application of the new theory and techniques. The current efforts are mainly directed to structures (faults), sedimentation, the relationship between reservoir evolution and oil accumulation, and some other relevant fields. However, the application of the new theory and techniques seems to be insufficient such as petroleum system, pool-forming dynamics, fluid pressure compartment, and basin simulation, etc. So is the dynamic and integrated research. As a result, incomplete knowledge and understandings derived from the research on pool-forming mechanism and reservoir prediction often do not accord with rea-lity of the basin. The study and exploration under the guidance of this knowledge will inevitably lead to errors and failure. This paper, based on the previous study of the other geologists on reformed basins, with emphasis on "wholeness or systematic, dynamic and integrated" research, presents a reverse thinking of beginning from conserved units in the basin and the combination of qualitative and quantitative study with new theory and technique by building a geological model. The paper also puts forward a new thought for studying the oil & gas accumulation and reservoir prediction , and establishes a new research system for reformed basin. It is verified by the known reservoir and oil accumulation area in the basin and has a practical value for use and reference. The new ideas and achievements in this research are as following: 1.This is the first time that the system for studying the reformed basin and its pool-forming mechanism and reservoir prediction is presented. A reverse thinking and combination of qualitative & quantitative are applied here with emphasis on "wholeness or systematic, dynamic and integrated" research, new theory, techniques & methods comprehensive use and geologic models building. 2. Identifying criterion and methods, classifying schemes, and denominating principles for the conserved units of reformed basins are presented in this paper. The geologic model of conserved units of Yingen-Ejinaqi Basin has been built. It is a practical method when combined with the traditional way for basin survey and the conserved units study. 3.The dynamic sources of basin deformation are believed to be stress, gravity and thermodynamics. The stress and gravity are key factors in basin deformation and pool forming, especially stress. Scientific proof is provided by classifying the functional type, style and range of the stress, gravity and thermodynamics. 4.The pool forming and reservoir distribution of Yingen-Ejinaqi Basin are controlled by multiple factors or geologic conditions or/and co-controlled by both of them. The qualitative and quantitative research on petroleum system and basin modeling will help us understand and determine the pool-forming period of the conserved unit (timing), the oil migrating direction (orientation), the oil accumulating region (location), the oil distributing border (bordering) and the size of oil accumulation (quantification). Thus the pool-forming and distribution zones can be predicted. 5.Three generating modes (reform-succession type, reform-destroyed type and reform-regenerating type or reform-newborn type) of pool forming for reformed basins are presented here, together with the inner relationships between basin deformation type, overlapping style and pool-forming modes. The pool-forming modes are determined by deformation type and overlapping style. Reservoir distribution will be predicted based on the modes and other concrete pool-forming conditions. 6.The evaluation methods of the conserved units and zones and the parameter selection are reliable in optimal selecting of target zones. The technical terms, new ideas and methods for the study of reformed basins, the pool-forming mechanism and reservoir prediction are presented in this paper. The concepts and terms, the identifying criterion, the denominating principles, the generating modes for pool forming, the methods of reservoir prediction, and the evaluation techniques for conserved units and zones can be used for reference in studies on the petroleum exploration of reformed basins in China and abroad. It serves as a typical example for further research of the reformed basins and the geologic regulations of oil accumulation. It has a practical value of use and reference. The future research in the field of pool-forming mechanism of the reformed basins may well be on the process simulation of pool-forming dynamics of the reformed basins. Experimental work has been conducted to simulate the processes by using quantitative and qualitative methods combined. The further study in this field calls for more efforts.

基于工程地质力学的岩土工程加固理论、技术与实践

In the engineering reinforcement of-rock and soil mass, engineers must consider how to obtain better reinforcing effect at the cost of less reinforcing expense, which, in fact, is the aim of reinforcement design. In order to accomplish the purpose, they require not only researching the material used to reinforce and its structure, but also taking into account of several important geological factors, such as the structure and property of rock and soil mass. How to improve the reinforcing effect according to engineering geomechanical principle at the respect of the reinforcement of engineering soil and rock mass is studied and discussed in this paper. The author studies the theory, technology and practice of geotechnical reinforcement based on engineering geomechanics, taking example for the soil treatment of Zhengzhou Airport, the effect analysis of reinforcement to the slope on the left bank of Wuqiangxi Hydropower Station and the reinforcing design of the No. 102 Landslide and unique sand-slide slope on the Sichuan-Tibet Highway. The paper is comprised of two parts for the convenience of discussion. In the first part, from the first chapter to the fifth chapter, trying to perform the relevant research and application at the viewpoint of soil mass engineering geomechanics, the author mainly discusses the study of reinforcing soft ground soil through dynamical consolidation and its application. Then, in the second part, from the sixth chapter to the eleventh chapter, the study of new technologies in the rock slope reinforcement and their application are discussed. The author finds that not only better reinforcing effect can be gained in the research where the principle and method of rock mass engineering geomechanics is adopted, but also new reinforcing technologies can be put forward. Zhengzhou Airport is an important one in central plains. It lies on Yellow River alluvial deposit and the structure of stratum is complex and heterogeneous. The area of airport is very large, which can result in differential settlement easily, damage of airport and aircraft accident, whereas, there are no similar experiences to dispose the foundation, so the foundation treatment become a principal problem. During the process of treatment, the method of dynamic compaction was adopted after compared with other methods using the theory of synthetic integration. Dynamic compaction is an important method to consolidate foundation, which was successfully used in the foundation of Zhengzhou Airport. For fill foundation, controlling the thickness of fill so as to make the foundation treatment can reach the design demand and optimum thickness of the fill is a difficult problem. Considering this problem, the author proposed a calculation method to evaluate the thickness of fill. The method can consider not only the self-settlement of fill but also the settlement of the ground surface under applied load so as to ensure the settlement occurred during the using period can satisfy the design demand. It is proved that the method is correct after using it to choose reasonable energy of dynamic compaction to treat foundation. At the same time, in order to examine the effect of dynamic compaction, many monitor methods were adopted in the test such as static loading test, modulus of resilience test, deep pore pressure -test, static cone penetration test and the variation of the pore volume measurement. Through the tests, the author summarized the discipline of the accumulation and dissipation of pore pressure in Yellow River alluvial deposit under the action of dynamic compaction, gave a correct division of the property change of silt and clay under dynamic compaction, determined the bearing capacity of foundation after treatment and weighted the reinforcing effect of dynamic consolidation from the variation of the soil particle in microcosmic and the parameter of soil mass' density. It can be considered that the compactness of soil is in proportion to the energy of dynamic compaction. This conclusion provided a reference to the research of the "Problem of Soil Structure-the Central Problem of Soil Mechanics in 21 Century ". It is also important to strengthen rock mass for water conservancy and electric power engineering. Slip-resistance pile and anchoring adit full of reinforced concrete are usually adopted in engineering experience to strengthen rock mass and very important for engineering. But there also some deficiency such as the weakest section can't be highlighted, the monitor is inconvenient and the diameter of pile and adit is very large etc. The author and his supervisor professor Yangzhifa invented prestressed slip-resistance pile and prestressed anchoring adit full of reinforced concrete, utilizing the advantage that the prestressed structure has better anti-tensile characteristic (this invention is to be published). These inventions overcome the disadvantages of general slip-resistance pile and anchoring adit full of reinforced concrete and have the functions of engineering prospecting, strengthening, drainage and monitor simultaneous, so they have better strengthened effect and be more convenient for monitor and more economical than traditional methods. Drainage is an important factor in treatments of rock mass and slop. In view of the traditional drainage method that drainage pore often be clogged so as to resulted in incident, professor Yangzhifa invented the method and setting of guide penetration by fiber bundle. It would take good effect to use it in prestressed slip-resistance pile and anchoring adit full of reinforced concrete. In this paper, the author took example for anchoring adit full of reinforced concrete used to strengthen Wuqiangxi left bank to simulate the strengthened effect after consolidated by prestressed slip-resistance pile, took example for 102 landslide occurred along Sichuan-Tibet highway to simulate the application of slip-resistance pile and the new technology of drainage. At the same time the author proposed the treatment method of flowing sand in Sichuan-Tibet highway, which will benefit the study on strengthening similar engineering. There are five novelties in the paper with the author's theoretical study and engineering practice: 1. Summarizing the role of pore water pressure accumulation and dissipation of the Yellow River alluvial and diluvial soil under the action of dynamical consolidation, which has instructive significance in the engineering construction under the analogical engineering geological conditions in the future. It has not been researched by the predecessors. 2. Putting forward the concept of density D in microcosmic based on the microcosmical structure study of the soil sample. Adopting D to weight the reinforcing effect of dynamic consolidation is considered to be appropriate by the means of comparing the D values of Zhengzhou Airport's ground soil before with after dynamically consolidating reinforcement, so a more convenient balancing method can be provided for engineering practice. 3. According to the deep research into the soil mass engineering geology, engineering rock and soil science, soil mechanics, as well as considerable field experiments, improving the consolidating method in airport construction, from the conventional method, which is dynamically compactmg original ground surface firstly, then filling soil and dynamically layer-consolidating or layer-compacting at last to the upgraded method, which is performing dynamical consolidation after filling soil to place totally at the extent of the certain earth-filling depth. The result of the dynamical consolidation not only complies with the specifications, but also reduces the soil treatment investment by 10 million RMB. 4. Proposing the method for calculating the height of the filled soil by the means of estimating the potential displacement produced in the original ground surface and the filled earth soil under the possible load, selecting the appropriate dynamically-compacting power and determining the virtual height of the filled earth soil. The method is proved to be effective and scientific. 5. According to the thought of Engineering Geomechanics Metal-Synthetic Methodology (EGMS), patenting two inventions (to the stage of roclamation, with Professor Yang Zhi-fa, the cooperative tutor, and etc.) in which multi-functions, engineering geological investigation, reinforcement, drainage and strength remedy, are integrated all over in one body at the viewpoint of the breakage mechanism of the rock slope.

鄂尔多斯盆地苏里格气田储层次生孔隙性质与分布预测

Permian reservoir in Sulige area of Ordos Basin, on which this paper focused, belongs to fluvial-delta lithofacies. The majority formations in this area are complicated channel sand deposit with serious inhomogeneity which makes natural gas exploration be very tough in this area. This inhomogeneity can be found everywhere both in large horizontal area and vertical profile of inner and interbedded formations.This paper studied the inhomogeneity characteristic of Permian formation in sulige area of Ordos Basin according to the logging data.Correlating with core data, a criterion to distinguish different type of reservoirs by using logging data is determined after the study of logging response is done considering the diverse conditions of deposit environments, lithology and reservoir space. The characteristic relationships between the various type formations and logging responses fully and systemically are established.It investigated reservoir parameter calculation methods amply. Combining the conventional and special logging data, basing on the feature of low porosity -permeability formation of sulige area, a set of methods to calculate reservoir parameters was formed including primary porosity, secondary porosity, fracture porosity, permeability and water saturation under the conditions of both low porosity-permeability and inhomogenous reservoirs. One thing should be pay close attention is the parameter M for calculating saturation. It is found that the M in low porosity -permeability formation decreases as the porosity decrease, which is opposite to the law that M increases as the porosity decrease in the formation with intermediate to high porosity and permeability. This view has innovated the traditional theory and offered theory basis for the logging interpretation of low porosity - permeability reservoir. Meanwhile it also improved the Arqi formula theoretically and enhanced the logging interpretation accuracy and rescued a number of formations which has been thought to be hopeless according to the old theory.By using advantage logging interpretation procedure, a territorial synthetic geology evaluation to the inhomogeneous reservoir was completed basing on the single well interpretation. All the reservoir evaluation parameters including sand formation thickness, primary porosity, secondary porosity were calculated and evaluated. The rules of changing and development for sand formation thickness, sand physical properties and secondary porous were found at different formations of upper part of the Member 8 of Shihezi, lower part of the Member 8 of Shihezi, the Member 1 of shanxi and the Member 2 of shanxi individually. Evaluation and Correlation of these five formations were also completed and one conclusion was arrived: upper part of the Member 8 of Shihezi formation has the best performance followed by the lower part of the Member 8 of Shihezi, the Member 1 of shanxi and the Member 2 of shanxi formation.After studied the relationship between reservoir deposition characteristic and the natural gas richness, it is regarded that reservoir inhomogeneity is the key issue of the impaction on the natural gas. Natural gas in Sulige gas field was mainly accumulated in sands of channel bar, distributary channel and debouchure bar. Especially, the quartz sand with rich of secondary porous space has obvious better physical properties than other reservoir and usually can forms the concentration of natural gas.

Screening and analysis of bioactive compounds with biofingerprinting chromatogram analysis of traditional Chinese medicines targeting DNA by microdialysis/HPLC

Biofingerprinting chromatogram, analysis, which is defined as the comparison of fingerprinting chromatograms of the extract of traditional Chinese medicines (TCMs) before and after the interaction with biological systems (DNA, protein. cell. etc.), was proposed for screening and analysis of the multiple bioactive compounds in TCMs. A method of microdialysis sampling combined with high performance liquid chromatography (HPLC) was applied to the study of DNA-binding property for the extracts of TCMs. Seven compounds were found to bind to calf thymus DNA (ct-DNA) from the TCMs of Coptis chinensis Franch (Coptis), but only three ones from Phellodendron amurense Rupr. (Phellodendron) and none from Sophoraflavescens Ait. (Sophora) to bind to ct-DNA. respectively. Three of them were identified as berberine, palmatine and jatrorrhizine and their association constants (K) to ct-DNA were determined by microdialysis/HPLC. Competitive binding behaviors of them to ct-DNA were also investigated. © 2005 Elsevier B.V. All rights reserved.

Improved performance of comprehensive two-dimensional HPLC separation of traditional Chinese medicines by using a silica monolithic column and normalization of peak heights

Performance of comprehensive two-dimensional liquid chromatography system is greatly improved than we reported previously by using a silica monolithic column as for the second dimensional separation. Due to the increase of the elution speed on the second dimensional monolithic column, the first dimensional column efficiency and analysis rate can be greatly improved as comparing with conventionally second dimensional column. The developed system was applied to analysis of methanol extraction of two umbelliferae herbs Ligusticum chuanxiong Hort. and Angelica sinensis (Oliv.) Diels by using CN column as for the first dimensional separation and a silica monolithic ODS column for the second dimensional separation, and the obtained three-dimensional chromatograms were treated by normalization of peak heights with the value of the highest peak or setting a certain value using a software written in-house. It was observed that much more peaks for low-abundant components in TCM extract can clearly be detected here than we reported before, due to the large difference for the amount of components in TCMs' extract. With the above improvements in separation performance and data treatment, totally about 120 components in methanol extraction of Rhizoma chuanxiong and 100 in A. sinensis were separated with UV detection within 130 min. This result meant that both the number of peaks detected increase twice but the analysis time decease twice if comparing with the previously reported result. (c) 2005 Published by Elsevier B.V.