[Discovery and application of traditional Chinese medicine efficacy markers based on systematic traditional Chinese medicine].

The quality marker(Q-marker) of traditional Chinese medicine(TCM) is a new concept of TCM quality control proposed in recent years. It is a hot issue in the research of modern Chinese medicine. The TCM efficacy is a high-level summary of the TCM therapeutic effect under the guidance of TCM theory. On this basis, it is of considerable significance to explore the TCM efficacy marker for the TCM modernization. However, the traditional research strategy based on the single herb and decoction piece in macro TCM level, or the drug research strategy based on the biological effect of the targets, is quite different from the characteristics of multiple components of TCM, as well as the weak and low-selective effect of Chinese medicine ingredients on targets. Therefore, how to select representative ingredients to characterize the TCM overall efficacy is a problematic point in establishing TCM efficacy markers. In this paper, the concept and method of Q-marker were introduced into the study of Chinese medicine efficacy. The research method for systematic TCM was used to systematically discuss the connotation of TCM efficacy markers, the principles of discovery and determination, common research ideas and techniques by taking the representative research results as an example. This study provides new ideas for the research and discovery of TCM efficacy markers.

Application of lipidomics on research of traditional Chinese medicine / 中草药

Lipidomics is a newly emerged discipline that systematically studies the types, functions and metabolic pathways of lipids in organisms, which can identify biomarkers and molecular mechanisms of diseases or drugs by analyzing changes in lipid metabolism networks from different physiological or pathological states. As an important component of systems biology, lipidomics has the research mentality which is similar to the overall concept as well as the system theory of traditional Chinese medicine (TCM). Therefore, lipidomics becomes a powerful tool to promote the modernization of TCM. Lipidomics technology is briefly described in this paper, and the application of lipidomics technology on research of TCM including pharmacodynamics, action mechanism and toxicity are summed up, and at the same time the experience of other disciplines in the research of Chinese materia medica property theory and quality control of TCM are combined to expand the application ideas of lipidomics in TCM research, as well as provide references for the organic integration between TCM theory and modern technology.

Establishment of quality control method for biological activity of Honghua injection / 中国药理学通报

Aim To establish a method for determining the antithrombotic biological activity of Honghua injection, which can be used to evaluate and control its quality. Methods Collagen-adrenalin was used to induce mouse acute cerebral thrombosis model and hemiplegic protection rate of Honghua injection in mice as an index to investigate the antithrombotic activity of Honghua injection. The experimental conditions of the administration dosage, inducer dosage, and different strains of mice were investigated, and the experimental conditions were optimized by orthogonal design. Results Honghua injection which was made by 5. 0 g crude drug · kg-1and continuously ip for 3 d, had obvious protective effect on collagen-adrenalin-induced acute cerebral thrombosis in mice. The established biological activity limit method showed a good repeatability, intermediate precision, and reproducibility. Each sample showed different degrees of differences in biological activity. Conclusions The thrombus test in mice can be used as a method for measuring the biological activity of Honghua injection. It is recommended to use 5. 0 g crude drug · kg-1dose as the limit for bioactive process optimization and product quality controlment of Honghua injection.

Discovery and application of traditional Chinese medicine efficacy markers based on systematic traditional Chinese medicine / 中国中药杂志

The quality marker(Q-marker) of traditional Chinese medicine(TCM) is a new concept of TCM quality control proposed in recent years. It is a hot issue in the research of modern Chinese medicine. The TCM efficacy is a high-level summary of the TCM therapeutic effect under the guidance of TCM theory. On this basis, it is of considerable significance to explore the TCM efficacy marker for the TCM modernization. However, the traditional research strategy based on the single herb and decoction piece in macro TCM level, or the drug research strategy based on the biological effect of the targets, is quite different from the characteristics of multiple components of TCM, as well as the weak and low-selective effect of Chinese medicine ingredients on targets. Therefore, how to select representative ingredients to characterize the TCM overall efficacy is a problematic point in establishing TCM efficacy markers. In this paper, the concept and method of Q-marker were introduced into the study of Chinese medicine efficacy. The research method for systematic TCM was used to systematically discuss the connotation of TCM efficacy markers, the principles of discovery and determination, common research ideas and techniques by taking the representative research results as an example. This study provides new ideas for the research and discovery of TCM efficacy markers.

[Establishment of QAMS method with dehydroandrographolide as internal reference substance for quality control of Andrographis Herba].

To solve the problems of the poor resolution of chromatographic separation,the weak durability of the relative correction factors,and the low accuracy of content determination results in the quantitative analysis of multi-components by single-marker( QAMS) method with andrographolide as the internal reference substance in the existing research of Andrographis Herba,a new QAMS method using dehydroandrographolide as the internal reference substance was established for the first time in this study. This new method can be used to simultaneously determine four diterpene lactones,including andrographolide( A),neoandrographolide( B),14-deoxyandrographolide( C),and dehydroandrographolide( S) through the optimization of chromatographic conditions and systematic investigation of methodology. At the present HPLC chromatographic conditions,four components could be well separated( R > 1. 5),and the methodology validations could satisfy the requirement of quantitative analysis. The relative correction factors( RCFs) of fA/S,fB/S,fC/S were determined as 0. 65,0. 54,0. 78,respectively. The relative standard deviations( RSDs) of their RCFs ranged between 1. 3%-5. 1%,0. 25%-0. 33%,0. 070%-0. 15%,0. 070%-0. 22%,respectively with three brands of HPLC instruments,five brands of C18 column,different flow rates( 0. 9,1. 0,1. 1 m L·min~(-1)),and different column temperatures( 25,30,35 ℃),indicating good durability of the RCFs. The relative retention value( RRV) method was used to locate the chromatographic peak of the components to be determined.The RRVs of rA/S,rB/S,and rC/Swere 0. 44,0. 86,0. 97,respectively. The RSDs of the RRVs ranged between 0. 030%-1. 6% with different HPLC instruments and columns,showing accurate peak location. The present QAMS method and the external standard method( ESM)were both used to determine the contents of four diterpene lactones from Andrographis Herba( 6 batches of medicinal materials and 18 batches of cut crude drugs). The relative errors of the determined content results between two methods were less than 2. 0%. It demonstrated that there was no significant difference in content results between these two methods,indicating good accuracy of the present QAMS method. Therefore,in this study,an accurate and highly durable QAMS method using dehydroandrographolide as the internal reference substance was established for simultaneous determination of four diterpene lactones. This method could be used to effectively control the quality of Andrographis Herba and provide technical basis for the formulation of traditional Chinese medicine industry standard and improvement of the Chinese Pharmacopoeia standard of Andrographis Herba.

Establishment of QAMS method with dehydroandrographolide as internal reference substance for quality control of Andrographis Herba / 中国中药杂志

To solve the problems of the poor resolution of chromatographic separation,the weak durability of the relative correction factors,and the low accuracy of content determination results in the quantitative analysis of multi-components by single-marker( QAMS) method with andrographolide as the internal reference substance in the existing research of Andrographis Herba,a new QAMS method using dehydroandrographolide as the internal reference substance was established for the first time in this study. This new method can be used to simultaneously determine four diterpene lactones,including andrographolide( A),neoandrographolide( B),14-deoxyandrographolide( C),and dehydroandrographolide( S) through the optimization of chromatographic conditions and systematic investigation of methodology. At the present HPLC chromatographic conditions,four components could be well separated( R > 1. 5),and the methodology validations could satisfy the requirement of quantitative analysis. The relative correction factors( RCFs) of fA/S,fB/S,fC/S were determined as 0. 65,0. 54,0. 78,respectively. The relative standard deviations( RSDs) of their RCFs ranged between 1. 3%-5. 1%,0. 25%-0. 33%,0. 070%-0. 15%,0. 070%-0. 22%,respectively with three brands of HPLC instruments,five brands of C18 column,different flow rates( 0. 9,1. 0,1. 1 m L·min~(-1)),and different column temperatures( 25,30,35 ℃),indicating good durability of the RCFs. The relative retention value( RRV) method was used to locate the chromatographic peak of the components to be determined.The RRVs of rA/S,rB/S,and rC/Swere 0. 44,0. 86,0. 97,respectively. The RSDs of the RRVs ranged between 0. 030%-1. 6% with different HPLC instruments and columns,showing accurate peak location. The present QAMS method and the external standard method( ESM)were both used to determine the contents of four diterpene lactones from Andrographis Herba( 6 batches of medicinal materials and 18 batches of cut crude drugs). The relative errors of the determined content results between two methods were less than 2. 0%. It demonstrated that there was no significant difference in content results between these two methods,indicating good accuracy of the present QAMS method. Therefore,in this study,an accurate and highly durable QAMS method using dehydroandrographolide as the internal reference substance was established for simultaneous determination of four diterpene lactones. This method could be used to effectively control the quality of Andrographis Herba and provide technical basis for the formulation of traditional Chinese medicine industry standard and improvement of the Chinese Pharmacopoeia standard of Andrographis Herba.

[New qualities control strategy of Scutellaria baicalensis by isothermal titration calorimetry].

It has been focused on that there will be precipitates when decoction of Scutellariat Radix mixed with Coptidis Rhizoma. Precipitation was derived from interaction between acidic and basic compounds. This study was based on the interaction between active ingredients after compatibility, strived to explore whether it was feasible to judge the qualities of different Scutellariat Radix by isothermal titration calorimetry (ITC), build a new method established to characterize the qualities of traditional Chinese medicine by taking a series of active ingredients as index. We selected Scutellariat Radix (including three batches of different Scutellariat Radix bought from market and immature Scutellariat Radix which usually was used as adulterant) in different batches as the samples. First, we used ITC to determine the binding heat of the reactions between berberine and the decoctions of different Scutellariat Radix. The test showed that the binding heat of berberine titrated Scutellariat Radix was Scutellariat Radix A (－317.20 µJ), Scutellariat Radix B (－292.83 µJ), Scutellariat Radix C (－208.95 µJ) and immature Scutellariat Radix (－21.53 µJ), respectively. We chose deionized water titrated by berberine (2.51 µJ) as control. The heat change of berberine titrated immature Scutellariat Radix was much less than berberine titrated Scutellariat Radix. Then we determined the absorbance of different decoctions of Scutellariat Radix by UV Spectrophotometry on the maximum absorption wavelength, and the result is： Scutellariat Radix A (0.372), Scutellariat Radix B (0.333), Scutellariat Radix C (0.272), immature Scutellariat Radix (0.124). The absorbance of immature Scutellariat Radix was also less than Scutellariat Radix. The result of ITC assay was corresponded to UV spectrophotometry test. In conclusion, ITC could be used to characterize the quality of Scutellariat Radix. The new method to characterize the qualities of traditional Chinese medicine by taking a kind of active ingredients as index building by ITC was simple, scientific and feasible.

[A novel pattern for predicting the quality of Chinese herba preparation intelligently by high performance liquid chromatographic formula fingerprints].

In order to build the fusion models, the high performance liquid chromatographic (HPLC) fingerprints of scutellariae radix (SR), rhei radix et rhizoma (RRR), coptidis rhizoma (CR) and their synthesizing fingerprints were developed in this study. After exploring the consistency between the fingerprints of compound synthesizing fingerprints (CSF) and the sample, the quality of traditional Chinese medicine preparation was predicted intelligently using CSF. HPLC coupled with diode array detector was used to obtain chromatograms of SR, RRR, CR and Yi Qing Tablet (YQT) samples at 268 nm. Meanwhile, the quality of CSF and the 15 batches of YQT samples was evaluated by systematically quantified fingerprint method (SQFM) qualitatively and quantitatively. The chromatograms showed that CSF covered the main fingerprints' information of each herb and the 55 common peaks of CSF covered the main information of the 50 common peaks in YQT sample. The evaluation results showed that among the 15 batches of YQT samples, only YQT-S01 was grade 5 and the others were all above grade 3. Most of the CSFs were grade 2 or grade 1 except CSF-2 which was grade 6. The fingerprints of Chinese herba preparation could be replaced by CSF to achieve a novel pattern for predicting the quality of TCM preparation intelligently by studying the relationship between the standard fingerprints and the CSF, and simultaneously developing first-class evaluation software.

[Application of DOSC combined with SBC in batches transfer of NIR quantitative model].

Near infrared model established under a certain condition can be applied to the new samples status, environmental conditions or instrument status through the model transfer. Spectral background correction and model update are two types of data process methods of NIR quantitative model transfer, and orthogonal signal regression (OSR) is a method based on spectra background correction, in which virtual standard spectra is used to fit a linear relation between master batches spectra and slave batches spectra, and map the slave batches spectra to the master batch spectra to realize the transfer of near infrared quantitative model. However, the above data processing method requires the represent activeness of the virtual standard spectra, otherwise the big error will occur in the process of regression. Therefore, direct orthogonal signal correction-slope and bias correction (DOSC-SBC) method was proposed in this paper to solve the problem of PLS model's failure to predict accurately the content of target components in the formula of different batches, analyze the difference between the spectra background of the samples from different sources and the prediction error of PLS models. DOSC method was used to eliminate the difference of spectral background unrelated to target value, and after being combined with SBC method, the system errors between the different batches of samples were corrected to make the NIR quantitative model transferred between different batches. After DOSC-SBC method was used in the preparation process of water extraction and ethanol precipitation of Lonicerae Japonicae Flos in this paper, the prediction error of new batches of samples was decreased to 7.30% from 32.3% and to 4.34% from 237%, with significantly improved prediction accuracy, so that the target component in the new batch samples can be quickly quantified. DOSC-SBC model transfer method has realized the transfer of NIR quantitative model between different batches, and this method does not need the standard samples. It is helpful to promote the application of NIR technology in the preparation process of Chinese medicines, and provides references for real-time monitoring of effective components in the preparation process of Chinese medicines.

New qualities control strategy of Scutellaria baicalensis by isothermal titration calorimetry / 中国中药杂志

It has been focused on that there will be precipitates when decoction of Scutellariat Radix mixed with Coptidis Rhizoma. Precipitation was derived from interaction between acidic and basic compounds. This study was based on the interaction between active ingredients after compatibility, strived to explore whether it was feasible to judge the qualities of different Scutellariat Radix by isothermal titration calorimetry (ITC), build a new method established to characterize the qualities of traditional Chinese medicine by taking a series of active ingredients as index. We selected Scutellariat Radix (including three batches of different Scutellariat Radix bought from market and immature Scutellariat Radix which usually was used as adulterant) in different batches as the samples. First, we used ITC to determine the binding heat of the reactions between berberine and the decoctions of different Scutellariat Radix. The test showed that the binding heat of berberine titrated Scutellariat Radix was Scutellariat Radix A (－317.20 μJ), Scutellariat Radix B (－292.83 μJ), Scutellariat Radix C (－208.95 μJ) and immature Scutellariat Radix (－21.53 μJ), respectively. We chose deionized water titrated by berberine (2.51 μJ) as control. The heat change of berberine titrated immature Scutellariat Radix was much less than berberine titrated Scutellariat Radix. Then we determined the absorbance of different decoctions of Scutellariat Radix by UV Spectrophotometry on the maximum absorption wavelength, and the result is： Scutellariat Radix A (0.372), Scutellariat Radix B (0.333), Scutellariat Radix C (0.272), immature Scutellariat Radix (0.124). The absorbance of immature Scutellariat Radix was also less than Scutellariat Radix. The result of ITC assay was corresponded to UV spectrophotometry test. In conclusion, ITC could be used to characterize the quality of Scutellariat Radix. The new method to characterize the qualities of traditional Chinese medicine by taking a kind of active ingredients as index building by ITC was simple, scientific and feasible.

Application of DOSC combined with SBC in batches transfer of NIR quantitative model / 中国中药杂志

Near infrared model established under a certain condition can be applied to the new samples status, environmental conditions or instrument status through the model transfer. Spectral background correction and model update are two types of data process methods of NIR quantitative model transfer, and orthogonal signal regression (OSR) is a method based on spectra background correction, in which virtual standard spectra is used to fit a linear relation between master batches spectra and slave batches spectra, and map the slave batches spectra to the master batch spectra to realize the transfer of near infrared quantitative model. However, the above data processing method requires the represent activeness of the virtual standard spectra, otherwise the big error will occur in the process of regression. Therefore, direct orthogonal signal correction-slope and bias correction (DOSC-SBC) method was proposed in this paper to solve the problem of PLS model's failure to predict accurately the content of target components in the formula of different batches, analyze the difference between the spectra background of the samples from different sources and the prediction error of PLS models. DOSC method was used to eliminate the difference of spectral background unrelated to target value, and after being combined with SBC method, the system errors between the different batches of samples were corrected to make the NIR quantitative model transferred between different batches. After DOSC-SBC method was used in the preparation process of water extraction and ethanol precipitation of Lonicerae Japonicae Flos in this paper, the prediction error of new batches of samples was decreased to 7.30% from 32.3% and to 4.34% from 237%, with significantly improved prediction accuracy, so that the target component in the new batch samples can be quickly quantified. DOSC-SBC model transfer method has realized the transfer of NIR quantitative model between different batches, and this method does not need the standard samples. It is helpful to promote the application of NIR technology in the preparation process of Chinese medicines, and provides references for real-time monitoring of effective components in the preparation process of Chinese medicines.