[Material basis and mechanism of Bletillae Rhizoma for melasma, gastrointestinal hemorrhage, lung cancer and bronchoplumonary inflammation as "homotherapy for heteropathy" based on UHPLC-Q-Exactive Orbitrap HRMS coupled with network pharmacology and molecular docking].

Based on UHPLC-Q-Exactive Orbitrap HRMS coupled with the network pharmacology and molecular docking, the common material basis and molecular mechanisms of Bletillae Rhizoma for melasma, gastrointestinal hemorrhage, lung cancer and bronchoplumonary inflammation as "homotherapy for heteropathy" were explored. The fingerprint of 17 batches of Bletillae Rhizoma from different areas was established using HPLC, and the similarity analysis was carried out. The common chemical components of the 17 batches of Bletillae Rhizoma were identified using UHPLC-Q-Exactive Orbitrap HRMS. Depending on the bioavailability and drug-like properties of the common components, the active chemical components were screened, and then their protein targets were collected using the Traditional Chinese Medicine Database and Analysis Platform(TCMSP) and SwissTargetPrediction database. The protein targets related to diseases were retrieved from the databases DrugBank, TTD and GeneCards to produce a Venn diagram. The shared targets were obtained between drugs and diseases as "homotherapy for heteropathy" targets. The protein-protein interaction(PPI) was analyzed with the STRING database, and KEGG and GO analyses of the "homotherapy for heteropathy" targets were performed using the Bioconductor database. Cytoscape 3.7.2 software was employed to construct the "chemical components of Bletillae Rhizoma-homotherapy for heteropathy targets" network and PPI network, and topological analysis was conducted to screen out the key active chemical components and core targets. Finally, the affinity between the active components and core targets was evaluated using the molecular docking by AutoDock Vina 4.2.6, which verified the interaction between them. Thirteen common peaks were identified by fingerprint chromatography, and the similarity between different batches was 0.941-0.998. Fifty-three chemical components were identified by mass spectrometry in Bletillae Rhizoma, and 18 common chemical constituents were obtained in the 17 batches of Bletillae Rhizoma. Network pharmacologic screening showed that the pharmacodynamic substances of Bletillae Rhizoma for melasma, gastrointestinal hemo-rrhage, lung cancer and bronchoplumonary inflammation with "homotherapy for heteropathy" were 11 compounds, such as polysaccharides, biphenanthrenes, dihydrophenanthrenes and bibenzyls. There were 42 common targets identified for the treatment of different diseases. These targets were involved in biological processes such as cell response to chemical stress, reactive oxygen species and positive regulation of protein kinase B signal transduction. They were also involved in 121 signaling pathways, encompassing vital pathways such as PI3K-Akt, ErbB, Rap1, FoxO, MAPK and estrogen. Molecular docking results showed a strong affinity between the key active components and the core targets. This study provides a preliminary explanation of how Bletillae Rhizoma exerts its therapeutic effect on chloasma, gastrointestinal hemorrhage, lung cancer, and bronchopneumonic lesions as "homotherapy for heteropathy" through a combined action involving multiple components, targets, and pathways. These findings offer a certain theoretical basis for the further deve-lopment and application of Bletillae Rhizoma.

Studies on fingerprints and efficacy-related substance of classical prescription Zhuru Decoction / 中国中药杂志

Based on fingerprint and network pharmacology,the whole process quality control of Zhuru Decoction was conducted and efficacy-related substances were predicted.The fingerprints of raw materials,decoction pieces and Zhuru Decoction were established,and 25 common peaks were identified,including 9 common chromatographic peaks of 3'-hydroxy puerarin,puerarin,3'-methoxy puerarin,puerarin,aperioside,daidzin,daidzein,liquiritin,glycyrrhizic acid and 6-gingerol, with similarity all greater than 0.95.The main groups of pharmacodynamic substances can be transferred from raw materials,decoction pieces to Zhuru Decoction step by step,with a clear affiliation relationship.Based on the testability and traceability,the active ingredients were screened,and the network relationship of "component-target-pathway" was constructed and analyzed for the nine chemical components screened by network pharmacology.The enriched pathways included energy metabolism,alcoholism,and smooth muscle contraction and relaxation-related pathways.The nine active components of Zhuru Decoction may achieve the effects of clearing heat, alleviating a hangover, harmonizing stomach and stopping vomiting through these signaling pathways.Based on transitive and traceable properties of the above 9 components as well as their close relationship to the efficacy of Zhuru Decoction,these 9 components can be identified as potential efficacy-related substances and provide basis for the overall quality control of Zhuru Decoction.

[Studies on fingerprints and efficacy-related substance of classical prescription Zhuru Decoction].

Based on fingerprint and network pharmacology,the whole process quality control of Zhuru Decoction was conducted and efficacy-related substances were predicted.The fingerprints of raw materials,decoction pieces and Zhuru Decoction were established,and 25 common peaks were identified,including 9 common chromatographic peaks of 3'-hydroxy puerarin,puerarin,3'-methoxy puerarin,puerarin,aperioside,daidzin,daidzein,liquiritin,glycyrrhizic acid and 6-gingerol, with similarity all greater than 0.95.The main groups of pharmacodynamic substances can be transferred from raw materials,decoction pieces to Zhuru Decoction step by step,with a clear affiliation relationship.Based on the testability and traceability,the active ingredients were screened,and the network relationship of "component-target-pathway" was constructed and analyzed for the nine chemical components screened by network pharmacology.The enriched pathways included energy metabolism,alcoholism,and smooth muscle contraction and relaxation-related pathways.The nine active components of Zhuru Decoction may achieve the effects of clearing heat, alleviating a hangover, harmonizing stomach and stopping vomiting through these signaling pathways.Based on transitive and traceable properties of the above 9 components as well as their close relationship to the efficacy of Zhuru Decoction,these 9 components can be identified as potential efficacy-related substances and provide basis for the overall quality control of Zhuru Decoction.

Influence of Gamma-rays Radiation Sterilization on the Content and Fingerprint Chromatography of Effec-tive Constituents in Simiao Junyi Ointment / 中国药师

Objective:To explore the degradation effect of gamma-rays radiation sterilization on the effective constituents in Simiao Junyi ointment and study the changes of fingerprint chromatography before and after the sterilization to provide basis for the feasibility of gamma-rays radiation sterilization for Simiao Junyi ointment. Methods:The contents of tetrahydropalmatine and PNS ( Panax Notogin-seng saponins) in Simiao Junyi ointment were determined by HPLC, and the fingerprint chromatography was established. The content changes of tetrahydropalmatine and PNS in Simiao Junyi ointment before and after the gamma-rays radiation sterilization were compared among the same batch and various batches, and the relative retention time and relative peak area in the fingerprint chromatography were also compared. Results:The content of tetrahydropalmatine had no change basically before and after the gamma-rays radiation steriliza-tion(P >0. 05), and there was no change in the total content of PNS (P >0. 05). Comparing the HPLC fingerprint chromatography at 280 nm, the relative retention time had statistically significant change after the gamma-rays radiation sterilization ((P 0. 05). The number of characteristic peaks reduced by one, namely the C8 characteristic peak disappeared in the pasteurized chromatography, and the areas of C6, C9 and C14 peak decreased significantly, while that of C12 increased. Conclusion:Gamma-rays radiation sterilization have no notable effect on the content of tet-rahydropalmatine and PNS in Simiao Junyi ointment, it can be used for the sterilization of Simiao Junyi ointment.

Fingerprint of Xinan Capsules by Principal Component Analysis and Cluster Analysis / 中国药师

Objective:To study the HPLC fingerprint of Xinan capsules from different manufacturers, and establish the chemical pattern recognition method by using principal component analysis and cluster analysis in order to provide reference for the quality con-trol of Xinan capsules. Methods:The HPLC chromatographic column was Agilent ZORBAX SB-C18 (250 mm × 4. 6 mm, 5 μm);the mobile phase was 0.1% formic acid(A)-acetonitrile(B)– tetrahydrofuran(C) with gradient elution, the flow rate was 1.0 ml· min-1;the detection wavelength was 350 nm and the column temperature was 30 ℃. Totally 15 batches of samples were analyzed by the Evaluation System of Traditional Chinese Medicine Chromatographic Fingerprint Similarity (2004A version) and SPSS 19. 0 statisti-cal software. Results:According to the results of cluster analysis and principal component analysis, 10 batches of Xinan capsules were screened out, and the fingerprint common pattern was established. Conclusion:The method is accurate and reliable, and can be used to control the quality of Xinan capsules.

HPLC Fingerprint of the Ingredients in Simiao Junyi Ointment / 中国药师

Objective:To establish an analytical method for HPLC fingerprint chromatography of Simiao Junyi ointment to provide basis for the quality control standard. Methods:The separation conditions were established to obtain the HPLC fingerprint chromatog-raphy of the main ingredients in Simiao Junyi ointment. The conditions were as follows:the chromatographic column was Ultimate C18-ODS(250 mm ×4.6 mm,5 μm), the mobile phase was acetonitrile-0.1% phosphate solution, the flow rate was 1.0 ml·min-1, the detection wavelength was 280 nm, and the column temperature was 35℃. The common peaks in the chromatography were analyzed for their belongings. Results:Gradient elution was performed under the above optimal separation conditions, the constituents in Simiao Ju-nyi ointment were separated from each other perfectly, and the optimal fingerprint chromatography was obtained. Though the methodolo-gy examination, the indicators such as precision, stability and repeatability of the method were all promising, and the fingerprint chro-matography could be seen clearly and was easy to be analyzed. The relationships between Simiao Junyi ointment and the common peaks of four medicinal materials in the fingerprint chromatography were preliminary determined, which provided important basis for the quali-ty control of Simiao Junyi ointment. Conclusion:The HPLC fingerprint chromatography of Simiao Junyi ointment can be used as an a-nalysis method for the quality control of Simiao Junyi ointment, which provides reference for the quality control standard for the finished product.

Quality assessment of Shuangyu Granules based on simultaneous determination by HPLC fingerprint and multi-components / 中草药

Objective: To establish an HPLC fingerprint chromatography and determine seven compounds of multi-components in Shuangyu Granules (SG). Methods: The Kromasil C18 (250 mm × 4.6 mm, 3.5 μm) column was used with a mobile phase of acetonitrile-0.05% trifluoroacetic acid gradient elution. The flow rate was 0.8 mL/min, the column temperature was 30℃, and the detection wavelengths were 230 and 327 nm. The common peaks were identified by Q-TOF/MS. Results: The fingerprint chromatography included 17 mutual peaks, and the similarity was more than 0.95. Fourteen common peaks had been identified by LC-Q-TOF/MS, seven of which were unequivocally identified via comparing the retention times and mass spectra data with those of the standard compounds. Then the seven marker components were quantified. The developed quantitative method was validated in terms of accuracy (the recoveries ranged from 97.8% to 101.8% with RSDs less than 2%). Conclusion: The method is rapid, simple, and accurate and can be used for the quality control of SG.

Simultaneous determination of six active components by a single standard to determine multicomponents combined with fingerprint analysis for the quality control of Rhizoma Chuanxiong.

To control the quality of Rhizoma Chuanxiong, a method based on high-performance liquid chromatography method coupled with diode array detection was developed for the quantitative analysis of six active ingredients using a single standard to determine multi-components and chemical fingerprint analysis for the first time. The separation was performed on an Agilent Zorbax SB-C18 column by gradient elution with methanol and aqueous phase (containing 0.5% glacial acetic acid) at a flow rate of 1.0 mL/min. The UV wavelength was set at 274 nm. This assay was fully validated with respect to precision, repeatability, and accuracy. All calibration curves showed good linearity (R(2) > 0.9994) within test ranges. The limit of detection and limit of quantification were lower than 0.01 and 0.03 µg/mL, respectively. The relative standard deviation for repeatability and the intermediate precision of six analytes were less than 1.6 and 2.5%, respectively, the overall recovery was 96.1-103.1%. In addition, fingerprint chromatography using hierarchical clustering analysis and similarity analysis was performed to differentiate and classify the samples. The method described here could provide a more comprehensive and reasonable scientific assessment of the quality of Rhizoma Chuanxiong. Therefore, the strategy is feasible, credible, and is easily and effectively adapted for evaluating the quality control of Rhizoma Chuanxiong.

Nine components in Shiwei Penan Granule and their fingerprint chromatography / 中草药

Objective: To determine the nine components in Shiwei Penan Granule and establish the fingerprnt analysis for the quality control of Shiwei Penan Granule. Methods: The method was performed on a Thermo C18 column (250 mm × 4.6 mm, 5 μm); The gradient mobile phase consisted of acetonitrile (A)-0.1% phosphoric acid (B) with a flow rate of 1.0 mL/min (0-16 min, 5%-14% A; 16-20 min, 14%-17% A, 20-35 min, 17%-19% A; 35-60 min, 19%-35% A; 60-62 min, 35%-75% A; 62-78 min, 75% A); The detection wavelength was set at 230 nm (for quantitative analysis and fingerprint). Results: Gallic acid, sodium danshensu, chlorogenic acid, paeoniflorin, polydatin, benzoic acid, salvianolic acid B, emodin, and physcion were baseline seperated with good linearity relationships (r > 0.999 9) between concentration and peak areas over the linear ranges. The average recoveries of the compounds were 100.15% (RSD = 1.39%), 99.89% (RSD = 1.71%), 99.92% (RSD = 0.67%), 99.28% (RSD = 0.60%), 99.89% (RSD = 0.80%), 99.72% (RSD = 1.83%), 99.91% (RSD = 0.79%), 100.06% (RSD = 0.94%), and 99.97% (RSD = 1.36%). Using Traditional Chinese Medicine Fingerprint Similarity Evaluation System (2012 Edition) to analyze the fingerprint of 15 batches of Shiwei Penan Granule, the similarity values between the reference fingerprint and the 15 batches were higher than 0.977. Conclusion: The method is simple, rapid, and accurate, and can be used as an effective method to evaluate the quality of Shiwei Penan Granule.

Investigation of research method of HPLC specific chromatography of biochemical injection with multiple components / 中国生化药物杂志

The existing literature on fingerprint chromatography at home and abroad and the quality control of biochemical injection with multiple components were reviewed in this article.Combined with the laboratory research, It is proposed that the strategy for HPLC specific chromatography of biochemical injection with multiple components in order to provide the basis for effectively promoting the establishment and development of HPLC specific chromatography of biochemical injection with multiple components.

Exploration and classification of chromatographic fingerprints as additional tool for identification and quality control of several Artemisia species.

The World Health Organization accepts chromatographic fingerprints as a tool for identification and quality control of herbal medicines. This is the first study in which the distinction, identification and quality control of four different Artemisia species, i.e. Artemisia vulgaris, A. absinthium, A. annua and A. capillaris samples, is performed based on the evaluation of entire chromatographic fingerprint profiles developed with identical experimental conditions. High-Performance Liquid Chromatography (HPLC) with Diode Array Detection (DAD) was used to develop the fingerprints. Application of factorial designs leads to methanol/water (80:20 (v/v)) as the best extraction solvent for the pulverised plant material and to a shaking bath for 30 min as extraction method. Further, so-called screening, optimisation and fine-tuning phases were performed during fingerprint development. Most information about the different Artemisia species, i.e. the highest number of separated peaks in the fingerprint, was acquired on four coupled Chromolith columns (100 mm × 4.6 mm I.D.). Trifluoroacetic acid 0.05% (v/v) was used as mobile-phase additive in a stepwise linear methanol/water gradient, i.e. 5, 34, 41, 72 and 95% (v/v) methanol at 0, 9, 30, 44 and 51 min, where the last mobile phase composition was kept isocratic till 60 min. One detection wavelength was selected to perform data analysis. The lowest similarity between the fingerprints of the four species was present at 214 nm. The HPLC/DAD method was applied on 199 herbal samples of the four Artemisia species, resulting in 357 fingerprints. The within- and between-day variation of the entire method, as well as the quality control fingerprints obtained during routine analysis, were found acceptable. The distinction of these Artemisia species was evaluated based on the entire chromatographic profiles, developed by a shared method, and visualised in score plots by means of the Principal Component Analysis (PCA) exploratory data-analysis technique. Samples of different quality could be indicated on the score plots. No multi-component analysis was required to reach the goal. Furthermore, differences related to the origin of some of the not-certified samples were shown. The importance of the specific herbal part used for its identification was also presented. In addition, no differences were observed among fingerprints of lyophilised or conditioned-air dried samples. Finally, a classification technique, Soft Independent Modelling by Class Analogy (SIMCA), was successfully evaluated as identification technique for unknown samples. Six additional Artemisia species (29 herbal samples) were identified as not belonging to any of the four modelled classes. The developed chromatographic fingerprints and the evaluation of the entire profiles provide an added value to the distinction, identification and quality control of the simultaneously investigated Artemisia species.

Research for GC fingerprint chromatography of Yujin Injection / 中成药

AIM: To establish the fingerprint chromatography of Yujin Injection (Herba Houttuyniae, Flos Lonicerae) by GC. In the paper the authors examine the similarity among standard and sample chromatogra-phies. METHODS: GC was used to analyze the volatile ingredients, SGE 30QC3 colume (30m? 0.32mm? 0.5?m) was used with column temperature from 100℃ to 170℃ with 2℃?min -1 below 150℃ and 5℃?min -1 above 150℃, flow rate at 1.0mL?min -1 and detector temperature at 200℃. RESULTS: Standard fingerprint consisted of 17 marker peaks, the comparison of similarity's RSD to the injection of different batch had no more than 2%. CONCLUSION: According to the selected chromatographic conditions, a good fingerprint of the injection has been described. The method is simple, accurate with good reproducibility. It may be practical for the quality control of Yujin Injection.

Fingerprint chromatography of Shengmai Injection by HPLC / 中成药

AIM: To establish the fingerprint chromatography of Shengmai Injection(Radix et Rhizoma Ginseng Rubra,Radix Ophiopogonis,Fructus Schisandrae Chinensis) METHODS: A Waters symmetryshield TMRP 18 column(4.6 mm?250 mm;5 ?m) was adopted,acetonitrille-water as a mobile phase;The detection wavelength was set at 203 nm. RESULTS: According to the selected chromatographic conditions,standard fingerprint chromatography which was obtained consisted of 20 communal peaks. CONCLUSION: The method is simple,accurate with good reproducibility,and can be applied to the quality control of Shengmai Injection.

Affirming marker peaks for fingerprint chromatography of Shengmai Injection / 中成药

AIM: To affirm marker peaks for the fingerprint chromatography of Shengmai Injection. METHODS: LC-MS/MS method was used, with a Waters symmetryshield TM RP_ 18 column(4.6 mm?250 mm; 5.0 ?m), acetonitrile-water as a mobile phase, The detection wavelength was at 203 nm. Ion trap mass spectrum. RESULTS: Affirming marker peaks for fingerprint chromatography of Shengmai Injection and 10 marker peaks were affirmed. CONCLUSION: The method can affirm marker peaks for the fingerprint chromatography of Shengmai Injection. It is simple, accurate, and has practicality.