Study on Quality Markers of Poria cocos Based on UPLC-Q-TOF-MS and Network Pharmacology Technology

[Objectives]To analyze the main chemical components of traditional Chinese medicine(TCM)Poria cocos by liquid chromatography-mass spectrometry,and explore the active components for P.cocos in the treatment of primary dysmenorrhea(PD)by network pharmacology to predict its quality markers(Q-marker).[Methods]Ultra performance liquid chromatography-quadrupole tandem time-of-flight mass spectrometry(UPLC-Q-TOF-MS)in positive and negative ion mode was used to collect high quality MS and MS/MS data of Poria cocos,and qualitative characterization of the components in Poria cocos was performed using Analyst TF 1.7.1 and PeakView 2.2 software with reference to internal databases and literature.Taking the above identified chemical components as the research object,we used network pharmacology to discover the potential effective components and their key targets of PD,and metabolic pathway enrichment analysis of the core targets was performed to screen the Q-marker of P.cocos based on the five principles of Q-marker of TCM.[Results]UPLC-Q-TOF-MS technique was used to identify 41 chemical components of P.cocos,including 3 amino acids,26 triterpenoids,4 lactones,7 organic acids and 1 adenosine.It was more likely to lose H 2O and CO 2 during cleavage and break at the carbonyl group.The triterpenoids were mainly in the form of[M-H]-peaks in negative ion mode,which was easy to lose some structural fragments such as H 2O,CH 3COOH,CH 4,CO 2,etc.Further network pharmacological analysis showed that 302 targets of chemical components of P.cocos,518 targets of PD,28 common targets of component and disease,and 27 core targets such as PTGS2,ESR1,TNF,IL1B were observed by PPI interactions network analysis.451 biological processes such as hormone response and inflammatory response regulation were obtained by GO enrichment analysis.KEGG enrichment analysis showed that 89 pathways including PI3K/Akt signaling pathway,IL-17 signaling pathway and TNF signaling pathway were obtained.The connectivity value of components was analyzed.The core components with the connectivity value greater than 10,including poricoic acid A,polyporenic acid,polyporenic acid C,and 25-hydroxy-3-epidehydrotumoric acid were selected,while the key targets with the connectivity value greater than 15 included TNF,PTGS2,IL1B and CASP3.Molecular docking between core components and key targets was performed,and most of the docking energy was less than-5 kcal/mol,indicating that the binding between the active components and target proteins of P.cocos was relatively stable,so 23 active components of P.cocos were determined.Following the five principles of Q-marker,four possible Q-markers of P.cocos were predicted,including poricoic acid A,pachymic acid,polyporenic acid C,and 25-hydroxy-3-epidehydrotumoric acid.[Conclusions]P.cocos was mainly composed of triterpenoids,its effect on the treatment of PD may be achieved mainly by poricoic acid A,pachymic acid,polyporenic acid C,and 25-hydroxy-3-epi-dehydrotumoric acid acting on PTGS2,ESR1,TNF,IL1B and other targets to regulate PI3K/Akt signaling pathway,IL-17 signaling pathway,TNF signaling pathway,etc.Based on these active components,poricoic acid A,pachymic acid,polyporenic acid C,and 25-hydroxy-3-epi-dehydrotumoric acid could be taken as Q-markers of P.cocos,which provided a solid basis for further improving the quality standard of P.cocos.

Research progress and quality markers prediction analysis of Dysosma versipellis

Dysosma versipellis is the rhizome of Bajiaolian in Berberaceae.It is a unique medicinal plant in China and it was the first to be recorded in Shennong’s Classic of Materia Medica(unknown author,25–220 C.E.).It has complex chemical components as well as complex,pharmacological and toxicological effects are extensive.Based on the recent research progress,the chemical constituents,pharmacological activities and toxicological effects of Dysosma versipellis were summarized.Combined with the core concept of quality markers of traditional Chinese medicine,the prediction and analysis were carried out from the aspects of phytogenetics and the source pathway of specific chemical constituents,traditional medicinal properties,traditional efficacy,chemical composition measurability,blood components,and pharmacokinetics,in order to provide reference for further study of Dysosma versipellis.

Multimodal integrated strategy for the discovery and identification of quality markers in traditional Chinese medicine

With the modernization and internationalization of traditional Chinese medicine(TCM),the requirement for quality control has increased.The quality marker(Q-marker)is an important standard in this field and has been implemented with remarkable success in recent years.However,the establishment of Qmarkers remains fragmented and the process lacks systematicity,resulting in inconsistent quality control and insufficient correlation with clinical efficacy and safety of TCM.This review introduces four multimodal integrated approaches that contribute to the discovery of more comprehensive and accurate Qmarkers,thus aiding in the establishment of new quality control patterns based on the characteristics and principles of TCM.These include the whole-process quality control strategy,chemical-activity-based screening method,efficacy,safety,and consistent combination strategy,and TCM theory-guided approach.Furthermore,methodologies and representative examples of these strategies are described,and important future directions and questions in this field are also proposed.

Clinical metabolic analysis combined with traceability of biosynthetic pathway:a new approach to quality marker of Chinese materia medica

Quality marker(Q-marker)of Chinese materia medica(CMM)plays an important role in quality control of CMM products.However,its research strategy and technique remain unclear.Based on the fact that quality standard of CMM should be associated with clinical efficacy,taking Jinqi Jiangtang tablet treating type 2 diabetes as an example,the Q-marker related to activity via the reverse analysis of drug metabolism in clinic and traceability of botanic biosynthetic pathways is discovered and validated.Therefore,we proposed a new research strategy of Q-marker of CMM with"Discovery of clinical active constituents as guidance,Reverse analysis of metabolic transformations as link,and Traceability of biosynthesis pathways as key",to improve quality control of CMM products.

Research Progress on Rodgersia and Predictive Analysis on its Quality Markers

Rodgersia is a traditional Chinese medicine that contains a variety of chemical constituents,including flavonoids,terpenoids,phenylpropanoids,gallic acid derivatives,steroids,volatile oils,and tannins,with anti-inflammatory,antioxidant,antitumor,antibacterial,antivirus,hepatoprotective,and other properties.In this paper,the main chemical constituents and pharmacological effects of Rodgersia are summarized.On this basis,the quality markers of this genus were predicted based on chemical composition,traditional medicinal properties,traditional efficacy,and measurable components.This review provides the basis for in-depth research,utilization,and quality control of Rodgersia.

A New Concept on Quality Marker for Quality Assessment and Process Control of Chinese Medicines

Chinese medicine （CM） is the most typical conventional therapy compared with any other traditional or alternative medicine systems. The active components of CMs are either primary or secondary metabolites generated by metabolic and biosynthetic enzymes in plants, protecting the plants from environmental stress. The characteristics of these metabolites are diverse, complicated and unique. In this paper, current approaches for quality assessment were extensively reviewed, a new concept of quality marker （Q-marker） was then proposed for CM quality assessment. Additionally, definition of the Q-marker, as well as the relevant methods, were discussed, on the basis of the biosynthetic pathways of secondary metabolites and source of biological active components. Study design of Q-marker is complex system for quality assessment and production process control of CM products with transitivity and traceability. Therefore, the system with characteristics of transmission and traceability is expected to be established for regulation of quality. Upon the concept which the transitivity and traceability in the quality assessment and production process control covered the entire process, such as raw materials, decoction slices, processing, extraction and production can be further enhanced. The transitivity and traceability will inevitably require close attention to ＂who, what, where, when, and why＂ details at each stage of Q-markers of CM production form raw materials to patent product. The establishing quality standards are enablers of many and various transitivity and traceability solutions, not a solution in them. It means that the transitivity and traceability system is readily link between products and across borders in quality. According to the thinking mode and methods of investigation on quality assessment of CM product, we focus on the entire process, in terms of safety and effectiveness and quality control. The standard preparation of CM or CM decoction is not only the basis for study of Q-marker, but also the basis for transmission and traceability of the quality of CM product.

Quality Marker Concept Inspires the Quality Research of Traditional Chinese Medicines

Traditional Chinese medicines （TCMs） have been used in China for thousands of years for the treatment of various diseases, highly treated as the Chinese cultural heritage. It has played the pivotal role in maintaining the health of Chinese people and even the entire globe.

Research on Quality Markers of Moutan Cortex: Quality Evaluation and Quality Standards of Moutan Cortex

Objective To identify the quality markers of Moutan Cortex（MC） and establish the quality evaluation methods for multi-component assay and fingerprinting of MC. Methods The chemical constituents in MC were identified by HPLC-QTOF-MS. UPLC was employed for the multi-component assay and fingerprinting of MC. Furthermore, text mining was carried out to review the biosynthesis pathways and pharmacological and pharmacokinetic studies related to MC, and in silico target fishing was conducted to construct compound-target networks for MC. Results Sixteen compounds were clearly identified in MC and their structures were confirmed through comparison with literature data. In addition, the biosynthetic pathways and component specificities of the identified compounds were summarized and confirmed by text mining.Pharmacological activities, including traditional usage and modern pharmacological studies were summarized. A total of 282 targets from Homo sapiens were fished for 13 compounds. In addition, pharmacokinetic studies of different compounds were synopsized. Finally, multi-component assay and fingerprint of MC were established. Conclusion Eight major components are selected as quality markers of MC, such as oxypaeoniflorin, apiopaeonoside, albiflorin, paeonolide, paeoniflorin, 1,2,3,4,6-penta-O-galloyl-β-D-glucose, mudanpioside C and paeonol. These eight quality markers are successfully applied to the quality evaluation of MC, and could be useful in improving the current quality standards of MC.

Study on Quality Markers and Action Mechanisms of Inulae Flos on Anti-Hepatitis Through Network Pharmacology and High-Performance Liquid Chromatography Fingerprints

Objective:The objective of the study is to combine network pharmacology with high-performance liquid chromatography(HPLC)to screen for quality markers(Q-markers)of Inulae Flos and predict mechanism on anti-hepatitis.Materials and Methods:Active ingredient library of Inulae Flos is structured using databases and the literature.“Compound-target-pathway”network on anti-hepatitis and protein–protein interaction(PPI)network are constructed using network pharmacology.Next,chromatographic fingerprints of Inulae Flos in 7 origins are obtained through HPLC,and chemometric analysis is implemented to identify chemical markers,which is combined with network pharmacology to identify Q-markers and detect content.Results:1,6-O,O-Diacetylbritannilactone,Ivangustin,and Inulanolide A are key ingredients of Inulae Flos to interact with 82 potential targets related to anti-hepatitis.Furthermore,signal transducer and activator of transcription 3,tumor necrosis factor,interleukin-6,and transcription factor AP-1 are the core targets in the PPI network.Chromatographic fingerprints of the Inulae Flos define 20 common peaks and identify 8 peaks using reference substances.Through partial least square discriminant analysis,7compounds including caffeic acid,chlorogenic acid,and 1,6-O,O-Diacetylbritannilactone were main chemical markers for variability.1,6-O,O-Diacetylbritannilactone is both a key ingredient and exclusive chemical marker.Therefore,1,6-O,O-diacetylbritannilactone is a Q-marker of Inulae Flos,and the average content is 1.82 mg/g.Conclusion:1,6-O,O-diacetylbritannilactone is determined to be a Q-marker of Inulae Flos.

Molecular Markers in Improvement of Fiber Quality Traits in Cotton

Cotton is the worlds leading natural fiber crop,and it is the cornerstone of textile industries worldwide.The cotton industry is confronted with problems in cost of production and

Novel assays for quality evaluation of XueBiJing:Quality variability of a Chinese herbal injection for sepsis management

XueBiJing is an intravenous five-herb injection used to treat sepsis in China.The study aimed to develop a liquid chromatography-tandem mass spectrometry(LC-MS/MS)-or liquid chromatography-ultraviolet(LC-UV)-based assay for quality evaluation of XueBiJing.Assay development involved identifying marker constituents to make the assay therapeutically relevant and building a reliable one-point calibrator for monitoring the various analytes in parallel.Nine marker constituents from the five herbs were selected based on XueBiJing's chemical composition,pharmacokinetics,and pharmacodynamics.A selectivity test(for“similarity of response”)was developed to identify and minimize interference by nontarget constituents.Then,an intercept test was developed to fulfill“linearity through zero”for each analyte(absolute ratio of intercept to C response,<2%).Using the newly developed assays,we analyzed samples from 33 batches of XueBiJing,manufactured over three years,and found small batch-to-batch variability in contents of the marker constituents(4.1%-14.8%),except for senkyunolide I(26.5%).

Integrated approach of network pharmacology and chemometrics for quality control of Chinese patent medicine:A case study on Huo-Luo-San

Traditional Chinese medicine(TCM)has garnered significant global interest owing to its multi-component and multi-target theoretical framework and extensive therapeutic efficacy.However,the identification of quality markers(Q-markers)remains a formidable challenge in TCM.Hence,this study aimed to integrate network pharmacology and chemometrics to identify Q-markers in Chinese patent medicine,with a focus on Huo-Luo-San(HLS)as a case study.HLS,a widely used powdered Chinese patent medicine in China,comprises a complex formula of 10 herbs,initially formulated during the Qing dynasty for treating fractures.Initially,13 components,chlorogenic acid,typhaneoside,isorhamnetin-3-O-neohesperidoside,cynaroside,notoginsenoside R_1,ginsenoside Rg_1,baicalin,berberine hydrochloride,ginsenoside Rb_1,dehydrocostus lactone,dioscin,imperatorin,and costunolide,were selected as phytochemical markers for each herb based on the Chinese Pharmacopoeia(2020 version),forming the“Herbs-Compounds-targets”network of HLS using network pharmacology.Subsequently,employing network pharmacology,the 13 HLS components were quantified using UPLC-QqQ-MS.Chromatographic conditions were optimized on a Waters Cortecs C_(18)column(2.1 mm×100 mm,1.6μm)with a gradient elution comprising 0.1%formic acid in water and acetonitrile.Analyte detection was performed in the multiple-reaction monitoring mode,and the method underwent validation for linearity,detection limit,precision,repeatability,stability,and accuracy.The validated method was then utilized to analyze the 13 components in 15 batches of HLS samples.Chemometric techniques,including hierarchical cluster analysis,principal component analysis,orthogonal partial least squares projection discriminant analysis,and box map analyses,were subsequently employed to identify the Q-markers.Ultimately,six components,baicalin,notoginsenoside R_1,berberine hydrochloride,dioscin,imperatorin,and chlorogenic acid,were selected as Q-markers for HLS.The integration of network pharmacology with chemometrics represented a novel approach for selecting Q-markers in Chinese patent medicine.

Predicting the grades of Astragali radix using mass spectrometrybased metabolomics and machine learning

Astragali radix(AR,the dried root of Astragalus)is a popular herbal remedy in both China and the United States.The commercially available AR is commonly classified into premium graded(PG)and ungraded(UG)ones only according to the appearance.To uncover novel sensitive and specific markers for AR grading,we took the integrated mass spectrometry-based untargeted and targeted metabolomics approaches to characterize chemical features of PG and UG samples in a discovery set(n=16 batches).A series of five differential compounds were screened out by univariate statistical analysis,including arginine,calycosin,ononin,formononetin,and astragalosideⅣ,most of which were observed to be accumulated in PG samples except for astragalosideⅣ.Then,we performed machine learning on the quantification data of five compounds and constructed a logistic regression prediction model.Finally,the external validation in an independent validation set of AR(n=20 batches)verified that the five compounds,as well as the model,had strong capability to distinguish the two grades of AR,with the prediction accuracy>90%.Our findings present a panel of meaningful candidate markers that would significantly catalyze the innovation in AR grading.

Imaging the crystal orientation of 2D transition metal dichalcogenides using polarization-resolved second-harmonic generation

We use laser-scanning nonlinear imaging microscopy in atomically thin transition metal dichalcogenides(TMDs)to reveal information on the crystalline orientation distribution,within the 2D lattice.In particular,we perform polarization-resolved second-harmonic generation(PSHG)imaging in a stationary,raster-scanned chemical vapor deposition(CVD)-grown WS2 flake,in order to obtain with high precision a spatially resolved map of the orientation of its main crystallographic axis(armchair orientation).By fitting the experimental PSHG images of sub-micron resolution into a generalized nonlinear model,we are able to determine the armchair orientation for every pixel of the image of the 2D material,with further improved resolution.This pixel-wise mapping of the armchair orientation of 2D WS2 allows us to distinguish between different domains,reveal fine structure,and estimate the crystal orientation variability,which can be used as a unique crystal quality marker over large areas.The necessity and superiority of a polarization-resolved analysis over intensity-only measurements is experimentally demonstrated,while the advantages of PSHG over other techniques are analysed and discussed.

Research progress on the pharmacological effects and chemical constituents of Pien Tze Huang and its potential Q-markers

Pien Tze Huang(PTH)was documented as an imperial prescription composed of Notoginseng Radix,Calculus Bovis,Snake Gallbladder,and Musk.It is famous in China and Asian countries due to its excellent effects in heat clearing,detoxifying,swelling reduction,and pain relieving.Modern pharmacological studies demonstrate that PTH shows excellent effects against various inflammatory diseases,liver diseases,and cancers.This review summaries the pharmacological effects,clinical applications,and mainchemical components of PTH.More importantly,its potential quality markers(Q-markers)were then analyzed based on the“five principles”of Q-markers under the guidance of Traditional Chinese Medicine theory,including transfer and traceability,specificity,efficacy,compatibility,and measurability.As a result,ginsenosides Rb1,ginsenoside Rg1,ginsenoside Rd,ginsenoside Re,notoginsenoside R1,dencichine,bilirubin,biliverdin,taurocholic acid,and muscone are considered as the Q-markers of PTH.These findings will provide guidance and assistance for the construction of a quality control system for PTH.

QUALITY CONTROL MARKERS OF DU-HUO-JI-SHENG-TANG IDENTIFIED ACCORDING TO ANTI-INFLAMMATORY EFFECTS

The Du-Huo-Ji-Sheng-Tang(DHJST)is the top commercial concentrated aqueous extract used in clinics for treating osteoarthritis.However,choosing the quality control markers for DHJST is an important problem for herbal medicine industry.In this study,DHJST was divided into three groups according to the medical philosophy of traditional Chinese medicine(TCM),such as Si-Wu-Tang(SWT),wind-dampness herbs(WDH)and tonifying

Study of biological markers in skin quality treatment by subcutaneous injection of a stabilized composition of 26 mg/mL of high molecular weight HA

Aim:Hyaluronic acid(HA)injectables have gained rapid acceptance for the treatment of skin rejuvenation.A novel HA/sorbitol composition intended for skin quality improvement containing 2.6%of high molecular weight HA stabilized by sorbitol was recently designed to be injected subcutaneously.The aim of this study was to assess the expression of biological markers of skin quality after administration of the composition.Methods:The HA/sorbitol composition was evaluated after injection into the superficial adipose tissue with ex vivo cultured human skin explants versus a product comparator to study the general morphology of the skin tissues and the expression of HA,elastin,collagen type Ⅰ,collagen type Ⅲ,and fibrillin-1 in the dermal layer.Results:The results demonstrate that the HA/sorbitol composition is able to boost the production of HA,elastin,collagen type Ⅰ,collagen type Ⅲ,and fibrillin-1 in the dermis while providing a proper quality of skin morphology.Conclusion:The HA/sorbitol composition improved biological markers of skin quality in the dermis after product injection into the superficial adipose tissue.This novel composition can be considered as an attractive solution to treat skin aging by injecting a specific HA/sorbitol formulation to strategically target the subcutaneous tissue to improve the quality of the different layers of the skin.

ETCM v2.0:An update with comprehensive resource and rich annotations for traditional Chinese medicine

Existing traditional Chinese medicine(TCM)-related databases are still insufficient in data standardization,integrity and precision,and need to be updated urgently.Herein,an Encyclopedia of Traditional Chinese Medicine version 2.0(ETCM v2.0,http://www.tcmip.cn/ETCM2/front/#/)was constructed as the latest curated database hosting 48,442 TCM formulas recorded by ancient Chinese medical books,9872 Chinese patent drugs,2079 Chinese medicinal materials and 38,298 ingredients.To facilitate the mechanistic research and new drug discovery,we improved the target identification method based on a two-dimensional ligand similarity search module,which provides the confirmed and/or potential targets of each ingredient,as well as their binding activities.Importantly,five TCM formulas/Chinese patent drugs/herbs/ingredients with the highest Jaccard similarity scores to the submitted drugs are offered in ETCM v2.0,which may be of significance to identify prescriptions/herbs/ingredients with similar clinical efficacy,to summarize the rules of prescription use,and to find alternative drugs for endangered Chinese medicinal materials.Moreover,ETCM v2.0 provides an enhanced Java Script-based network visualization tool for creating,modifying and exploring multi-scale biological networks.ETCM v2.0 may be a major data warehouse for the quality marker identification of TCMs,the TCM-derived drug discovery and repurposing,and the pharmacological mechanism investigation of TCMs against various human diseases.

Ultrahigh-resolution nonlinear optical imaging of the armchair orientation in 2D transition metal dichalcogenides

We used nonlinear laser scanning optical microscopy to study atomically thin transition metal dichalcogenides(TMDs)and revealed,with unprecedented resolution,the orientational distribution of armchair directions and their degree of organization in the two-dimensional(2D)crystal lattice.In particular,we carried out polarization-resolved second-harmonic generation(PSHG)imaging for monolayer WS2 and obtained,with high-precision,the orientation of the main crystallographic axis(armchair orientation)for each individual 120×120 nm^(2) pixel of the 2D crystal area.Such nanoscale resolution was realized by fitting the experimental PSHG images,obtained with sub-micron precision,to a new generalized theoretical model that accounts for the nonlinear optical properties of TMDs.This enabled us to distinguish between different crystallographic domains,locate boundaries and reveal fine structure.As a consequence,we can calculate the mean orientational average of armchair angle distributions in specific regions of interest and define the corresponding standard deviation as a figure-of-merit for the 2D crystal quality.