Chemical profiling of principle active and toxic constituents in herbs containing aristolochic acids

Objective:To clear the amounts of the principal active/toxic components in herbs containing aristolochic acids(HCAAs),which are still used as medicine and/or seasoning in many ethnic minority areas of China.Methods:In this study,six major active and toxic components in HCAAs were extracted with ultrasonic extraction.With 6-O-methyl guanosine as internal standard,the target compounds were analyzed qualitatively and quantitatively by using ultrahigh performance liquid chromatography-electrospray ionization-tandem mass spectrometry(UPLC-ESI-MS/MS)with multiple reaction monitoringinformation dependent acquisition-enhanced production ion scanning mode(MRM-IDA-EPI)combined with dynamic background subtraction(DBS)function.Results:The method showed good linearity in the linear range of the six analytes.The limit range of detection was from 0.01 ng/mL to 0.27 ng/mL.All of the detection repeatability,extraction repeatability and accuracy of the method were good.After extraction,the samples remained stable at 15℃ within 24 h.Six analytes were all found in samples except aristolactam(AL)in sample 2,and the contents varied greatly.The contents of these compounds decreased in fruits,leaves and stems of Aristolochia delavayi successively.Conclusion:This method has the advantages of less sample dosage,simple operation,short analysis cycle,high sensitivity,specificity and accuracy.It laid a good foundation for guiding the safety of HCAAs,the indepth study of pharmacological and toxicological effects and the scientific and standardized processing and compatibility of HCAAs.

Comprehensive chemical profiling and quantitative analysis of ethnic Yi medicine Miao-Fu-Zhi-Tong granules using UHPLC-MS/MS

Developing analytical methods for the chemical components of natural medicines remains a challenge due to its diversity and complexity.Miao-Fu-Zhi-Tong(MFZT)granules,an ethnic Yi herbal prescription,comprises 10 herbs and has been clinically applied for gouty arthritis(GA)therapy.Herein,a series of chemical profiling strategies including in-house library matching,molecular networking and MS/MS fragmentation behavior validation based on ultra-high performance liquid chromatography-tandem mass spectrometry(UHPLC-MS/MS)were developed for qualitative analysis of MFZT granules.A total of 207 compounds were identified or characterized in which several rare guanidines were discovered and profiled into alkyl substituted or cyclic subtypes.Moreover,network pharmacology analysis indicated that MFZT’s anti-gout mechanism was mostly associated with the nuclear factor kappa-B(NF-κB)signaling,nucleotide oligomerization domain(NOD)-like signaling and rheumatoid arthritis pathways,along with the synergistic effect of 84 potential active compounds.In addition,a quantitative analytical method was developed to simultaneously determine the 29 potential effective components.Among them,berberine,pellodendrine,3-feruloylquinic acid,neoastilbin,isoacteoside and chlorogenic acid derivatives at higher concentrations were considered as the chemical markers for quality control.These findings provide a holistic chemical basis for MFZT granules and will support the development of effective analytical methods for the herbal formulas of natural medicines.

Chemical profiling of Qixue Shuangbu Tincture by ultra-performance liquid chromatography with electrospray ionization quadrupole-time-of-flight high-definition mass spectrometry (UPLC-QTOF/MS)

The present study was designed to develop and validate a sensitive and reliable ultra high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(UPLC-QTOF/MS) method to separate and identify the chemical constituents of Qixue Shuangbu Tincture(QXSBT), a classic traditional Chinese medicine(TCM) prescription. Under the optimized UPLC and QTOF/MS conditions, 56 components in QXSBT, including chalcones, triterpenoids, protopanaxatriol, flavones and flavanones were identified and tentatively characterized within a running time of 42 min. The components were identified by comparing the retention times, accurate mass, and mass spectrometric fragmentation characteristic ions, and matching empirical molecular formula with that of the published compounds. In conclusion, the established UPLC-QTOF/MS method was reliable for a rapid identification of complicated components in the TCM prescriptions.

Chemical profiling and histochemical analysis of Bupleurum marginatum roots from different growing areas of Hubei province

Bupleuri Radix has been widely used in traditional Chinese medicine.In the current herbal market,the species Bupleurum marginatum Wall.ex DC.is the main source of Bupleuri Radix.Although Bupleuri Radix from the roots of B.marginatum grown wild in the North West of Hubei province has higher quality compared with those from other regions according to the previous investigations,the exhaustive exploitation driven by increasing demand has drastically reduced the wild resource.As a result,germplasm evaluation and quality resource exploration are important for the sustainable utilization and cultivation of B.marginatum.A preliminary study indicated differences in the tissue structure of B.marginatum grown in different areas of North Western Hubei province.In the current study,various tissues of the roots of B.marginatum grown in different areas of North Western Hubei were subjected to laser microdissection and analyzed by microscopy and ultra-high performance liquid chromatography quadrupole time-of-flight mass spectrometry(UHPLC–Q-TOF-MS).The results show that wild plants from Maqiao Town,Baokang County contain the most saikosaponins distributed mainly in cork,cortex and phloem.This study provides key chemical information for evaluating the quality of B.marginatum roots.

Characteristic chemical profile of Juhe Fang extract with lipid-lowering properties

Objective:The objective of this study was to verify the lipid-lowering effect of Juhe Fang extract(JHFE)and to determine its characteristic chemical profile in vitro and in vivo.Methods:A hyperlipidemia model was established by feeding mice a high-fat diet(HFD).After treatment for 30 days,serum total cholesterol(TC),triglyceride(TG),high-density lipoprotein cholesterol(HDL-C)and low-density lipoprotein cholesterol(LDL-C)levels were measured with an automatic biochemistry analyzer.The components from JHFE obtained from in vivo and in vitro experiments were investigated using an UPLC-Q Exactive-Orbitrap MS/MS.Results:The TC,TG,and LDL-C in the serum significantly decreased and the HDL-C significantly increased after JHFE treatment.A total of 95 compounds from JHEF including 15 phenolic acids(PA),4 phenylethanoid glycosides(PG),24 flavonoids(F),14 triterpenoids(T),10 diterpenoid glycosides(D),18 alkaloids(A)and 10 others(O)were identified.Trigonelline was discovered for the first time in a herbal medicine of Juhe Fang.Furthermore,68 compounds were identified in vivo including 28 prototype compounds and 40 metabolites.The metabolic characteristics of these components were revealed including identification of new metabolites of 4-hydroxyphenyl ethyl-8-O-[a-L-arabinopyranosyl-(1/6)]-b-D-glucopyranoside(PEG)and lirinidine.A total of 43 components from JHFE were absorbed and/or metabolized.The contribution rate of each type of chemical component from JHFE to its lipidlowering effect from high to low were A,F,PG,PA,D and T.Conclusion:The results of this study showed that JHFE demonstrated a significant lipid-lowering effect in a high-fat diet(HFD)-induced hyperlipidemia mouse model.Specific types of PA,PG,F,D,T and A formed the pharmaceutical architecture of the lipid-lowering effect of JHFE.This study should prove useful for clarifying the components responsible for the lipid-lowering effect of JHFE and provide a basis for precision quality control research.

Deciphering chemical and metabolite profiling of Chang-Kang-Fang by UPLC-Q-TOF-MS/MS and its potential active components identification

Chang-Kang-Fang(CKF)formula,a Traditional Chinese Medicine(TCM)prescription,has been widely used for the treatment of irritable bowel syndrome(IBS).However,its potential material basis and underlying mechanism remain elusive.Therefore,this study employed an integrated approach that combined ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-Q/TOF-MS)with network pharmacology to systematically characterize the phytochemical components and metabolites of CKF,as well as elucidating its underlying mechanism.Through this comprehensive analysis,a total of 150 components were identified or tentatively characterized within the CKF formula.Notably,six N-acetyldopamine oligomers from Cicadae Periostracum and eight resin glycosides from Cuscutae Semen were characterized in this formula for the first time.Meanwhile,149 xenobiotics(58 prototypes and 91 metabolites)were detected in plasma,urine,feces,brain,and intestinal contents,and the in vivo metabolic pathways of resin glycosides were elaborated for the first time.Furthermore,network pharmacology and molecular docking analyses revealed that alkaloids,flavonoids,chromones,monoterpenes,N-acetyldopamine dimers,p-hydroxycinnamic acid,and Cus-3/isomer might be responsible for the beneficial effects of CKF in treating IBS,and CASP8,MARK14,PIK3C,PIK3R1,TLR4,and TNF may be its potential targets.These discoveries offer a comprehensive understanding of the potential material basis and clarify the underlying mechanism of the CKF formula in treating IBS,facilitating the broader application of CKF in the field of medicine.

Biotechnology of the multipurpose tree species Arbutus unedo:a review

Arbutus unedo L.(strawberry tree,Ericaceae)is a woody species with a circum-Mediterranean distribu-tion.It has considerable ecological relevance in southern European forests due to its resilience to abiotic and biotic stressors.Its edible red berries are used in the production of traditional products,including an expensive spirit.Several compounds extracted from the species have bioactive prop-erties used by the cosmetic and pharmaceutical industries.The strawberry tree has gone from a neglected species to become a highly valuable crop with large cultivated areas in southern European and North African countries.Due to an increasing demand from farmers for plants with improved features,researchers have been trying to improve this spe-cies through conventional and biotechnological tools,focus-ing mainly on population analysis using molecular markers,in vitro cloning,tolerance to biotic and abiotic stresses,and intraspecific crosses to obtain genotypes with new char-acteristics.The objective of this review is to gather and update information about the species and make it available to researchers and stakeholders.Future research areas that are considered a priority for this species are highlighted.

Rabdosia serra alleviates dextran sulfate sodium salt-induced colitis in mice through anti-inflammation,regulating Th17/Treg balance,maintaining intestinal barrier integrity,and modulating gut microbiota

Rabdosia serra(R.serra),an important component of Chinese herbal tea,has traditionally been used to treat hepatitis,jaundice,cholecystitis,and colitis.However,the chemical composition of R.serra and its effect against colitis remain unclear.In this study,the chemical composition of the water extract of R.serra was analyzed using ultra performance liquid chromatography coupled with a hybrid linear ion trap quadrupole-orbitrap mass spectrometer(UPLC-LTQ-Orbitrap-MS).A total of 46 compounds,comprising ent-kaurane diterpenoids,flavonoids,phenolic acids,and steroids,were identified in the water extract of R.serra,and the extract could significantly alleviate dextran sulfate sodium salt-induced colitis by improving colon length,upregulating anti-inflammatory factors,downregulating proinflammatory factors,and restoring the balance of T helper 17/T regulatory cells.R.serra also preserved intestinal barrier function by increasing the level of tight junction proteins(zonula occludens 1 and occludin)in mouse colonic tissue.In addition,R.serra modulated the gut microbiota composition by increasing bacterial richness and diversity,increasing the abundance of beneficial bacteria(Muribaculaceae,Bacteroides,Lactobacillus,and Prevotellaceae_UCG-001),and decreasing the abundance of pathogenic bacteria(Turicibacter,Eubacterium_fissicatena_group,and Eubacterium_xylanophilum_group).Gut microbiota depletion by antibiotics further confirmed that R.serra alleviated colitis in a microbiota-dependent manner.Overall,our findings provide chemical and biological evidence for the potential application of R.serra in the management of colitis.

Comprehensive profiling and characterization of the absorbed components and metabolites in mice serum and tissues following oral administration of Qing-Fei-Pai-Du decoction by UHPLC-Q-Exactive-Orbitrap HRMS

Qing-Fei-Pai-Du decoction(QFPDD)is a Chinese medicine compound formula recommended for combating corona virus disease 2019(COVID-19)by National Health Commission of the People's Republic of China.The latest clinical study showed that early treatment with QFPDD was associated with favorable outcomes for patient recovery,viral shedding,hospital stay,and course of the disease.However,the effective constituents of QFPDD remain unclear.In this study,an UHPLC-Q-Orbitrap HRMS based method was developed to identify the chemical constituents in QFPDD and the absorbed prototypes as well as the metabolites in mice serum and tissues following oral administration of QFPDD.A total of 405 chemicals,including 40 kinds of alkaloids,162 kinds of flavonoids,44 kinds of organic acids,71 kinds of triterpene saponins and 88 kinds of other compounds in the water extract of QFPDD were tentatively identified via comparison with the retention times and MS/MS spectra of the standards or refereed by literature.With the help of the standards and in vitro metabolites,195 chemical components(including 104 prototypes and 91 metabolites)were identified in mice serum after oral administration of QFPDD.In addition,165,177,112,120,44,53 constituents were identified in the lung,liver,heart,kidney,brain,and spleen of QFPDD-treated mice,respectively.These findings provided key information and guidance for further investigation on the pharmacologically active substances and clinical applications of QFPDD.

The main strategies for soil pollution apportionment:A review of the numerical methods

Nowadays,a large number of compounds with different physical and chemical properties have been determined in soil.Environmental behaviors and source identification of pollutants in soil are the foundation of soil pollution control.Identification and quantitative analysis of potential pollution sources are the prerequisites for its prevention and control.Many efforts have made to develop methods for identifying the sources of soil pollutants.These efforts have involved the measurement of source and receptor parameters and the analysis of their relationships via numerical statistics methods.We have comprehensively reviewed the progress made in the development of source apportionment methodologies to date and present our synthesis.The numerical methods,such as spatial geostatistics analysis,receptor models,and machine learning methods are addressed in depth.In most cases,however,the effectiveness of any single approach for source apportionment remains limited.Combining multiple methods to address soil quality problems can reduce uncertainty about the sources of soil pollution.This review also constructively highlights the key strategies of combining mathematical models with the assessment of chemical profiles to provide more accurate source attribution.This review intends to provide a comprehensive summary of source apportionment methodologies to help promote further development.

Artificial Bear Bile:A Novel Approach to Balancing Medical Requirements and Animal Welfare

Bear bile has been a valuable and effective medicinal material in traditional Chinese medicine(TCM)for over 13 centuries.However,the current practice of obtaining it through bear farming is under scrutiny for its adverse impact on bear welfare.Here,we present a new approach for creating artificial bear bile(ABB)as a high-quality and sustainable alternative to natural bear bile.This study addresses the scientific challenges of creating bear bile alternatives through interdisciplinary collaborations across various fields,including resources,chemistry,biology,medicine,pharmacology,and TCM.A comprehensive efficacy assessment system that bridges the gap between TCM and modern medical terminology has been established,allowing for the systematic screening of therapeutic constituents.Through the utilization of chemical synthesis and enzyme engineering technologies,our research has achieved the environmentally friendly,large-scale production of bear bile therapeutic compounds,as well as the optimization and recomposition of ABB formulations.The resulting ABB not only closely resembles natural bear bile in its composition but also offers advantages such as consistent product quality,availability of raw materials,and independence from threatened or wild resources.Comprehensive preclinical efficacy evaluations have demonstrated the equivalence of the therapeutic effects from ABB and those from commercially available drained bear bile(DBB).Furthermore,preclinical toxicological assessment and phase I clinical trials show that the safety of ABB is on par with that of the currently used DBB.This innovative strategy can serve as a new research paradigm for developing alternatives for other endangered TCMs,thereby strengthening the integrity and sustainability of TCM.

Biochemical reactions in metabolite-protein interaction

Active endogenous metabolites regulate the viability of cells. This process is controlled by a series ofinteractions between small metabolites and large proteins. Previously, several studies had reported thatmetabolite regulates the protein functions, such as diacylglycerol to protein kinase C, lactose regulationof the lac repressor, and HIF-1α stabilization by 2-hydroxyglutarate. However, decades old traditionalbiochemical methods are insufficient to systematically investigate the bio-molecular reactions for a high-throughput discovery. Here, we have reviewed an update on the recently developed chemical proteomicscalled activity-based protein profiling （ABPP）. ABPP is able to identify proteins interacted eithercovalently or non-covalently with metabolites significantly. Thus, ABPP will facilitate the characteriza-tion of specific metabolite regulating; proteins in human disease progression.

Nuclear magnetic resonance based metabolomic differentiation of different Astragali Radix

Astragali Radix(AR) is one of the most popular herbal medicines in traditional Chinese medicine(TCM). Wild AR is believed to be of high quality, and substitution with cultivated AR is frequently encountered in the market. In the present study, two types of ARs(wild and cultivated) from Astragalus membranaceus(Fisch.) Bge. and A. membranaceus var. mongholicus(Bge.) Hsiao, growing in different regions of China, were analyzed by NMR profiling coupled with multivariate analysis. Results showed that both could be differentiated successfully and cultivation patterns or growing years might have greater impact on the metabolite compositions than the variety; the metabolites responsible for the separation were identified. In addition, three extraction methods were compared and the method(M1) was used for further analysis. In M1, the extraction solvent composed of water, methanol, and chloroform in the ratio of 1 : 1 : 2 was used to obtain the aqueous methanol(upper layer) and chloroform(lower layer) fractions, respectively, showing the best separation. The differential metabolites among different methods were also revealed. Moreover, the sucrose/glucose ratio could be used as a simple index to differentiate wild and cultivated AR. Meanwhile, the changes of correlation pattern among the differential metabolites of the two varieties were found. The work demonstrated that NMR-based non-targeted profiling approach, combined with multivariate statistical analysis, can be used as a powerful tool for differentiating AR of different cultivation types or growing years.

Identification and differentiation of major components in three different “Sheng-ma” crude drug species by UPLC/Q-TOF-MS

Cimicifugae Rhizoma(Sheng ma) is a Ranunculaceae herb belonging to a composite family and well known in China,has been widely used in traditional Chinese medicine.The Pharmacopoeia of the People's Republic of China contains three varieties(Cimicifuga dahurica(Turcz.),Cimicifuga foetida L.and Cimicifuga heracleifolia Kom.) which have been used clinically as "Sheng-ma".However,the chemical constituents of three components of "Sheng-ma" have never been documented.In this study,a rapid method for the analysis of the main components of "Sheng-ma" was developed using ultra-high performance liquid chromatography with quadrupole-time-of-flight mass spectrometry(UPLC/Q-TOF-MS).The present study reveals the major common and distinct chemical constituents of C.dahurica,C.foetida and C.heracleifolia and also reports principal component and statistical analyses of these results.The components were identified by comparing the retention time,accurate mass,mass spectrometric fragmentation characteristic ions and matching empirical molecular formula with that of the published compounds.A total of 32 common components and 8 markers for different "Sheng-ma" components were identified.These findings provide an important basis for the further study and clinical utilities of the three "Sheng-ma" varieties.

Evidence Based Validation of Indian Traditional Medicine–Way Forward

Evidence based validation of the ethno-pharmacological claims on traditional medicine(TM) is the need of the day for its globalization and reinforcement. Combining the unique features of identifying biomarkers that are highly conserved across species, this can offer an innovative approach to biomarker-driven drug discovery and development. TMs are an integral component of alternative health care systems. India has a rich wealth of TMs and the potential to accept the challenge to meet the global demand for them. Ayurveda, Yoga, Unani, Siddha and Homeopathy(AYUSH) medicine are the major healthcare systems in Indian Traditional Medicine. The plant species mentioned in the ancient texts of these systems may be explored with the modern scientific approaches for better leads in the healthcare. TM is the best sources of chemical diversity for finding new drugs and leads. Authentication and scientific validation of medicinal plant is a fundamental requirement of industry and other organizations dealing with herbal drugs. Quality control(QC) of botanicals, validated processes of manufacturing, customer awareness and post marketing surveillance are the key points, which could ensure the quality, safety and efficacy of TM. For globalization of TM, there is a need for harmonization with respect to its chemical and metabolite profiling, standardization, QC, scientific validation,documentation and regulatory aspects of TM. Therefore, the utmost attention is necessary for the promotion and development of TM through global collaboration and co-ordination by national and international programme.

A comprehensive classification method for VOC emission sources to tackle air pollution based on VOC species reactivity and emission amounts

In China, volatile organic compound（VOC） control directives have been continuously released and implemented for important sources and regions to tackle air pollution. The corresponding control requirements were based on VOC emission amounts（EA）, but never considered the significant differentiation of VOC species in terms of atmospheric chemical reactivity. This will adversely influence the effect of VOC reduction on air quality improvement. Therefore,this study attempted to develop a comprehensive classification method for typical VOC sources in the Beijing–Tianjin–Hebei region（BTH）, by combining the VOC emission amounts with the chemical reactivities of VOC species. Firstly, we obtained the VOC chemical profiles by measuring 5 key sources in the BTH region and referencing another 10 key sources, and estimated the ozone formation potential（OFP） per ton VOC emission for these sources by using the maximum incremental reactivity（MIR） index as the characteristic of source reactivity（SR）. Then, we applied the data normalization method to respectively convert EA and SR to normalized EA（NEA） and normalized SR（NSR） for various sources in the BTH region.Finally, the control index（CI） was calculated, and these sources were further classified into four grades based on the normalized CI（NCI）. The study results showed that in the BTH region,furniture coating, automobile coating, and road vehicles are characterized by high NCI and need to be given more attention; however, the petro-chemical industry, which was designated as an important control source by air quality managers, has a lower NCI.