Liquid Chromatography Quadrupole Research Articles

Flavonoid Profiles in the Pulp of Different Lemon Cultivars and Their Antioxidant Activity Based on UPLC–Q–TOF–MS

Previous studies have indicated that there may be differences among the varieties of lemon flavonoids, but the details have not yet been made clear, which limits the comprehensive use of different cultivated lemon varieties. In this study, ultra-performance liquid chromatography–quadrupole–time-of-flight–mass spectrometry (UPLC–Q–TOF–MS) and ultraviolet–visible spectroscopy (UV–Vis) were used to investigate the types and contents of flavonoids in the flesh of the main cultivated variety (Eureka) and five common lemon varieties, as well as their in vitro antioxidant activity. A total of 21 compounds were identified, five of which were common compounds. Among them, Verna, Lisbon, and Bearss each have characteristic components that can serve as potential criteria for variety identification. Each of the six varieties of lemon has strong antioxidant activity. The antioxidant activity of different lemon varieties is related to flavonoids. Therefore, Eureka and the other five varieties of lemon are good natural antioxidants, and the cultivation and industrial production of lemons should consider the needs and selection of suitable varieties.

Parkia javanica Edible Pods Reveal Potential as an Anti-Diabetic Agent: UHPLC-QTOF-MS/MS-Based Chemical Profiling, In Silico, In Vitro, In Vivo, and Oxidative Stress Studies.

According to the World Health Organization, over 422 million people worldwide have diabetes, with the majority residing in low- and middle-income countries. Diabetes causes 1.5 million fatalities a year. The number of diabetes cases and its prevalence have progressively increased over the last few decades. This study aims to determine the phytochemicals in the edible part of Perkia javanica, predict their α-glucosidase inhibitory potential, one of the promising targets for diabetes, and then carry out in vitro and in vivo studies. The phytochemicals present in the n-butanol fraction of the methanol extract of P. javanica pods were analyzed using UHPLC-QTOF-MS/MS (Ultra-High-Performance Liquid Chromatography-Quadrupole Time-of-Flight Mass Spectrometry). The UHPLC-QTOF analysis revealed the presence of 79 different compounds in the n-butanol fraction. Among these, six compounds demonstrated excellent binding affinities with α-glucosidase, surpassing the performance of two standard inhibitors, Miglitol and Voglibose. In vitro α-glucosidase inhibitory activities were assessed by the n-butanol fraction, followed by in vivo studies. According to the in vitro study, the inhibitory efficiency against α-glucosidase was determined to have an IC50 value of 261.9 µg/mL. The in vivo findings revealed a significant reduction in blood glucose levels in Swiss albino mice treated with the same extract, decreasing from 462.66 mg/dL to 228.66 mg/dL. Additionally, the extract significantly increased the activity of the enzymes catalase and superoxide dismutase (SOD) and decreased the amount of malondialdehyde (MDA) in the liver and kidney tissue. The predicted physicochemical parameters indicated that most of the compounds would be excreted from the body after inhibition in the small intestine without being absorbed. Considering the low cost and wide availability of raw materials, P. javanica pods can serve as a good food supplement that may help prevent type 2 diabetes management.

Immunoaffinity Intact-Mass Spectrometry for the Detection of Endogenous Concentrations of the Acetylated Protein Tumor Biomarker Neuron Specific Enolase.

Intact-mass spectrometry has huge potential for clinical application, as it enables both quantitative and qualitative analysis of intact proteins and possibly unlocks additional pathophysiological information via, e.g., detection of specific post-translational modifications (PTMs). Such valuable and clinically useful selectivity is typically lost during conventional bottom-up mass spectrometry. We demonstrate an innovative immunoprecipitation protein enrichment assay coupled to ultrahigh performance liquid chromatography quadrupole time-of-flight high resolution mass spectrometry (UPLC-QToF-HRMS) for the fast and simple identification of the protein tumor marker Neuron Specific Enolase Gamma (NSEγ) at low endogenous concentrations in human serum. Additionally, using the combination of immunoaffinity purification with intact mass spectrometry, the presence of NSEγ in an acetylated form in human serum was detected. This highlights the unique potential of immunoaffinity intact mass spectrometry in clinical diagnostics.

In-vivo metabolic profiling of the Natural products Emodin and Emodin-8-O-β-D-glucoside in rats using liquid chromatography quadrupole Orbitrap mass spectrometry

Emodin (EM) and emodin-8-O-β-D-glucoside (EG) were found in various medicinal plants such as Rheum palmatum, Aloe vera, Polygonum multiflorum, and Polygonum cuspidatum. They have different pharmacological properties and are considered potentially toxic substances. It is necessary to identify the metabolites and their distribution in the body. In this study, a comprehensive analytical strategy was developed to characterize the metabolites of EM and EG in vivo using UHPLC-Q-Exactive Orbitrap MS. 190 metabolites were identified in the bio-samples, in addition to the commonly reported glycoside hydrolysis, hydrogenation, hydroxylation, glucuronide conjugation, and sulfation conjugation. We also discovered new pathways such as formylation, acetylation, glycol acylation, lactation, glycerolization, malonylation, glycerol acid acylation, hydroxyvalerylation, erythrosylation, glutaric acidfication, hydroxybenzoylation, glutamylation, hydroxyglutamylation, ascorbylation, aspartylglycylation, dihydroxyphenylglycylation, trihydroxyphenylglycylation, dihydroxymethoxyphenylglycylation, and ring-opening of EM. Cluster analysis revealed that each tissue in the EM and EG groups exhibited a high degree of similarity in their metabolic pathway preferences and bodily distributions, as they clustered together. Interestingly, the presence of glycol acylation, glycerolization, and glycerol acid acylation in the liver may be related to the lipid-lowering effects of EM and EG. These findings offer valuable insights for a more comprehensive understanding of the safety and efficacy of EM and EG, as well as valuable methods for metabolic characterization.

Retrospective screening of new psychoactive substances (NPS) in post mortem samples from 2014 to 2021

Systematic retrospective processing of previously analysed biological samples has been proven to be a valuable tool in the search for new drugs (e.g. new psychoactive substances (NPS)) and for quality assessment in clinical and forensic toxicology. In a previous study, we developed a strategy for retrospective data-analysis using a personalized library of synthetic cannabinoids, designer benzodiazepines and synthetic opioids obtained from the crowdsourced database HighResNPS (https://highresnps.com). In this study, the same strategy was employed for the compounds within the groups of NPS that were not previously included such as synthetic cathinones, phenethylamines, aminoindanes, arylalkylamines, piperazine derivates, piperidines, pyrrolidines, indolalkylamines and arylcyclohexylamines. Synthetic opioids and designer benzodiazepines, which were not part of the previous study, were also included. To enhance the effectiveness of the retrospective analysis, a predicted retention time was included for all entries. Data files from the analysis of 2186 forensic post mortem samples with an Agilent Technologies 6540 ultra-high pressure liquid chromatography quadrupole time-of-flight mass spectrometry (UHPLC-QTOF-MS) performed in the laboratory from January 2014 to December 2021 were retrospectively processed with the up-to-date library. Tentative findings were classified in two groups: The findings where MS/MS data was acquired for library match (category 1) and the less certain findings where such data lacked (category 2). Five compounds of category 1 (three synthetic cathinones and two indolalkylamines) were identified in 12 samples. Only one of the findings, 4-MEAPP (4-methyl-α-ethylaminopentiophenone), was deemed plausible after reviewing case information. As many as 501 presumably positive category 2 findings were detected. Using the predicted retention time as an additional criterion the number was significantly reduced but still too high for a manual review. This work has demonstrated that the strategy developed in the previous study can be applied to other NPS groups. However, it is important to note the limitations such a method may have in detecting compounds at very low concentrations.

Cai's herbal tea enhances mitochondrial autophagy of type 1 diabetic mellitus β cells through the AMPK/mTOR pathway and alleviates inflammatory response.

This study investigates the therapeutic mechanisms of Cai's Herbal Tea in Type 1 Diabetes Mellitus (T1DM) mice, focusing on its effects on mitochondrial change and autophagy via the AMP-activated protein kinase (AMPK)-mammalian target of rapamycin (mTOR) pathway. The composition of Cai's Herbal Tea was analyzed by Ultra-High Performance Liquid Chromatography-Quadrupole Time of Flight Mass Spectrometry (UHPLC-Q/TOF-MS). C57BL/6 mice and Min6 pancreatic beta cells were divided into control, diabetic mellitus (DM)/high glucose (HG), and treatment groups (low, medium, and high doses of Cai's Tea, and Metformin). Key physiological parameters, pancreatic islet health, Min6 cell morphology, viability, and insulin (INS) secretion were assessed. Small Interfering RNA-AMPK (si-AMPK) was utilized to confirm the pathway involvement. Cai's Herbal Tea improved body weight, pancreatic islet pathological injury, and INS secretion whereas reduced total triglycerides, fasting blood sugar, and Interferon gamma (INF-γ) in T1DM mice, particularly at higher doses. In Min6 cells, Cai's Tea mitigated HG-induced damage and proinflammatory response, enhancing cell viability and INS secretion. Notably, it reduced swelling and improved cristae structure in treated groups of mitochondria and promoted autophagy via the AMPK-mTOR pathway, evidenced by increased LC3II/LC3I and P-AMPK/AMPK ratios, and decreased P-mTOR/mTOR and P62 expressions in pancreatic islet β-cells. Furthermore, these effects were converted by si-AMPK interference. Cai's Herbal Tea exhibits significant therapeutic efficacy in T1DM mice by improving mitochondrial health and inducing autophagy through the AMPK-mTOR pathway in pancreatic islet β-cells. These findings highlight its potential as a therapeutic approach for T1DM management.

LC-MS and GC–MS based metabolomics analysis revealed the impact of tea trichomes on the chemical and flavor characteristics of white tea

To explore the influence of tea trichomes on the quality of white tea, liquid chromatography quadrupole time-of-flight mass spectrometry (LC-Q-TOF-MS), and headspace solid phase microextraction gas chromatography-mass spectrometry (HS-SPME-GC–MS) were used to identify non-volatile and volatile compounds white tea without trichomes (WTwt) and pure trichomes (PT). It was found that the bitter and astringent compounds, caffeine (CAF), epigallocatechin gallate (EGCG), epicatechin gallate (ECG) and flavonol glycosides, were mainly enriched in the WTwt, with 16.3-fold, 47.1-fold and 28.7-fold decrease in CAF and EGCG and ECG, respectively, and the content of these compounds in PT were lower than the taste thresholds. In PT, kaempferol-3-O-(p-coumaroyl)-glucoside and kaempferol-3-O-(di-p-coumaroyl)-glucoside were non-volatile marker compounds, and decanal was significant aroma contributor with rOAV = 250.86. Moreover, the compounds in trichomes mainly contributed to the fruity and floral aroma of white tea, among which benzyl alcohol, (E)-geranylacetone, decanal, dodecanal and 6-methyl-5-hepten-2-one were the crucial aroma components, which were 2.1, 1.7, 1.8, 1.4 and 2.2 times as much as the WTwt in the PT, respectively. In conclusion, trichomes can improve the quality of white tea by reducing the bitterness and astringency, increasing the umami, as well as enhancing the fruity and floral aromas.

LC-MS and GC–MS based metabolomics analysis revealed the impact of tea trichomes on the chemical and flavor characteristics of white tea

To explore the influence of tea trichomes on the quality of white tea, liquid chromatography quadrupole time-of-flight mass spectrometry (LC-Q-TOF-MS), and headspace solid phase microextraction gas chromatography-mass spectrometry (HS-SPME-GC–MS) were used to identify non-volatile and volatile compounds white tea without trichomes (WTwt) and pure trichomes (PT). It was found that the bitter and astringent compounds, caffeine (CAF), epigallocatechin gallate (EGCG), epicatechin gallate (ECG) and flavonol glycosides, were mainly enriched in the WTwt, with 16.3-fold, 47.1-fold and 28.7-fold decrease in CAF and EGCG and ECG, respectively, and the content of these compounds in PT were lower than the taste thresholds. In PT, kaempferol-3-O-(p-coumaroyl)-glucoside and kaempferol-3-O-(di-p-coumaroyl)-glucoside were non-volatile marker compounds, and decanal was significant aroma contributor with rOAV = 250.86. Moreover, the compounds in trichomes mainly contributed to the fruity and floral aroma of white tea, among which benzyl alcohol, (E)-geranylacetone, decanal, dodecanal and 6-methyl-5-hepten-2-one were the crucial aroma components, which were 2.1, 1.7, 1.8, 1.4 and 2.2 times as much as the WTwt in the PT, respectively. In conclusion, trichomes can improve the quality of white tea by reducing the bitterness and astringency, increasing the umami, as well as enhancing the fruity and floral aromas.

Retrospective screening of new psychoactive substances (NPS) in post mortem samples from 2014 to 2021

Systematic retrospective processing of previously analysed biological samples has been proven to be a valuable tool in the search for new drugs (e.g. new psychoactive substances (NPS)) and for quality assessment in clinical and forensic toxicology. In a previous study, we developed a strategy for retrospective data-analysis using a personalized library of synthetic cannabinoids, designer benzodiazepines and synthetic opioids obtained from the crowdsourced database HighResNPS (https://highresnps.com). In this study, the same strategy was employed for the compounds within the groups of NPS that were not previously included such as synthetic cathinones, phenethylamines, aminoindanes, arylalkylamines, piperazine derivates, piperidines, pyrrolidines, indolalkylamines and arylcyclohexylamines. Synthetic opioids and designer benzodiazepines, which were not part of the previous study, were also included. To enhance the effectiveness of the retrospective analysis, a predicted retention time was included for all entries. Data files from the analysis of 2,186 forensic post mortem samples with an Agilent Technologies 6540 ultra-high pressure liquid chromatography quadrupole time-of-flight mass spectrometry (UHPLC-QTOF-MS) performed in the laboratory from January 2014 to December 2021 were retrospectively processed with the up-to-date library. Tentative findings were classified in two groups: The findings where MS/MS data was acquired for library match (category 1) and the less certain findings where such data lacked (category 2). Five compounds of category 1 (three synthetic cathinones and two indolalkylamines) were identified in 12 samples. Only one of the findings, 4-MEAPP (4-methyl-α-ethylaminopentiophenone), was deemed plausible after reviewing case information. As many as 501 presumably positive category 2 findings were detected. Using the predicted retention time as an additional criterion the number was significantly reduced but still too high for a manual review. This work has demonstrated that the strategy developed in the previous study can be applied to other NPS groups. However, it is important to note the limitations such a method may have in detecting compounds at very low concentrations.

Integrating UPLC-Q-TOF-MS and Network Pharmacology to Explore the Potential Mechanisms of Paeonia lactiflora Pall. in the Treatment of Blood Stasis Syndrome.

Paeonia lactiflora Pall. (PLP) is thought to promote blood circulation and remove blood stasis. This study used blood component analysis, network pharmacology, and molecular docking to predict the mechanism of PLP in the treatment of blood stasis syndrome (BSS). PLP was processed into Paeoniae Radix Alba (PRA) and Paeoniae Radix Rubra (PRR). PRA and PRR could significantly reduce whole blood viscosity (WBV) at 1/s shear rates and could increase the erythrocyte aggregation index (EAI), plasma viscosity (PV), and erythrocyte sedimentation rate (ESR) of rats with acute blood stasis. They prolonged the prothrombin time (PT), and PRR prolonged the activated partial thromboplastin time (APTT). PRA and PRR increased the thrombin time (TT) and decreased the fibrinogen (FBG) content. All the results were significant (p < 0.05). Ten components of Paeoniflorin, Albiflorin, Paeonin C, and others were identified in the plasma of rats using ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS). A protein-protein interaction network (PPI) analysis showed that AKT1, EGFR, SRC, MAPK14, NOS3, and KDR were key targets of PLP in the treatment of BSS, and the molecular docking results further verified this. This study indicated that PLP improves BSS in multiple ways and that the potential pharmacological mechanisms may be related to angiogenesis, vasoconstriction and relaxation, coagulation, and the migration and proliferation of vascular cells.

Exploring the Potential Lipid-Lowering and Weight-Reducing Mechanisms of FH06 Fermented Beverages Based on Non-Targeted Metabolomics and Network Pharmacology

Investigating the intricate pathways through which FH06 fermentation broth exerts lipid-lowering and weight-loss effects is pivotal for advancing our comprehension of metabolic regulation and therapeutic interventions. Ultrahigh-performance liquid chromatography quadrupole electrostatic field orbit trap mass spectrometry (UHPLC-QE-MS) detection and the ChEMBL database were used to determine the effective compounds in the FH06 fermentation broth and predict their targets. The TTD database and DisGeNET database were used to query obesity-related targets. The STRING database was used to construct protein interaction information. The Gene Ontology (GO) database and the Kyoto Encyclopedia of Genes and Genomes (KEGG) database were used to perform biological function annotation (GO) and KEGG pathway enrichment analyses of the targets. Results: A total of 85 effective compounds were screened from the fermentation broth of FH06; these compounds may act on TP53, PPARG, TNF, and other targets through 10 signaling pathways, such as the chemical carcinogenesis-receptor activation and lipid and atherosclerosis pathways, and exert pharmacological effects, such as hypoglycemic effects and weight loss. They also have anti-inflammatory, antioxidant, antitumor, and immunoregulatory effects. These findings reveal the active ingredients of FH06 fermentation broth and its multi-target and multi-channel characteristics in lipid lowering and weight loss. This study has positive implications for the clinical treatment of obesity using FH06, providing a theoretical and scientific basis for further developing of FH06-assisted lipid-lowering products.

A strategy integrating GC–MS, UPLC profiling, and DNA metabarcoding for characterization and discrimination of the medicinal and culinary pieces from four Curcuma species

Curcumae pieces prepared in ready-to-use forms, consumed extensively for their significant medicinal and culinary benefits, present a challenge in discrimination due to their heterogeneous origins, analogous morphologies, and varied components. This study explored the metabolites of nine Curcumae pieces (NCPs) by Gas Chromatography-Mass Spectrometry (GC–MS) and Ultra-Performance Liquid Chromatography/Ultra High-Performance Liquid Chromatography–Quadrupole Time of Flight–Mass Spectrometry (UPLC/UHPLC–QTOF–MS) technologies, augmented by multivariate statistical analysis (Principal Component Analysis, PCA; Orthogonal Partial Least Squares Discriminant Analysis, OPLS–DA) and machine learning algorithms (Random Forest, RF). Twenty-six key differential markers in volatile and non-volatile metabolites each were found subsequently. Additionally, DNA metabarcoding was employed to develop specific Single Nucleotide Polymorphism (SNP) markers for the precise identification of Curcuma species in single and mixed samples, and for CRa even contained in Chinese patent prescriptions. The integration of these three methodologies facilitates a more reliable authentication of NCPs, enhancing the overall strategy for their global discrimination. This research contributes to the advancement of quality control and the development of functional foods or products derived from Curcumae pieces, tailored to meet specific health and dietary requirements.

Chemical characterization and comparative analysis of different parts of Cocculus orbiculatus through UHPLC-Q-TOF-MS.

Cocculus orbiculatus (L.) DC. (C. orbiculatus) is a medicinal herb valued for its dried roots with anti-inflammatory, analgesic, diuretic, and other therapeutic properties. Despite its traditional applications, chemical investigations into C. orbiculatus remain limited, focusing predominantly on alkaloids and flavonoids. Furthermore, the therapeutic use of C. orbiculatus predominantly focuses on the roots, leaving the stems, a significant portion of the plant, underutilized. This study employed ultra-high performance liquid chromatography quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF-MS/MS) with in-house and online databases for comprehensive identification of components in various plant parts. Subsequently, untargeted metabolomics was employed to analyze differences in components across different harvest periods and plant sections of C. orbiculatus, aiming to screen for distinct components in different parts of the plant. Finally, metabolomic analysis of the roots and stems, which contribute significantly to the plant's weight, was conducted using chemometrics, including principal component analysis (PCA), partial least squares discriminant analysis (PLS-DA), orthogonal partial least squares discriminant analysis (OPLS-DA), and heatmaps. A total of 113 components, including alkaloids, flavonoids, and organic acids, were annotated across the root, stem, leaf, flower, and fruit, along with numerous previously unreported compounds. Metabolomic analyses revealed substantial differences in components between the root and stem compared to the leaf, flower, and fruit during the same harvest period. PLS-DA and OPLS-DA annotated 10 differentiating components (VIP > 1.5, P < 0.05, FC > 2 or FC < 0.67), with 5 unique to the root and stem, exhibiting lower mass spectrometric responses. This study provided the first characterization of 113 chemical constituents in different parts of C. orbiculatus, laying the groundwork for pharmacological research and advocating for the enhanced utilization of its stem.

Comprehensive profiling of the chemical constituents in Dayuanyin decoction using UPLC-QTOF-MS combined with molecular networking

Context Dayuanyin decoction is a traditional Chinese medicine formulation that is commonly used in modern clinical practice to treat viral infections such as viral pneumonia, viral fever, influenza, and hepatitis. Although the usage rate of Dayuanyin decoction is gradually increasing in clinical practice, its pharmacological constituents are still unclear. Objective This study comprehensively characterized the chemical constituents in Dayuanyin decoction using ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS) and molecular networking. Materials and methods The overall strategy involved retrieving structural information, such as fragment ions and precursor ion masses, from self-built databases to identify the target constituents of the Dayuanyin decoction extract. The identification of non-targeted constituents was achieved by analyzing different categories, fragment pathways, mass spectrometry data, and the relationships between clusters of structures in molecular networking. Unannotated constituents were inferred from secondary mass spectrometry similarity and molecular weight differences and annotated constituents in the same constituent cluster. A few predicted constituents were selected and validated by comparing them to reference standards under identical mass spectrometry conditions. Results This study preliminarily identified 216 constituents, including flavonoids, amino acids, alkaloids, triterpenes, steroidal saponins, phenylpropanoids, and other constituents. Conclusions This integrated strategy using UPLC-QTOF-MS and molecular networking lays the foundation for clinical research on pharmacologically active substances in Dayuanyin decoction and could be popularized for the comprehensive profiling of chemical constituents of other traditional Chinese medicines.

Urine as Biobased Fertilizer: The Netherlands as Case Study

Humankind has grown ever more dependent on mineral fertilizer to feed a growing world population. However, nutrients are rarely recovered and recycled after consumption, leading to environmental waste issues now and potential supply issues in the future. Urine is rich in these nutrients and therefore a prime candidate as precursor for biobased fertilizers. This work presents a case study to remove contaminants – predominantly pharmaceuticals – from human urine using activated carbon adsorbents in an effort to adhere to Dutch fertilizer legislation. In the urine, three pharmaceuticals were identified by target analysis and 11 other contaminants by suspect screening using liquid chromatography quadrupole time of flight mass spectrometry (LC-QTOF/MS). These were successfully removed by treatments with both granulated and powdered activated carbon, albeit with a loss of 16-17% and 2-4% of dissolved nutrients, respectively. Further screening of inorganic contaminants and persistent organic pollutants by inductively coupled plasma mass spectrometry (ICP-MS) and gas chromatography quadrupole-orbitrap mass spectrometry (GC-Q-Orbitrap/MS) showed that all prerequisites for fertilizer status are met, paving the way for its future legal use as biobased fertilizer in the Netherlands.

Rapid Classification and Identification of Chemical Compounds and Semi-Quantitative Metabolism of Huangkui Capsules and the Protective Effects of Its Quercetin Derivatives against Tacrolimus-induced HK-cell Reduction

Abstract Objective: The study aimed to establish an effective strategy for systematically characterizing and verifying compounds in Huangkui capsules (HKCs). Materials and Methods: An ultrahigh-performance liquid chromatography quadrupole time-of-flight mass spectrometry (MS) method was effectively established and utilized for the chemical compound characterization in HKC, with the support of MS-DIAL, MS-FINDER, and Global Natural Product Social Molecular Network. Multiple rat samples were analyzed after the oral administration. Metabolites were identified based on specific cleavage behaviors, and metabolic pathways were predicted. Semi-quantitative analysis of the metabolome profiles was conducted using post-data processing. High concentrations in vivo were investigated for their role in tacrolimus-induced death of HK-2 cells. Results: In total, 129 compounds were identified in HKC, of which 74 were reported for the first time. In this study, we successfully identified and uncovered 19 prototypes and 123 metabolites from the biosamples. The concentrations of glucuronidation and methylation metabolites of quercetin were the highest in the kidney and intestinal tissues. In contrast, significant glucuronidation of quercetin metabolites was observed with high blood concentrations. Notably, quercetin glucuronidation and methylation metabolites protected HK-cell against tacrolimus-induced injury in a dose-dependent manner. Conclusion: This study successfully established a reliable and efficient strategy for comprehensive characterization of traditional Chinese medicine compounds. This strategy harnessed the power of various intelligent postprocessing technologies to provide a more thorough understanding of active components and their metabolic transformations in the body. These results suggest that quercetin metabolites should be evaluated for their protective capacity against kidney disease.

Integrated metabolomic and transcriptomic analysis of triterpenoid accumulation in the roots of Codonopsis pilosula var. modesta (Nannf.) L.T.Shen at different altitudes.

Codonopsis Radix is a beneficial traditional Chinese medicine, and triterpenoid are the major bioactive constituents. Codonopsis pilosula var. modesta (Nannf.) L.T.Shen (CPM) is a precious variety of Codonopsis Radix, which is distributed at high mountain areas. The environment plays an important role in the synthesis and metabolism of active ingredients in medicinal plants, but there is no report elaborating on the effect of altitude on terpenoid metabolites accumulation in CPM. This study aims to analyse the effects of altitude on triterpenoid biosynthetic pathways and secondary metabolite accumulation in CPM. The untargeted metabolomics based on liquid chromatography-tandem mass spectrometry (LC-MS/MS) and 10 triterpenoids based on ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) method were analysed at the low-altitude (1480 m) and high-altitude (2300 m) CPM fresh roots. The transcriptome based on high-throughput sequencing technology were combined to analyse the different altitude CPM triterpenoid biosynthetic pathways. A total of 17,351 differentially expressed genes (DEGs) and 55 differentially accumulated metabolites (DAMs) were detected from the different altitude CPM, and there are significant differences in the content of the 10 triterpenoids. The results of transcriptome study showed that CPM could significantly up-regulate the gene expression levels of seven key enzymes in the triterpenoid biosynthetic pathway. The CPM at high altitude is more likely to accumulate triterpenes than those at low altitude, which was related to the up-regulation of the gene expression levels of seven key enzymes. These results expand our understanding of how altitude affects plant metabolite biosynthesis.

Study on the material basis of Zhujing pill in treating fundus lesions through component analysis and network pharmacology.

Zhujing pill (ZP) is a famous Chinese herbal formula that has been widely used to treat diabetic retinopathy, macular degeneration, retinitis pigmentosa and other fundus lesions. In this study, the material basis and mechanism of ZP in the treatment of fundus lesions were evaluated via the high-performance liquid chromatography fingerprint, ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry, network pharmacology and molecular docking. A total of 32 common components were found and 31 components were identified in 15 batches of ZP samples. Moreover, 134 common key targets and 17 putative active components that are connected to fundus lesions were identified. Molecular docking revealed that quercetin, kaempferol, isorhamnetin, 5-O-feruloylquinic acid, plantagoside and 2'-acetylacteoside have the ability to interact with the core targets such as AKT1, TP53, TNF, IL-6 and Jun. Our findings revealed that the therapeutic effects of ZP on fundus lesions are mediated by multiple components, targets and pathways, including at least six active ingredients and 11 targets. The study provides new ideas for further research on the material basis and mechanisms of traditional Chinese medicine prescriptions.

An efficient method for rapid screening of triterpenoid saponins in three Glycyrrhiza species using rapid resolution liquid chromatography quadrupole time-of-flight mass spectrometry combined with mass defect filtering

Triterpenoid saponins, a major bioactive component of liquorice, possess high hydrophilicity and often co-occur with other impurities of similar polarity. Additionally, subtle structural differences of some triterpenoid saponins bring challenges to comprehensive characterisation. In this study, triterpenoid saponins of three Glycyrrhiza species were systematically analysed using rapid resolution liquid chromatography quadrupole time-of-flight mass spectrometry (RRLC-Q-TOF-MS) coupled with mass defect filtering (MDF). Firstly, comprehensive date acquisition was achieved using RRLC-Q-TOF-MS. Secondly, a polygonal MDF method was established by summarizing known and speculated substituents and modifications based on the core structure to rapidly screen potential triterpenoid saponins. Thirdly, based on the fragmentation patterns of reference compounds, an identification strategy for characterisation of triterpenoid saponins was proposed. The strategy divided triterpenoid saponins into three distinct classes. By this strategy, 98 triterpenoid saponins including 10 potential new ones were tentatively characterised. Finally, triterpenoid saponins of three Glycyrrhiza species were further analysed using principle component analysis (PCA) and orthogonality partial least squares discriminant analysis (OPLS-DA). Among these, 18 compounds with variable importance in projections (VIP) > 1.0 and P values < 0.05 were selected to distinguish three Glycyrrhiza species. Overall, our study provided a reference for quality control and rational use of the three species.

Mechanism of action of Sambucus williamsii Hance var. miquelii in the treatment of osteoporosis analyzed by UHPLC-HRMS/MS combined network pharmacology and experimental validation

Sambucus williamsii Hance var. miquelii(SWH) is a precious wild Chinese herb whose fruit, rhizome, leaves and root bark can be used as medicine. Sambucus Linn has pharmacological effects such as anti-osteoporosis, promoting fracture healing, anti-viral and anti-inflammatory. In this study, the main chemical components of the alcoholic extracts from SWH were rapidly identified by ultra-high performance liquid chromatography-quadrupole orbit trap high-resolution mass spectrometry (UHPLC- HRMS MS), and a total of 42 compounds were characterized from the alcoholic extracts of SWH. The results of network pharmacological validation showed that kaempferol, quercetin, luteolin, isorhamnetin and morroniside were the main active components, and KEGG enrichment demonstrated that SWH mainly affected the signaling pathways such as PI3K-Akt, TNF and FoxO by modulating the related targets such as AKT1, PIK3R1, EGFR, RELA SRC and PTGS2. The molecular docking results showed binding solid activity between the main active components of SWH and the targets. The network pharmacology was validated by establishing an animal model of osteoporosis (OP) in rats by gavage administration of vitamin A acid. The results of the pharmacological experiments showed that SWH could improve the degree of bone loss in the femur of osteoporotic rats, increase the number of trabeculae and decrease trabeculae porosity, up-regulate the Ca and P content in the serum of OP rats, down-regulate the scope of ALP and BGP in the serum, and promote the calcification of the bone matrix, and then exert the anti-OP efficacy. In this study, network pharmacology and pharmacological experiments verified the pharmacological mechanism of SWH in anti-OP rats. This provides a theoretical basis for the research and development of anti-OP drugs and a reference for the application of other traditional Chinese medicines in treating OP diseases.