Ultra-high Performance Liquid Chromatography Quadrupole Research Articles

Identification, Quantitation, and Sensory Evaluation of Thiols in Bordeaux Red Wine with Characteristic Aging Bouquet.

Great Bordeaux red wines are known for their distinctive aging bouquet. However, the nature of volatile chemicals underpinning this sensory expression is not fully understood. This work investigated the empyreumatic aging bouquet of a collection of premium Bordeaux red wines using silver-ion (Ag+) solid-phase extraction, cryogenic heart-cutting multidimensional gas chromatography mass spectrometry/olfactometry, and comprehensive two-dimensional gas chromatography time-of-flight mass spectrometry. In doing so, a substantial number of "meaty" odors were revealed. Three detected "meaty" notes were tentatively or unequivocally attributed to furan thiols. Among them, 2-methyltetrahydrofuran-3-thiol (1) with a pleasant "meaty" aroma was reported in wine for the first time. Its trans isomer (trans-1a) was resolved from its racemate by chemical modification, which confirmed its presence in wine. The odor detection threshold of trans-1a in the model wine was determined at 55 ng/L. Moreover, an additive effect between 1 and literature-known 2-methyl-3-furanthiol was observed. By a new ultra high-performance liquid chromatography quadrupole Orbitrap high-resolution mass spectrometry method, the concentration of trans-1a, in addition to those of 2-methyl-3-furanthiol and 2-furfuryl thiol, was measured in the wines at ng/L levels.

Urinary Paraben Concentrations and Associations with the Periconceptional Urinary Metabolome: Untargeted and Targeted Metabolomics Analyses of Participants from the Early Pregnancy Study.

Parabens, found in everyday items from personal care products to foods, are chemicals with endocrine-disrupting activity, which has been shown to influence reproductive function. This study investigated whether urinary concentrations of methylparaben, propylparaben, or butylparaben were associated with the urinary metabolome during the periconceptional period, a critical window for female reproductive function. Changes to the periconceptional urinary metabolome could provide insights into the mechanisms by which parabens could impact fertility. Urinary paraben concentrations were measured in paired pre- and postconception urine samples from 42 participants in the Early Pregnancy Study, a prospective cohort of 221 women attempting to conceive. We performed untargeted and targeted metabolomics analyses using ultrahigh-performance liquid chromatography quadrupole time-of-flight mass spectrometry. We used principal component analysis, orthogonal partial least-squares discriminant analysis, and permutation testing, coupled with univariate statistical analyses, to find metabolites associated with paraben concentration at the two time points. Potential confounders were identified with a directed acyclic graph and used to adjust results with multivariable linear regression. Metabolites were identified using fragmentation data. Seven metabolites were associated with paraben concentration (variable importance to projection score , false discovery rate-corrected ). We identified four diet-related metabolites to the Metabolomics Standards Initiative (MSI) certainty of identification level 2, including metabolites from smoke flavoring, grapes, and olive oil. One metabolite was identified to the class level only (MSI level 3). Two metabolites were unidentified (MSI level 4). After adjustment, three metabolites remained associated with methylparaben and propylparaben, two of which were diet-related. No metabolomic markers of endocrine disruption were associated with paraben concentrations. This study identified novel relationships between urinary paraben concentrations and diet-related metabolites but not with metabolites on endocrine-disrupting pathways, as hypothesized. It demonstrates the feasibility of integrating untargeted metabolomics data with environmental exposure information and epidemiological adjustment for confounders. The findings underscore a potentially important connection between diet and paraben exposure, with applications to nutritional epidemiology and dietary exposure assessment. https://doi.org/10.1289/EHP12125.

A practical method for rapid discrimination of constituents in Psoraleae Fructus by UPLC-Q-Orbitrap MS.

Psoraleae Fructus (PF) is one of the most frequently used traditional Chinese medicine, which has good efficacy in warming kidney to activate yang, promoting inspiration to relieve asthma and warming spleen to stop diarrhea. However, the chemical composition of PF is complex, which makes it difficult to determine its active and toxic components. In order to rapidly classify and identify the chemical components of the extracts from PF, this research was processed with CNKI, PubMed, and PubChem databases and data post-processing technique basing on ultrahigh performance liquid chromatography quadrupole orbitrap mass spectrometry (UPLC-Q-Orbitrap MS) technique. Finally, 73 chemical components were discriminated, including 44 flavonoids, 18 coumarins, and 11 terpenoids, with the cleavage rules of each chemical component summarized. This study established a UPLC-Q-Orbitrap MS method for the separation and discrimination of the chemical constituents of PF, which can lay a foundation for the further study of its medicinal substances and quality control.

6-Oxofurostane and (iso)Spirostane Types of Saponins in Smilax sieboldii: UHPLC-QToF-MS/MS and GNPS-Molecular Networking Approach for the Rapid Dereplication and Biodistribution of Specialized Metabolites.

Identifying novel phytochemical secondary metabolites following classical pharmacognostic investigations is tedious and often involves repetitive chromatographic efforts. During the past decade, Ultra-High Performance Liquid Chromatography-Quadrupole Time of Flight-Tandem Mass Spectrometry (UHPLC-QToF-MS/MS), in combination with molecular networking, has been successfully demonstrated for the rapid dereplication of novel natural products in complex mixtures. As a logical application of such innovative tools in botanical research, more than 40 unique 3-oxy-, 3, 6-dioxy-, and 3, 6, 27-trioxy-steroidal saponins were identified in aerial parts and rhizomes of botanically verified Smilax sieboldii. Tandem mass diagnostic fragmentation patterns of aglycones, diosgenin, sarsasapogenin/tigogenin, or laxogenin were critical to establishing the unique nodes belonging to six groups of nineteen unknown steroidal saponins identified in S. sieboldii. Mass fragmentation analysis resulted in the identification of 6-hydroxy sapogenins, believed to be key precursors in the biogenesis of characteristic smilaxins and sieboldins, along with other saponins identified within S. sieboldii. These analytes' relative biodistribution and characteristic molecular networking profiles were established by analyzing the leaf, stem, and root/rhizome of S. sieboldii. Deducing such profiles is anticipated to aid the overall product integrity of botanical dietary supplements while avoiding tedious pharmacognostic investigations and helping identify exogenous components within the finished products.

Analysis of Antibiotics in Bivalves by Ultra-High Performance Liquid Chromatography-Quadrupole Time-of-Flight Mass Spectrometry.

The presence of pharmaceuticals in aquatic ecosystems mostly originates from wastewater treatment plants (WWTPs) and such a situation can be responsible for significant negative impacts on natural ecosystems, such as estuarine and coastal areas. Bioaccumulation of pharmaceuticals, namely antibiotics, in exposed organisms is known to have remarkable effects on different trophic levels of non-target organisms such as algae, invertebrates and vertebrates, including the emergence of bacterial resistance. Bivalves are a highly appreciated seafood product, as they are fed by filtering water, and can bioconcentrate chemicals, being ideal for biomonitoring environmental health hazards in coastal and estuarine ecosystems. To use this sentinel species, an analytical strategy was developed to be used in accessing antibiotics, from human and veterinary medicine, and evaluate their occurrence as emerging pollutants in aquatic environments. The optimized analytical method was fully validated according to the European requirements defined by the Commission Implementing Regulation 2021/808. The validation comprised the following parameters: specificity, selectivity, precision, recovery, ruggedness, linearity, and the decision limit CCα, as well as the limit of detection (LoD) and limit of quantification (LoQ). The method was validated for 43 antibiotics to allow their quantification in both contexts, environmental biomonitoring and food safety.

Comprehensive characterization of narirutin metabolites in vitro and in vivo based on Analogous-Core recursion analysis strategy using UHPLC-Q-Exactive Orbitrap MS/MS

Narirutin, extracted from the dried ripe pericarp of Rutaceae orange and its cultivated varieties, is a dihydroflavone compound with various biological activities and pharmacological effects. So far as we know, the metabolism profiling of narirutin has been insufficient until now. In the present study, an efficient method was employed to perform rapid analysis and identification of narirutin metabolites by using ultra-high performance liquid chromatography quadrupole exactive orbitrap MS/MS (UHPLC-Q-Exactive Orbitrap MS/MS) combined with an analogous core recursion (ACR) analysis strategy. Firstly, according to the basic core structure of the dihydroflavonoid, the cleavage mode and fragmentation ions of narirutin were summarized. Secondly, based on the difference of the substituent groups, the fragment ions of narirutin were preliminarily inferred and verified by the secondary mass spectra of its standard compound. Thirdly, the fragment ions of narirutin in positive and negative ion modes were summarized to provide a basis for the identification of metabolites. Fourthly, candidate metabolites were accurately and tentatively identified according to the cleavage law of mass spectrometry, literature reports, comparison of reference substances, and especially the diagnostic fragment ion clusters (DFICs) and characteristic suggestive ions (CSIs) deduced preliminarily. Finally, a total of 46 metabolites (30 in vivo and 19 in vitro), including prototype drugs were identified based on the ACR analysis strategy, chromatographic retention behavior of metabolites, corresponding ClogP value and accurate molecular weight. Among them, 22 metabolites were discovered in rats plasma, 12 in urine, 4 in liver tissue, 19 in liver microsomes, respectively. Additionally, metabolites relative contents were also studied by extracted ion chromatography (EIC) method. Our result also illustrated that narirutin primarily underwent glucuronidation, sulfation, glutathione binding, deglycosylation, oxidation, reduction, methylation, hydroxylation, ring-opening and their composite reactions. A novel strategy was constructed to comprehensively elucidate the biotransformation pathways of narirutin in vitro and in vivo, and a systematic metabolic profile of narirutin was generated. This study provided a great reference for further understanding of the biotransformation pathways and guiding for clinical application of narirutin.

Role of simulated in vitro gastrointestinal digestion on biotransformation and bioactivity of astragalosides from Radix Astragali

Radix Astragali (RA) is commonly used in Asian herbal therapy or food supply, and astragalosides and flavonoids are its major components with diverse pharmaceutical effects. To provide new information on the potential cardiovascular benefits of RA administered orally, the bioaccessibility of these compounds with relevant in vitro digestion parameters was determined for four digestion phases (oral, gastric, small and large intestines) by ultrahigh-performance liquid chromatography quadrupole time-of-flight-mass spectrometry (UPLC-Q-TOF/MS). Meanwhile, we compared the effects of digestion products on advanced glycation end products (AGEs)-induced intracellular reactive oxygen species (ROS) levels in a human arterial endothelial cells (HAECs) model, and studied the potential of RA against oxidative stress-related cardiovascular disease. The changes of saponins and flavonoids composition and antioxidant activity after digestion in intestines were mainly due to the astragaloside IV (AS-IV) biosynthesis involving saponins acetyl isomerization and deacetylation, and the flavonoid glycosides converted to aglycone by deglycosylation processes. All these results suggest that acetyl biotransformation of RA in small intestine directly influenced the response to oxidative stress, and might provide a reference for elucidation of the multi-component action after oral RA in cardiovascular health care.

Simultaneous screening and quantitation of stimulant drugs in dried blood spot (DBS) samples using ultra-high performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS)

Dried blood spots (DBS) have seen recent development as an alternative to whole blood samples for drug testing. The ease of use of DBS makes the technique suitable and attractive for drug testing applications. Quantitation of drug concentrations in the ng/mL range can be achieved from DBS samples using 10 to 20 µL of blood. The establishment of drug per se limits for drivers requires timely methods of sampling blood for accurate drug concentration measurements. Method validation for the analysis of forensically relevant stimulant drugs and their metabolites in DBS samples by ultra-performance liquid chromatography-quadrupole time-of-flight high-resolution mass spectrometry (UPLC-QTOF-MS) is presented. DBS blood samples were prepared by spiking 20 µL of blood-containing analytes ranging from 10 to 1000 ng/mL onto Whatman® 903 Protein Saver cards. Analytes were extracted from whole DBS samples punched from protein saver cards by sonication in water followed by PRiME® MCX μElution solid phase extraction. An LOQ of 10 ng/mL was achieved for all analytes. Precision (0.09% to 14.67%), bias (–17.15% to 13.91%) and matrix effects (–18.7% to 23.4%) were suitable for validation in 10/14 analytes. DBS drug stability varied by analyte. DBS sampling may assist in overcoming forensic challenges associated with blood sampling, and accurate drug quantitation even at low concentrations is possible. The results of this work suggest that DBS microsampling is suitable for drug-impaired driving cases in jurisdictions where per se limits for drugs exist, particularly for zero-tolerance drug limits.

Comprehensive Profiling of Mangiferin Metabolites In Vivo and In Vitro Based on the "Drug Metabolite Clusters" Analytical Strategy.

Mangiferin, a natural flavonoid compound with multiple biological activities (e.g., anti-inflammatory, anti-oxidant, anti-diabetic, and anti-tumor), has gained increased research interest in recent years. Nevertheless, the metabolic processing of mangiferin has not been fully investigated. In this study, a rapid and efficient analytical strategy named "Drug Metabolite Clusters" was applied for comprehensive profiling of mangiferin metabolites in rat plasma, urine, and feces samples in vivo following oral administration and liver microsomes in vitro. First, the biological samples were pretreated with methanol, acetonitrile, and solid phase extraction (SPE) for further liquid chromatography-mass spectrometry (LC-MS) analysis. Second, the raw data were acquired using ultra-high performance liquid chromatography quadrupole exactive orbitrap high-resolution mass spectrometry (UHPLC-Q-Exactive Orbitrap HRMS) under the positive and negative full-scan/dd MS2 modes. Third, mangiferin and its basic metabolites (norathyriol, trihydroxyxanthone, and dihydroxyxanthone) were selected as mangiferin metabolite cluster centers by referring to the relevant literature. Subsequently, according to the pyrolysis law of mass spectrometry, literature reports, and reference material comparison, especially the diagnostic product ions (DPIs), the candidate metabolites were accurately preliminarily identified, and mangiferin metabolite clusters based on metabolite cluster center changes were formed. As a result, a total of 67 mangiferin metabolites (mangiferin included) were detected, including 29 in plasma, 48 in urine, 12 in feces, and 6 in liver microsomes. Among them, trihydroxyxanthones were first detected in rat urine samples after oral mangiferin. We found that mangiferin mainly underwent deglucosylation, dehydroxylation, methylation, glucuronidation, sulfation, and other composite reactions in rats. Herein, we have elucidated the metabolites and metabolic pathways of mangiferin in vivo and in vitro, which provided an essential theoretical basis for further pharmacological studies of mangiferin and a comprehensive research method for the identification of drug metabolites.

Analytical Platform for the Study of Metabolic Pathway of Propyl Propane Thiosulfonate (PTSO) from Allium spp.

The present work is focused on the development of an analytical platform to elucidate the metabolic pathway of PTSO from onion, an organosulfur compound well-known for its functional and technological properties and its potential application in animal and human nutrition. This analytical platform consisted of the use of gas chromatography-mass spectrometry (GC-MS) and ultra-high performance liquid chromatography quadrupole with time-of-flight MS (UHPLC-Q-TOF-MS) in order to monitor volatile and non-volatile compounds derived from the PTSO. For the extraction of the compounds of interest, two different sample treatments were developed: liquid-liquid extraction (LLE) and salting-out assisted liquid-liquid extraction (SALLE) for GC-MS and UHPLC-Q-TOF-MS analysis, respectively. Once the analytical platform was optimised and validated, an in vivo study was planned to elucidate PTSO metabolisation, revealing the presence of dipropyl disulfide (DPDS) in liver samples with concentrations between 0.11 and 0.61 µg g-1. The DPDS maximum concentration in the liver was observed at 0.5 h after the intake. DPDS was also present in all plasma samples with concentrations between 2.1 and 2.4 µg mL-1. In regard to PTSO, it was only found in plasma at times above 5 h (0.18 µg mL-1). Both PTSO and DPDS were excreted via urine 24 h after ingestion.

An efficient method for qualitation and quantitation of multi-components of the herbal medicine Qingjin Yiqi Granules

Qingjin Yiqi Granules (QJYQ) is a Traditional Chinese Medicines (TCMs) prescription for the patients with post-COVID-19 condition. It is essential to carry out the quality evaluation of QJYQ. A comprehensive investigation was conducted by establishing deep-learning assisted mass defect filter (deep-learning MDF) mode for qualitative analysis, ultra-high performance liquid chromatography and scheduled multiple reaction monitoring method (UHPLC-sMRM) for precise quantitation to evaluate the quality of QJYQ. Firstly, a deep-learning MDF was used to classify and characterize the whole phytochemical components of QJYQ based on the mass spectrum (MS) data of ultra-high performance liquid chromatography quadrupole time of flight tandem mass spectrometry (UHPLC-Q-TOF/MS). Secondly, the highly sensitive UHPLC-sMRM data-acquisition method was established to quantify the multi-ingredients of QJYQ. Totally, nine major types of phytochemical compounds in QJYQ were intelligently classified and 163 phytochemicals were initially identified. Furthermore, fifty components were rapidly quantified. The comprehensive evaluation strategy established in this study would provide an effective tool for accurately evaluating the quality of QJYQ as a whole.

UPLC-QTOF-MS-Based Metabolomics and Antioxidant Capacity of Codonopsis lanceolata from Different Geographical Origins.

Codonopsis lanceolata (C. lanceolata) has been commonly utilized as a therapeutic plant in traditional medicine. In this study, we examined variations in metabolites in C. lanceolata roots grown in different regions using ultra-high performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS). Multivariate analysis showed that the metabolite profiles of plants grown in Hoengseong and Jeongseon were more similar to each other than to that of C. lanceolata grown in Jeju. Most primary metabolites were present at higher levels in C. lanceolata grown in Jeju. In contrast, C. lanceolata grown in Hoengseong and Jeongseon had high levels of secondary metabolites such as phenylpropanoids and triterpenoid saponins, respectively. In addition, the bioactive compound content and antioxidant capacity of in C. lanceolata grown in Hoengseong and Jeongseon were observed to be higher than those of C. lanceolata grown in Jeju. This study suggests that metabolomics is an effective approach to investigate the difference of metabolite profiling in C. lanceolata from different geographical origins, and is useful for evaluating its pharmacological potential.

Periodate activation with copper oxide nanomaterials for the degradation of ciprofloxacin - A new insight into the efficiency and mechanisms

In this study, CuO was synthesized using a facile precipitation method and was characterized using various techniques for the efficient activation of periodate (PI). Compared to systems consisting of PI alone (0.5 mM) and CuO alone (0.5 g/L), the CuO (0.5 g/L)/PI (0.5 mM) process exhibited improved degradation performance for ciprofloxacin (CIP), a widely used refractory antibacterial drug, with 98% of this pollutant (20 mg/L) removed in a reaction time of 30 min. Both radical and nonradical degradation pathways were identified to play a significant role in the developed CuO/PI system, where the nonradical pathway (holes and electrons) played the most important role in the degradation of CIP. Under both acidic (pH = 3) and near-neutral (pH = 6) conditions, the system showed an improved efficiency for the degradation of CIP in comparison with alkaline conditions (pH = 10). This was attributed to a more facile adsorption of the PI ions onto the surface of the CuO nanoparticles, induced by the electrostatic attraction between the PI ions and the CuO nanomaterial. Analysis of the degraded samples with ultrahigh-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UHPLC Q-TOF MS) demonstrated that cleavage of the piperazine ring and defluorination were the primary degradation pathways of CIP under this oxidation process. The CuO/PI system also showed an excellent removal efficiency for other refractory organic contaminants, such as sulfamethoxazole (SMX) and methylene blue (MB), in various water matrices, including real wastewater. Moreover, respiration tests revealed that the toxicity of the effluents was reduced after the treatment process. In conclusion, this research presents a novel, promising periodate activation technique for the removal of refractory organic contaminants in wastewater treatment.

Analysis of chemical constituents of Sabia parviflora by ultrahigh performance liquid chromatography quadrupole time of flight tandem mass spectrometry

Effective and targeted identification of chemical components of the Chinese herbal medicine Sabia parviflora remains a major challenge. Herein, we used ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry to analyze the chemical composition of S.parviflora. Its chemical components were rapidly identified using the characteristic ion filtration method, which involves these steps: (1) summarize the characteristic ions based on similar skeletons and compounds with substitution patterns, and establish a database; (2) screen and classify different types of compounds in S. parviflora based on the characteristic ions; and (3) identify the compounds based on molecular weight, secondary fragments, and the database. In the present study, the characteristic ions in S. parviflora were grouped into five major classes. A total of 104 components were identified, including 12 potentially novel compounds. This rapid and accurate method provides an important basis for basic chemical research in S. parviflora.

High-Yield Recovery of Antioxidant Compounds from Bambusa chungii Culms Using Pressurized Hot Water Extraction.

A large amount of waste from Bambusa chungii culms is generated from the bamboo pulping industry, causing disposal problems. Nevertheless, bamboo culms are a suitable source of functional ingredients, such as antioxidant compounds. However, because of the high compactness and tightness in their material structure, obtaining phytochemicals from bamboo culms using conventional organic solvent extraction methods can be inefficient. In this research, we developed a pressurized hot water extraction (PHWE) method to recover 19 target phenolic compounds from Bambusa chungii culms. The extracted compounds were determined by ultra-high performance liquid chromatography-quadrupole time-of-flight-mass spectrometry (UPLC-QTOF-MS). The antioxidant potential of the extracts was evaluated by 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), and ferric reducing antioxidant power (FRAP) assays. We investigated the effects of temperature, extraction time, and the material-to-liquid ratio on PHWE, and these parameters were optimized with a Box-Behnken design experiment and response surface tool. The optimal extraction condition was found at 170 °C, with a 1:30 g/mL material-to-liquid ratio and a 14 min extraction time. Following these optimal parameters, the total yield of target phenolic compounds (TYPC) reached 3.85 mg/g of raw material, and the half-maximal inhibitory concentrations (IC50) for the DPPH and ABTS tests were 94.7 mg/L and 21.88 mg/L, respectively. The FRAP value was 1.23 μmol FSE/mg of dried extract. A strong correlation between TYPC and the antioxidant activity of the extract was confirmed. The TYPC and antioxidant capacity of the optimal PHWE extract of the Bambusa chungii culms were both considerably higher than those of extracts obtained from conventional solvent extractions. These results indicated that PHWE is an excellent green technique for recovering phenolic compounds from bamboo culms, and the PHWE extracts of Bambusa chungii culms may be a good source of natural antioxidants.

SmDXS5, acting as a molecular valve, plays a key regulatory role in the primary and secondary metabolism of tanshinones in Salvia miltiorrhiza.

Red sage, the dry root and rhizome of the herbaceous plant Salvia miltiorrhiza Bunge, is widely used for treating various diseases. The low content of tanshinones (terpenoids) has always restricted development of the S. miltiorrhiza industry. Here, we found that SmDXS5, a rate-limiting enzyme-coding gene located at the intersection of primary and secondary metabolism, can effectively change the transcription level and secondary metabolome profile of hairy roots of S. miltiorrhiza, and significantly increase the content of tanshinones. Agrobacterium rhizogenes was used to infuse S. miltiorrhiza explants, and hairy roots of S. miltiorrhiza expressing the SmDXS5 gene were obtained successfully. We identified 39 differentially accumulated metabolites (DAMs) by metabolomics based on ultra-high performance liquid chromatography quadrupole exactive mass spectrometry and multivariate statistics. These DAMs might be key metabolites of SmDXS5 gene regulation. RNA sequencing was used to compare gene expression between the hairy roots of the SmDXS5 overexpressing group and the blank control (BC) group. Compared with the BC group, 18,646 differentially expressed genes were obtained: 8994 were upregulated and 9,652 downregulated. The combined transcriptome and metabolome analyses revealed that the mevalonate and methylerythritol phosphate pathways and synthase gene expression levels in the SmDXS5 overexpressing group were upregulated significantly, and the accumulation of tanshinone components was increased significantly, which promoted the process of glycolysis and promoted the transformation of carbohydrates to secondary metabolism. Moreover, the expression of SmPAL, the first rate-limiting enzyme gene of the phenylpropane pathway, decreased, reducing the accumulation of phenolic acid, another secondary metabolite. Therefore, SmDXS5 can be defined as a 'valve' gene, mainly responsible for regulating the distribution of primary and secondary metabolic flow of tanshinones in S. miltiorrhiza, and for other secondary metabolic pathways. The discovery of SmDXS5 and its molecular valve function in regulating primary and secondary metabolism will provide a basis for the industrial production of tanshinone components, and cultivation of high quality S. miltiorrhiza.

Ultraviolet (UV) light-induced geometrical isomerization of cinnamic acid containing molecules: A plausible, non-enzymatic approach to modify metabolite composition of plant extracts

• UV light can be used to enhance the cinnamic acid containing-metabolite composition of a plant. • LC-MS in combination with multivariate statistics can be used to probe for the presence of photo-induced metabolites in plant extracts. • UV-irradiated plant extracts have a coherent metabolite composition as compared to naturally growing plants. Cinnamic acid containing molecules are widely distributed in plants, and they are known to possess several pharmacological properties. Structurally, these molecules consist of C 6 C 3 skeletal backbone, with an olefin functional group on the C 3 part . Naturally, plants produce a broad spectrum of cinnamic acid derivatives with a trans configuration, however, cis isomers are formed due to excessive UV-light exposure, thereby increasing the already complex metabolomes of plants containing these compounds. In high sunlight intensive areas, the prevalence of the cis isomers of cinnamic acid containing molecules is becoming more prevalent and synonymous with extracts of many plants. It is therefore imperative to develop analytical and data handling strategies to account for the presence of these molecules, which can serve as a template for rational design of pharmacologically active plant extracts or smart photo-switchable drugs. Herein, we show that a combination of ultra-high performance liquid chromatography quadrupole time of flight mass spectrometry (UHPLC-QTOF-MS) and principal component analysis (PCA) allows differentiation of plants extracts of which metabolome modification through UV-radiation has been carried out. The results of the current study show that geometrical isomerism is a feasible, non-enzymatic approach to modify metabolite composition of plants. This simple method can be used to create “super” extracts of plants containing structurally diverse metabolites to be evaluated for various pharmacological properties in the future.

Multi-omics analyses reveal the specific changes in gut metagenome and serum metabolome of patients with polycystic ovary syndrome.

The purpose of this study was to investigate the specific alterations in gut microbiome and serum metabolome and their interactions in patients with polycystic ovary syndrome (PCOS). The stool samples from 32 PCOS patients and 18 healthy controls underwent the intestinal microbiome analysis using shotgun metagenomics sequencing approach. Serum metabolome was analyzed by ultrahigh performance liquid chromatography quadrupole time-of-flight mass spectrometry. An integrative network by combining metagenomics and metabolomics datasets was constructed to explore the possible interactions between gut microbiota and circulating metabolites in PCOS, which was further assessed by fecal microbiota transplantation (FMT) in a rat trial. Fecal metagenomics identified 64 microbial strains significantly differing between PCOS and healthy subjects, half of which were enriched in patients. These changed species showed an ability to perturb host metabolic homeostasis (including insulin resistance and fatty acid metabolism) and inflammatory levels (such as PI3K/Akt/mTOR signaling pathways) by expressing sterol regulatory element-binding transcription factor-1, serine/threonine-protein kinase mTOR, and 3-oxoacyl-[acyl-cattier-protein] synthase III, possibly suggesting the potential mechanisms of gut microbiota underlying PCOS. By integrating multi-omics datasets, the panel comprising seven strains (Achromobacter xylosoxidans, Pseudomonas sp. M1, Aquitalea pelogenes, Porphyrobacter sp. HL-46, Vibrio fortis, Leisingera sp. ANG-Vp, and Sinorhizobium meliloti) and three metabolites [ganglioside GM3 (d18:0/16:0), ceramide (d16:2/22:0), and 3Z,6Z,9Z-pentacosatriene] showed the highest predictivity of PCOS (AUC: 1.0) with sensitivity of 0.97 and specificity of 1.0. Moreover, the intestinal microbiome modifications by FMT were demonstrated to regulate PCOS phenotypes including metabolic variables and reproductive hormones. Our findings revealed key microbial and metabolite features and their interactions underlying PCOS by integrating multi-omics approaches, which may provide novel insights into discovering clinical diagnostic biomarkers and developing efficient therapeutic strategies for PCOS.

Quantification and identification analysis of Ziziphus jujuba Mill. peel pigmentation at different developmental stages.

The fruit peel of a color mutant jujube cultivar, 'Sanbianhong' (SBF), was investigated using an ultra-high performance liquid chromatography quadrupole Orbitrap mass spectrometry (UHPLC-Q-Orbitrap MS) at five ripening stages (S1, Young fruit stage; S2, swelling stage; S3, white-mature stage; S4, pre-mature stage and S5, mature stage). Lutein, β-carotene, chlorophyll a, chlorophyll b, and 13 anthocyanins were identified. Chlorophyll a and cyanidin 3-O-galactoside were considered key color metabolites in S1 with the content of 1.083mg/g of fresh weight (FW) and 4.585mg/g of FW, respectively. Delphinidin (0.488mg/g FW) and cyanidin (6.259mg/g FW) were identified as the key pigments in S3. Delphinidin 3-O-glucoside (0.256mg/g FW) was identified as the key anthocyanin in maturity S5. Herein, the identification and quantitation of pigment-related metabolites of SBF were studied for the first time, and the results provide a theoretical basis for understanding the pigment changes of jujube fruit during ripening.

Identification of the chemical components of ethanol extract of Chenopodium ambrosioides and evaluation of their in vitro antioxidant and anti tumor activities

Purpose: To determine the characteristic chemical components of the ethanol extract of Chenopodium ambrosioides and evaluate their antioxidant and anti-tumor effects in vitro.
 Methods: The plant powder (5 g) was extracted with 1 L of 80 % ethanol at room temperature for 45 min, and then placed at 60 oC at varying microwave power and duration to obtain optimal extraction conditions. Characteristic chemical components were detected using ultra-high performance liquid chromatography quadrupole time of flight mass spectrometry (UPLC-Q-TOF-MS/MS). Kaempferitrin was isolated from the 80 % ethanol extract using a D101 macroporous resin column, and its content was assessed by high performance liquid chromatography (HPLC). The antioxidant effect of kaempferitrin was evaluated by its ability to scavenge 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azinobis-(3-ethylbenzthiazoline-6-sulphonate) (ABTS) radicals, while its anti-proliferation activity in human liver cancer cells SMMC-7721 was determined using cell counting kit-8 (CCK-8) reagent.
 Results: Three characteristic components of ethanol extract of C. ambrosioides were obtained, namely, kaempferitrin, kaempferol-3-O-apigenin-7-O-rhamnoside and kaempferol-3-O-acetylapigenin-7-O-rhamnoside. Kaempferitrin was shown to possess strong DPPH radical and moderate ABTS radical scavenging activities. Kaempferitrin significantly inhibited the proliferation of SMMC-7721 cells at doses of 4 and 8 μg/mL, with half-maximal concentration (IC50) of 0.38 μM (p < 0.05).
 Conclusion: Kaempferitrin extracted from C. ambrosioides has antioxidant and anti-tumor activities. The results reported here indicate that C. ambrosioides may have potential use in herbal medicine practice.