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Fungal endophytes, symbiotic microorganisms residing within plants, are renowned for producing bioactive secondary metabolites with diverse beneficial properties. We investigated the antimicrobial potential of fungal endophytes isolated from Solanum mauritianum, an invasive weed, against clinically significant bacterial pathogens. Selected fungal endophytes (Penicillium chrysogenum, Fusarium sp., and Paracamarosporium leucadendri) were isolated from the plant's leaves and fruits. Their crude extracts were tested against various referenced strains, such as Mycobacterium species (M. smegmatis ATCC 607 and M. bovis ATCC 27290), Staphylococcus aureus ATCC 6571, Bacillus subtilis ATCC 11774, Klebsiella species (K. pneumoniae ATCC 10031 and K. oxytoca ATCC 8724), Escherichia coli ATCC 10536, and Pseudomonas aeruginosa ATCC 10145, using the Kirby-Bauer disk diffusion method. Resazurin Microtiter Assay was used for the determination of the minimum inhibitory concentration. The chemical nature of the secondary metabolites in the crude extracts produced by fungal endophytes was evaluated using high-resolution liquid chromatography–mass spectrometry (LC-MS) using water and acetonitrile gradient. Liquid chromatography quadrupole time-of-flight mass spectrometry (LC-QTOF-MS/MS) was employed for untargeted metabolomics. LC-QTOF-MS/MS identified 63 bioactive compounds across the three endophytes. P. chrysogenum had the highest activity against S. aureus and M. smegmatis (1.15 mg/mL and 0.02 mg/mL, respectively), while P. leucadendri demonstrated moderate activity against M. smegmatis (2.91 mg/mL) and E. coli (1.16 mg/mL). Fusarium sp. exhibited the broadest spectrum of antibacterial activity, with MIC values ranging from 0.03 mg/mL (B. subtilis) to 10 mg/mL (M. smegmatis). P. leucadendri produced 29 metabolites, Fusarium sp. had 23 identified metabolites, and a total of 11 metabolites were identified from P. chrysogenum. The fruits of the plant, accounting for 60%, appeared to be the most abundant in the endophyte diversity when compared to the stems and leaves. This study highlights the potential of fungal endophytes from S. mauritianum as a source of novel bioactive compounds, particularly against multidrug-resistant pathogens, contributing to the ongoing efforts to combat antimicrobial resistance. [ABSTRACT FROM AUTHOR]

The overuse of antibiotics and antitumor drugs has resulted in more and more extensive pollution of water bodies with organic drugs, causing detrimental ecological effects, which have attracted attention towards effective and sustainable methods for antibiotics and antitumor drug degradation. Here, the hybrid nanomaterial (g-C 3 N 4 @Fe/Pd) was synthesized and used to remove a kind of both an antibiotic and antitumor drug named mitoxantrone (MTX) with 92.0% removal efficiency, and the MTX removal capacity is 450 mg/g. After exposing to the hybrid material the MTX aqueous solution changed color from dark blue to lighter progressively, and LC-UV results of residual solutions show that a new peak at 3.0 min (MTX: 13.2 min) after removal by g-C 3 N 4 @Fe/Pd appears, with the simultaneous detection of intermediate products indicating that g-C 3 N 4 @Fe/Pd indeed degrades MTX. Detailed mass spectrometric analysis suggests that the nuclear mass ratio decreased from 445.2 (M+1H) to 126.0 (M+1H), 169.1 (M+1H), 239.2 (M+1H), 267.3 (M+1H), 285.2 (M+1H), 371.4 (M+1H) and 415.2 (M+1H), and the maximum proportion (5.63%) substance of all degradation products (126.0 (M+1H)) is 40-100 times less toxic than MTX. A mechanism for the removal and degradation of mitoxantrone was proposed. Besides, actual water experiments confirmed that the maximum removal capacity of MTX by g-C 3 N 4 @Fe/Pd is up to 492.4 mg/g (0.02 g/L, 10 ppm). [ABSTRACT FROM AUTHOR]

BACKGROUND: Mangifera indica L. (mango), a medicinal plant rich in biologically active compounds, has potential to be used in disease‐preventing and health‐promoting products. The present investigation reveals and uncovers bioactive metabolites with remarkable therapeutic efficiency from mango (family: Anacardiaceae) seeds. RESULTS: Biological activity was determined by antimicrobial, antioxidant and anticancer assays, and metabolite profiling was performed on gas chromatography coupled to quadrupole time‐of‐flight mass spectrometry (GC‐QTOF‐MS) and liquid chromatography coupled to quadrupole time‐of‐flight mass spectrometry (LC‐QTOF‐MS) platforms. Validation of active metabolites was carried out by in silico molecular docking (Molinspiration Cheminformatics Server and PASS). Extracted and identified metabolites were screened; 54 compounds associated with various groups were selected for the in silico interaction study. CONCLUSIONS: Molecular docking revealed lead molecules with a potential binding energy score, efficacy and stable modulation with a selected protein domain. Investigation, directed by in vitro and in silico analysis, confirms mango seeds as an excellent source of potential metabolites as a therapeutic agent. © 2024 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Due to the emergence of drug-resistant microorganisms, the search for broad-spectrum antimicrobial compounds has become extremely crucial. Natural sources like plants and soils have been explored for diverse metabolites with antimicrobial properties. This study aimed to identify microorganisms from agricultural soils exhibiting antimicrobial effects against known human pathogens, and to highlight the chemical space of the responsible compounds through the computational metabolomics-based bioprospecting approach. Herein, bacteria were extracted from soil samples and their antimicrobial potential was measured via the agar well diffusion method. Methanolic extracts from the active bacteria were analyzed using the liquid chromatography quadrupole time-of-flight mass spectrometry (LC-QTOF-MS) technique, and the subsequent data was further analyzed through molecular networking approach which aided in identification of potential anti-microbial compounds. Furthermore, 16S rRNA gene sequencing enabled identification of the active bacterial isolates, where isolate 1 and 2 were identified as strains of Bacillus pumilus, whilst isolate 3 was found to be Bacillus subtilis. Interestingly, isolate 3 (Bacillus subtilis) displayed wide-ranging antimicrobial activity against the tested human pathogens. Molecular networking revealed the presence of Diketopiperazine compounds such as cyclo (D-Pro-D-Leu), cyclo (L-Tyr-L-Pro), cyclo (L-Pro-D-Phe), and cyclo (L-Pro-L-Val), alongside Surfactin C, Surfactin B, Pumilacidin E, and Isarrin D in the Bacillus strains as the main anti-microbial compounds. The application of the molecular networking approach represents an innovation in the field of bio-guided bioprospection of microorganisms and has proved to be an effective and feasible towards unearthing potent antimicrobial compounds. Additionally, the (computational metabolomics-based) approach accelerates the discovery of bioactive compounds and isolation of strains which offer a promising avenue for discovering new clinical antimicrobials. Finally, soil microbial flora could serve an alternative source of anti-microbial compounds which can assist in the fight against emergence of multi-drug resistance bacterial pathogens. [ABSTRACT FROM AUTHOR]

As part of a long-term campaign to document, study, and conserve the Heroes tapestries from The Cloisters collection at The Metropolitan Museum of Art, organic colorant analysis of Julius Caesar (accession number 47.101.3) was performed. Analysis with liquid chromatography–quadrupole time-of-flight mass spectrometry (LC-qToF-MS) revealed the presence of several multiply chlorinated xanthones produced only by certain species of lichen. Various lichen dye sources have been documented in the literature for centuries and are classified as either ammonia fermentation method (AFM) or boiling water method (BWM) dyes based on their method of production. However, none of these known sources produce the distinctive metabolites present in the tapestry. LC-qToF-MS was also used to compare the chemical composition of the dyes in the tapestry with that of several species of crustose lichen. Lichen metabolites, including thiophanic acid and arthothelin, were definitively identified in the tapestry based on comparison with lichen xanthone standards and a reference of Lecanora sulphurata, confirming the presence of a lichen source. This finding marks the first time that lichen xanthones have been identified in a historic object and the first evidence that BWM lichen dyes may have been used prior to the eighteenth century.

Abstract Leontodon hispidulus Boiss is a wild annual plant growing in Egypt. The present study aims for the first time, to evaluate the phytochemical profile of the main secondary metabolites of the optimized ethanolic extract of the plant using Quadrupole Time-of-Flight Liquid chromatography-mass spectrometry and Gas chromatography-mass spectrometry. It also aims to assess the anticancer activity of its different fractions against the prostate carcinoma cell line. Moreover, an in-silico docking study was performed using the Hexokinase-two enzyme. LC-qToF-MS analysis revealed the tentative identification of 36 phenolic compounds including the glycosides of (luteolin, quercetin, kaempferol, apigenin, isorhamnetin, and daidzein), coumarines (esculin, esculetin, and daphnetin), and phenolic acids (chlorogenic, caffeic, quinic, P-coumaric, and rosmarinic). GC–MS/MS analysis revealed the presence of 18 compounds where palmitic acid, myristic acid, alpha-amyrin, and beta-amyrin were the major ones. The cytotoxic activity results revealed that methylene chloride and ethyl acetate fractions showed the highest cytotoxic activity against the PC3 cell line, with IC50 values of 19, and 19.6 μg/ml, respectively. Interestingly, the docking study demonstrated that apigenin-7-O-glucoside, luteolin-7-O-glucoside, kaempferol-3-O-glucuronide, quercetin-4′-O-glucoside, esculin, rosmarinic acid, chlorogenic acid, and α-amyrin exhibited high affinity to the selected target, HEK-2 enzyme.

As part of a long-term campaign to document, study, and conserve the Heroes tapestries from The Cloisters collection at The Metropolitan Museum of Art, organic colorant analysis of Julius Caesar (accession number 47.101.3) was performed. Analysis with liquid chromatography–quadrupole time-of-flight mass spectrometry (LC-qToF-MS) revealed the presence of several multiply chlorinated xanthones produced only by certain species of lichen. Various lichen dye sources have been documented in the literature for centuries and are classified as either ammonia fermentation method (AFM) or boiling water method (BWM) dyes based on their method of production. However, none of these known sources produce the distinctive metabolites present in the tapestry. LC-qToF-MS was also used to compare the chemical composition of the dyes in the tapestry with that of several species of crustose lichen. Lichen metabolites, including thiophanic acid and arthothelin, were definitively identified in the tapestry based on comparison with lichen xanthone standards and a reference of Lecanora sulphurata, confirming the presence of a lichen source. This finding marks the first time that lichen xanthones have been identified in a historic object and the first evidence that BWM lichen dyes may have been used prior to the eighteenth century. [ABSTRACT FROM AUTHOR]

• High altitude connected with high UV radiation may limit the production of secondary metabolites by Cyanobacteria. • Strains isolated from two microbial mats in which no secondary metabolites were detected produced them in laboratory conditions. Ever-new cyanotoxins are being discovered, with planktic Cyanobacteria being the most studied communities, although records of cyanotoxins from benthic communities are becoming increasingly common. ​​Thus, the latter also started threatening users of water for recreational or drinking purposes. However, vast areas of the globe, i.a. Central Asia, are still understudied in this respect. Our recent study of benthic Cyanobacteria in the mountains of Eastern Pamir (Tajikistan) suggested that cyanotoxin production in high mountain environments is very limited. Here we present further study of Cyanobacteria, cyanotoxins, and other biologically active compounds such as geosmin and 2-Methylisoborneol (MIB) in microbial mats and water above them. Samples were collected in Eastern Pamirs, in UV (7–11) and altitude gradient (1000–4000 m a.s.l.). In the study, we used microscopic and genetic identification of Cyanobacteria based on NGS of V3-V4 16S rRNA amplicon and toxin assays using LC-QTOF-MS. The analyses demonstrated that the studied microbial mats contained potentially toxic Cyanobacteria (Anabaena, Lyngbya, Nostoc, Oscillatoria, and Phormidium). The production of cyanotoxins and taste and odor compounds (T&O) in natural environments was restricted to altitudes up to 3000 m a.s.l. Three water samples at 1000, 2000 and 3000 m a.s.l. contained MIB and/or geosmin, while debromoaplysiatoxin was noted at 2000 m a.s.l. Additionally, two strains (Hillbrichtia pamiria gen. nov. sp. nov. and Nostoc paludosum) isolated from sites at about 4000 m, in which no cyanotoxins or T&O were identified, produced debromoaplysiatoxin and microginin in laboratory conditions. The results suggest that in a stressful environment Cyanobacteria do not produce toxins. [ABSTRACT FROM AUTHOR]

Leontodon hispidulus Boiss is a wild annual plant growing in Egypt. The present study aims for the first time, to evaluate the phytochemical profile of the main secondary metabolites of the optimized ethanolic extract of the plant using Quadrupole Time-of-Flight Liquid chromatography-mass spectrometry and Gas chromatography-mass spectrometry. It also aims to assess the anticancer activity of its different fractions against the prostate carcinoma cell line. Moreover, an in-silico docking study was performed using the Hexokinase-two enzyme. LC-qToF-MS analysis revealed the tentative identification of 36 phenolic compounds including the glycosides of (luteolin, quercetin, kaempferol, apigenin, isorhamnetin, and daidzein), coumarines (esculin, esculetin, and daphnetin), and phenolic acids (chlorogenic, caffeic, quinic, P-coumaric, and rosmarinic). GC–MS/MS analysis revealed the presence of 18 compounds where palmitic acid, myristic acid, alpha-amyrin, and beta-amyrin were the major ones. The cytotoxic activity results revealed that methylene chloride and ethyl acetate fractions showed the highest cytotoxic activity against the PC3 cell line, with IC50 values of 19, and 19.6 μg/ml, respectively. Interestingly, the docking study demonstrated that apigenin-7-O-glucoside, luteolin-7-O-glucoside, kaempferol-3-O-glucuronide, quercetin-4′-O-glucoside, esculin, rosmarinic acid, chlorogenic acid, and α-amyrin exhibited high affinity to the selected target, HEK-2 enzyme. [ABSTRACT FROM AUTHOR]

Due to the emergence of drug-resistant microorganisms, the search for broad-spectrum antimicrobial compounds has become extremely crucial. Natural sources like plants and soils have been explored for diverse metabolites with antimicrobial properties. This study aimed to identify microorganisms from agricultural soils exhibiting antimicrobial effects against known human pathogens, and to highlight the chemical space of the responsible compounds through the computational metabolomics-based bioprospecting approach. Herein, bacteria were extracted from soil samples and their antimicrobial potential was measured via the agar well diffusion method. Methanolic extracts from the active bacteria were analyzed using the liquid chromatography quadrupole time-of-flight mass spectrometry (LC-QTOF-MS) technique, and the subsequent data was further analyzed through molecular networking approach which aided in identification of potential anti-microbial compounds. Furthermore, 16S rRNA gene sequencing enabled identification of the active bacterial isolates, where isolate 1 and 2 were identified as strains of Bacillus pumilus, whilst isolate 3 was found to be Bacillus subtilis. Interestingly, isolate 3 (Bacillus subtilis) displayed wide-ranging antimicrobial activity against the tested human pathogens. Molecular networking revealed the presence of Diketopiperazine compounds such as cyclo (D-Pro-D-Leu), cyclo (L-Tyr-L-Pro), cyclo (L-Pro-D-Phe), and cyclo (L-Pro-L-Val), alongside Surfactin C, Surfactin B, Pumilacidin E, and Isarrin D in the Bacillus strains as the main anti-microbial compounds. The application of the molecular networking approach represents an innovation in the field of bio-guided bioprospection of microorganisms and has proved to be an effective and feasible towards unearthing potent antimicrobial compounds. Additionally, the (computational metabolomics-based) approach accelerates the discovery of bioactive compounds and isolation of strains which offer a promising avenue for discovering new clinical antimicrobials. Finally, soil microbial flora could serve an alternative source of anti-microbial compounds which can assist in the fight against emergence of multi-drug resistance bacterial pathogens.

Fungal endophytes, symbiotic microorganisms residing within plants, are renowned for producing bioactive secondary metabolites with diverse beneficial properties. We investigated the antimicrobial potential of fungal endophytes isolated from Solanum mauritianum, an invasive weed, against clinically significant bacterial pathogens. Selected fungal endophytes (Penicillium chrysogenum, Fusarium sp., and Paracamarosporium leucadendri) were isolated from the plant’s leaves and fruits. Their crude extracts were tested against various referenced strains, such as Mycobacterium species (M. smegmatis ATCC 607 and M. bovis ATCC 27290), Staphylococcus aureus ATCC 6571, Bacillus subtilis ATCC 11774, Klebsiella species (K. pneumoniae ATCC 10031 and K. oxytoca ATCC 8724), Escherichia coli ATCC 10536, and Pseudomonas aeruginosa ATCC 10145, using the Kirby-Bauer disk diffusion method. Resazurin Microtiter Assay was used for the determination of the minimum inhibitory concentration. The chemical nature of the secondary metabolites in the crude extracts produced by fungal endophytes was evaluated using high-resolution liquid chromatography–mass spectrometry (LC-MS) using water and acetonitrile gradient. Liquid chromatography quadrupole time-of-flight mass spectrometry (LC-QTOF-MS/MS) was employed for untargeted metabolomics. LC-QTOF-MS/MS identified 63 bioactive compounds across the three endophytes. P. chrysogenum had the highest activity against S. aureus and M. smegmatis (1.15 mg/mL and 0.02 mg/mL, respectively), while P. leucadendri demonstrated moderate activity against M. smegmatis (2.91 mg/mL) and E. coli (1.16 mg/mL). Fusarium sp. exhibited the broadest spectrum of antibacterial activity, with MIC values ranging from 0.03 mg/mL (B. subtilis) to 10 mg/mL (M. smegmatis). P. leucadendri produced 29 metabolites, Fusarium sp. had 23 identified metabolites, and a total of 11 metabolites were identified from P. chrysogenum. The fruits of the plant, accounting for 60%, appeared to be the most abundant in the endophyte diversity when compared to the stems and leaves. This study highlights the potential of fungal endophytes from S. mauritianum as a source of novel bioactive compounds, particularly against multidrug-resistant pathogens, contributing to the ongoing efforts to combat antimicrobial resistance.

In this study, 31 household spray-type cleaning products from 10 categories were examined to identify the substances contained in these products and evaluate their resulting toxicities. After substance detection via LC-QTOF-MS, suspect and non-target screenings were performed using UNIFI software. A total of 48 substances were identified during suspect screening. Among them, surfactant (freq. 22/31(71.0%)), plasticizer (freq. 23/31(74.2%)), antioxidant (freq. 20/31(64.5%)) were detected with high frequencies. Among the 48 compounds, tributyl citrate acetate was the most frequently detected (frequency 13/31, 41.9%). In addition, 51 substances were identified via non-target analysis. Among them, oleamide, a type of surfactant, showed a high frequency of detection (22/31(71.0%)). The toxicities of the 98 substances (suspect: 47 (except for confidence level 5) and non-target: 51) identified were evaluated using the European Chemicals Agency (ECHA) database and Toxtree. Among them, 20 substances were classified as a substance of caution. The risk assessment for 3 substances identified with references standards was also performed. The suspect and non-target screening technique proved to be a useful method that can be applied when there are many non-specific chemicals and their components in the consumer products are not well known. [ABSTRACT FROM AUTHOR]

Targeted analysis of contaminants in water are often focused on a narrow range of chemicals, falling short of the true pollution status of water bodies. Non targeted screening presents several advantages in identifying less prioritized pollutants, entirely unknown compounds and transformation products. In this study we employed a non-targeted screening workflow established on a high-resolution quadrupole time-of-flight mass spectrometer coupled to an ultrahigh performance liquid chromatograph (UHPLC-QTOF-MS) to identify known and unknown pollutants in South African waste and surface waters. Level two confidence identification of 315 compounds was achieved based on mass accuracy, isotope patterns and MS/MS spectra match. Pharmaceuticals, drugs, and metabolites made up 40% of the detected compounds, biological compounds and industrial chemicals along with their metabolites constituted 24 and 18% respectively, while personal care products, pesticides and food additives made up approximately 5, 4 and 4% respectively. Several antiretroviral drugs were confirmed with level one confidence using isotope labelled standards. A wide range of “new” pharmaceuticals, pesticides and metabolites were documented in South African waters for the first time. Seventeen (17) pharmaceuticals were reported for the first time in South African waters, of which four are reported for the first time ever in surface water.The result of this exploratory study highlights the presence of several contaminants of public health concern that have hitherto received little to no attention in previous wastewater-based epidemiological studies. We provide a detailed list of priority contaminants for future studies on targeted-analysis.

A high-throughput screening and quantification method for 479 pesticides in green tea was established based on solid-phase extraction combined with liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (LC-QTOF-MS). Pesticides were extracted from samples using an optimized SPE (TPT cartridges) procedure. LC-QTOF-MS in All Ions MS/MS scan mode acquired full MS data for quantification and product ion spectra for identification. LC-QTOF-MS quantification was achieved using matrix-matched standard calibration curves to achieve the optimal method accuracy. The method performance characteristics included the linearity, overall recovery, precision, and measurement uncertainty being evaluated. The validation results exhibited a good sensitivity with the LOQs of 5–55 µg/kg, which was satisfactory for their MRLs in China or the EU. The recoveries of more than 92.7% of the 479 pesticides in green tea were 70–120% at the three spiked levels with a precision of ≤20%. Finally, this method was employed to analyze 479 pesticides in 95 tea samples from markets in China. The test results of the tea samples showed that tolfenpyrad, buprofezin, and pyridaben were found with lower concentrations. The method has effectively improved the determination efficiency of pesticide residue screening by high-resolution mass spectrometry in green tea. [ABSTRACT FROM AUTHOR]

BACKGROUND: Nanoencapsulation has opened promising fields of innovation for pesticides. Conventional pesticides can cause side effects on plant metabolism. To date, the effect of nanoencapsulated pesticides on plant phenolic contents has not been reported. RESULTS: In this study, a comparative evaluation of the phenolic contents and metabolic profiles of strawberries was performed for plants grown under controlled field conditions and treated with two separate active ingredients, azoxystrobin and bifenthrin, loaded into two different types of nanocarriers (Allosperse® polymeric nanoparticles and SiO2 nanoparticles). There were small but significant decreases of the total phenolic content (9%) and pelargonidin 3‐glucoside content (6%) in strawberries treated with the nanopesticides. An increase of 31% to 125% was observed in the levels of gallic acid, quercetin, and kaempferol in the strawberries treated with the nanoencapsulated pesticides compared with the conventional treatments. The effects of the nanocarriers on the metabolite and phenolic profiles was identified by principal component analysis. CONCLUSION: Overall, even though the effects of nanopesticides on the phenological parameters of strawberry plants were not obvious, there were significant changes to the plants at a molecular level. In particular, nanocarriers had some subtle effects on plant health and fruit quality through variations in total and individual phenolics in the fruits. Further research will be needed to assess the impact of diverse nanopesticides on other groups of plant metabolites. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

The repurposing of expired drugs through bioconversion remains one of the most crucial research milestones, as this practice reduces drug contamination while producing compounds of significance. The present study investigated the bioconversion of ganoderic acid A (GAA) using Penicillium brevicompactum over a period of 3, 6 and 9 days. The GAA intensity reduced from 22,099 cps on day 0 to 11,040, 4700 and 18,126 cps on day 3, 6 and 9, respectively, thus demonstrating the degradation of GAA over time. The produced metabolites that were recovered using ethyl acetate as a solvent were determined using LC-QTOF-MS. P. brevicompactum produced a variety of compounds in the absence of GAA, while in its presence, it was observed that P. brevicompactum was able to convert GAA and produced ganomastenol A/B/D, vitamin E succinate, and aminopregnane on day 3, while on day 6, armillaripin and ganolucidic acid A were produced. After 9 days of operation, vitamin E succinate, ganolucidic acid A and lucilactaene were produced. The present study is the first report on the ability of P. brevicompactum to bioconvert GAA. The identified metabolites have been established to possess bioactivity against various ailments, thus contributing to the discovery of new drugs. [ABSTRACT FROM AUTHOR]

Yangzheng Mixture is a traditional Chinese medicine used in clinical practice as an adjuvant therapy for tumors. However, little is known about its active components in tumor treatment. The purpose of this study was to explore the potential anti‐tumor components of Yangzheng Mixture to better promote its clinical application. Using LC–MS/MS, 43 components were detected in concentrated Yangzheng Mixture. Six components, comprising astragaloside, calycosin, formononetin, isoquercitrin, ononin, and calycosin‐7‐O‐β‐D‐glucoside, were identified in rat plasma. The cancer cell absorption assay showed that the intracellular concentration of four components, calycosin, calycosin‐7‐O‐β‐D‐glucoside, formononetin, and ononin, increased with extended incubation time and demonstrated potential anti‐tumor effects. The MTT assay results confirmed that Yangzheng Mixture inhibited different tumor cells proliferation. Additionally, the colony formation assay, flow cytometry analysis and wound healing displayed that Yangzheng Mixture and a combination of four components could inhibit colony formation, arrest the cell cycle and impair cell migration of tumor cells, including HCT‐116, MHCC‐97L, MCF‐7 and NCI‐H1299. In summary, our study highlighted the plausible application of Yangzheng Mixture as a potential adjuvant treatment for tumors. Furthermore, it identified effective anti‐tumor components and provided evidences for the further clinical application of Yangzheng Mixture. [ABSTRACT FROM AUTHOR]

According to the World Health Organization, women's breast cancer is among the most common cancers with 7.8 million diagnosed cases during 2016–2020 and encompasses 15 % of all female cancer-related mortalities. These mortality events from triple-negative breast cancer are a significant health issue worldwide calling for a continuous search of bioactive compounds for better cancer treatments. Historically, plants are important sources for identifying such new bioactive chemicals for treatments. Here we use high-throughput screening and mass spectrometry analyses of extracts from 100 plant species collected in Chinese ancient forests to detect novel bioactive breast cancer phytochemicals. First, to study the effects on viability of the plant extracts, we used a MTT and CCK-8 cytotoxicity assay employing triple-negative breast cancer (TNBC) MDA-MB-231 and normal epithelial MCF-10A cell lines and cell cycle arrest to estimate apoptosis using flow cytometry for the most potent three speices. Based on these analyses, the final most potent extracts were from the Amur honeysuckle (Lonicera maackii) wood/root bark and Nigaki (Picrasma quassioides) wood/root bark. Then, 5 × 106 MDA-MB-231 cells were injected subcutaneously into the right hind leg of nude mice and a tumour was allowed to grow before treatment for seven days. Subsequently, the four exposed groups received gavage extracts from Amur honeysuckle and Nigaki (Amur honeysuckle wood distilled water, Amur honeysuckle root bark ethanol, Nigaki wood ethanol or Nigaki root bark distilled water/ethanol (1:1) extracts) in phosphate-buffered saline (PBS), while the control group received only PBS. The tumour weight of treated nude mice was reduced significantly by 60.5 % within 2 weeks, while on average killing 70 % of the MDA-MB-231 breast cancer cells after 48 h treatment (MTT test). In addition, screening of target genes using the Swiss Target Prediction, STITCH, STRING and NCBI-gene database showed that the four plant extracts possess desirable activity towards several known breast cancer genes. This reflects that the extracts may kill MBD-MB-231 breast cancer cells. This is the first screening of plant extracts with high efficiency in 2 decades, showing promising results for future development of novel cancer treatments.

Six sartan substances including telmisartan, losartan, valsartan, irbesartan, olmesartan, and candesartan were selected in this work. A simple and sensitive liquid chromatography–tandem mass spectrometry method was developed and validated to simutaniously detect six drugs. Linearity, LOD, and LOQ were satisfied for routine quantitative and qualitative detection. The linearity ranged from 2 to 1000 ng/mL with the determination coefficient more than 0.98. The intra-day and inter-day precisions were all below 27.5%. All the recoveries were above 80%, and matrix effects were between 25 and 120%. Profiling detection of six sartan substances in human urine was also developed after administration. Irbesartan urinary metabolic pathway was firstly investigated by liquid chromatography quadruple time-of-flight mass spectrometry using full scan and targeted tandem mass spectrometry techniques. Fifteen urinary metabolites of irbesartan were identified and characterized, and ten of them were firstly reported. [ABSTRACT FROM AUTHOR]

Foreign amylase addition to honey in an effort to disguise diastase activity has become a widespread form of food fraud. However, since there is no report on the investigation in Japan, we investigated foreign amylases in 67 commercial honeys in Japan. First, the α-glucosidase and diastase activities of honeys were measured, which revealed that only α-glucosidase activity was significantly low in several samples. As both enzymes are secreted from honeybee glands, it is unlikely that only one enzyme was inactivated during processing. Therefore, we suspected the presence of foreign amylase. α-Amylase in honey were assigned using protein analysis software based on LC-QTOF-MS. As a result, α-amylases from Aspergillus and Geobacillus were detected in 13 and 6 out of 67 honeys, respectively. To detect foreign amylases easily, we developed a cost-effective method using native PAGE. Conventional native PAGE failed to separate the α-amylase derived from honeybee and Geobacillus. However, when native PAGE was performed using a gel containing 1 % maltodextrin, the α-amylase from honeybee did not migrated in the gel and the α-amylase could be separated from the other two α-amylases. The results from this method were consistent with those of LC-QTOF-MS method, suggesting that the novel native PAGE method can be used to detect foreign amylases. [ABSTRACT FROM AUTHOR]

Tri-Yannarose is a Thai traditional herbal medicine formula composed of Areca catechu, Azadirachta indica, and Tinospora crispa. It possesses antipyretic, diuretic, expectorant, and appetite-stimulating effects. This study aimed to evaluate the antioxidant activities, cytotoxicity, and chemical constituents of an aqueous extract following a Tri-Yannarose recipe and its plant ingredients. The phytochemical analysis was performed using LC-QTOF-MS. Antioxidant activities were determined using DPPH, ABTS, TPC, TFC, FRAP, NBT, MCA, and ORAC assays. Cytotoxicity was investigated using a methyl thiazol tetrazolium (MTT) assay. In addition, the relationship between the chemical composition of Tri-Yannarose and antioxidant activities was investigated by examining the structure–activity relationship (SAR). The results of the LC-QTOF-MS analysis revealed trigonelline, succinic acid, citric acid, and other chemical constituents. The aqueous extract of the recipe showed significant scavenging effects against ABTS and DPPH radicals, with IC50 values of 1054.843 ± 151.330 and 747.210 ± 44.173 µg/mL, respectively. The TPC of the recipe was 92.685 mg of gallic acid equivalent/g of extract and the TFC was 14.160 mg of catechin equivalent/g of extract. All extracts demonstrated lower toxicity in the Vero cell line according to the MTT assay. In addition, the SAR analysis indicated that prenyl arabinosyl-(1–6)-glucoside and quinic acid were the primary antioxidant compounds in the Tri-Yannarose extract. In conclusion, this study demonstrates that Tri-Yannarose and its plant ingredients have potent antioxidant activities with low toxicity. These results support the application of the Tri-Yannarose recipe for the management of a range of disorders related to oxidative stress. [ABSTRACT FROM AUTHOR]

To analyze the amino acid differences in the metabolism of the fruiting body and mycelium of Taiwanofungus camphoratus, the LC-QTOF-MS system spectrometry was used to analyze the metabolome of three groups of samples, including the fruiting body and mycelium of T. camphoratus, and the SIMCA software（v16.0.2）was used to screen and identify the main amino acid metabolites, seek important differential metabolites, analyze important metabolic pathways, and tap potential markers for diversity analysis. Principal component analysis（PCA）and orthogonal projections to latent structures- discriminant analysis（OPLS-DA）among the samples demonstrated that there were differences among samples under negative ion conditions. Under the condition of t-test P < 0.05 and VIP > 1, 288 metabolites were significantly different between the fruiting body（ACFB）and the mycelium cultured in liquid for 14 d（ACM-14）, 15 of them were the main amino acids. There were 332 significantly different metabolites between in ACFB and the mycelium cultured in liquid for 28 d (ACM-28), 11 of them were the main amino acids. There were 283 significantly different metabolites between ACM-14 and ACM-28, of which 14 were the main amino acids. The difference of amino acid metabolism between the fruiting body and mycelia of Taiwanofungus camphoratus was greater than that between the mycelia, especially the difference of L-arginine. All above results may provide a theoretical basis for studying the differences of amino acid metabolism in fruiting body and mycelium of T. camphoratus, mining potential marker metabolites and developing products. [ABSTRACT FROM AUTHOR]

Recent research reported that lurasidone degrades in unpreserved ante‐mortem human whole blood inoculated with microorganisms known to dominate postmortem blood specimens. In vitro degradation occurred at a similar rate to risperidone, known to degrade in authentic postmortem specimens until below analytical detection limits. To identify the lurasidone degradation products formed, an Agilent 6520 liquid chromatograph quadrupole‐time‐of‐flight mass spectrometer (LC‐QTOF‐MS) operating in auto‐MS/MS mode was used. Numerous degradation products not previously reported in prior in vitro or in vivo pharmacokinetic studies or forced degradation studies were detected. Accurate mass data, mass fragmentation data, acetylation experiments, and a proposed mechanism of degradation analogous to risperidone supports initial identification of the major degradation product as N‐debenzisothiazole‐lurasidone (calculated m/z [M + H]+ = 360.2646). A standard was unavailable to conclusively confirm this identification. Retrospective data analysis of postmortem cases involving lurasidone identified the presence of the major degradation product in four of six cases where lurasidone was also detected. This finding is significant for toxicology laboratories screening for this drug in postmortem casework. The major postmortem lurasidone degradation product has consequently been added to the LC‐QTOF‐MS drug screen at Forensic Science SA (FSSA) to indicate postmortem lurasidone degradation in authentic postmortem blood specimens and as a marker of lurasidone administration in the event lurasidone is degraded to concentrations below detection limits. [ABSTRACT FROM AUTHOR]

Two filamentous endophytic ascomycete fungi with different phenotypes were isolated and characterized from the vegetative tissue of Rheum spiciforme, a threatened and endemic trans-Himalayan medicinal herb. The visible morphological characteristics and microscopic features allowed the recognition of the endophytes as Fusarium proliferatum and Aureobasidium pullulans. Molecular identification involved partial DNA sequence analyses of internal transcribed spacer (ITS), elongation factor 1α (EF-1α), and calmodulin (CAL) regions of F. proliferatum DNA, and ITS and large subunit (LSU) of the nuclear ribosomal DNA (rDNA) of A. pullulans. The respective phylogenetic analyses by the Maximum Likelihood Method based on the combined ITS-EF-1α-CAL and ITS-LSU sequences showed that the former isolate is closest to F. proliferatum and the latter to A. pullulans. Moreover, using the LC-QTOF-MS technique, selected populations of R. spiciforme (one population with endophytes and another showing no apparent colonization), as well as the isolated endophytes, were evaluated for various specialized chemical constituents belonging to anthraquinone (rhein, physcion, emodin, and aloe-emodin), stilbenoid (piceatannol and resveratrol), and flavonoid (rutin, quercetin, and naringenin) class of compounds. Except for rutin, the results demonstrated that the identified endophytes possibly play an important role in sharing the metabolite synthesis machinery of the host. The current study may contribute to a better comprehension of the host-endophyte relation and open the door to the cost-effective synthesis of highly-valuable bioactive chemicals from microbial sources. [ABSTRACT FROM AUTHOR]

The presence of 220 emerging contaminants belonging to different classes (artificial sweeteners, personal care products, coffee and tobacco-related compounds, and industrial chemicals) was investigated in hospital wastewater for the first time. Twenty samples were collected within two sampling periods from two points of a Greek General Hospital. Target compounds were analyzed using a solid-phase extraction protocol followed by UHPLC-ESI-QToF-MS analysis. Analytical results showed that 23 micropollutants were detected at least once in hospital wastewater samples in Period 1, while 27 compounds were detected at least once in Period 2. The coffee and tobacco-related compounds were the most frequently detected substances, followed by artificial sweeteners, parabens, and industrial chemicals. The highest mean concentrations were recorded for the artificial sweeteners cyclamic acid (377 μg/L) and saccharine (295 μg/L), followed by caffeine (193 μg/L), nicotine (162 μg/L), and the industrial chemical lauryl diethanolamide (153 μg/L). The group of artificial sweeteners contributed up to 55.1% (Point A/Period 1) to the total concentration of studied chemicals. The detection of high concentrations of artificial sweeteners in hospital effluents reveals that hospitals should be considered as important point-sources of these contaminants. [ABSTRACT FROM AUTHOR]

TOPSIS, known as Technique for Order of Preference by Similarity to Ideal Solution, is an example of a multi-criteria decision analysis (MCDA) tools useful for decision making for ranking or selecting alternatives based on multiple criteria. In this study, rankings of dispersive-extraction solvents used in DLLME for the extraction of mifepristone in tap water samples was performed to assess their greenness. Two TOPSIS methods based on the objective mean weight (TOPSIS-MW) and entropy (TOPSIS-Entropy) weighting methods were proposed. Criteria were divided into three main categories describing different analytical perspectives: analytical performance, solvents greenness, and combination of analytical performance and solvent greenness. The properties and safety data of the solvents, mean detector response and coefficient of variance were considered as attributes to rank the greener solvent pairs from the combination of sixteen different extraction and disperser solvents as alternatives. The best overall analytical performance for mifepristone was for dichloroethane-acetone and chlorobenzene-tetrahydrofuran pair of solvents, weighted by mean and entropy weights, respectively. The assessment of solvents greenness showed that chloroform-acetone was the most favourable option in terms of green chemistry. The first rank in the combination of the two group of assessment criteria was also achieved by chloroform-acetone solvents. The performance of TOPSIS-MW and TOPSIS-Entropy was evaluated against that of simple additive weighting (SAW) weighted by mean and entropy in terms of the performance of LC-QTOF-MS. The methods were found to be closely correlated with some leading precisely to similar ranking observations.

A systematic study was performed into the degradation of ziprasidone in simulated postmortem blood. Fifteen potential degradation products not previously reported in the literature were observed. Four resulted from degradation in human blood, whereas the remaining products resulted from reaction with solvents: four from alkaline degradation, four from reaction with acetaldehyde, and three from reaction with acetone. To identify possible degradation products, a liquid chromatograph‐diode array detector (LC‐DAD) and liquid chromatograph quadrupole‐time‐of‐flight mass spectrometer (LC‐QTOF‐MS) operating in auto‐MS/MS mode were used. It was indicated from red‐shifted UV–Vis spectra, accurate mass data, mass fragmentation data, and a deuteration experiment that the site of ziprasidone degradation, in the in vitro blood experiments, was the methylene carbon of the oxindole moiety. The major in vitro blood degradation products were proposed to be E/Z isomers of 3‐ethylidene‐ziprasidone. Further, another in vitro degradation product in microbially inoculated blood specimens was proposed to be 3‐ethyl‐ziprasidone. 3‐Ethylidene‐ziprasidone was hypothesized to form from the reaction of ziprasidone with acetaldehyde derived from the ethanol used to spike ziprasidone into the in vitro blood experiments. Data from two postmortem investigations were available for retrospective reanalysis. Attempts were made to detect degradation products of ziprasidone, but none were found. [ABSTRACT FROM AUTHOR]

• Bidens pilosa plant contains three HQT genes and one HCT gene responsible for chlorogenic acids biosynthesis. • The SMRT sequencing approach allowed for full length transcripts of HQT/HCT genes. • Bidens pilosa produce chlorogenic acids of differing structural hierarchies. • The HQT.HCT genes from B. pilosa shares sequence homology with other orthologues from the Compositae family. Bidens pilosa L. is an underutilised plant that serves as an ingredient for traditional herbal medicine and as food. This plant has received considerable attention because of its chemistry, which comprises of chemicals from various classes, with those from the phenylpropanoid pathway (e.g., chlorogenic acids) existing in abundance. B. pilosa L. produces chemically diverse chlorogenic acids (CGAs), characterised mainly by isomers thereof. The enzymes involved in the biosynthesis of chlorogenic acids have been decoded and characterised in many plants. However, genes that play a role in the biosynthesis of chlorogenic acids in B. pilosa L. are not yet described and, as such, the aim of the current study was to identify different isoforms of the Hydroxycinnamoyl-CoA: quinate/shikimate acid hydroxycinnamoyl transferase (HQT/HCT) genes that play a role in the diversification of chlorogenic acids in Bidens pilosa L. Herein, a robust gene sequencing technology through Single Molecule Real Time (SMRT) approach was applied to establish the gene sequences encoding transferases responsible for CGA production in B. pilosa L. Sequence homology of the established genes was evaluated by means of multiple sequence alignment against already published orthologous genes from closely related plants. Thereafter, phylogeny trees were constructed to graphically display the taxonomical relationship with other chlorogenic acid producing plants. To further demonstrate the functionality of these genes in plants, chlorogenic acid profiling was carried out using Liquid chromatography quadrupole time of flight mass spectrometer (LC-QTOF-MS). B. pilosa L. plants produced isomeric forms of mono, di and tri-acylated CGAs. A total of three HQT genes and one HCT gene have been identified in this study and have been shown through homology to be related with other related genes from other CGAs producing plants. Future work should involve expression of the identified genes in other non-chlorogenic acid producing plants to enhance their nutraceutical value. [ABSTRACT FROM AUTHOR]

Novel synthetic opioid (NSO) continue to emerge in the United States in the midst of an opioid crisis. The NSO 2F‐viminol was identified in casework at the Center for Forensic Science Research and Education through its NPS Discovery program in 2019. Little information and published literature were available for this new opioid at the time. To address this, human liver microsomes (HLMs) were used to perform in vitro metabolism studies with a drug standard. The goal was to predict in vivo metabolism. Experimental samples were prepared using HLMs, NADPH, phosphate buffer (pH 7.4), and a 2F‐viminol standard. Standard samples were prepared containing only drug, control samples were prepared with drug and HLMs but no NADPH cofactor, and metabolism reaction mixtures contained drug, HLMs and NADPH. The subsequent mixtures were incubated with light shaking to allow metabolism to occur. After cleanup, metabolite mixtures were analyzed via a SCIEX TripleTOF 5600+ liquid chromatograph quadrupole‐time‐of‐flight mass spectrometer (LC‐QTOF‐MS). The generated metabolic structures were elucidated using SCIEX MetabolitePilot software (version 2.0). In addition to remaining parent drug, seven metabolites of 2F‐viminol were discovered, including N‐dealkylated and hydroxylated species. The proposed primary metabolites of 2F‐viminol were N‐dealkylation (sec‐butyl) + hydroxylation and N‐dealkylation (sec‐butyl); however, they should be confirmed in authentic samples, and forensic laboratories should consider adding 2F‐viminol and its metabolites to screening protocols to help in extending the window of detection for the parent drug in toxicological samples. As NSOs continue to appear, forensic laboratories must continue metabolism experiments to generate information about pharmacokinetics. [ABSTRACT FROM AUTHOR]

The purpose of this study is to investigate the difference of in vitro–in vivo correlation of α-amanitin from clearance perspectives as well as to explore the possibility of extra-hepatic metabolism of α-amanitin. First, a liquid chromatography-quadrupole-time-of-flight-mass spectrometric (LC-qTOF-MS) method for α-amanitin in rat plasma was developed and applied to evaluate the in vitro liver microsomal metabolic stability using rat and human liver microsomes and the pharmacokinetics of α-amanitin in rat. The predicted hepatic clearance of α-amanitin in rat liver microsomes was quite low (5.05 mL/min/kg), whereas its in vivo clearance in rat (14.0 mL/min/kg) was close to the borderline between low and moderate clearance. To find out the difference between in vitro and in vivo metabolism, in vitro and in vivo metabolite identification was also conducted. No significant metabolites were identified from the in vivo rat plasma and the major circulating entity in rat plasma was α-amanitin itself. No reactive metabolites such as GSH-adducts were detected either. A glucuronide metabolite was newly identified from the in vitro liver microsomes samples with a trace level. A semi-mass balance study was also conducted to understand the in vivo elimination pathway of α-amanitin and it showed that most α-amanitin was mainly eliminated in urine as intact which implies some unknown transporters in kidney might play a role in the elimination of α-amanitin in rat in vivo. Further studies with transporters in the kidney would be warranted to figure out the in vivo clearance mechanism of α-amanitin. [ABSTRACT FROM AUTHOR]

A high-throughput screening and quantification method for 479 pesticides in green tea was established based on solid-phase extraction combined with liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (LC-QTOF-MS). Pesticides were extracted from samples using an optimized SPE (TPT cartridges) procedure. LC-QTOF-MS in All Ions MS/MS scan mode acquired full MS data for quantification and product ion spectra for identification. LC-QTOF-MS quantification was achieved using matrix-matched standard calibration curves to achieve the optimal method accuracy. The method performance characteristics included the linearity, overall recovery, precision, and measurement uncertainty being evaluated. The validation results exhibited a good sensitivity with the LOQs of 5–55 µg/kg, which was satisfactory for their MRLs in China or the EU. The recoveries of more than 92.7% of the 479 pesticides in green tea were 70–120% at the three spiked levels with a precision of ≤20%. Finally, this method was employed to analyze 479 pesticides in 95 tea samples from markets in China. The test results of the tea samples showed that tolfenpyrad, buprofezin, and pyridaben were found with lower concentrations. The method has effectively improved the determination efficiency of pesticide residue screening by high-resolution mass spectrometry in green tea.

In the present study, the bioactive substance geraniol was tested in vitro and in planta against B. cinerea on cucumber plants, and the changes in the metabolic profile of cucumber plants inoculated with the pathogen and/or treated with geraniol were monitored by a novel LC-QTOF-MS method employing target and suspect screening. The aforementioned treatments were also studied for their impact on membrane lipid peroxidation calculated as malondialdehyde (MDA) content. Additionally, geraniol-loaded nanoemulsions (GNEs) were synthesized and tested against B. cinerea as an integrated formulation mode of geraniol application. The EC50 values calculated for geraniol and GNEs against B. cinerea were calculated at 235 μg/mL and 105 μg/mL, respectively. The in planta experiment on cucumber plants demonstrated the ability of geraniol and GNEs to significantly inhibit B. cinerea under greenhouse conditions. The LC-QTOF-MS analysis of the metabolic profile of the cucumber plants treated with geraniol demonstrated an increase in the concentration levels of myricetin, chlorogenic acid, and kaempferol rhamnoside, as compared to control plants and the presence of B. cinerea caused an increase in sinapic acid and genistein. These compounds are part of important biosynthetic pathways mostly related to responses against a pathogen attack. [ABSTRACT FROM AUTHOR]

The combination of high-resolution LC-MS untargeted metabolomics with stable isotope-resolved tracing is a promising approach for the global exploration of metabolic pathway activities. In our established workflow we combine targeted isotopologue feature extraction with the non-targeted X13CMS routine. Metabolites, detected by X13CMS as differentially labeled between two biological conditions are subsequently integrated into the original targeted library. This strategy enables monitoring of changes in known pathways as well as the discovery of hitherto unknown metabolic alterations. Here, we demonstrate this workflow in a PTEN (phosphatase and tensin homolog) null breast cancer cell line (MDA-MB-468) exploring metabolic pathway activities in the absence and presence of the selective PI3Kβ inhibitor AZD8186. Cells were fed with [U-13C] glucose and treated for 1, 3, 6, and 24 h with 0.5 µM AZD8186 or vehicle, extracted by an optimized sample preparation protocol and analyzed by LC-QTOF-MS. Untargeted differential tracing of labels revealed 286 isotope-enriched features that were significantly altered between control and treatment conditions, of which 19 features could be attributed to known compounds from targeted pathways. Other 11 features were unambiguously identified based on data-dependent MS/MS spectra and reference substances. Notably, only a minority of the significantly altered features (11 and 16, respectively) were identified when preprocessing of the same data set (treatment vs. control in 24 h unlabeled samples) was performed with tools commonly used for label-free (i.e. w/o isotopic tracer) non-targeted metabolomics experiments (Profinder´s batch recursive feature extraction and XCMS). The structurally identified metabolites were integrated into the existing targeted isotopologue feature extraction workflow to enable natural abundance correction, evaluation of assay performance and assessment of drug-induced changes in pathway activities. Label incorporation was highly reproducible for the majority of isotopologues in technical replicates with a RSD below 10%. Furthermore, inter-day repeatability of a second label experiment showed strong correlation (Pearson R2 > 0.99) between tracer incorporation on different days. Finally, we could identify prominent pathway activity alterations upon PI3Kβ inhibition. Besides pathways in central metabolism, known to be changed our workflow revealed additional pathways, like pyrimidine metabolism or hexosamine pathway. All pathways identified represent key metabolic processes associated with cancer metabolism and therapy.

Tri-Yannarose is a Thai traditional herbal medicine formula composed of Areca catechu, Azadirachta indica, and Tinospora crispa. It possesses antipyretic, diuretic, expectorant, and appetite-stimulating effects. This study aimed to evaluate the antioxidant activities, cytotoxicity, and chemical constituents of an aqueous extract following a Tri-Yannarose recipe and its plant ingredients. The phytochemical analysis was performed using LC-QTOF-MS. Antioxidant activities were determined using DPPH, ABTS, TPC, TFC, FRAP, NBT, MCA, and ORAC assays. Cytotoxicity was investigated using a methyl thiazol tetrazolium (MTT) assay. In addition, the relationship between the chemical composition of Tri-Yannarose and antioxidant activities was investigated by examining the structure–activity relationship (SAR). The results of the LC-QTOF-MS analysis revealed trigonelline, succinic acid, citric acid, and other chemical constituents. The aqueous extract of the recipe showed significant scavenging effects against ABTS and DPPH radicals, with IC50 values of 1054.843 ± 151.330 and 747.210 ± 44.173 µg/mL, respectively. The TPC of the recipe was 92.685 mg of gallic acid equivalent/g of extract and the TFC was 14.160 mg of catechin equivalent/g of extract. All extracts demonstrated lower toxicity in the Vero cell line according to the MTT assay. In addition, the SAR analysis indicated that prenyl arabinosyl-(1–6)-glucoside and quinic acid were the primary antioxidant compounds in the Tri-Yannarose extract. In conclusion, this study demonstrates that Tri-Yannarose and its plant ingredients have potent antioxidant activities with low toxicity. These results support the application of the Tri-Yannarose recipe for the management of a range of disorders related to oxidative stress.

The Smilacaceae family has been used as a food source and herbal medicine for a long time. This study aims to identify the phytochemicals extracted from Smilax glabra and Smilax corbularia by using LC-QTOF/MS analysis and determine their bioactive potential. Compounds were identified from S. glabra and S. corbularia extracts by LC–QTOF-MS and it was found that longistylin A and CAY10435 have higher degrees of matching compounds (99.66% and 99.87%). Smilax glabra showed antioxidant capacity, i.e., DPPH and ABTS at percentage inhibitions of 71.94 ± 1.46% and 59.84 ± 4.80%, respectively, and FRAP at 730.69 ± 33.62 mg AAE/100 g sample. The total phenolic compound contents of the ethanol, methanol, and water extracts were 0.017 ± 0.001, 0.015 ± 0.001, and 0.016 ± 0.001 mg GAE/g, respectively, while the total flavonoid contents were 0.043 ± 0.002, 0.033 ± 0.002, and 0.006 ± 0.003 mg QE/g, respectively. The anti-inflammatory capacity showed 97.26% protection and 2.74% hemolysis. The antimicrobial activity can inhibit Gram-positive bacteria with a minimum inhibitory concentration (MIC) of 62.5 mg/mL and a minimum bactericidal concentration (MBC) of 500 mg/mL. Smilax corbularia showed antioxidant capacity, i.e., DPPH and ABTS at percentage inhibitions of 72.24 ± 0.64% and 39.87 ± 2.37%, respectively, and FRAP at 208.33 ± 50.80 mg AAE/100 g sample. The total phenolic compound contents of the ethanol, methanol, and water extracts were 0.006 ± 0.000, 0.007 ± 0.002, and 0.002 ± 0.001 mg GAE/g, respectively, while the total flavonoid contents of the ethanol and methanol extracts were 0.012 ± 0.001 and 0.008 ± 0.000 mg QE/g, respectively. The anti-inflammatory capacity showed 96.64% protection and 3.36% hemolysis. The antimicrobial activity of the extracts can inhibit Gram-positive bacteria with a MIC of 31.25 mg/mL and MBC of 125 mg/mL for the ethanol extract and a MIC of 125 mg/mL and MBC of 62.5 mg/mL for the methanol extract. In conclusion, Smilax glabra and Smilax corbularia were found to contain several phytochemicals that can be used for further study. Both Smilax species can also be used as sources of antioxidants and herbal medicines for killing Gram-positive bacteria. [ABSTRACT FROM AUTHOR]

Epaltes mexicana is a plant widely used in traditional medicine and as a food in Mexico; however, its phytochemical and pharmacological studies are limited. This study aimed to identify the active secondary metabolites of Epaltes mexicana and determine its cytotoxic activity on cancer cell lines. Three organic extracts were obtained by maceration using n -hexane, dichloromethane, and methanol. The n -hexane extract was fractioned by simple column chromatography. Eight terpenes were annotated in collection 6 (C6) by LC-QTOF-MS using a gradient elution and Electrospray Ionization (ESI) in positive ion mode: 1) Gibberellin A15, 2) farfugin A, 3) dehydromyodesmone, 4) eremopetasitenin A1, 5) hydroxyisonobilin, 6) anhydrocinnzeylanine, 7) nigakilactone H and 8) taxodione. On the other hand, C6 showed a concentration-dependent cytotoxic effect on cancer cell lines MCF-7 (E max = 74.69 ± 6.19 % and IC 50 = 6.31 μg/mL), MDA-MB-231 (E max = 79.28 ± 12.12 % and IC 50 = 124.21 μg/mL), and SiHa (E max = 82.96 ± 6.02 % and IC 50 = 124.31 μg/mL). The C6 did not show a cytotoxic effect against DU-145 and non-cancerous cells from the mammary glands MCF-10A. These results indicate cytotoxic specificity on cancer cell lines and support the hypothesis that terpenes identified in E. mexicana must be investigated and developed for non-clinical and clinical trials as potential anti-cancer drugs. [Display omitted] • Eight terpenes were annotated in the active collection of Epaltes mexicana. • Eight terpenes were found in positive ion mode by LC-QTOF-MS analysis. • The terpenes showed cytotoxic activity on MCF-7, MDA-MB-231 and SIHa. • E. mexicana showed selective cytotoxic activity in breast and cervical cancer cell lines. [ABSTRACT FROM AUTHOR]

[Display omitted] • A LC-MS/MS method with microscale palytoxin oxidation was successfully validated. • High instrumental sensitivity and extract stability was achieved. • The quantitative method includes six palitoxin-like molecules in its scope. • In silico prediction provided solubility information about the main analytes. • Palytoxin ions were detected and confirmed in Haff disease-related samples. Haff disease typically develops after eating contaminated marine or freshwater species, especially fish. Despite still having an unknown etiology, recent reports have suggested its possible correlation with palytoxins. Therefore, the present work aimed to optimize and perform a validation of a sensitive method using liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) for the analysis of palytoxin and some of its analogs, with the main purpose of investigating their presence in marine and freshwater food samples associated with Haff disease in Brazil. The method optimization was performed using a central composite rotatable design and fish samples fortified with the palytoxin standard. Then, the optimized method was validated for different food matrices, including freshwater and marine fish, mollusks, and crustaceans. The sample preparation involved a solid–liquid extraction using methanol and water, solid-phase extraction using Strata-X cartridges, and on-column palytoxin oxidation. The detection of the main oxidized fragments (amino and amide aldehydes) was achieved by LC-MS/MS with electrospray ionization in positive mode, using a C18 column, as well as acetonitrile and water as mobile phases, both acidified with 0.1 % of formic acid. After optimization and validation, the etiological investigation involved the analysis of 16 Brazilian Haff disease-related food samples (in natura and leftover meals) from 2022. The method was demonstrated to be appropriate for quantitative analysis of freshwater and marine species. So far, it has proven to be one of the most sensitive methods related to palytoxin detection (LOD 10 μg/kg), being able to work in a range that includes the provisional ingestion limit (30 μg/kg). Regarding the Haff disease-related samples analysis, there is a strong indication of palytoxin contamination since the amino aldehyde (common fragment for all palytoxins) was detected in 15 of the 16 samples. Selected results were confirmed using liquid chromatography coupled with high-resolution mass spectrometry (LC-HRMS). [ABSTRACT FROM AUTHOR]

[Display omitted] • A new LC-QTOF-MS method for 4 PDE5Is was developed and validated. • 49 food supplements for enhancing sexual performance were analyzed. • 65% of samples contained PDE5Is and their structural analogs. • 2 PDE5Is and 8 PDE5Is analogs were identified in food supplements. • Undeclared PDE5Is and their analogs in food supplements pose risks to human health. This paper presents the development and validation of an analytical method using liquid chromatography-time of flight mass spectrometry (LC-QTOF-MS) suitable for screening and identifying suspect PDE5I analogs and quantifying four target PDE5Is in various food supplements. The method validation comprised of selectivity, linearity, accuracy, the limit of detection (LOD) and quantification (LOQ), and evaluation of matrix effects. The method showed good linearity with coefficients of determination higher than 0.992 for all analytes. LODs and LOQs ranged from 0.2 to 2.2 ng/mL and 2 to 20 ng/mL, respectively. The accuracy and precision of the method were deemed satisfactory, with values ranging from 88.5 to 115.9 % for intra- and 86.9 to 111.2 % for inter-day experiments, and RSDs ranged from 0.4 to 4.5 % and 0.9 to 5.2 %, respectively. Out of 49 analyzed food supplements collected on the Croatian market, 65 % were not in compliance with EU regulations and pose possible risks to human health. [ABSTRACT FROM AUTHOR]

Pitongshu (PTS) is a clinically effective empirical formula for the treatment of FD. The efficacy and safety of PTS have been demonstrated in randomized, controlled, double-blind trials, but there is a lack of understanding of the systematic evaluation of the efficacy of PTS and its material basis. To investigate the efficacy of PTS in Functional dyspepsia (FD) mice and possible Q-markers. In this study, we used "irregular feeding + chronic unpredictable chronic stimulation" to establish a mice model of FD with hepatogastric disharmony. The efficacy of PTS was assessed from hair condition, behavioral, pain, gastrointestinal function, and serum 5-HT, GAS, MTL levels in mice by instillation of different doses of PTS. In addition, the composition of drugs in blood was analyzed by LC-QTOF-MS and potential Q-markers were selected by combining network pharmacology, molecular docking and actual content. Our study showed that different doses of PTS increased pain threshold and writhing latency, decreased the number of writhings, increased gastric emptying rate and small intestinal propulsion rate, decreased total acidity of gastric contents and gastric acid secretion, and increased serum levels of 5-HT, GAS, and MTL in mice to different degrees. Enrichment analysis showed that PTS may be anti-FD through multiple pathways such as Serotonergic synapse, thyroid hormone signaling pathway, cholinergic synapse, and dopaminergic synapse. In addition, potential active ingredient substances were explored by LC-QTOF-MS combined with bioinformatics. Combined with the actual contentselected six constituents, hesperidin, neohesperidin, naringin, paeoniflorin, magnolol and honokiol, possible as Q-markers. PTS may exert its anti-FD effects through multi-component, multi-target and multi-pathway". Constituents, hesperidin, neohesperidin, naringin, paeoniflorin, magnolol and honokiol may be the Q-markers of its anti-FD effects. [Display omitted] [ABSTRACT FROM AUTHOR]

The Echinococcus granulosus sensu lato species complex is responsible for the neglected zoonotic disease known as cystic echinococcosis (CE). Humans and livestock are infected via fecal-oral transmission. CE remains prevalent in Western China, Central Asia, South America, Eastern Africa, and the Mediterranean. Approximately one million individuals worldwide are affected, influencing veterinary and public health, as well as social and economic matters. The infection causes slow-growing cysts, predominantly in the liver and lungs, but can also develop in other organs. The exact progression of these cysts is uncertain. This study aimed to understand the survival mechanisms of liver and lung CE cysts from cattle by determining their metabolite profiles through metabolomics and multivariate statistical analyses. Non-targeted metabolomic approaches were conducted using quadrupole-time-of-flight liquid chromatography/mass spectrometry (LC-QTOF-MS) to distinguish between liver and lung CE cysts. Data processing to extract the peaks on complex chromatograms was performed using XCMS. PCA and OPLS-DA plots obtained through multiple statistical analyses showed interactions of metabolites within and between groups. Metabolites such as glutathione, prostaglandin, folic acid, and cortisol that cause different immunological reactions have been identified both in liver and lung hydatid cysts, but in different ratios. Considering the differences in the metabolomic profiles of the liver and lung cysts determined in the present study will contribute research to enlighten the nature of the cyst and develop specific therapeutic strategies. [Display omitted] [ABSTRACT FROM AUTHOR]

Daporinad (FK866) is one of the highly specific inhibitors of nicotinamide phosphoribosyl transferase (NAMPT) and known to have its unique mechanism of action that induces the tumor cell apoptosis. In this study, a simple and sensitive liquid chromatography–quadrupole-time-of-flight–mass spectrometric (LC-qTOF-MS) assay has been developed for the evaluation of drug metabolism and pharmacokinetics (DMPK) properties of Daporinad in mice. A simple protein precipitation method using acetonitrile (ACN) was used for the sample preparation and the pre-treated samples were separated by a C18 column. The calibration curve was evaluated in the range of 1.02~2220 ng/mL and the quadratic regression (weighted 1/concentration2) was used for the best fit of the curve with a correlation coefficient ≥ 0.99. The qualification run met the acceptance criteria of ±25% accuracy and precision values for QC samples. The dilution integrity was verified for 5, 10 and 30-fold dilution and the accuracy and precision of the dilution QC samples were also satisfactory within ±25% of the nominal values. The stability results indicated that Daporinad was stable for the following conditions: short-term (4 h), long-term (2 weeks), freeze/thaw (three cycles). This qualified method was successfully applied to intravenous (IV) pharmacokinetic (PK) studies of Daporinad in mice at doses of 5, 10 and 30 mg/kg. As a result, it showed a linear PK tendency in the dose range from 5 to 10 mg/kg, but a non-linear PK tendency in the dose of 30 mg/kg. In addition, in vitro and in vivo metabolite identification (Met ID) studies were conducted to understand the PK properties of Daporinad and the results showed that a total of 25 metabolites were identified as ten different types of metabolism in our experimental conditions. In conclusion, the LC-qTOF-MS assay was successfully developed for the quantification of Daporinad in mouse plasma as well as for its in vitro and in vivo metabolite identification. [ABSTRACT FROM AUTHOR]

This study presents the immunomodulatory potential of Abrus precatorius Linn. (Indian wild licorice) leaves. A hydroalcoholic extract of A. precatorius leaves (EAPL) was prepared by maceration. Thirty male mice were divided into five groups as follows: control group, model control group (cyclophosphamide-treated), and three treatment groups (treated with EAPL at doses of 100, 200 and 300 mg/kg, per os, daily for 14 days). Parameters, including hematological, biochemical, organ indices, hemagglutination test (HA titer), delayed-type hypersensitivity (DTH), interleukin-2 (IL-2) level, splenocyte proliferation and liquid chromatography quadrupole time-of-flight mass spectrometry (LC-QTOF-MS) analysis were evaluated. Histopathological examination was carried out for the spleen, kidney and liver. Cyclophosphamide (CPMD)-induced changes in white blood cells, lymphocytes and platelets were improved in the treatment groups. Total protein and albumin levels in the treatment groups were significantly higher. EAPL treatment significantly stimulated splenic T-lymphocytemediated proliferation. Neutrophil adhesion was insignificantly decreased in the model control group compared to the normal control group, which was slightly improved by EAPL treatment. EAPL treatment significantly improved the HA titer and cell-mediated immunity, which is an indication of antibody production. The IL-2 level was significantly higher in the treatment groups that received 200 and 300 mg/kg EAPL. LC-QTOF-MS analysis of EAPL showed the presence of flavonoids, lignans, iridoids and phenolic glycosides. These results suggest that A. precatorius leaves are a good candidate for a new immunomodulatory herbal formulation.

High resolution mass spectrometry (HR-MS) offers the opportunity to track large numbers of non-target analytes through water treatment processes, providing a more comprehensive view of reactor performance than targeted evaluation. Both approaches were used to evaluate the performance of a pilot scale advanced oxidation process (AOP) employing ultraviolet light and hydrogen peroxide (UV/H2O2) to treat municipal wastewater effluent. Twelve pharmaceuticals and personal care products were selected as target compounds and added to reactor influent. Target compound removal over a range of flow rates and hydrogen peroxide addition levels was assessed using a liquid chromatograph combined with a quadrupole time-of-flight mass spectrometer (LC-qTOF-MS). Target compound removals were used to determine hydroxyl radical concentrations and UV fluence under pilot scale conditions. The experiments were also analyzed using a nontarget approach, which identified "molecular features" in either reactor influent or effluent. Strong correlation (r = 0.94) was observed between target compound removals calculated using the targeted and non-targeted approaches across the range of reactor conditions tested. The two approaches also produced consistent rankings of the performance of the various reactor operating conditions, although the distribution of compound removal efficiencies was usually less favorable with the broader, nontarget approach. For example, in the UV only treatment 8.3% of target compounds and 2.2% of non-target compounds exhibited removals above 50%, while 100% of target compounds and 74% of non-target compounds exhibited removals above 50% in the best condition tested. These results suggest that HR-MS methods can provide more holistic evaluation of reactor performance, and may reduce biases caused by selection of a limited number of target compounds. HR-MS methods also offer insights into the composition of poorly removed compounds and the formation of transformation products, which were widely detected.

In this study, the antidiabetic, antiobesity, antioxidant, and antihyperlipidemic effects potential of Pistacia atlantica Desf. leaves were evaluated by in vitro methods. The effects of the leaves of the plant on pancreatic lipase, pancreatic cholesterol esterase, and PTP1B enzymes were investigated for the first time and it was observed that leaf methanol extract (IC50: 123.67 ± 0.40 μg/ml) and n -hexane sub-extract (IC50: 61.03 ± 0.11 μg/ml) had much stronger effects on pancreatic cholesterol esterase enzyme than simvastatin (IC50: 142.30 ± 5.67 μg/ml). The methanolic extract of P. atlantica leaves exerted strong inhibitory effect on the enzymes (α -amylase and α -glucosidase) effective on carbohydrate digestion. It was thought that the methanol extract could provide significant benefits against oxidative stress in diabetes mellitus since it showed antioxidant activities (DPPH radical scavenging activity and reducing power) as strong as reference compounds (ascorbic acid and quercetin). Qualitative and quantitative analyzes of rutin (0.328 ± 0.000 g/100 g dry extract), methyl gallate (5.245 ± 0.014 g/100 g dry extract), quercetin-3- O- glucoside (0.231 ± 0.000 g/100 g dry extract), and gallic acid (0.528 ± 0.127 g/100 g dry extract) in methanol extract were performed by RP-HPLC. The phytochemical content of the active sub-fraction obtained from the leaf methanol extract by activity-guided fractionation and column chromatography studies was characterized by LC-QTOF-MS. The presence of trigalloylglucose, digalloylglucose, and methyl gallate in the G6 coded sub-fraction obtained by chromatographic techniques from the ethyl acetate sub-extract, which has the highest inhibitory effect on α -amylase and α -glucosidase enzymes, was determined by LC-QTOF-MS. In addition to the G5 coded subfraction, a strong α -glucosidase enzyme inhibitory activity was also observed in the G6 coded sub-fraction, and methyl gallate, methyl digallate, 2″- O -galloyl-quercetin-3- O -hexoside, and myricetin-3- O -hexoside were identified in this sub-fraction. This study displayed that the methanol extract of P. atlantica leaves could be a potential source for bioactive compounds with antidiabetic effects by showing inhibitory effects on enzymes involved in carbohydrate digestion. [ABSTRACT FROM AUTHOR]

Plants produce thousands of compounds, collectively called the metabolome, which mediate interactions with other organisms. The metabolome of an individual plant may change according to the number and nature of these interactions. We tested the hypothesis that tree diversity level affects the metabolome of four subtropical tree species in a biodiversity–ecosystem functioning experiment, BEF‐China. We postulated that the chemical diversity of leaves, roots and root exudates increases with tree diversity. We expected that the strength of this diversity effect differs among leaf, root and root exudates samples. Considering their role in plant competition, we expected to find the strongest effects in root exudates.Roots, root exudates and leaves of four tree species (Cinnamomum camphora, Cyclobalanopsis glauca, Daphniphyllum oldhamii and Schima superba) were sampled from selected plots in BEF‐China. The exudate metabolomes were normalized over their non‐purgeable organic carbon level. Multivariate analyses were applied to identify the effect of both neighbouring (local) trees and plot diversity on tree metabolomes. The species‐ and sample‐specific metabolites were assigned to major compound classes using the ClassyFire tool, whereas potential metabolites related to diversity effects were annotated manually.Individual tree species showed distinct leaf, root and root exudate metabolomes. The main compound class in leaves was the flavonoids, whereas carboxylic acids, prenol lipids and specific alkaloids were most prominent in root exudates and roots. Overall, plot diversity had a stronger effect on metabolome profiles than the local diversity. Leaf metabolomes responded more often to tree diversity level than exudates, whereas root metabolomes varied the least. We found no uniform or general pattern of alterations in metabolite richness or diversity in response to variation in tree diversity. The response differed among species and tissues.Synthesis. Classification of metabolites supported initial ecological interpretation of differences among species and organs. Particularly, the metabolomes of leaves and root exudates respond to differences in tree diversity. These responses were neither linear nor uniform and individual metabolites showed different dynamics. More controlled interaction experiments are needed to dissect the causes and consequences of the observed shifts in plant metabolomes. [ABSTRACT FROM AUTHOR]

Bacillus spp. have been used as a biocontrol strategy to eliminate/reduce toxic fungicides in viticulture. Furthermore, the presence of fungi that are resistant to commonly used products is frequent, highlighting the need for new biocontrol strains. Aspergillus carbonarius can produce ochratoxins, including ochratoxin A (OTA), which has a regulatory maximum allowable limit for grape products. The purpose of this study was to assess the ability of four Amazonian strains of Bacillus (P1, P7, P11, and P45) to biocontrol A. carbonarius and various forms of ochratoxins in grapes. Berries treated with strain P1 presented no fungal colonies (100% reduction), while P7, P11 and P45 strains caused a reduction of 95, 95 and 61% on fungal counts, respectively. Six forms of ochratoxin were found in the grapes inoculated with A. carbonarius, including ochratoxin α, ochratoxin β, ochratoxin α methyl-ester, ochratoxin α amide, N-formyl-ochratoxin α amide, and OTA. Four of these ochratoxin forms (ochratoxin β, ochratoxin α methyl-ester, ochratoxin α amide, N-formyl-ochratoxin α amide) are reported for the first time in grapes. These ochratoxins were identified using liquid chromatography coupled to quadrupole time-of-flight mass spectrometry (LC-QToF-MS). All Bacillus strains inhibited the synthesis of OTA, which is the most toxic form of ochratoxin. No ochratoxin form was found when P1 and P7 were used. Although some forms of ochratoxin were detected in grapes treated with Bacillus spp. P11 and P45, the levels decreased by 97%. To our knowledge, this is the first report on the inhibition of Aspergillus carbonarius-derived ochratoxin by Bacillus species. P1 strain, identified as Bacillus velezensis, was found to be the most promising for completely inhibiting fungal growth and production of all ochratoxins. [ABSTRACT FROM AUTHOR]

Abstract Background Ganoderma spore is a minuscule germ cell ejected from Ganoderma gills during its growth maturity period, it has been considered with high exploitable potential in health-care products manufacture. Methods After testing sporoderm-broken rate, the triterpenoids in 12 batches of broken and unbroken Ganoderma spore powder (GSP) samples were compared with Ganoderma lucidum fruiting body (GL) by high performance thin-layer chromatography (HPTLC) and further verified by liquid chromatography coupled with electrospray ionization quadrupole-time-of-flight mass spectrometry (LC–QTOF–MS); meanwhile, the dissolution of triterpenoids after bionic extraction was also investigated by HPTLC. Results The sporoderm-broken rate of all the broken GSP samples was over 85%. The relative peak area of triterpenoids in GSP samples were lower than 50% of that in fruiting body, and the dissolution of triterpenoids in artificial gastrointestinal fluid was lower than in methanol. Conclusions This study demonstrated that there were little triterpenoids in GSP. Triterpenoids in GSP also seldom be dissolved in artificial gastrointestinal fluid.