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The Papaveraceae plant family serves as a botanical reservoir for a variety of medicinal compounds that have been traditionally utilized in Chinese medicine for numerous generations. Growing attention towards the pharmaceutical potential of Papaveraceae has resulted in the identification of many alkaloids, which have attracted significant attention from the scientific community because of their structural complexity and wide range of biological activities, such as analgesic, antihypertensive, antiarrhythmic, anti-inflammatory, antibacterial, anti-tumor, anti-cancer, and other activities, making them potential candidates for medical use. The primary objective of this review is to analyze the existing literature on the historical use of Papaveraceae plants, focusing on their alkaloid structures and relationship with pharmacological effects, as well as provide a theoretical basis for their clinical application, with the goal of unveiling the future potential of Papaveraceae plants. [ABSTRACT FROM AUTHOR]

Rationale: The chemical composition of the new compound aloe capsule (NCAC) has not been fully elucidated to date, which poses challenges for pharmacology, scientific, and rational drug use. In this study, a rapid and sensitive method was established to comprehensively study the constituents in the NCAC. The contents of the NCAC were analyzed by UPLC-Q-Orbitrap-MS and data postprocessing technology., Methods: Firstly, the extract was separated by UPLC system, and the fragment information was obtained under positive and negative ions and then compared with the characteristic fragments described in the literature, so as to achieve the purpose of rapid identification of the compound of NCAC., Results: A total of 56 compounds including 21 chromones, 6 pyranones, 7 anthraquinones, 7 anthrone, 5 alkaloids, 4 amino acids, and 6 other components were detected in NCAC by comparing the retention time and mass spectrometry information and retrieving the reference literature., Conclusions: The UPLC-Q-Orbitrap-MS method was developed and utilized successfully to identify the major constituents in NCAC and would be helpful for improving the quality control standard of NCAC., (© 2024 John Wiley & Sons Ltd.)

This study aims to realise rapid detecting of tropane alkaloids (TAs) in food. For this purpose, a broad-spectrum single-chain fragment variable was fused with horseradish peroxidase to create an antibody-enzyme complex (AEC) with antigen recognition and catalytic activity. A multi-signal immunosensor platform based on AEC in the direct competitive reaction mode was constructed using 3,3',5,5'-tetramethylbenzidine and 10-acetyl-3,7-dihydroxyphenoxazine as substrates. The sensitivity of TAs in the immunosensor platform ranged from 0.25 μg/kg to 7912.46 μg/kg. Honey was selected as a representative food sample, and the limit of detection of TAs in honey ranged from 0.02 μg/kg to 409.11 μg/kg, with a recovery rate of 65.7 %-117.1 % and a coefficient of variation less than 21.4 %. Results showed that the immunosensor platform possesses satisfactory accuracy and precision, which highlights its potential for practical applications and its suitability as an ideal tool for rapid screening of TAs in food., Competing Interests: Declaration of competing interest The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Carpaine, a major alkaloid present in Carica papaya leaves, has been shown to increase platelet counts in patients suffering from thrombocytopenia. Numerous commercial papaya leaf products are available, but few provide proper bioactive ingredient information. We present herein a technique for rapid screening of carpaine in these products using DART-MS. The results indicate that carpaine was detected in various forms (powder, solution) of papaya leaves. Its presence was confirmed by examining the mass pattern when conducted on a standard solution at both low and high voltages (+10 V and +90 V), using MS 1 and MS 2 data obtained from LC-QTOF-MS/MS. The protonated molecule was identified at m/z = 479.38, with a fragment ion at m/z = 240.20. LOD for identifying carpaine in powder and solution matrices were 5.0 × 10 -5 %w/w and 0.05 μg/mL, respectively. The proposed method has been successfully validated with the AOAC International standards and can be used to identify carpaine in papaya leaf products., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Large-scale production of tobacco generates significant amounts of waste, making the reuse and recycling of tobacco waste crucial for sustainability. However, the high diversity of tobacco metabolites requires a comprehensive analysis of tobacco waste extracts, so as to optimize extraction selectivity and recovery. In this study, we thoroughly analyzed the metabolic composition of 168 tobacco extracts using nontarget analysis. The extracts were derived from 56 tobacco wastes covering all four major tobacco types: flue-cured, dark air-cured, Burley, and Oriental tobaccos. From each sample, we obtained three types of extracts, such as concretes, absolutes, and supercritical fluid extraction (SFE) oils, through various extraction protocols. By matching deconvolved experimental MS/MS spectra with databases, 587 metabolites were putatively annotated, which were categorized into 24 groups, including alkaloids, indoles, and flavor compounds. The metabolic compositions of extracts from dark air-cured and Burley tobaccos were found to be similar, whereas flue-cured and Oriental tobaccos contained more diverse flavor components. Notably, concretes of various tobaccos prepared with 70 %-ethanol contained similar amounts of addictive alkaloids. However, the absolutes obtained by sequential extraction of these concretes with 95 %-ethanol effectively removed alkaloids while modestly maintaining the flavor compounds. Conversely, extraction protocols incorporating supercritical fluid extraction followed by 95 %-ethanol extraction resulted in the loss of most flavor compounds, likely due to the incompatibility between the solvents used in the two extraction steps. In conclusion, our study demonstrates that absolutes derived through secondary extraction with varying ethanol-water ratios holds significant potential for utilization., Competing Interests: Declaration of competing interest The authors declare no conflicts of interest., (Copyright © 2024 Elsevier B.V. All rights reserved.)

A rapid and sensitive enrichment technique, field enhanced sample injection-dynamic pH junction-sweeping (FESI-DypH-sweeping) was successfully developed for the simultaneous separation and concentration of alkaloids and stereoisomers of Uncariae ramulus cum uncis (UR) by cyclodextrin electrokinetic chromatography (CDEKC) with diode array detection system. The sample was prepared in a low-conductivity (FESI), low-pH (DypH) sample matrix (4 mM phosphate buffer, 3% methanol, pH=3), and the background electrolyte (BGE) consisted of a high-conductivity, high-pH buffer (40 mM phosphate buffer, pH 7.0, 8 mg/mL carboxymethyl-β-cyclodextrin, and 30% methanol). The crucial parameters influencing separation and enrichment efficiency were systematically optimized using single variable method. Under the optimum conditions, the method provided satisfactory resolution for seven alkaloids within 18 minutes, achieving enrichment factors ranging from 25 to 78 folds. The limits of detections (LODs) ranged from 0.04 to 0.12 μg/mL and limits of quantifications (LOQs) ranged from 0.10 and 0.40 μg/mL respectively. Intra-day and inter-day precision, expressed as relative standard deviations, were all below 3.5%., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

The determination of isoquinoline alkaloids (IQAs) is crucial for maximizing the pharmacological benefits of Rhizoma coptidis (RC). In this study, the dendritic silica sphere (dSiO 2 ) was covalently modified with cyanuric chloride (CNCl) to synthesize a porous dNSiO 2 -CNCl NP. The fluorescence of berberine (Ber), palmatine (Pal), coptisine (Cop), and jatrorrhizine (Jat) was remarkably enhanced by dNSiO 2 -CNCl NP for at least 38-fold, inspiring the accurate determination of the isoquinoline total alkaloids (IQTAs) in RC using fluorometry. The method exhibited good linearity within the concentration range of 0.005 to 1 μM, with a limit of detection (LOD) and quantitation (LOQ) of 0.001 and 0.004 μM. Specially, Jat was discovered to form a stable covalent linkage with dNSiO 2 -CNCl, showing an innovative and enduring fluorescent nanoparticle derived from a natural product. Furthermore, using dNSiO 2 -CNCl as a substrate for fluorescence enhancement and a dispersive solid-phase extraction (d-SPE) sorbent, a fluorescence-mediated HPLV-UV method was developed to monitor Ber, Pal, and Cop in serum. Rapid qualitative analysis using fluorescence was initially performed. In chromatography analysis, the linear range for Cop was 0.20 to 10 μM, with LOD and LOQ at 0.035 and 0.110 μM, while Pal and Ber showed good linearity in the range of 0.05 to 10 μM, with LODs and LOQs at 0.013 and 0.040 μM. The spiked recovery rates ranged from 90.4 % to 99.8 % for the three IQAs, with intra-day precision below 7.9 % and inter-day precision below 9.9 %. The dual-functional dNSiO 2 -CNCl NP exhibited promising prospect in fluorescence sensing and separation applications for detecting drugs., Competing Interests: Declaration of competing interest We declare that we do not have any commercial or associative interest that represents a conflict of interest in connection with the manuscript submitted., (Copyright © 2024 Elsevier B.V. All rights reserved.)

The adulteration of honey is a globally growing issue due to its medicinal benefits and health-promoting properties. This problem primarily involves the addition of sugars and other substances. To address these concerns, a comparative study was conducted to investigate the dielectric and spectroscopic properties of pure, sugar solution added honey, and commercially available honey. Quantitative analysis was performed using a microwave X-band bench, while qualitative assessments were carried out using Fourier-transform infrared (FTIR), Ultraviolet (UV), and NMR spectroscopy. Dielectric properties, including dielectric constant, dielectric loss, and loss tangent, were measured at a constant frequency of 8.73 GHz under ambient conditions. Pure honey exhibits distinct dielectric properties due to its low moisture content, in comparison to adulterated and commercial honey. FTIR spectroscopy was used to identify various functional groups within the wavenumber range of 4000-400 cm⁻¹, allowing for the examination of glucose and fructose contents. Hydroxymethylfurfural (HMF) content, a key differentiator between pure and commercial honey, was assessed using UV spectroscopy. Additionally, ¹H NMR spectroscopy revealed the presence of antioxidants such as tyrosine and trigonelline in pure honey, which were notably absent in commercial honey samples. The integration of results from microwave X-band, FTIR, UV, and ¹H NMR techniques effectively highlighted the distinct properties of pure honey, thereby enabling the detection of sugar solution adulteration. This approach demonstrated the potential of these methods in accurately identifying honey adulteration, showcasing their effectiveness in distinguishing between the pure honey and adulterated samples., (© 2025 Institute of Food Technologists.)

Introduction: Corydalis DC., the largest genus of Papaveraceae, comprises numerous species known for their abundant alkaloid content and historical use in clinical medicine. Recently, a new species of genus Corydalis named Corydalis huangshanensis Lu Q. Huang & H. S. Peng was discovered in the Huangshan Mountains of Anhui Province, China., Objective: To compare the chemical characteristics of C. huangshanensis and other 13 Corydalis species, aiming to elucidate the potential medicinal value of this new species., Materials and Methods: The chemical constituents of C. huangshanensis and other 13 medicinal plants of genus Corydalis were analyzed using ultra-high-performance liquid chromatography Q-Exactive Plus hybrid quadrupole-Orbitrap mass spectrometer (Q-Orbitrap) mass technology. The differences in the alkaloids in the 14 species were distinguished by chemometrics., Results: The mass spectrometry fragmentation information and relative content of 72 alkaloids were obtained. Orthogonal partial least squares discriminant analysis (OPLS-DA) and cluster heat mapping analysis showed that these 14 species were divided into two groups. The clustering relationship between C. huangshanensis and C. decumbens (Thunb.) Pers. was similar, exhibiting similar chemical compositions and characteristics. These results indicate the potential pharmacological effects of C. huangshanensis., Conclusion: This study enhances our understanding of the chemical classification of Corydalis and provides a basis for speculations on the medicinal value of C. huangshanensis., (© 2024 John Wiley & Sons Ltd.)

Introduction: Stemona tuberosa Lour. (ST) is a significant traditional Chinese medicine (TCM) renowned for its antitussive and insecticidal properties. ST is commonly subjected to processing in clinical practice before being utilized as a medicinal substance. Currently, the customary technique for processing ST is honey-fried. Nevertheless, the specific variations in chemical constituents of ST before and after honey-fried remain unclear., Objective: This work aimed to analyze the variations in chemical constituents of ST before and after honey-fried and to study the distribution of differential markers in the roots., Methods: UPLC-Orbitrap Fusion MS combined with molecular network analysis was used to analyze the metabolome of ST and honey-fried ST (HST) and to screen the differential metabolites by multivariate statistical analysis. Spatial metabolomics was applied to study the distribution of differential metabolites by desorption electrospray ionization mass spectrometry imaging (DESI-MSI)., Results: The ST and HST exhibited notable disparities, with 56 and 61 chemical constituents found from each, respectively. After processing, the types of alkaloids decreased, and 12 differential metabolites were screened from the common compounds. The notable component variations were epibisdehydro-tuberostemonine J, neostenine, tuberostemonine, croomine, neotuberostemonine, and so forth. MSI visualized the spatial distribution of differential metabolites., Conclusions: Our research provided a rapid and effective visualization method for the identification and spatial distribution of metabolites in ST. Compared with the traditional method, this method offered more convincing data supporting the processing mechanism investigations of Stemona tuberosa from a macroscopic perspective., (© 2024 John Wiley & Sons Ltd.)

Background and Aims: Epichloë endophytes are vertically transmitted via grass seeds and chemically defend their hosts against herbivory. Endophyte-conferred plant defence via alkaloid biosynthesis might occur independently of costs for host plant growth. However, fitness consequences of endophyte-conferred defence and transgenerational effects on herbivore resistance of progeny plants are rarely studied. The aim of this study was to test whether severe defoliation in mother plants affects their seed production, seed germination rate and the endophyte-conferred resistance of progeny plants., Methods: In a field study, we tested the effects of defoliation and endophyte symbiosis (Epichloë uncinata) on host plant (Festuca pratensis) performance, loline alkaloid concentrations in leaves and seeds, seed biomass and seed germination rates. In a subsequent greenhouse study, we challenged the progeny of the plants from the field study to aphid herbivory and tested whether defoliation of mother plants affects endophyte-conferred resistance against aphids in progeny plants., Key Results: Defoliation of the mother plants resulted in a reduction of alkaloid concentrations in leaves and elevated the alkaloid concentrations in seeds when compared with non-defoliated endophyte-symbiotic plants. Viability and germination rate of seeds of defoliated endophyte-symbiotic plants were significantly lower compared with those of non-defoliated endophyte-symbiotic plants and endophyte-free (defoliated and non-defoliated) plants. During 6 weeks of growth, seedlings of defoliated endophyte-symbiotic mother plants had elevated alkaloid concentrations, which was negatively correlated with aphid performance., Conclusions: Endophyte-conferred investment in higher alkaloid levels in seeds, elicited by defoliation, provided protection from herbivores in progenies during the first weeks of plant establishment. Better protection of seeds via high alkaloid concentrations was negatively correlated with seed germination, indicating a trade-off between protection and viability., (© The Author(s) 2024. Published by Oxford University Press on behalf of the Annals of Botany Company.)

Harmful algal biotoxins in the marine environment are a threat to human food safety due to their bioaccumulation in bivalve shellfish. Whilst official control monitoring provides ongoing risk management for regulated toxins in live bivalve molluscs, no routine monitoring system is currently in operation in the UK for other non-regulated toxins. To assess the potential presence of such compounds, a systematic screen of bivalve shellfish was conducted throughout Great Britain. A rapid dispersive methanolic extraction was used with UHPLC-MS/MS analysis to test for fifteen cyclic imines and seven brevetoxins in 2671 shellfish samples taken from designated shellfish harvesting areas around Great Britain during 2018. Out of the 22 toxins incorporated into the method, only pinnatoxin G, 13-desmethyl spirolide C and 20-methyl spirolide G were detected, with maximum concentrations of 85.4 µg/kg, 13.4 µg/kg and 51.4 µg/kg, respectively. A follow up study of pinnatoxin G-positive samples examined its potential esterification to fatty acids and concluded that following hydrolysis, pinnatoxin G concentration increased by an average of 8.6%, with the tentative identification of these esters determined by LC-HRMS. This study highlights the requirement for ongoing monitoring of emerging threats and the requirement for toxicological and risk assessment studies.

Solid-state cholesteric liquid crystals (CLC solid ) with one-dimensional photonic structure offer a promising platform for constructing chemical and biological optical sensors, owing to their facile fabrication, signal readout, and sensitive and selective responsiveness to target analytes. In this study, we designed a CLC solid photonic structure intertwined with an interpenetrating polymeric network (IPN) immobilized with a cylindrospermopsin aptamer (CY9) for the selective detection of the cylindrospermopsin toxin (CYT) in water. Upon exposure to CYT, it induced a blue shift in the color of the IPN CY9 biosensor chip. This shift occurred because the CY9 aptamer selectively bound to the CYT, reducing the polarity of the IPN hydrogel, leading to water release and shrinkage of the photonic structure. The IPN CY9 biosensor chips demonstrated the ability to detect CYT within a linear range of 4.2-120 nM, with a limit of detection of 2.55 nM. This innovative biosensor chip not only provides a new strategy for designing targeted toxin biosensors by immobilizing different receptors but also exhibits significant potential for use in portable kits for remote areas.

Cyanobacteria produce diverse classes of toxins including microcystins, nodularins, anatoxins, cylindrospermopsins and saxitoxins, encompassing a range of chemical properties and mechanisms of toxicity. Comprehensive analysis of these toxins in cyanobacterial, environmental and biological samples generally requires multiple methods of extraction and analysis. In this work, a method was developed for the major classes of cyanotoxins, which comprised of a three-step liquid-solid extraction method using 75 % CH 3 CN with 0.1 % HCOOH and a hydrophilic interaction liquid chromatography (HILIC) elution gradient that provided retention of the less polar microcystins through to the highly polar saxitoxins. Detection was performed by tandem mass spectrometry in selected reaction monitoring mode with positive and negative polarity switching. Identification criteria included matching retention times and product ion ratios with available standards. In-house validation demonstrated good performance of the method including precision ranging from 1.5 (microcystin-LA) to 5.8 (gonyautoxin-2) % RSDs, and detection limits ranging from 0.01 (cylindrospermopsin) to 0.99 (gonyautoxin-3) µg/g in freeze dried material cyanobacteria. Recovery was assessed using spiked non-toxic cyanobacterial samples (Aphanizomenon sp.) and ranged from 83 (neosaxitoxin) to 107 % ([Dha 7 ]microcystin-LR). As a demonstration of application, toxin profiles in cyanobacterial cultures, benthic and planktonic cyanobacteria field samples, and shellfish reference materials were successfully evaluated. The procedure is also amenable for extension to other polar toxin classes including domoic acid and guanitoxin. With increasing reports of cyanobacterial blooms globally, the method represents a powerful quantitative screening tool for measuring cyanotoxins across a broad range of samples., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Crown Copyright © 2024. Published by Elsevier B.V. All rights reserved.)

Fibraurea tinctoria Lour. (Menispermaceae) is a medicinal plant used by many local tribes in Indonesia and other countries. This species has pharmacological potential, such as antimalaria, antibacterial, antitumor, antioxidant, anticancer, antidiabetic, anti-inflammatory, and hepato-protector. However, the secretory structures and sites of secondary metabolite accumulation still need to be studied. This study aimed to explore the secretory structures and reveal the accumulation sites of the main classes of secondary metabolites in the root, stem, and leaves of F. tinctoria. Histochemical analysis using various specific reagents was performed on freehand sections of the fresh organs to detect alkaloids, phenolics, terpenoids, and lipophilic compounds. Observations of secretory structures were carried out using light microscopy. The result showed that idioblast was the predominant secretory structure in the plant organs and contained all the chemical groups investigated, while laticifer accumulated alkaloids were found only in the stem. Some secondary metabolites were localized in the xylem, intercellular space, and cuticle layer. Crystals were observed in the root, stem, and leaf of this species. These results might be helpful as guide information for extracting the particular plant parts to obtain the desired secondary metabolites. It suggested that leaves are a potential alternative source of medicinal raw material of this species, in addition to the stem. RESEARCH HIGHLIGHTS: The first study on secretory structures of Fibraurea tinctoria. Secondary metabolites in F. tinctoria organs accumulate in specialized and unspecialized structures. The idioblast is the primary secretory structure in the organs of F. tinctoria spreading over the root, stem, and leaves., (© 2024 Wiley Periodicals LLC.)

The exponential number of food alerts about concerning levels of some plant-alkaloids, such as pyrrolizidine, tropane and opium alkaloids, have stressed the need to monitor their occurrence in foods to avoid toxic health effects derived from their intake. Therefore, analytical strategies to simultaneously monitor the occurrence of these alkaloids should be developed to ensure food safety an comply with regulations. Accordingly, this work proposes an efficient multicomponent analytical strategy for the simultaneous extraction of these alkaloids from commercial bakery products. The analytical method was validated and applied to the analysis of 15 samples, revealing that 100% of them contained at least one of the target alkaloids, in some cases exceeding the maximum limits legislated. Moreover, in two samples the 3 different alkaloid families were detected. These results confirm the importance of simultaneously monitoring these alkaloids in food and highlight also considering some opium alkaloids in current legislation., Competing Interests: Declaration of competing interest I, Isabel Sierra Alonso, hereby declare that I have read and understood the policies of the Food Research International regarding competing interests. I wish to inform the editorial team and readers about any potential conflicts of interest associated with the submitted manuscript entitled " Improving the food safety of bakery products by simultaneously monitoring the occurrence of pyrrolizidine, tropane and opium alkaloids"., (Copyright © 2024. Published by Elsevier Ltd.)

In this work, electromembrane extraction (EME) was used for the first time to separate aconitine (AC), mesaconitine (Mes-AC) and hypaconitine (Hyp-AC) from biological samples and Chinese herbal medicines. Efficient EME of polar and high molecular weight aconitine alkaloids from different sample matrices was achieved with the solvent of 1-ethyl-2-nitrobenzene (ENB). Under the optimal EME conditions, EME provided recoveries for all targets in the range of 72%-74 %, 85%-103 % and 92%-94 % for whole blood, urine and aqueous samples. The proposed EME systems combined with LC-MS/MS and HPLC-UV were evaluated using different sample matrices, and the methods displayed satisfactory analytical characteristic including negligible matrix effect. The LOD and LOQ of AC, Mes-AC, and Hyp-AC by EME-LC-MS/MS were in the range of 0.002-0.068 ng/mL and 0.005-0.228 ng/mL respectively. The LOD and LOQ of AC, Mes-AC, and Hyp-AC by EME-HPLC-UV were in the range of 0.06-0.26 μg/mL and 0.20-0.86 μg/mL, respectively. The coefficient of determination, R 2 -value was ≥0.9926 for all cases, and the accuracy in the linear ranges was in the range of 91%-111 %. Finally, the method was successfully applied for the qualitative and quantitative analysis of AC and Mes-AC in the whole blood and herbal medicine dreg samples from an actual forensic case, and poisoning by aconitum alkaloids was identified as the cause of death. Therefore, we believe that EME could be a powerful tool to identify poisoning, and EME has great potential for efficient separation of polar and high molecular weight substances. These are of great importance in the fields of but not limited to forensic science, Traditional Chinese Medicine and clinics., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Background: Despite the widespread implementation of analytical hardware capable of recording large-scale datasets for botanical natural products, the data processing procedures for compound annotation remain a bothersome obstacle that demand a tremendous amount of time and expert knowledge., Methods: Herein, an automatic LC-MS/MS data analysis workflow with AutoAnnotatoR was introduced for the compound annotation of plant derived natural products, which has the merits of great efficiency, high accuracy, saving time and simplified process. This procedure enabled automatic matching of MS 2 data with characteristic fragment ions, as well as MS 1 data with compound libraries, which improves the accuracy of structural elucidation. Notably, the optimization of collision energy for each target ion was successfully performed for the first time, facilitating the acquisition of comprehensive fragmentation information., Results: The automatic analysis workflow with AutoAnnotatoR was successfully applied for the annotation of alkaloids from 10 botanical origins of Fritillaria species. Consequently, a total of 2684 chemical constituents were tentatively characterized, with 23 components being unambiguously validated by reference standards and 2434 being probable novel chemicals., Conclusion: The entire data analysis procedure takes only a few hours, vastly improving analysis speed while assuring high accuracy. This method provides a powerful tool for the rapid and precise annotation of complex natural products. The workflow is publicly accessible on Github as an open-source R package called AutoAnnotatoR (https://github.com/anyaling2022/AutoAnnotatoR)., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier GmbH.)

Shenfu Qiangxin (SFQX) pills are proprietary traditional Chinese medicine used for the treatment of chronic heart failure with significant clinical effects. However, the systematic identification and quantification of complexed components in SFQX have not been reported yet. In this work, a reliable and comprehensive method for a rapid identification of chemical components was developed by data dependent acquisition using ultra performance liquid chromatography coupled with high-resolution mass spectrometry (UHPLC-Q Exactive-Orbitrap-MS). A total of 104 compounds mainly including flavonoids, saponins, alkaloids, anthraquinones, coumarins, and phenolic acids were identified through database searching. The identified compounds were further ascribed to herb species in SFQX pills, such as Ginseng Radis et Rhizoma, Aconiti Lateralis Radix Praeparata (processed), Mori Cortex, Lepidii Semen, Rhei Radix et Rhizoma, and Polyporus. Thirty-two representative compounds were elaborated with cleavage pathways and characteristic fragments in MS/MS spectrum of commercially available standards. A quantification method based on parallel reaction monitoring technique was established, and absolute quantification of 28 components in SFQX pills was then carried out. This work constitutes a basis and methodological reference for the quality control, consistency evaluation, and standardization of similar proprietary Chinese medicines., (© 2024 Wiley‐VCH GmbH.)

Higenamine (HG) is a β 2 receptor agonist and was explicitly added to the Prohibited List of the World Anti-Doping Agency in 2017. This compound is prohibited in both in- and out-of-competition athletes and falls under the category of nonthreshold substances. Because of HG presence in numerous plants, as evidenced by a growing body of research data, an exception was made for HG in the TD2017MRPL document, in which adverse analytical findings (AAFs) were not reported if the urinary HG concentration was less than 10 ng/mL. In this study, a comprehensive and systematic analysis of the HG content in five batches of samples from each of the 48 natural spices selected for this investigation was conducted using UPLC-MS/MS technology. Method validation was carried out in accordance with the ICH Analytical Procedures and Methods Validation for Drugs and Biologics Guidance, and the experimental results demonstrated that the method provided appropriate sensitivity, precision, stability, linearity, and accuracy. HG was detected for the first time in Houttuynia cordata, Zingiber officinale, Cinnamomum cassia, Stevia rebaudiana, Piper nigrum, Siraitia grosuenorii, Platycodon grandiflorus, and Myristica fragrans. Furthermore, the content of HG was found to vary significantly among the different plant parts of Nelumbo nucifera, such as rhizomes, leaves, seeds, and plumules. This paper provides systematic and comprehensive data to support the safe use of spices in athletes' diets, thereby reducing the risk of food-sourced doping violations., (© 2024 John Wiley & Sons Ltd.)

As β-carboline (βC) alkaloids, posing potential health risks, are present in a wide variety of foods, determining the exposure degrees of food to these alkaloids from dietary activity is key to ensuring food safety. Here, we developed a rapid and sensitive simultaneous analytical method for six βC alkaloids in food. We optimized the buffered QuEChERS method, which includes a clean-up process through dispersive solid phase extraction, to extract the target compounds from food matrices; then, these compounds were detected via liquid chromatography-tandem mass spectrometry. We established calibration ranges for each target compound and matrix within the range of 0.05-250 μg/kg, and verified linearity (R 2  ≥ 0.99) and limit of quantitation (≤1.63 μg/kg). Furthermore, we validated trueness (85.8%-118.8%) and precision (≤18.7%) at three levels within the calibration range, including the lowest and highest concentrations. Finally, we employed the developed method to determine the βC alkaloid contents in 304 samples of 41 food items and dietary exposure of six βC alkaloids resulting from daily intake. Although βC alkaloids were detected in 86.2% of the samples, exposure level to the 41 food items was insufficient to cause toxicity., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Foothill death camas (Z. paniculatus) grows on the foothill ranges of western North America and is acutely toxic to livestock grazing these ranges. The toxic alkaloids in foothill death camas are zygadenine and a series of zygadenine esters, with zygacine, the 3-acetyl ester of zygadenine, being the most abundant. In this study, earwax was evaluated as a specimen to determine livestock exposure to foothill death camas. Death camas alkaloids were detected in the earwax of sheep administered oral doses of foothill death camas alkaloids. In addition, death camas alkaloids were detected in the earwax of sheep that grazed rangeland with abundant death camas. This study demonstrates the potential of earwax as a noninvasive specimen for chemical analyses to aid in the diagnosis of livestock that may have been exposed to and poisoned by death camas. The results from this study indicate that diagnosticians should analyze for zygacine and zygadenine in the earwax of livestock suspected to have been poisoned by foothill death camas., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Published by Elsevier Ltd.)

Cathinones are often sold as "legal" alternatives to controlled stimulants such as amphetamine, MDMA and cocaine. Cathinones are the second largest group of new psychoactive substances (NPS), with close to 170 monitored by the European Monitoring Centre for Drugs and Drug Addiction (EMCDDA). Although all cathinones are related to the parent compound cathinone, one of the psychoactive principles in khat, assigning consistent, informative and user-friendly common names to these substances is challenging. Over time different naming approaches have been applied, leading to cathinones being known by several names. This work provides a framework and practical examples for the consistent naming of cathinones which is easy to understand and can be applied by the forensic community, researchers, clinical practitioners, and policy makers. The scope of the issue and rationale for earlier naming approaches are also discussed. The new naming framework has been developed based on established naming approaches and centered around the common "cathinone," and "phenone" motifs/scaffolds. The proposed framework establishes clear rules to derive the EMCDDA framework names for cathinones. Each name is, in turn, composed by a principal name containing a parent letter, derived after the "cathinone" or the "phenone" scaffold. Additional substitutions are prepended to the principal name. The framework also provides for exceptions for several cathinones and structural analogs scheduled under UN and EU legislation., (© 2024 The Authors. Drug Testing and Analysis published by John Wiley & Sons Ltd.)

Benzylisoquinoline alkaloids are significant phyto-nutraceuticals with a bitter flavor. However, due to their complex structures, the sensing and identification of benzylisoquinoline alkaloids is challenging. Benzylisoquinoline alkaloids are commonly extracted from plants such as Coptidis Rhizoma (CR), while the adulteration and dyeing of CR are prevalent. Herein, we innovatively developed a colorimetric sensor array based on the inhibitory effect of alkaloids on urease to identify benzylisoquinoline alkaloids and CR. Urea and pH indicators are used to convert the inhibitory effect of alkaloids on urease into visible signals. Different benzylisoquinoline alkaloids have different inhibition abilities to urease, resulting in their own unique colorimetric response pattern. Combined with linear discriminant analysis (LDA), the colorimetric sensor array has successfully identified different types and concentrations of benzylisoquinoline alkaloids, as well as benzylisoquinoline alkaloid mixtures. The array was further employed to identify CR, encompassing its authenticity, variety, adulteration, and dyeing., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Dendrobine is a sesquiterpene alkaloid primarily used in the treatment of inflammatory diseases, immune system disorders, and conditions related to oxidative stress. To understand the possible degradation pathways of dendrobine for its quality control, we conducted an in-depth investigation of its degradation products using forced degradation methods. The separation of dendrobine and its degradation products was achieved on a Shim-pack XR-ODS III (75 mm × 2 mm, 1.6 µm) column with a methanol-water mixture as the mobile phase under isocratic conditions, the isolated compounds were examined in positive ion mode with an ion trap-time of flight mass spectrometer (IT-TOF). In order to obtain in-depth structural information about the degradation products, mass spectrometry was performed using a five-stage fragmentation approach. This method allowed for thorough structural clarification via several rounds of selective fragmentation and high-resolution detection. System control and data acquisition were managed using LCMSsolution 3.81 software. The results showed that dendrobine undergoes significant degradation under oxidative, acidic, hydrolytic and thermal conditions, resulting in the formation of several degradation products with notable structural changes. Under oxidative conditions, dendrobine primarily generates two degradation products with mass increases of 16 Da and 32 Da, indicating mono-oxidation and di-oxidation reactions. Acidic degradation led to the identification of three degradation products, including a novel compound with an 18 Da mass increase, suggesting potential hydrolysis or dehydration reactions. Hydrolytic and thermal conditions resulted in the formation of two and three degradation products, respectively, with structural changes indicating possible molecular cleavage and reorganization mechanisms. In contrast, dendrobine exhibited strong stability under alkaline and photolytic conditions, with no significant degradation products detected. Detailed characterization of the degradation products via multi-stage mass spectrometry revealed key reaction pathways and mechanisms involved in dendrobine's degradation, providing critical insights for assessing its chemical stability and optimizing storage conditions., Competing Interests: Declaration of Competing Interest The authors declare that there is no conflict of interest regarding the publication of this paper., (Copyright © 2025 Elsevier B.V. All rights reserved.)

Exploring the type and amounts of the secondary metabolites (SMs) in a given fodder species was considered as a meaningful act for safe and profitable utilization of that particular feedstuff in the livestock industry. This study was conducted in the Konso zone, southern Ethiopia, to explore the secondary metabolite composition of Commelina species in two seasons and at two different altitudes. Samples were collected from the two altitudes and seasons. A completely randomized design was used in a factorial arrangement (five species (C. benghalensis, C. imberbis, C. diffusa, C. albescens, and C. africana), two altitudes, and two seasons) with three repetitions per treatment. The SM contents of the Commelina species were reasonably influenced by both seasonal and altitudinal changes. The mean alkaloid (3.67%), total phenols (9.76 mg GAE/g), flavonoid (3.81 mg CE/g) and condensed tannin (1.10 mg CE/g) values for the herb species in wet season inclined (p < 0.001) to 7.02%, 14.07 mg GAE/g, 7.68 mg CE/g and 2.38 mg CE/g, respectively, in dry season. The wet season saponin concentration of the species (2.65 g/Kg) significantly decreased (p < 0.001) to 1.28 g/Kg in the dry season. Similarly, the lowland saponin (2.26 g/Kg), alkaloid (3.70%), total phenols (10.89 mg GAE/g), flavonoid (4.71 mg CE/g), and condensed tannin (0.98 mg CE/g) contents were increased (p < 0.01) to 3.03 g/Kg, 5.47%, 13.61 mg GAE/g, 6.37 mg CE/g, and 1.81 mg CE/g, respectively, in the midlands. Alkaloids, total phenols, flavonoids and condensed tannin concentrations showed positive correlations with each other (P<0.05) and with seasonal (P<0.001) and altitudinal changes (P<0.001) as well. The findings of this study suggested that the SM concentrations of Commelina species were within the limits tolerable for ruminants. In conclusion, Commelina species could serve as a safe and beneficial forage herb to boost nutrient intake, improve nutrient use efficiency and hinder methane emissions, for animals consuming them, in areas where they are available in abundance., Competing Interests: The authors have declared that no competing interests exist, (Copyright: © 2024 Gelgelo et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Background: In the past three decades, liquid chromatography (LC) has been recognized as a significant environmental, health, and safety burden due to its heavy reliance on toxic organic solvents. Various chromatographic modes are in vogue today for complex analyses, such as sub/supercritical fluid chromatography (SFC) and enhanced fluidity liquid chromatography (EFLC). These modes are often advertised as "universally green" compared to the traditional allliquid reversed (RPLC) and normal phases (NPLC). Quantitative greenness evaluations must be done to validate or invalidate this assumption and allow separation scientists to make educated choices when deciding on what mode to use., Results: In this work, we modify the Analytical Method Greenness Score (AMGS) to include the cycle time of the instrument, and with the help of the first-order optimality condition (by setting the AMGS gradient = 0), we show that SFC and EFLC are not always the greenest option as they are often thought to be. Most of the greenness metrics have ignored the cycle time of instruments, yet this key component changes the entire AMGS response to flow rate. The complex case of separating tobacco alkaloid enantiomers (nicotine, nornicotine, anabasine, and anatabine) was selected as an illustrative example for comparing and contrasting separation modes using the modified greenness metric. These enantiomers have been selected due to their notorious difficulty in separation over the past 30 years. Using this family of molecules, four unique retention patterns were observed covering a wide variety of retention phenomena seen in small molecule enantioseparations., Significance: The modified AMGS metric will assist practicing analytical chemists in assessing the environmental impact of their separation methods from a single run in a given chromatographic mode. The proposed methodology identifies the minimum AMGS score corresponding to the greenest separation for routine chemical analysis., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Daniel W. Armstrong has a potential research conflict of interest due to a financial interest with the company AZYP, LLC. A management plan has been created to preserve objectively in research in accordance with UTA policy., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Cyanobacteria proliferate in warm, nutrient-rich environments and release toxic secondary metabolites into natural waters. Using cyanotoxin-contaminated water to irrigate crops could expose humans and biota, but the risk may be low if agricultural soils can sorb and retain cyanotoxins. In this report, we compared the sorption and desorption capacities of multi-class cyanotoxins/anabaenopeptins in soils of variable properties with a batch sorption procedure. The target compounds were anabaenopeptin-A, anabaenopeptin-B, anatoxin-a, cylindrospermopsin, and microcystins -LR, -RR, -LA, -LY, -LW, and -LF. Based on solid-liquid distribution coefficients (K d ), we classified cylindrospermopsin and microcystin-LA/-LY as "very low sorptivity", anabaenopeptin-A, -B and microcystin-LR, -LF, and -LW as "low sorptivity", and anatoxin-a and microcystin-RR as "medium sorptivity". We remain concerned about irrigating agricultural land with water contaminated with high levels of CYN and MC-LA/-LY because of their relatively low affinity and high desorption proportion in soils. The results also suggest that soil sorption can be an effective immobilization pathway for anatoxin-a and microcystin-RR. The generated data will be useful for prioritizing research on the most bioavailable cyanotoxins/anabaenopeptins that are likely to be released by the soil matrix, for environmental risk assessment., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Crown Copyright © 2024. Published by Elsevier B.V. All rights reserved.)

Honey, a widely consumed food, is susceptible to contamination by various toxic substances during production. Tropane alkaloids, with their potent neurotoxicity, are frequently found in honey. Hence, there is an acute need for rapid and effective detection methods to monitor these alkaloids. Lateral flow immunoassay (LFIA), known for its simple operation, low cost, and reliable results, holds great promise. In this study, we developed an efficient and user-friendly analytical method for the simultaneous detection of six tropane alkaloids (atropine, L-hyoscyamine, scopolamine, anisodamine, homatropine, and apoatropine) in honey based on an AuNPs lateral flow immunoassay (AuNPs-LFIA) with broad-spectrum antibodies. Under optimal conditions, the calculated detection limits were 0.22, 0.29, 0.51, 6.34, 0.30, and 0.94 ng/mL, respectively. By diluting the honey sample five times, the contaminants can be readily detected using LFIA. Semi-quantitative and quantitative analyses can be completed within 17 min. This innovative method fills the void in LFIA for detecting tropane alkaloids and serves as a valuable reference for LFIA detection of honey samples, providing a crucial strategy for the accurate detection of these important compounds.

Histolocalization and quantification of secondary metabolites established the occurrence of alkaloids, phenols and acetogenins in various plant organs of A. muricata at different stages of plant growth. Annona muricata L. possesses broad assemblage of ethno-pharmaceutical and therapeutic upsides ascribed to biosynthesis of secondary metabolites. Currently, the bioactivity is characterized by the production of acetogenins (ACGs), and also by the biosynthesis of alkaloids, primarily benzylisoquinolines derived from tyrosine and phenolic compounds. As a result of high variability of metabolite production in A. muricata, the present study evaluated the histochemical analysis of various plant parts at different developmental stages of growth. Presences of phytometabolites were determined using in-situ histochemical localization of alkaloids, phenolic compounds and acetogenins, while its estimation employing spectroscopic quantification techniques to understand structure, development, time-course deposition, content and distribution of secondary phytoconstituents. Study revealed distinct presence of secondary metabolites in cells and tissues of all plant organs in various intensities and patterns. Alkaloids and acetogenins occurred mostly in secretory cavities or idioblast cells while sparsely associated with other cells or tissues. Phenolic compounds emerged widely, with no confined distribution patterns occurring in storage cells and tissues. Quantitative analysis revealed highest accumulation of alkaloids, phenols and acetogenins in mature root-barks, mature unripe fruit rind and mature leaf respectively. Secondary metabolites generally occurred in young stages at low concentrations as compared to matured growth stages of plant parts. Such in-situ histochemical and biochemical approach will help in identifying the cells and tissues responsible for synthesis, storage and biological effects of potential secondary metabolites which could be further used for culturing and enhancing its production with biotechnological aids., Competing Interests: Declarations Competing interests The authors declare that they have no conflicts of interest to disclose. Data availability All data generated or analyzed during this study are included in the published article. In case, one wished to consult further information, it will be made available through the corresponding author on reasonable request., (© 2024. The Author(s).)

Qing-Kai-Ling oral liquid is commonly used clinically for the treatment of fever and upper respiratory tract infection. Moreover, studies have shown that Qing-Kai-Ling oral liquid has an anti-pneumonia effect. However, owing to its complex pharmacodynamic material basis, its pharmacological research and clinical application are limited. To address this problem, the chemical constituents of Qing-Kai-Ling oral liquid were identified by ultra-high performance liquid chromatography quadrupole-Exactive Orbitrap mass (UHPLC-Q-Exactive Orbitrap MS) and network pharmacology methods, which were used to predict its potential anti-pneumonia target and signalling pathway. A total of 150 compounds were identified and tentatively characterized, including 35 amino acids and their derivatives, 36 organic acids, 20 terpenoids, 20 alkaloids, 12 glycosides, 7 flavonoids, and 20 others. Among them, 14 compounds were accurately identified by comparing their retention time and mass spectrum data with those of reference substances. Additionally, we performed molecular simulation calculations via Density Function Theory to determine the plausibility of the compound cleavage reactions and further confirm compound structures. Furthermore, 90 key targets were screened through network pharmacology, with the particular focus on the PI3K-AKT, MAPK and TNF signalling pathways. This method achieved the first comprehensive identification of the chemical composition of Qing-Kai-Ling oral liquid and elucidated its potential mechanism of anti-pneumonia. The results provide valuable reference and data support for pharmacodynamic substance research and quality control., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

In this report, we describe a fluorescent assay for the detection of six marine toxins in water. The mechanism of detection is based on a duplex-to-complex structure-switching approach. The six aptamers specific to the targeted cyanotoxins were conjugated to a fluorescent dye, carboxyfluorescein (FAM). In parallel, complementary DNA (cDNA) sequences specific to each aptamer were conjugated to a fluorescence quencher BHQ1. In the absence of the target, an aptamer-cDNA duplex structure is formed, and the fluorescence is quenched. By adding the toxin, the aptamer tends to bind to its target and releases the cDNA. The fluorescence intensity is consequently restored after the formation of the complex aptamer-toxin, where the fluorescence recovery is directly correlated with the analyte concentration. Based on this principle, a highly sensitive detection of the six marine toxins was achieved, with the limits of detection of 0.15, 0.06, 0.075, 0.027, 0.041, and 0.026 nM for microcystin-LR, anatoxin-α, saxitoxin, cylindrospermopsin, okadaic acid, and brevetoxin, respectively. Moreover, each aptameric assay showed a very good selectivity towards the other five marine toxins. Finally, the developed technique was applied for the detection of the six toxins in spiked water samples with excellent recoveries.

In this paper, we examine intraspecific variation in the quantity of alkaloid chemical defence in field collected individuals of the polymorphic ladybird beetle Adalia decempunctata (10-spot ladybird). Like its more widely studied relative Adalia bipunctata (2-spot ladybird), A. decempunctata possesses the alkaloids adaline and adalinine, which are, respectively, the major and minor alkaloids of A. bipunctata. We focused especially on alkaloid concentration in relation to colour pattern morph, sex, and the relationship between female and egg parameters. There was a marked sexual dimorphism in the balance of the two alkaloids, with adaline predominating in females and adalinine predominating in males: in males, on average, over 70% of total alkaloid was adalinine. Females had a lower proportion of adalinine (< 10%) than their eggs (> 15%) and relationships between egg alkaloid and female alkaloid or fecundity were weak or non-existent. Colour pattern morph had a borderline (although not) significant relationship with adaline concentration and total alkaloid concentration, which could be further explored with laboratory reared individuals. The sexual dimorphism in alkaloid content, which seems likely due to differences in synthesis, might be related to their relative costs to the two sexes and might provide insight into the evolution of alkaloid diversity in ladybirds., (© 2024. The Author(s).)

Arabica coffee is one of the most consumed beverages in the world. The chemical components present in raw Arabica coffee beans (RACB) are directly related to the cup quality of the beverage. Environmental and genetic factors influence the content and profile of these components. Then, this work aimed to evaluate different chemical parameters of RACB from 3 varieties planted in 3 different experimental farms located within the "Alta Mogiana" terroir harvested in 2021 and 2022 to identify a better variety for each farm to produce a high cup quality Arabica coffee. The harvest period had a strong influence on most of the studied parameters because atypical weather conditions occurred in the 2021 harvest. The RACB harvested in 2022 yielded better results and supposedly will produce a beverage of high cup quality. Samples harvested in this period presented mainly average moisture levels closer to optimum (11.02 against 8.56 % in 2021); low total titratable acidity (98.00 against 169.75 mL 0.1 M NaOH/100 g in 2021); high amounts of free amino acids (0.96 against 0.93 g GAE/100 g in 2021), low amounts of CGA (4.27 against 4.85 g/100 g in 2021) and caffeine (1.08 against 1.76 g/100 g in 2021) and high amounts of trigonelline (1.12 against 0.96 g/100 g in 2021). The Rome Sudan variety had the best combination of chemical results, mainly when cultivated in Farm 2 in 2022, presenting high amounts of protein content (15.24 %) and free amino acids (0.96 g GAE/100 g), low total titratable acidity (98.3 mL 0.1 M NaOH/100 g), low amounts of CGA (4.55 g/100 g) and caffeine (1.29 g/100 g) and high amounts of trigonelline (1.11 g/100 g). The analysis of chemical compounds could predict the best farm to cultivate each variety studied and was a guide to foresee a higher cup quality of RACB beverages., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The liquid chromatography-high resolution mass spectrometry (LC-HRMS) technique enables the detection of phytochemicals present in the extracts. LC-HRMS-generated mass list showed abundant compounds of interest, artifacts, and primary metabolites. The identification of a secondary metabolite of interest within the extract is very challenging. We hypothesized that identifying the "new metabolite" in the whole metabolome is more challenging than identifying it within the class of metabolites. The proposed prioritization strategy focused on the elimination of unknown and prioritizing the known class of secondary metabolites to identify new metabolites. The prioritization strategy demonstrated on Murraya paniculata for the identification of new metabolites. LC-HRMS-generated information is used as a filter to target the secondary metabolite and the new metabolites. This strategy successfully annotated the new coumarin and coumarin alkaloids from the mass list of 1448 metabolites. Varanasine ( 3 ), schroffanone ( 4 ), schroffanene ( 5 ), and O -methylmurraol ( 9 ) are new compounds, and coumarin ( 1 , 2 , and 6-8 ) are known. Varanasine ( 3 ) is the first naturally occurring 7-aminocoumarin with additional N -formyl functionality. The isolates were screened for cytotoxicity against the panel of cancer cell lines. Varanasine ( 3 ) and minumicrollin ( 6 ) showed significant cytotoxicity and apoptosis-inducing potential. The immunoblot analysis confirmed inhibition of apoptotic protein PARP-1 and caspase-3 expression by 3 and 6 .

Background and Aims: Using ecological momentary assessment (EMA), we undertook a natural experiment wherein kratom-product variability was a tool to assess kratom dose-response relationships based on product form and alkaloid level., Methods: Between July-November 2022, 357 US kratom consumers (56.6 % male, 90.2 % non-Hispanic white) completed 15 days of EMA; 348 participants submitted samples of the products used most often during EMA. These were assayed for ten alkaloids using ultra-performance liquid chromatography-tandem mass spectrometry. Self-reported kratom effects were modeled as a function of kratom amount and alkaloid content., Results: Participants used over 220 brands. The most-reported product forms were loose powder (55.8 %) and encapsulated powder (26.8 %); extracts were used less (419 uses across 9.48 % of participants). Of the 12,244 use-event entries, 7726 had follow-up data (15-180minutes after use) on felt effects. Effects were stronger in participants with a higher average amount per use. Within-person dose-response relationships were obscured by highly-consistent within-person dosages. Effects of loose powder decreased over three hours; effects of extracts started higher but decreased more rapidly. Dose-response relationships for specific alkaloids could not be reliably established because total alkaloid content and relative levels of specific alkaloids showed limited variability between products. Higher levels of corynoxine alkaloids were associated with slightly stronger effects, possibly an artifact of modeling data with low alkaloid variability., Conclusions: Alkaloid content was surprisingly consistent across kratom products, and participants were consistent in the amount they used across events. Firm conclusions about alkaloid-effect relationships for kratom will require experimenter-controlled manipulations of agent and dose., Competing Interests: Declaration of Competing Interest KES serves as a paid scientific advisor to the International Plant and Herbal Alliance, and as an expert witness in legal cases involving kratom. CRM serves as an expert witness in legal cases involving kratom. All other authors have nothing to declare., (Copyright © 2024 The Author(s). Published by Elsevier B.V. All rights reserved.)

Among the emerging investigative fields, forensic medicine and toxicology lead to analyzing fatalities in medico-legal expert opinion formulating. While discussing the problem, the authors have selected 96 fatal cases from their expert practice including the period from 2010 to 2023, in which deaths were connected with taking new psychoactive substances (NPS's) belonging to various chemical categories, mainly synthetic cathinones (SC), synthetic cannabinoids (SCan) and non-medical synthetic opioids (NSO). In the investigated cases, toxicological analysis revealed 37 NPS's and their 9 metabolites. The cases involved the use of SC's (64 cases - 67 %), Scan's, including their metabolites (10 cases - 10 %) and NSO's, including their metabolites (6 cases - 6 %). The remaining cases involved the simultaneous use of NSO with SC and/or SCan, including their metabolites (8 cases - 8 %), or SC with SCan (5 cases - 5 %). In three cases (3 %), compounds belonging to other groups were taken. In twenty-five cases, more than one NPS was found. Moreover, in twenty-seven cases, ethyl alcohol was also detected at the concentration range of 0.6-3.6 ‰. The concentration of xenobiotics determined in blood represented extensive ranges of concentration. The victims were at the age of 16-58 years of life. The group included eleven women (11 %). Generally, the deaths related to NPS's were predominantly of an accidental character (81 %), while the manner of death in sixteen cases (17 %) was suicide, including hanging (5 cases), jumping from a great height (3 cases), self-injury and exsanguination (1 case), as well as acute drug intoxication (6 cases) and intoxication with central nervous system hypoxia after an hanging (1 case). Among the analyzed cases there were two victims of homicide (2 %), in one of which the perpetrator being under the influence of the mixture of the synthetic opioid U-47700 and synthetic cannabinoid AB-FUBINACA. In twenty-eight cases, medications used in psychiatry were found, which suggested that the victims were struggling with mental problems before death. As it was implied by the available information, more than 36 % of the victims had mental problems., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Aptamers are good affinity receptors for bio-assays, while colorimetric method is suitable for point-of-care sensing via direct visualization. But previously aptamers often need complex re-engineering for colorimetric measurement at the cost of affinity and performance. Here isoquinoline alkaloids are found to own unique light-activated oxidative capacity, which can be specifically triggered by unmodified aptamers. This feature is universal for two alkaloids to efficiently oxidize four chromogenic substrates with obvious color changes. Based on a dye-displacement process, we have developed a novel light-activated aptamer system for the colorimetric assay of estradiol. It shows a good sensitivity with a detection limit of 326 nM, and this homogeneous assay is reliable to avoid artifacts in previous heterogeneous scheme. Besides, it is proven to be a universal design to assay other two targets. Significantly, they do not employ any aptamers re-engineering but only simply use their parental aptamers. Therefore, this light-activated oxidative capacity of isoquinoline alkaloid can serve as an ideal tool for colorimetric assay of various targets based on aptamer's specific recognition., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Mitragyna speciosa is a perennial plant native to Asia, well known for its psychoactive properties. Its major alkaloid mitragynine is known to have sedative and euphoric effects. Hence, the plant has been a subject of abuse, leading to addiction, necessitating efficient analytical methods to detect its psychoactive constituents. However, current chromatography-based methods for detecting the alkaloids are time consuming and costly. Quantitative nuclear magnetic resonance (qNMR) spectroscopy emerges as a promising alternative due to its nondestructive nature, structural insights, and short analysis time. Hence, a rapid and precise qNMR method was developed to quantify selected major psychoactive alkaloids in various parts of M. speciosa. Mitragynine, specioliatine, and speciogynine were quantified in relation to the integral value of the -OCH 3 groups of the alkaloids and the internal standard 1,4-dinitrobenzene. The precision and reproducibility of the method gave a relative standard deviation (RSD) of 2%, demonstrating the reliability of the method. In addition, the method showed excellent specificity, sensitivity, high linearity range (R 2  = 0.999), and limits of detection (LOD) and quantification (LOQ) values. The analysis revealed that the red-veined M. speciosa leaves contained higher levels of mitragynine (32.34 mg/g), specioliatine (16.84 mg/g) and speciogynine (7.69 mg/g) compared to the green-veined leaves, stem bark, or fruits., (© 2024 John Wiley & Sons Ltd.)

Steroidal glycoalkaloids (GAs) are toxins produced by solanaceous plants. As there are no fully standardized methods for their extraction and determination in food, the research aimed to: (1) develop and critically compare methods based on gas (GC) and liquid (LC) chromatography, including their coupling with mass spectrometry, and (2) to develop and optimize a universal GA extraction method. Hyphenated techniques proved to be the most useful in GA analysis: LC-MS was the most sensitive one, while GC-MS offered the highest chromatographic resolution. It was proven that quantitative results obtained using different analytical techniques cannot be directly compared. New extraction method that is more efficient than the AOAC method (997.13) was then designed and optimized. It was characterized by higher absolute recovery (99% and 34%, respectively) and allowed to extract much more GAs from the same material (e.g. 21.2 ± 1.4 and 11.82 ± 0.97 mg g -1 of potato tubers, respectively)., Competing Interests: Declaration of competing interest The authors declare that they have no conflict of interest. The authors state that the research did not include human and/or animal participation., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Food safety is crucial to attaining food security and sustainability. Unsafe foods for human and animal consumption lead to product recalls and rejection, negatively impacting the global economy and trade. Similarly, climate change can adversely affect the availability of safe and nutritious food at the table. The changing climatic conditions and global food trade and transport can make the movement of toxic plants possible, resulting in food crops being increasingly invaded by some species of plants that produce toxic secondary metabolites, such as tropane alkaloids (TAs). Datura stramonium from the Solanaceae plant family is an invasive and virulent plant that produces high amounts of two TAs, atropine and scopolamine. Various food poisoning events following accidental or deliberate ingestion of foods contaminated by atropine and scopolamine from seeds of D. stramonium have been recorded in different locations globally. Due to these incidents, regulatory agencies require the development of plant toxin detection methods that can be used in the food chain as early as possible. This systematic review thus focuses on the TA determination techniques in food and feeds published between 2013 and 2023. A particular focus was given to the sample preparation methods, the improvements of each technique claimed, and data to support the performance of each method, especially the ability to measure at or below the maximum level. The review concludes with other technological advancements, including rapid spectroscopy, electrophoresis, and colorimetric methods, as well as the possibility of coupling with smartphones for use in on-farm detection and the challenges in applying them., (© 2024 The Author(s). Comprehensive Reviews in Food Science and Food Safety published by Wiley Periodicals LLC on behalf of Institute of Food Technologists.)

Specialized metabolites play important roles in plants and can, for example, protect plants from predators or pathogens. Alkaloids, due to their pronounced biological activity on higher animals, are one of the most intriguing groups of specialized metabolites, and many of them are known as plant defense compounds. Poison hemlock, Conium maculatum , is well-known for its high content of piperidine alkaloids, of which coniine is the most famous. The distribution, localization, and diversity of these compounds in C. maculatum tissues have not yet been studied in detail. The hemlock alkaloids are low molecular weight compounds with relatively high volatility. They are thus difficult to analyze on-tissue by MALDI mass spectrometry imaging due to delocalization, which occurs even when using an atmospheric pressure ion source. In this manuscript, we describe an on-tissue derivatization method that allows the subsequent determination of the spatial distribution of hemlock alkaloids in different plant tissues by mass spectrometry imaging. Coniferyl aldehyde was found to be a suitable reagent for derivatization of the secondary amine alkaloids. The imaging analysis revealed that even chemically closely related hemlock alkaloids are discretely distributed in different plant tissues. Additionally, we detected a yet undescribed hemlock alkaloid in Conium maculatum seeds.

A paper-based colorimetric sensor array mediated by a novel nanozyme (CuCo 2 O 4 ) was developed using a screen-printing technology. The aim was to facilitate the identification of different kinds of alkaloids. Typically, three chromogenic substrates (3,3',5,5'-tetramethylbenzidine, 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid), and o-phenylenediamine) were selected as sensing elements, which can be catalyzed by a CuCo 2 O 4 nanozyme with peroxidase-like activity to yield corresponding oxidized products, thereby inducing color changes. Owing to the varying inhibitory ability of different alkaloids on acetylcholinesterase (AChE), a decrease in choline (Ch) concentration occurs and subsequently results in the restoration of color within the units of sensor array. Color data can be transformed into hue information with a smartphone. The above color variations generated a unique "fingerprint" pattern on five alkaloids (berberine, palmatine, jatrorrhizine, eserine, and harmane), which can be successfully discriminated through linear discriminant analysis in the range 0.2 to 20 µM. Furthermore, the sensor arrays allowed successful discrimination of the above five alkaloids in Chinese herbal medicine samples and recognition of 22 blind samples. This work presents a novel nanozyme-based paper sensor array, which is a user-friendly and reliable platform for probing different alkaloids. In addition, the developed sensing strategy enables the identification of AChE-related diseases, positively contributing to the screening available of AD-associated drugs., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

Zuojin Pill (ZJP), a traditional Chinese medicine prescription composed of Rhizoma Coptidis and Euodiae Fructus in the ratio of 6:1 (w/w), has been widely used for the treatment of gastric disorders. However, an in-depth understanding of in vivo metabolism and distribution profiles of protoberberine alkaloids (PBAs) and indole alkaloids (IDAs) in ZJP is lacking. In this study, a method using ultra-high performance liquid chromatography coupled with quadruple time-of-flight mass spectrometry (UHPLC-Q-TOF-MS) was developed to systematically screen the alkaloids and their metabolites in rat plasma and various tissues after oral administration of ZJP. Furthermore, bioinformatics and molecular docking analyses were conducted to elucidate the contribution of the alkaloids and metabolites enriched in the stomach to the therapeutic effect of ZJP on gastritis. A total of 33 compounds, including 7 prototype alkaloids and 26 metabolites, were chemically defined or tentatively identified in this work. The metabolic pathways of PBAs (hydroxylation, oxidation, reduction, demethylation, demethylenation, glucuronide conjugation, sulfate conjugation) and IDAs (hydroxylation, glucuronide conjugation) were revealed. Notably, 7 prototype alkaloids and 18 metabolites were detected in the stomach, indicating their propensity for gastric distribution. These alkaloids and metabolites showed strong affinities with the 7 hub targets associated with gastritis, such as CCR7, CXCR4, IL6, IFNG, CCL2, TNF, and PTPRC, and could be considered the potential active substances of ZJP for treating gastritis. In conclusion, this study clarified the gastric distribution propensity of PBAs and IDAs and their metabolites, as well as their favorable binding interactions with gastritis-related targets, which could provide essential data for the further study of the pharmacodynamic material basis and gastroprotective mechanism of ZJP., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Codonopsis Radix (CR), a traditional tonic medicinal material in China, has been proven to possess a variety of bioactive functions. However, its chemical composition and in vivo metabolic pattern have not been fully elucidated. In this study, AB-8 macroporous resin column chromatography was employed for the enrichment of small molecular components in CR. Furthermore, a method combining ultra-performance liquid chromatography-quadrupole-orbitrap mass spectrometry with Acquire X intelligent data acquisition technology software was developed for the preliminary screening and identification of the chemical composition of CR in vitro and their metabolites in vivo. As a result, a total of 116 components were preliminarily characterized in the CR extract, including 28 polyacetylenes, 33 organic acids, 4 amino acids, 23 alkaloids, 9 phenylpropanoids, 6 terpenoids, 2 nucleosides, and 11 others. Additionally, a total of 84 compounds, including 37 prototype components and 47 metabolites, were identified in the plasma, urine, and feces of rats after oral administration of CR. Specifically, 11, 24, 19, 32, and 25 constituents were identified in the heart, liver, spleen, lung, and kidney, respectively. Of note, the lung and spleen are the organs with the highest distribution of CR compounds. These findings will serve as valuable data for future research on the correlation between the chemical composition and pharmacological effects of CR., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Sinomenii Caulis (SC), a commonly used traditional Chinese medicine for its therapeutic effects on rheumatoid arthritis, contains rich chemical components. At present, most studies mainly focus on sinomenine, with little research on other alkaloids. In this study, a comprehensive profile of compounds in SC extract, and biological samples of rats (including bile, urine, feces, and plasma) after oral administration of SC extract was conducted via ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC/Q-TOF-MS). The fragmentation patterns and potential biotransformation pathways of six main types of alkaloids in SC were summarized, and the corresponding characteristic product ions, relative ion intensity, and neutral losses were obtained to achieve rapid classification and identification of complex components of SC from in vitro to in vivo. As a result, a total of 114 alkaloid compounds were identified, including 12 benzyl alkaloids, 4 isoquinolone alkaloids, 32 aporphine alkaloids, 28 protoberberine alkaloids, 34 morphinan alkaloids and 4 organic amine alkaloids. After administration of SC extract to rats, a total of 324 prototypes and metabolites were identified from rat plasma, urine, feces and bile, including 81 aporphines, 95 protoberberines, 117 morphinans and 31 benzylisoquinolines. The main types of metabolites were demethylation, hydrogenation, dehydrogenation, aldehydation, oxidation, methylation, sulfate esterification, glucuronidation, glucose conjugation, glycine conjugation, acetylation, and dihydroxylation. In summary, this integrated strategy provides an additional approach for the incomplete identification caused by compound diversity and low abundance, laying the foundation for the discovery of new bioactive compounds of SC against rheumatoid arthritis., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

This study aimed to identify and quantify the primary components in lotus leaf and to explore the hypolipidemic components through spectral-effect relationships and chemometric methods. Utilizing a data-dependent acquisition-diagnostic fragment ion/characteristic neutral loss screening strategy (DFI-NLS), a reliable HPLC-Q-TOF-MS analysis was conducted, identifying 77 compounds, including 36 flavonoids, 21 alkaloids, 3 terpenoids, 11 organic acids, 4 phenols, 1 lignin and 1 unsaturated hydrocarbon. A straightforward HPLC-DAD method was developed for the simultaneous determination of seven major components in lotus leaf, and quercetin-3-O-glucuronide (Q3GA) was identified as the most abundant component. The HPLC fingerprints of 36 lotus leaf sample batches were assessed using chemometric approaches such as principal component analysis and hierarchical cluster analysis. The hypolipidemic effect of these samples was analyzed by measuring total cholesterol (TC) and total triglycerides (TG) levels in palmitic acid (PA) and oleic acid (OA)-induced lipid modeling in HepG-2 cells, employing partial least squares regression and grey relation analysis to investigate the spectral-effect relationship of the lotus leaf. The in vivo hypolipidemic effect of these compounds was assessed using an egg yolk powder-induced high-fat zebrafish model. The findings indicated that peak No.11 (Q3GA) in the chemical fingerprint was significantly associated with hypolipidemic activity, suggesting it as a potential hypolipidemic compound in lotus leaf. In summary, this study facilitates the exploration of the phytochemical compounds and their bioactive properties in the lotus leaf., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Bletilla striata has been used in traditional Chinese medicine for thousands of years to treat a variety of health diseases. Currently, metabolic causes of differences in medicinal values are unknown, due to the lack of a large-scale and comprehensive investigation of metabolites in Bletilla species. In order to gain a better understanding of the major chemical constituents responsible for the medicinal value, this study aimed to explore the metabolomic differences among three Bletilla species (Bletilla striata: Bs, Bletilla ochracea: Bo and Bletilla formosana: Bf). There were 258 different metabolites between 'Bo' and 'Bf', the contents of 109 metabolites had higher abundance, while 149 metabolites showed less accumulation. There were 165 different metabolites between the 'Bs' and 'Bf', content of 72 metabolites was increased and content of 93 metabolites was decreased. There were 239 different metabolites between the 'Bs' and 'Bo', content of 145 metabolites was increased and content of 94 metabolites was decreased. In the Bo&#95;vs&#95;Bf, Bs&#95;vs&#95;Bf and Bs&#95;vs&#95;Bo groups, the major differential categories were flavonoids, phenolic acids, organic acids and alkaloids. Moreover, the differential metabolites were clustered into clear and distinct profiles via K-means analysis. In addition, the major differential categories were flavonoids, phenolic acids, organic acids and alkaloids. The 'Flavonoid biosynthesis' (ko00941) and 'Phenylalanine metabolism' (ko00360) pathways were significantly enriched in Bo&#95;vs&#95;Bf, Bs&#95;vs&#95;Bf and Bs&#95;vs&#95;Bo comparisons. These results clarify the metabolomics in different Bletilla species, as well as providing basis for the phamaceutical value of novel species of Bletilla., (© 2024. The Author(s).)

Efficiently mining and identification of new compounds from the extensive MS/MS datasets of plant extracts poses a significant challenge due to the structural diversity and compositional complexity inherent in natural products (NPs). Various data post-processing techniques have been developed to simplify the interpretation of MS/MS data; however, they often suffer from limited specificity and precision. Meanwhile, structure annotation following data post-processing is particularly time-consuming. In this study, we introduced an innovative strategy named MS-SMART, which integrates three intelligent algorithms: automatic mining of diagnostic ions, rapid filtration of alkaloids from untargeted MS/MS data, and structural recommendations for filtered components. The feasibility of this approach for rapidly discovering novel compounds was demonstrated using berberine-type alkaloids as an example. Firstly, diagnostic ions were automatically extracted and validated using available reference data. Subsequently, berberine-type compounds were filtered from raw MS/MS data. Finally, the structures of the target components were recommended using building blocks derived from berberines reported in various plants. A total of 103, 198, 60, 80 and 51 berberines were efficiently identified in diverse families and genera, including Stephaniae Epigaeae Radix, Coptidis Rhizoma, Phellodendri Chinensis Cortex, Phellodendri Amurensis Cortex and Corydalis Decumbentis Rhizoma, with 99, 169, 50, 64 and 40 new compounds identified, respectively. Among these, 8, 14, 8, 7 and 12 berberines were confirmed by reference compounds. This strategy provides a new research paradigm for the rapid discovery and identification of different types of new compounds in complex samples., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

The identification of molecules within complex mixtures is a major bottleneck in natural products (NPs) research. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) has emerged as the main tool for the high-throughput characterization of NPs. The large amount of data sets by LC-MS/MS presents a challenge for data processing and interpretation, and the LC-MS/MS molecular network (MN) is one of the most prominent tools for analyzing large MS/MS data sets, widely used for rapid classification, identification, and structural speculation of unknown compounds. However, the existence of a large number of redundant nodes leads to false-positive results. To solve this problem, we proposed the in-depth analysis of MN. In this study, in-depth analysis of MN of five NPs representing the common structures of saponin, steroid, flavonoid, alkaloid, and phenolic acid revealed the presence of redundant nodes (including other adducts, isotope, and in-source fragmentation) in addition to the normal nodes, which can lead to false-positive identification results. Additionally, the reasons for different redundant nodes are discussed and experimentally verified, and it was found that the impact of redundant nodes can be mitigated by optimizing the experimental conditions and employing Feature-Based Molecular Networking. Furthermore, Ion Identity Molecular Networking can rapidly discover and screen redundant nodes, simplifying the in-depth analysis of MN and improving the network connectivity of structurally related molecules. Finally, a combination formulation of 7 NPs is used as an example to provide a guide for in-depth analysis of MN for comprehensive characterization of complex systems. This study highlights the importance of an in-depth analysis of MN for better understanding and utilization of MS/MS data in complex systems to reduce the false-positive rate of identification by screening and filtering redundant nodes.