Kava is a beverage prepared from the rhizomes and roots of the tropical evergreen plant Piper methysticum. Kava contains six major kavalactones. Legacy studies found that kavalactones did not inhibit human CYPs in vitro at relevant concentrations, and kava did not inhibit the major CYPs in humans in vivo and was devoid of clinically significant drug interactions. Nevertheless, CYP2B6 was never evaluated. Recent investigation identified that (+)-dihydromethysticin, one of the six natural kavalactones, was an inhibitor of CYP2B6 in vitro. This investigation evaluated kava extract and (+)-dihydromethysticin effects on expressed CYP2B6 activity- both wild-type CYP2B6.1 and the active allelic variant CYP2B6.4. Effects on human liver microsomal CYP2B6 were also evaluated. Two independent kava extracts inhibited expressed and human liver microsomal CYP2B6 activity, in a concentration-dependent manner, assessed using the CYP2B6 probe substrates 7-ethoxy-4-trifluoromethyIcoumarin O-deethylation and S-ketamine N-demethylation. Ki values for CYP2B6 inhibition by (+)-dihydromethysticin alone and in kava extracts were 0.01-0.05 μM. Kava and (+)-dihydromethysticin interaction with expressed CYP2B6.1 and CYP2B6.4 generated a difference spectrum consistent with formation of a metabolite-inhibitor complex. (+)-Dihydromethysticin underwent NADPH-dependent metabolism by expressed CYP2B6 and human liver microsomes. These results show that kava extracts inhibit CYP2B6, an effect attributed to (+)-dihydromethysticin, which appears to act as a mechanism-based inhibitor. Because kava consumption achieves plasma (+)-dihydromethysticin concentrations exceeding the inhibitory Ki, there is a potential risk of in vivo CYP2B6-mediated herb-drug interactions. Such potential merits clinical drug-herb interaction studies using standard CYP2B6 substrate probes.

The genus Oncosiphon (Asteraceae), consisting of aromatic herbs, is indigenous to southern Africa. Oncosiphon species have been documented in Khoi-San ethnobotany as herbal remedies for typhoid fever, pneumonia, and as diuretics. Research on the biological properties and comprehensive phytochemical profiling of these important Oncosiphon species is currently limited. This study was therefore undertaken to address the knowledge void in chemical profiling, through the application of various analytical techniques to analyse the volatile and non-volatile constituents of three South African Oncosiphon species. The aerial parts of Oncosiphon suffruticosus (n = 28), O. grandiflorus (n = 16), and O. africanus (n = 4) were collected from various locations in the Western Cape Province of South Africa. The stems and leaves (SL) were separated from the flowers (F) and analysed as distinct samples. The methanol: chloroform (1:1, v/v) extracts were prepared and analysed using ultra–high–performance liquid chromatography quadrupole time-of-flight time–of–flight mass spectrometry (UHPLC–QToF–MS) and a semi–automated high–performance thin–layer chromatography (HPTLC) system. Multivariate data analysis was performed on the UHPLC–QToF–MS data to determine interspecies chemical variation. Two-dimensional (2D) gas chromatography (GCxGC–ToF–MS) was used to determine the headspace volatile profiles of the intact aerial parts. The results show that the non-volatile profiles of the Oncosiphon species are characterised by amino acids, phenolic acids, flavonoids, sesquiterpene lactones, and fatty acid derivatives. The HPTLC profiles of O. grandiflorus and O. africanus are chemically more closely related, and O. suffruticosus has a distinct profile, which is supported by the chemometrics results of the flowers. The major headspace volatile compounds in Oncosiphon flowers are α-pinene, α-ocimene, eucalyptol, o-cymene, and artemisia alcohol, whereas the stems and leaves mainly consist of α-ocimene, eucalyptol, and yomogi alcohol.

To date, the phytochemical composition of the aerial parts of Nanophyton iliense U.P. Pratov has not been comprehensively investigated. In the present study, qualitative metabolite profiling of the methanolic extract of the aerial parts was performed using liquid chromatography coupled with diode-array detection and quadrupole time-of-flight mass spectrometry (LC-DAD-QToF-MS) operating in both positive and negative electrospray ionization modes. A total of 81 metabolites were tentatively identified based on accurate mass measurements, MS/MS fragmentation patterns obtained in all-ion MS/MS mode, and comparison with previously reported literature data. The detected compounds included hydroxycinnamic acid amides, phenolic acids, flavonoids (including glycosides), amino acids, organic acids, sulfated derivatives, and nucleosides. Among them, the flavonoid narcissin (isorhamnetin-3-O-rutinoside) was isolated from the extract, and its structure was confirmed by 1H and 13C NMR spectroscopy supported by COSY, HSQC, and HMBC experiments. Additionally, a compound with the molecular formula C17H14O5 was detected; however, its structure could not be conclusively established based on the available spectroscopic data and is therefore reported as an unidentified metabolite. The present study provides the first systematic qualitative characterization of the metabolite profile of N. iliense and establishes a foundation for future quantitative and bioactivity-oriented investigations of this species.

Castanopsis sieboldii is a phenolic-rich evergreen species in the family Fagaceae, yet comprehensive quantitative and tissue-specific metabolite profiling remains limited. In this study, an integrated analytical workflow comprising UHPLC-PDA quantification, LC-QToF-MS identification, and chemometric analysis was developed to characterize phenolic constituents in leaves, flowers, fruits, and stems. A validated UHPLC-PDA method enabled the simultaneous quantification of six major phenolics, with limits of detection ranging from 0.01 to 0.05 μg/mL and limits of quantification from 0.025 to 0.1 μg/mL. Among all tissues, 3‑O‑galloylshikimic acid (Compound 1) was the predominant metabolite (0.1-204 mg/g), followed by caffeoylquinic acids (Compounds 2-3), ellagic acid (Compound 4), and flavonoid glycosides (Compounds 5-6). LC-QToF-MS analysis facilitated the tentative annotation of 185 metabolites, including phenolic acids, ellagitannins, galloylshikimic acids, and flavonoid glycosides, based on accurate mass measurements and characteristic MS/MS fragmentation patterns. Chemometric evaluation using principal component analysis (PCA), partial least squares discriminant analysis (PLS-DA), and hierarchical clustering analysis (HCA) revealed clear tissue-specific clustering, with leaves exhibiting the highest chemical diversity and phenolic abundance, whereas fruits showed minimal levels. PCA captured 80% of total variance in the first two components, and the PLS-DA model showed strong predictive performance (R²Y ≈ 1.00; Q² ≈ 0.98) although interpretation should consider the limited sample size. This study provides a comprehensive, tissue-resolved phenolic profile of C. sieboldii, establishing a robust chemical foundation for future pharmacological, ecological, and quality-control applications.

Elymus repens (L.) Gould has a long history of use in traditional medicine across the British Isles, particularly among Gaelic and Anglo-Saxon communities, where it has been employed as a diuretic and anti-inflammatory agent. Despite its ethnopharmacological significance, comprehensive insights into its phytochemical composition and biological activities remain limited. The present study aimed to provide an integrated characterization of E. repens through macro- and microscopic analyses, advanced phytochemical profiling, and evaluation of the biological activities of the rhizome part. Microscopic examination revealed distinct anatomical features differentiating rhizome and stem, leaf tissues, supporting accurate identification and pharmacognostic standardization. Chemical profiling using Liquid Chromatography-Quadrupole Time-of-Flight Mass Spectrometry (LC-QToF-MS) enabled the tentative identification of 93 metabolites, including amino acids, in aerial and rhizome extracts. These compounds were primarily classified into polyamines (e.g., feruloylputrescine, hydroxycoumaroylagmatine), phenolic acids (gallic, vanillic, and ferulic acids), flavonoids (apigenin, tricin, saponarin analogues), amino acids (arginine, tyrosine, tryptophan), organic acids (malic, succinic acids), nucleosides (adenosine, thymidine), and phospholipids. Biological evaluation demonstrated that the hydroethanolic rhizome extract exhibited notable antifungal activity against Aspergillus fumigatus (IC50 = 34.9 µg/mL). Additionally, hydroethanolic extracts of both the aerial and rhizome parts showed no cytotoxicity in the Artemia salina lethality assay at concentrations up to 10 mg/mL, indicating a favorable preliminary safety profile. Of particular interest is tricin, which possesses anti-inflammatory, antioxidant, antimicrobial, and potential nephroprotective therapeutic effects; its mechanism of action is associated with the suppression of oxidative stress, the inhibition of pro-inflammatory mediators, and the disruption of metabolic processes in microbial cells. Overall, this study provides a comprehensive phytochemical and pharmacognostic characterization of E. repens, highlighting its potential as a source of bioactive compounds. The metabolite profile obtained directly from the biologically active extract, combined with its proven antifungal activity, serves as direct confirmation of the antimicrobial aspect of this plant’s traditional use, while its broader applications in ethnomedicine require targeted pharmacological testing. These findings contribute valuable data for chemotaxonomic classification and future pharmacological investigations.

Medicinal and aromatic plants are vital to the global herbal trade, with Panax species (ginseng) being highly valued for their health-promoting benefits. Ensuring the quality and safety of the plant raw material is essential; however, intentional or unintentional adulteration with morphologically similar species often comprises product authenticity, which is common in herbal products. Comparative morphoanatomy together with high-performance liquid chromatography coupled with ultraviolet detection (HPLC-UV) analyses were conducted on four Panax species (P. ginseng, P. quinquefolius, P. japonicus, and P. notoginseng) and two common adulterants (Codonopsis pilosula and Platycodon grandiflorus). Microscopic examination of intact roots, rhizomes, and their powders revealed critical diagnostic traits. The presence of secretory ducts and abundant starch grains were consistent in Panax species, but absent or scarce in adulterants. While xylem architecture effectively differentiated most species, the high anatomical similarity between P. ginseng and P. quinquefolius remained a challenge. The morphology of starch grains and xylem vessels also served as diagnostic features to differentiate the ginseng species from both adulterants. HPLC-UV profiles corroborated these findings, identifying eight major ginsenosides in Panax samples, while adulterants were entirely devoid of these compounds. These results confirm that anatomical analysis provides robust, practical criteria for the authentication of ginseng. Integrating microscopic techniques with chemical profiling offers a reliable, complementary approach to detecting adulteration, including economically motivated bulking, thereby ensuring the botanical integrity and safety of herbal products.

Bulbine Wolf (Asphodelaceae) comprises over 80 species, with five holding significant medicinal value in South Africa for treating skin and gastrointestinal ailments. While roots are traditionally prioritised, the chemical profiles of leaves and rhizomes remain underexplored although they are also medicinally important. This study used chromatographic and mass spectrometry-based analytical techniques (HPTLC, LC-QToF-MS, and UPLC-MS) combined with chemometric analysis to chemically profile the metabolites of five Bulbine species from the Eastern Cape. Chemical profiling tentatively identified 73 compounds, including flavonoids, phenolic acids, and anthraquinones. Phenylanthraquinones, specifically knipholone and knipholone 6 '-methyl ether, were ubiquitous across all taxa. Species-specific markers were identified, such as bulbine-knipholone in B. abyssinica, gaboroquinones in B. frutescens, and bulbnatalonosides in B. latifolia. Chemometric modelling revealed that while rhizome chemistry is relatively conserved, root profiles provide the most robust inter-species differentiation. Bulbine frutescens and B. latifolia displayed the highest chemical diversity, whereas B. asphodeloides was notably rich in anthocyanins. The significant overlap in bioactive constituents between leaves, roots and rhizomes suggests that aerial parts could serve as viable substitutes in traditional remedies. These findings establish a chemical basis for species authentication and support sustainable harvesting practices by reducing the destructive exploitation of rhizomes and roots.

7-Hydroxymitragynine (7-OH), a minor alkaloid in kratom (Mitragyna speciosa (Korth.) Havil.) (Rubiaceae) and an active metabolite of mitragynine, is a critical transient precursor to mitragynine pseudoindoxyl, which exhibits >2-fold μ-opioid receptor-biased agonism and improved isoform selectivity relative to its precursors. Although present only in trace amounts in kratom leaves, a recent survey identified hundreds of consumer products with semisynthetic 7-OH and mitragynine pseudoindoxyl. Their high potency, ease of access, and risk of addiction have prompted the US FDA to recommend scheduling 7-OH as a Schedule I controlled substance. To address this public health crisis and the associated analytical challenges, a comprehensive analysis of commercially available kratom products was performed. The results revealed substantial inconsistencies between the quantities claimed on the products' labels and the actual 7-OH amount detected in several of these products. Along with 7-OH and mitragynine pseudoindoxyl, several (over)oxidized byproducts were detected, providing crucial insights into their synthetic origins. Analysis of >98% of 7-OH-labeled products indicated a semisynthetic origin, with labeled doses ranging from 0.001 to 33.6 mg per serving. Furthermore, consistent with the prior report, 7-OH degrades into the potentially toxic byproduct 3-dehydromitragynine under simulated gastric conditions. These findings underscore significant safety concerns over oral 7-OH products and necessitate validated analytical monitoring to guide regulatory oversight and consumer protection.

Entada elephantina (Burch.) S.A. O'Donnell & G.P. Lewis (formerly Elephantorrhiza elephantina (Burch.) Skeels), a member of the Fabaceae, is an ethnomedicinal plant widely used in South Africa. Despite studies supporting its traditional applications, comprehensive phytochemical characterisation remains limited. In this study, chemical profiling and marker quantification were performed on methanolic root extracts of E. elephantina (n = 50) collected from the Northern Cape Province, South Africa. High-performance thin-layer chromatography coupled to mass spectrometry (HPTLC-MS) and LC-MS analyses confirmed the presence of catechin, epicatechin gallate (ECG), epigallocatechin gallate (EGCG), quercetin 3'-glucoside, and gallic acid across all samples. Ultra-high performance liquid chromatography-diode array detector-quadrupole time-of-flight mass spectrometry (UHPLC-DAD-QToF-MS) enabled the tentative identification of 93 metabolites, predominantly flavonoids and their acylated derivatives, alongside minor phenolic and organic acids. Detected flavonoids included (epi)catechin aglycone derivatives, galloylated flavonoids such as taxifolin galloylated hexoside, and dimeric and trimeric ECG derivatives. Quantification using a validated UPLC-PDA method demonstrated notable variation in quercetin 3'-glucoside (1.23-2.54 mg/kg), catechin (0.42-0.88 mg/kg), ECG (0.39-0.74 mg/kg), kaempferol (0.008-0.018 mg/kg), and EGCG (0.004-0.014 mg/kg). Molecular networking revealed two principal structural subclusters, reflecting spectral similarity and shared fragmentation patterns within flavonoid families. This comprehensive chromatography-based approach establishes a phytochemical reference framework to support quality control, standardisation, and potential commercial development of this medicinal species.

The psychedelic mushroom market has expanded rapidly due to changing regulations and increasing consumer demand. Product diversity now extends beyond traditional capsules and tablets to include gummies, powders, and confectionery items, complicating quality control efforts. To assess the quality and potential adulteration of Amanita musca-ria and Psilocybe cubensis-based products, a validated LC-QToF-MS method was developed. This method focused on five characteristic compounds: ibotenic acid (IBA), muscimol (MUS), muscarine, psilocin, and psilocybin that are constituents of A. muscaria and P. cubensis mushrooms. Method validation demonstrated satisfactory linearity, precision, and recovery of all five analytes. Psilocin and psilocybin levels ranged from 0.001-1.6% and 9.9-19.3%, respectively, in five Psilocybe species samples, while IBA, MUS, and muscarine levels in two samples of Amanita muscaria were 0.03-0.04%, 0.01- 0.02%, and 0.01-0.02%, respectively. By comparing commercial products to authentic samples, we evaluated the overall quality of 27 across various formulations. Our analysis included 14 gummies, three chocolates, six capsules, one tablet, and three powders. Although 11 of 14 gummies claimed to contain Amanita mushroom extracts, only MUS and muscarine were detected, without IBA. Interestingly, one gummy product indicated the presence of psilocin and psilocybin despite the labeling that claimed, "no psilocybin." Eleven products contained psilocin and psilocybin as anticipated, but five products lacked all target compounds. These findings underscore the need for standardized product specifications. Nevertheless, the established LC-QToF-MS approach could serve as a valuable tool for evaluating the quality of magic mushroom-based consumer products.

ETHNOPHARMACOLOGICAL RELEVANCE:Barbeya oleoides Schweinf. (Barbeyaceae) is traditionally employed to manage diverse ailments, including infections, fever, edema, and related inflammatory conditions. AIM OF THE STUDY:This study aimed to investigate the wound-healing potential of the total methanolic extract (BO-TM) from the aerial parts of B. oleoides, as well as its defatted and mother liquor fractions (BO-ML). In addition, chemical profiling of identified bioactive constituents underpinning the observed therapeutic activity. METHODS:LC-DAD-QToF analysis was performed for BO-TM phytochemical profiling. BO-TM was prepared and subsequently defatted to yield methylene chloride (BO-MC) and mother liquor (BO-ML) fractions. In vitro wound healing effects on human skin fibroblast (HSF) migration were assessed using a scratch assay, with effective doses selected based on prior MTT cytotoxicity assay results. For in vivo evaluation, male Wistar albino rats were divided into 7 groups: Normal, positive control without treatment, reference (Mebo®), and treatment groups receiving BO-TM and BO-ML (2.5 &5 % in Vaseline base, topically) once daily for 10 days. Histopathological examination and biochemical markers analyses (MIP-2, collagen, MDA, GSH, and FOXO1) were performed to elucidate underlying mechanisms related to inflammation, oxidative stress, and tissue remodeling. RESULTS:Chemical profiling annotated 69 metabolites, comprising 20 flavonoids, 42 gallotannins and phenolic acid derivatives, and 5 triterpenoids. BO-TM (5 %) enhanced wound closure to 96 % versus 84 % in the untreated group. In vivo, treatment with BO-TM and BO-ML has achieved a significant increase in GSH, FOXO1, collagen, and MIP-2 levels while showing a significant decrease in MDA level compared to the positive control and Mebo® groups. CONCLUSION:The methanolic extract and its fraction of B. oleoides exhibited significant-wound-healing potential in in vitro and in vivo models, likely through antioxidant, anti-inflammatory, and pro-regenerative mechanisms mediated by its rich polyphenolic and triterpenoid content. These findings provide scientific support for its traditional use and highlight its therapeutic potential as a natural wound-healing agent by Yemeni and Eritrean healers. However, further preclinical and clinical investigations are warranted to comprehensively evaluate its safety, efficacy, and translational potential in human applications.

Echinacea products are among the most widely used herbal supplements in the United States, primarily valued for their purported alleviation of common cold symptoms. Herein, we report the isolation and characterization of eight alkamides including two new ones (1 and 2) from the roots of two Echinacea species: E. purpurea (1, 3, and 4) and E. angustifolia (2, and 5‒8). This study provides the first comprehensive 13C nuclear magnetic resonance assignments of all isolated alkamides. The absolute configuration of 1 was established using Diastereomeric Parameter 4 plus probability analysis and specific rotation. The anti-inflammatory activity of the purified compounds and extracts was evaluated by measuring the inhibition of inducible nitric oxide synthase in lipopolysaccharide-stimulated mouse macrophages (RAW264.7), alongside cytotoxicity assessments. The purified alkamides and the crude extracts exhibited modest anti-inflammatory effects. The current study provides an understanding of the chemical composition of Echinacea and identifies potential markers for distinguishing E. purpurea from E. angustifolia.

Pharmacopeias are essential for ensuring the quality of botanical raw materials, yet their utility is limited by slow adaptation to rapid innovation in the dietary supplement industry. Challenges arise from differing standards (e.g., the British Pharmacopoeia (BP) and the United States Pharmacopoeia (USP)), as well as a critical lack of comparative data for globally sourced ingredients. The post-COVID surge in adaptogens, such as ashwagandha, underscores the urgency of these issues. The commercial use of aerial parts-despite traditional root-only use and phytochemical variability-raises concerns about quality and safety. The situation necessitates rigorous source verification and re-evaluation of current pharmacopeial methods. To address these deficiencies, we developed and compared an HPLC-PDA method utilizing BP and USP standards to quantify three steroidal lactones (withaferin A, withanolide A, and withanoside IV) in ashwagandha samples and commercial products. Our analysis revealed substantial variations in steroidal lactone content. Alarmingly, over 44% of products failed to meet BP standards, and 60% failed to meet USP standards. The high failure rate was attributed to the elevated presence of co-eluted dihydrowithaferin sulfate, primarily found in leaf samples, which led to an underestimation of withanolide A. Only 10 of 25 supplement products met both standards; only two were confirmed as root-derived, while the remainder contained varying proportions of (un)disclosed aerial parts. These findings highlight critical plant-part-specific chemical variations and the need for enhanced source verification and improved quality control. Developing robust analytical methods and revisiting existing pharmacopeial guidelines are crucial for assessing the quality and safety of ashwagandha products, including novel formulations.

Andrographis paniculata, a popular immune-boosting supplement, faces quality concerns due to potential adulteration and reliance on single-marker quality assurance. To address these, a comprehensive UHPLC-PDA and LC-QToF-MS method was developed and validated to quantify and identify three classes of compounds: phenolic acids, diterpenoid lactones (including their glycosides), and flavonoid glycosides. The method achieved LOD of 10300 ng/mL and LOQ of 251000 ng/mL for fourteen reference standards. Analysis of plant samples revealed total phenolic acids (1-4; 0.3-13 mg/g), flavonoid glycosides (6, 10, and 11; 0.2-2 mg/g), diterpene lactones (7, 13, and 14; 1.4-31 mg/g), and diterpene lactone glycosides (5, 8, 9, and 12; 0.6-17 mg/g). Furthermore, over 100 phytochemicals were tentatively identified based on characteristic mass spectral features in A. paniculata. Applying this comprehensive phytochemical profile to various supplement products revealed that over 25 % of commercial A. paniculata products failed to meet pharmacopeia standards. Detailed analysis demonstrated significant variations in the plant-part-specific distribution of diterpene lactone and its glycosides, as well as flavonoid glycosides. Some products exhibited elevated mean values for these three analyte groups, while others showed significantly lower values contributing to the observed failures. Moreover, the analytical data revealed a significant discrepancy in andrographolide (7) content, a standard quality assurance marker, between plant materials (0.4-23 mg/g) and dietary supplements (0-278 mg/serving size). Specifically, a 13 % andrographolide decrease and a corresponding increase in 14-deoxy-11,12-didehydroandrographolide (14) in A. paniculata capsules suggest processing-induced dehydration, necessitating optimized production to maintain biologically active andrographolide content. These findings underscore the necessity for rigorous quality control and comprehensive analytical methods to ensure the quality assurance of A. paniculata supplements.

Inonotus obliquus is widely recognized as the Chaga mushroom. Chaga contains various bioactive compounds, including polysaccharides, triterpenoids, polyphenols, and melanin. To address the characterization and quantitative analysis of triterpenoids and phenolics in Chaga, a multi-analytical approach has been developed combining LC-PDA-ELSD and LC-DAD-QToF. These methods were designed to quantify 11 compounds, comprising seven triterpenoids and four fatty acids, using LC-PDA-ELSD, and four phenolics using the LC-DAD-QToF method. Calibration curves for these compounds demonstrated excellent linearity within the tested range. The methods exhibited high precision, with intra- and inter-day relative standard deviations below 3% and recoveries ranged from 91% to 104%. The validated methods were applied to analyze eleven sclerotia samples, one mycelium sample, three grain-based samples, and eighteen dietary supplements. Results revealed that eight of the eighteen supplements (44%) contained ground mycelium, which primarily showed the presence of fatty acids but lacks detectable levels of triterpenoid and phenolic markers characteristic of Chaga. Triterpenoids and hispidin, identified as key bioactive compounds, were detected in eight (44%) of the eighteen supplements; however, these products also contained fatty acids and/or betulin. Two (11%) of the 18 supplements showed the presence of phenolic compounds only; no triterpenoids were detected. Additionally, untargeted metabolomic screening using LC-DAD-QToF tentatively identified 103 compounds from diverse chemical groups, including nine reference compounds. These findings provide valuable insights for the quality assessment of dietary or food supplements marketed as containing Chaga.

Legumes, a dietary staple for centuries, have seen an influx of conventional and unconventional varieties to cater to human care conscious consumers. These legumes often undergo pretreatments like baking, soaking, or boiling to mitigate the presence of non-proteinogenic amino acids (NPAAs) and reduce associated health risks. The recent tara flour health scare, linked to the NPAA baikiain, emphasizes the need for robust analytical methods to ensure the safety and quality of both traditional and novel plant-based protein alternatives. While traditional techniques provide insights into protein and non-proteinogenic amino acid profiles, modern liquid chromatography-mass spectrometry (LC-MS) offers superior sensitivity and specificity for NPAA detection. This study employed an LC-QToF method with MS/MS analysis to comprehensively map the distribution of free NPAAs and proteinogenic amino acids (PAAs) in various legume samples. A total of 47 NPAAs and 20 PAAs were identified across the legume samples, with at least 7–14 NPAAs detected in each sample. Sulfur-containing NPAAs, such as S-methyl-L-cysteine, γ-glutamyl-S-methyl cysteine, and S-methyl homoglutathione, were predominantly found in Phaseolus and Vigna species. Cysteine and methionine were the sulfur-containing PAAs identified. Gel electrophoresis and soluble protein quantification were also conducted to understand legume protein composition holistically. This orthogonal approach provides a valuable tool for ensuring the overall quality of plant-based proteins and may aid in investigating food poisoning or outbreaks related to such products.

Ethnopharmacological relevance Phyllanthus amarus is ethnomedicinally used to treat gallbladder stones, kidney stones and chronic liver diseases. P. amarus is gaining popularity as an ingredient in many botanical dietary supplements. Aim of the study To evaluate the interaction of P. amarus extract and its lignans with human xenobiotic sensing receptors (PXR and AhR) and their downstream genes. Materials and methods Activation of PXR and AhR was measured by reporter gene assays. Gene expression analysis was performed in hepatic (HepG2) and intestinal (LS174T) cells by RT-PCR. CYP inhibition assays were carried out in baculosomes. The inhibitory effect on the ABC transporters (P-gp and BCRP) was investigated via rhodamine-123 and Hoechst 33342 uptake assays in Caco-2 and MDR-MDCK cells. Effect on CYP3A4 and CYP1A2 enzyme activity was measured in primary human hepatocytes. Results P. amarus extract and its lignans activated AhR and PXR in respective reporter cells. Tested extract and lignans significantly increased CYP3A4 mRNA but inhibited CYP3A4 enzyme activity when tested in primary human hepatocytes and CYP3A4-specific baculosomes. In contrast, increased CYP1A2 mRNA was associated with increased CYP1A2 enzyme activity in hepatocytes. No inhibition of CYP1A2 activity was detected in baculosomes. A weak inhibitory effect on ABC-transporters was observed. Conclusions Results suggest that overconsumption of P. amarus or P. amarus-containing botanical supplements may change CYP homeostasis which could alter the pharmacokinetics of substrate drugs, thereby elevating the risk of herb-drug interactions (HDIs) when taken concomitantly with conventional medications. Further studies are warranted to strengthen the clinical relevance of these findings.

The Brassicaceae family, commonly referred to as cruciferous plants, is globally cultivated and consumed, with the Brassica genus being particularly renowned for its functional components. These vegetables are rich sources of nutrients and health-promoting phytochemicals, garnering increased attention in recent years. This study presents a comprehensive microscopic, chromatographic, and spectroscopic characterization of Brassica napus L. seeds from Kazakhstan aimed at elucidating their morphological features and chemical composition. Microscopic analysis revealed distinct localization of flavonoids, total lipids, and alkaloids. High-performance thin-layer chromatography (HPTLC) analysis of seed extracts demonstrated a complex chemical profile with significant quantities of non-polar compounds in the hexane extracts. Additionally, methanolic extracts revealed the presence of diverse chemical compounds, including alkaloids, flavonoids, and glucosinolates. The chemical composition exhibited varietal differences across different Brassica species, with B. napus L. seeds showing higher concentrations of bioactive compounds. Furthermore, liquid chromatography–quadrupole time-of-flight mass spectrometry (LC-QToF-MS) analysis provided insights into the chemical composition, with sinapine isomers, feruloyl, and sinapoyl choline derivatives as major compounds in the seeds. This study contributes to a better understanding of the chemical diversity and quality control methods’ approximations of B. napus L. seeds, highlighting their importance in functional food and nutraceutical applications.

Background/Objectives: Sixty-eight percent of service members are living with overweight or obesity, some who may not consult a healthcare provider when they decide to lose weight. Instead, they often turn to weight-loss dietary supplements for self-care solutions. The purpose of this case series study was to examine the label accuracy and quality of select weight-loss dietary supplements sold on or near US military bases across the country. Methods: Weight-loss dietary supplements (n = 44) were selected and purchased in GNCs, Exchanges, and Shoppettes across 12 military installations from March 2023 to January 2024. Liquid chromatography-mass spectrometry was used to verify the label accuracy according to the Supplement Facts listed ingredients and whether they contained any ingredients prohibited for use in the military. Product label claims were analyzed using the Operation Supplement Safety (OPSS) Risk Assessment Scorecard. Results: Thirty-six products (82%) had inaccurate labels. Twenty-seven (61%) had ingredients listed on the label not detected through analysis. Sixteen products (36%) contained hidden ingredients. The four products purchased within one mile from the base listed multiple prohibited ingredients on the labels, with all detected. Forty (91%) products scored a “no-go” according to the OPSS Scorecard and none contained a third-party certification seal on the label. Multiple stimulants were included in products such that the product safety was unknown. Conclusions: The majority of weight-loss dietary supplements analyzed in this case series study had inaccurate labels and were considered a “no-go” according to the Scorecard. Service members should only have access to safe, high-quality dietary supplement products. OPSS is collaborating with the Department of Defense stakeholders to determine the most effective ways for service members to have access to third-party certified products on all military establishments.