Brahmi essence, developed from Bacopa monnieri (L.) Wettst. standardized extract and mulberry juice, was proven to improve the memory speed of healthy participants aged 55–80 years old, following a 12-week dietary program. However, the metabolites have not yet been reported. Our objective was to characterize the altered metabolites in the plasma, urine, and feces of healthy volunteers after consumption of Brahmi essence for 12 weeks, using the LC-MS metabolomics approach. The altered metabolites were selected from OPLS-DA S-plots; 15 metabolites in the plasma, 7 in the urine, and 17 in the feces samples were tentatively identified by comparison with an online database and literature. The metabolites in the plasma samples were in the classes of amino acids, acylcarnitine, and phospholipids. Benzeneactamide-4-O-sulphate and 3-hydroxyhippuric acid were found in urine samples. The metabolites in the class of amino acids, together with jujubogenin and pseudojujubogenin, were identified in the fecal samples. The aminoacyl-tRNA, aromatic amino acids, and branched-chain amino acid biosynthetic pathways were mainly related to the identified metabolites in all three samples. It could be implied that those metabolites and their pathways might be linked with the effect of Brahmi essence on memory speed.
Hemp essential oils are rich in bioactive compounds, including terpenes and cannabinoids, yet standardized analytical methods for their simultaneous quality control are limited. This study aimed to (i) validate a static headspace gas chromatography–mass spectrometry (SHS-GC-MS) method for simultaneous quantification of 20 terpenes and 2 cannabinoids and (ii) apply it to fingerprint essential oils from four hemp strains, including local (HRDI2, HRDI5) and internationally cultivated (Charlotte’s Angel, Cherry Wine) varieties. The method met AOAC validation criteria, with detection limits of 0.025–0.5 µg/mL for terpenes and 1 µg/mL for cannabinoids. Quantitation limits ranged from 0.1–1 µg/mL for terpenes and 5 µg/mL for cannabinoids. Intraday precision (%RSD) ranged from 0.27–11.00%, while interday precision ranged from 3.14–13.89%. The method recoveries ranged from 85.12–115.47%. Precision and recovery confirmed the method’s reliability. Multivariate statistical analysis identified 82 metabolites, revealing distinct chemical fingerprints among strains, and emerged as newly identified chemotype markers, supporting chemotype classification. This work demonstrates, for the first time, a solvent-free, automatable SHS-GC-MS approach for simultaneous terpene and cannabinoid profiling in hemp essential oils, enabling both qualitative and quantitative characterization and supporting regulatory compliance for the development of standardized phytopharmaceutical products.
The imbalance of phosphodiesterase 5 (PDE5) enzyme in the male body, or excessive PDE5 enzyme levels, can occur due to factors such as aging, diseases (e.g., cardiovascular disease, diabetes, depressive disorder), and physical behaviors (e.g., alcoholism, smoking, stress). PDE5 is directly associated with erectile dysfunction disease. Currently, many studies aim to find natural PDE5 inhibitors as an alternative to commercial drugs. This study is the first to demonstrate that the ethanolic leaf extract of D. cochinchinensis exhibits potent PDE5-inhibitory activity. The PDE5-inhibitory activity of five plant parts was evaluated: leaf (IC50 = 1.53 ± 0.12 µg/mL), twig (3.37 ± 0.54), fruit (14.92 ± 2.85), heartwood (19.05 ± 5.60), and bark (16.03 ± 2.92). However, there is still uncertainty about which compounds in leaf extract are responsible for the PDE5 inhibition. Therefore, the purpose of this study is to identify the chemical constituents in the leaf of D. cochinchinensis, including determining which of these compounds may act as PDE5 inhibitors. This study was achieved using at-line LC-QTOF-MS2.
The extract of Tectona grandis L.f. leaves along with its major bioactive constituents, (+)-Eperua-8,13-dien-15-oic acid (1) and (+)-Eperua-7,13-dien-15-oic acid (2), has demonstrated moderate inhibitory activity against the steroid 5α-reductase enzyme (5α-R), a key factor in androgenetic alopecia (AGA) pathogenesis. In this study, molecular interactions between these compounds and 5α-reductase type 1 (SRD5A1) were investigated, revealing binding affinities of −8.01 kcal/mol for 1 and −8.81 kcal/mol for 2. Moreover, their anti-inflammatory properties were evaluated via in vitro assays assessing nitric oxide inhibition and the suppression of pro-inflammatory cytokines, interleukin-1β (IL-1β) and interleukin-6 (IL-6). The results showed that 2 exhibited a stronger binding affinity to endothelial nitric oxide synthase (eNOS) (−9.00 kcal/mol) and demonstrated more selective inhibition of IL-6 (IC50 = 9.30 ± 2.65 μM, binding affinity = −6.83 kcal/mol) due to the presence of attractive charge interactions. This integrated experimental and computational approach highlights T. grandis bioactive compounds as promising dual-action candidates, simultaneously targeting 5α-reductase activity and cytokine-mediated inflammation and offering new mechanistic insights relevant to AGA management.
Chronic stress impairs cognition through gut–brain axis dysregulation and metabolic imbalance. This study applied untargeted fecal metabolomics to investigate the cognitive and metabolic effects of Bacopa monnieri (L.) Wettst (Brahmi), mixed Thai berry, and their combined extracts in rats exposed to chronic unpredictable mild stress. Cognitive performance was evaluated using the novel object recognition test. Fecal metabolites were profiled using LC-ESI-QTOF-MS, followed by data preprocessing and multivariate statistical analysis. Orthogonal partial least squares regression was applied to identify metabolites associated with the recognition index, and pathway enrichment analysis was subsequently performed to interpret biological relevance. All interventions were associated with improved recognition performance and treatment-related metabolic modulation. Biosynthesis of unsaturated fatty acids was consistently enriched across treatment groups, indicating a shared involvement of lipid remodeling. Treatment-specific responses were also observed: Brahmi was associated with linoleic and alpha-linolenic acid metabolism; mixed Thai berry extract demonstrated dose-dependent modulation of lipid metabolism, with low-dose supplementation additionally yielding amino-acid-derived metabolites; and bile acid-related signaling was uniquely detected in the low-dose combined extract group. These findings demonstrate that fecal metabolomics can capture distinct metabolic signatures associated with herbal extract-induced cognitive improvement and highlight lipid remodeling as a shared metabolic feature across interventions under chronic stress.
Background/Objectives: Chronic stress contributes to anxiety disorders and cognitive impairment, while effective multi-target therapeutic strategies remain limited. This study investigated the effects of a standardized extract prepared from the aerial parts of Bacopa monnieri (L.) Wettst. (Brahmi) extract and an anthocyanin-rich mixed Thai berry extract, administered individually and in combination, in a chronic unpredictable mild stress (CUMS) rat model. Methods: Extracts derived from Morus alba L. (mulberry), Antidesma ghaesembilla Gaertn. (mamao), and Syzygium nervosum DC. (ma-kiang) were characterized for anthocyanin and phenolic contents, antioxidant activities, and cyanidin-3-O-glucoside levels using HPLC. Male Sprague Dawley rats were subjected to a 14-day CUMS protocol and treated with Brahmi extract, mixed Thai berry extract, or their combinations. Behavioral assessments included the open-field test, elevated plus maze, and novel object recognition test. Histopathological evaluation of the prefrontal cortex and hippocampus was also performed. Results: Brahmi extract and mixed Thai berry extract attenuated selected anxiety-related behaviors and improved recognition memory-related parameters in CUMS-exposed rats. The low-dose berry extract produced the most consistent behavioral improvements, whereas combination-treated groups showed greater histological preservation. Histopathological analysis revealed reduced neuronal degeneration and improved tissue organization in the prefrontal cortex and hippocampus of treated animals. Conclusions: These findings support the potential therapeutic relevance of Bacopa monnieri and anthocyanin-rich Thai berry extracts under chronic stress conditions, with differential effects observed between individual and combination treatments.