The supramolecular solvent (SUPRAS) has garnered significant attention as an innovative, efficient, and environmentally friendly solvent for the effective extraction and separation of bioactive compounds from natural resources. However, research on the use of a SUPRAS for the extraction of phenolic compounds from plants, which are highly valued in food products due to their exceptional antioxidant properties, remains scarce. The present study developed a green, ultra-sound-assisted SUPRAS method for the simultaneous determination of three phenolic acids in Prunella vulgaris using high-performance liquid chromatography (HPLC). The experimental parameters were meticulously optimized. The efficiency and antioxidant properties of the phenolic compounds obtained using different extraction methods were also compared. Under optimal conditions, the extraction efficiency of the SUPRAS, prepared with octanoic acid reverse micelles dispersed in ethanol–water, significantly exceeded that of conventional organic solvents. Moreover, the SUPRAS method demonstrated greater antioxidant capacity. Confocal laser scanning microscopy (CLSM) images revealed the spherical droplet structure of the SUPRAS, characterized by a well-defined circular fluorescence position, which coincided with the position of the phenolic acids. The phenolic acids were encapsulated within the SUPRAS droplets, indicating their efficient extraction capacity. Furthermore, molecular dynamics simulations combined with CLSM supported the proposed method’s mechanism and theoretically demonstrated the superior extraction performance of the SUPRAS. In contrast to conventional methods, the higher extraction efficiency of the SUPRAS can be attributed to the larger solvent contact surface area, the formation of more types of hydrogen bonds between the extractants and the supramolecular solvents, and stronger, more stable interaction forces. The results of the theoretical studies corroborate the experimental outcomes.

Supramolecular solvent (SUPRAS) has received increasing interest as an innovative, efficient, and green solvent for the effective extraction and separation of bioactive compounds from natural resources. However, studies on the use of SUPRAS for the extraction of phenolic components from plants, which are highly valued in food products due to their excellent antioxidant capacity, are still lacking. The present study developed a green ultrasound-assisted and efficient SU-PRAS method to simultaneously determine three phenolic acids in Prunella vulgaris by high-performance liquid chromatography (HPLC) and the experimental parameters were optimized in detail. The efficiency and antioxidant properties of the phenolic compounds obtained by different extraction methods were also compared. Under the optimal conditions, the extraction efficiency of SUPRAS, prepared with octanoic acid reverse micelles dispersed in ethanol-water, was substantially higher than conventional organic reagents. Moreover, this SUPRAS method also showed more significant antioxidant capacities. The confocal laser scanning microscopy (CLSM) images presented the spherical droplet structure of SUPRAS (A well-defined circle), which coincided with the position of the fluorescence position of phenolic acids. The phenolic acids were encapsulated in the vesicles by the SUPRAS droplets, which indicated the efficient extraction capacities of SUPRAS for the target compounds. Finally, combined with the CLSM method, the mechanism of the proposed method was further clarified, and the superior extraction performance of SUPRAS was theoretically demonstrated through molecular dynamics simulations. In comparison with conventional methods, the higher extraction efficiency of SUPRAS can be illustrated by the larger solvent contact surface area, more types of hydrogen bonds between extractants and supramolecular solvents, and stronger and more stable interaction forces. The results of the theoretical studies are in rational agreement with the experimental outcome.

In managing unique complexities associated with Chinese medicinal quality assessment, metabolomics serves as an innovative tool. This study proposes an analytical approach to assess differing qualities of Scrophularia ningpoensis （S. ningpoensis）Hemsl by identifying potential biomarker metabolites and their activity with the corresponding secondary metabolites. The methodology includes four steps; first, a GC-MS based metabolomics exploration of the Scrophularia ningpoensis Hemsl. Second, a multivariate statistical analysis (PCA, PLS-DA, OPLS-DA) for quality assessment and biomarker identification. Third, the application of ROC analysis and pathway analysis based on identified biomarkers. Finally, validation of the associated active ingredients by HPLC. The analysis showed distinct metabolite profiles across varying grades of S. ningpoensis Hemsl, establishing a grading dependency relationship. Select biomarkers (gluconic Acid, d-xylulose, sucrose, etc.) demonstrated robust grading performances. Further, the Pentose Phosphate Pathway, deemed as most influential in grading, was tied to the synthesis of key constituents (iridoids, phenylpropanoids). HPLC validation tests affirm a decreasing trend in harpagoside and cinnamic acid levels between first and third-grade samples. In conclusion, this GC-MS based metabolomics combined HPLC method offers a sound approach to assess and distinguish quality variations in S. ningpoensis Hemsl samples.

Background: Bovis Calculus (NBC) is widely used for the management of convulsive disorder, but its clinical application is constrained by limited availability and high cost. Bovis Calculus Artifactus (BCA) and Bovis Calculus Sativus (BCS) are commonly employed as substitutes. However, systematic evidence comparing their anticonvulsant efficacy and pharmacological characteristics remains limited. Methods: This study evaluated the substitution potential of BCA and BCS relative to NBC and characterized their distinct pharmacological profiles in a pentylenetetrazol (PTZ)-induced acute seizure mouse model. Seizure latency and duration were evaluated, hippocampal pathology was examined by hematoxylin–eosin and Nissl staining, neurotransmitters and neuroinflammatory factors were quantified. Network pharmacology and molecular docking analyses were used to explore the underlying mechanisms. Results: The results showed that preventive administration of NBC, BCA, and BCS significantly alleviated PTZ-induced acute seizures, although their efficacy patterns differed markedly. NBC exhibited the broadest anticonvulsant spectrum, significantly prolonging seizure latency at multiple seizure stages. BCS showed a more restricted effect, primarily delaying stage IV seizures, whereas BCA predominantly reduced seizure frequency without significantly affecting seizure latency, suggesting that seizure frequency may represent a sensitive endpoint for evaluating certain anticonvulsant agents. Histopathological analyses demonstrated that all three preparations exerted neuroprotective effects in hippocampal subregions. NBC provided the most extensive morphological preservation in H&E staining, while all treatments significantly improved neuronal survival as assessed by Nissl staining. Biochemical analyses revealed distinct regulatory characteristics among the three Bovis Calculus sources. All treatments partially restored PTZ-induced neurotransmitter disturbances and attenuated neuroinflammatory responses. BCA showed prominent regulation of neurotransmitters and IL-1β, NBC exhibited broader modulation of neurotransmitter homeostasis, whereas BCS displayed stronger suppression of IL-6 and TNF-α despite limited effects on 5-HT regulation. These findings indicate that comparable protective effects against PTZ-induced acute seizures may be achieved through different neurochemical and inflammatory regulatory patterns. Network pharmacology and molecular docking analyses further suggested the potential involvement of PI3K-AKT and MAPK-related signaling pathways in the effects of NBC and its substitutes. Conclusions: Collectively, this study provides a comparative evaluation of NBC, BCA, and BCS and indicates that their protective activities against PTZ-induced seizures are associated with distinct neurochemical and anti-inflammatory profiles. These findings offer experimental evidence supporting the rational evaluation and application of alternative Bovis Calculus resources.