Radix Rehmanniae (RR) is a kind of herb which is widely used in the clinical and food processing industry. There are four forms of RR used in traditional Chinese medicine practice, which include fresh RR (FRR), raw RR (RRR), processed RR (PRR), and another processed RR (APRR), in which the APRR was processed by nine cycles of repeated steaming and drying. There are a large number of saccharides in RR. However, the differences in content were shown by different processing methods. In this study, an effective method using high-performance liquid chromatography (HPLC) and high-performance liquid chromatography-mass spectrometry (LC-MS) coupled with multivariate statistical analysis to rapidly distinguish different RR samples and validate the proposed chemical conversion mechanism. The datasets of the content of saccharides were subjected to principal component analysis (PCA) and one-way analysis of variance. The results showed that there different changes occurred in the contents of saccharides corresponding to the different processing methods, in which the contents of monosaccharides—namely arabinose, glucose, mannose, and galactose—had an increasing trend or remained relatively stable. However, the contents of fructose and oligosaccharides, including manninotriose, melibiose, sucrose, and raffinose, first increased and then reduced, or gradually decreased, yet the content of stachyose gradually decreased. The MSⁿ data indicated that manninotriose, melibiose, and some monosaccharides were produced by the hydrolysis of oligosaccharides. In addition, the fragmentation pathways of 1-phenyl-3-methyl-5-pyrazolone (PMP) derivatization of monosaccharides were also found that its glycosidic bond was first broken and subsequently its inside ring broke, and the characteristic fragment ions were produced at m/z 511.22, 493.20, 373.16, and 175.08 in the PMP derivatization of monosaccharides. In conclusion, this study illustrates the change and chemical conversion mechanism of saccharides by processing in RR samples which might play a key role in further application of RR.

Introduction: Sanjin tablets (SJT) are a well-known Chinese patent drug that have been used to treat urinary tract infections (UTIs) for the last 40 years. The drug consists of five herbs, but only 32 compounds have been identified, which hinders the clarification of its effective substances and mechanism.Methods: The chemical constituents of SJT and their effective substances and functional mechanism involved in the treatment of UTIs were investigated by using high performance liquid chromatography-electrospray ionization-ion trap-time of flight-mass spectrometry (HPLC-ESI-IT-TOF-MSⁿ), network pharmacology, and molecular docking.Results: A total of 196 compounds of SJT (SJT-MS) were identified, and 44 of them were unequivocally identified by comparison with the reference compounds. Among 196 compounds, 13 were potential new compounds and 183 were known compounds. Among the 183 known compounds, 169 were newly discovered constituents of SJT, and 93 compounds were not reported in the five constituent herbs. Through the network pharmacology method, 119 targets related to UTIs of 183 known compounds were predicted, and 20 core targets were screened out. Based on the “compound–target” relationship analysis, 94 compounds were found to act on the 20 core targets and were therefore regarded as potential effective compounds. According to the literature, 27 of the 183 known compounds were found to possess antimicrobial and anti-inflammatory activities and were verified as effective substances, of which 20 were first discovered in SJT. Twelve of the 27 effective substances overlapped with the 94 potential effective compounds and were determined as key effective substances of SJT. The molecular docking results showed that the 12 key effective substances and 10 selected targets of the core targets have good affinity for each other.Discussion: These results provide a solid foundation for understanding the effective substances and mechanism of SJT.

Nardosinone, a sesquiterpene peroxide, is one of the main active constituents of the ethnomedicine Nardostachyos Radix et Rhizoma, and it has many bioactivities, such as antiarrhythmia and cardioprotection. To elucidate its in vivo existence forms, its metabolism is first studied using mice. All urine and feces are collected during the six days of oral dosing of nardosinone, and blood is collected at one hour after the last dose. Besides, to validate some metabolites, a fast experiment is performed, in which nardosinone was orally administered and the subsequent one-hour urine is collected and immediately analyzed by UHPLC-Q-TOF-MS. In total, 76 new metabolites are identified in this study, including 39, 51, and 12 metabolites in urine, plasma, and feces, respectively. Nardosinone can be converted into nardosinone acid or its isomers. The metabolic reactions of nardosinone included hydroxylation, hydrogenation, dehydration, glucuronidation, sulfation, demethylation, and carboxylation. There are 56 and 20 metabolites with the structural skeleton of nardosinone and nardosinone acid, respectively. In total, 77 in vivo existence forms of nardosinone are found in mice. Nardosinone is mainly excreted in urine and is not detected in the feces. These findings will lay the foundation for further research of the in vivo effective forms of nardosinone and Nardostachyos Radix et Rhizoma.

Chrysanthemum vestitum, a perennial plant in the Compositae family and closely related to Chrysanthemum morifolium, has long been used as both food and medicine. To explore the structural characteristics and potential anti-inflammatory mechanisms of polysaccharides derived from C. vestitum, a homogeneous polysaccharide, CVP_C, was isolated from C. vestitum using hot water extraction and alcohol precipitation. The crude extract was then purified via DEAE-52 cellulose chromatography and gel filtration system. Structural characterization of CVP_C was performed by Fourier-transform infrared (FT-IR) spectroscopy, molecular weight determination, scanning electron microscopy (SEM), monosaccharide composition analysis, thermogravimetric analysis (TGA), nuclear magnetic resonance (NMR) spectroscopy, and methylation analysis. An LPS-induced ALI mouse model was established to assess the protective effects of CVP_C. Inflammatory cytokine levels (IL-6 and TNF-α) in lung tissues and bronchoalveolar lavage fluid (BALF) were quantified, and histopathological changes were evaluated. In addition, gut microbiota profiling was employed to study the underlying mechanisms. Structural analysis revealed that CVP_C is a xylan-type polysaccharide and has a molecular weight of 25.049 kDa. It is primarily composed of mannose (Man), galactose (Gal), glucosamine (GlcN), glucose (Glc), glucuronic acid (GlcA), xylose (Xyl), rhamnose (Rha), and arabinose (Ara). The backbone structure consisted of → 4)-β-D-Xylp-(1 → residues. In vivo, CVP_C significantly reduced total protein content in BALF and suppressed the expression levels of TNF-α and IL-6 in both BALF and lung tissues, effectively attenuating pulmonary inflammation in the murine model. Histopathological examination and lung injury score further demonstrated inhibition of inflammatory cell infiltration and alleviation of lung tissue injury following CVP_C treatment. Gut microbiota analysis revealed that CVP_C reversed LPS-induced dysbiosis by increasing the relative abundance of Firmicutes while decreasing that of Bacteroidota, thereby ameliorating ALI. CVP_C is a novel acidic xylan polysaccharide that protects against ALI by reducing pro-inflammatory cytokine secretion and inhibiting inflammatory cell infiltration. Its therapeutic mechanism is likely linked to modulation of gut microbiota composition, promoting beneficial bacteria and suppressing pathogenic taxa. These findings indicate that CVP_C has potential for use as a natural anti-inflammatory agent and functional food ingredient for the promotion of respiratory and gastrointestinal health.