Citri Reticulatae Pericarpium Viride (CRPV) is the processed product of Citrus reticulata Blanco. We systematically analyzed two CRPV types, Geqingpi (GQP) and Sihuaqingpi (SHQP), based on powder color, microscopic characteristics, and chemical composition. In addition, we characterized their constituents via ultra-high-performance liquid chromatography with hybrid quadrupole-orbitrap mass spectrometry (UHPLC-Q-Exactive Orbitrap-MS). Both showed significant differences in their powder color and microscopic characteristics. Fourier-transform infrared (FT-IR) spectroscopic analysis results showed that the C=O peak absorption of carboxylic acids and their carbonyl esters in SHQP was higher than that of GQP, while the C-OH and C-H plane bending peaks of polysaccharides were lower than those of GQP. We analyzed these data via similarity analysis, PCA, and OPLS-DA. GQP and SHQP had large distinct differences. Based on the mass measurements for molecular and characteristic fragment ions, we identified 44 main constituents from CRPV, including different flavonoid glycosides and flavonoid aglycones in SHQP and GQP, respectively. We found luteolin-6-C-glucoside, orientin, rhoifolin, and pilloin solely in SHQP, and naringenin and hesperetin only in GQP. The peak area measurements showed GQP having a higher flavonoid glycoside (narirutin, hesperidin, etc.) content, whereas SHQP had a higher polymethoxyflavone (nobiletin, tangeretin, etc.) content. Since we holistically analyzed two CRPV types, the results can not only support future pharmacological research, but also provide a scientific basis for formulating more reasonable CRPV quality standards and guide its clinical potential as a precision medicine.

The roots of Salvia miltiorrhiza (Danshen) is a precious herbal medicine used to treat cardiovascular diseases. This study establishes a high-performance liquid chromatography-tandem mass spectrometric (HPLC-MS/MS) method to quantify seven bioactive constituents from Danshen in rat plasma simultaneously. Chromatographic separation is performed on an Agilent Eclipse Plus C18 column (150 mm × 2.1 mm, 5 μm), utilizing a gradient of acetonitrile and 0.2% formic acid aqueous solution as the mobile phase, at a flow rate of 0.6 mL/min. We conduct a tandem mass spectrometric detection with electrospray ionization (ESI) interface via multiple reaction monitoring (MRM) in both positive and negative ionization mode. Our results show that a linear relationship is established for each analyte of interest over the concentration range of 0.5–300 ng/mL with r ≥ 0.9976. The validated method is successfully used to compare the pharmacokinetic properties of crude and wine-processed Danshen extract orally administered to rats. C_max of tanshinone IIA, C_max, and AUC_0-t of dihydrotanshinone I decrease significantly (p < 0.05) in the wine-processed group. No significant changes for other compounds are observed. These results might provide meaningful information for the further application of wine-processed Danshen and understanding of wine-processing mechanisms.

Matrix-assisted diffusion-ordered spectroscopy (MA DOSY) technology enables efficient the virtual separation of components in a mixture according to their coefficients (D). In the current research, MA DOSY technology was applied for the analysis of a flavonoid mixture. To establish the method, five representative active flavonoid ingredients, including baicalein, baicalin, quercetin, puerarin and rutinum, were selected for the mixture model. The effects of the type and concentration of the matrix, solvent polarity, and nuclear magnetic resonance (NMR) experimental conditions on the resolution of the DOSY spectrum were investigated. It turned out that sodium dodecyl sulfate (SDS) showed the best performance in increasing the resolution of different analytes, which initially increased to the peak below the added amount of 9 mg, and decreased upon the addition of more SDS. In addition, the five flavonoids showed higher resolution in DMSO–d₆ than in MeOD. Experimental parameters of DOSY, including the number of scans (NS), dummy scans (DS), and value of the FID data points (TD), were also optimized. Finally, the above optimized method was used for the qualitative analysis of the total flavonoid mixture extracted from Scutellaria baicalensis Georgi. A total of nine compounds were identified and confirmed by comparing them with mass spectrometry data, which further verifies the practical value of this method upon analyzing flavonoid mixtures and provides some reference significance for the follow-up research.

Owing to their endocrine disruption effect, the hormone levels in animal-source food and medicines need to be efficiently and accurately quantified by a reliable analytical method. In the current study, by using a homonuclear decoupling and heteronuclear single quantum correlation (HSQC) experiment, coupled with non-uniform sampling (NUS) that was used to shorten the experimental time, we developed a method to quantify 10 hormone residues in animal-source products. This method was validated following the guidelines of USP–NF 2022. The application of the homonuclear decoupling (HD) technique to conventional HSQC yielded 2D spectra that exhibited excellent signal separation and specificity. For all the tested hormones, good linearity with correlation coefficients of more than 0.99 was observed in the linear range of 0.2–6 mg/0.6 mL. Satisfactory precision and recoveries of spiked animal samples were also obtained. Finally, the method was applied in residue determination of 10 hormones in real animal-source samples at the ug/g level.

Background/Objectives: Rauvolfia vomitoria alkaloids (RVA) exhibited excellent activity against colorectal cancer (CRC) growth in the heterotopic model. However, their antitumor effects in the orthotopic models, as well as the underlying mechanisms related to gut microbiota and serum metabolomics, remain poorly understood. Methods: We performed 16S rRNA sequencing and serum metabolomics analysis in orthotopic CRC-bearing mice following different treatments. Univariate and multivariate logistic regression analyses were used to screen key gut microbes and metabolites associated with RVA treatments. KEGG enrichment analysis was conducted to identify the metabolic pathways modulated by RVAs in vivo. Results: RVA, either alone or in combination with 5-Fluorouracil (5-FU), exhibited a potent inhibitory effect on CRC growth, comparable to intraperitoneally administered 5-FU. In terms of body weight and organ indices, RVA showed lower toxicity compared to 5-FU. Both treatments induced measurable shifts in the gut microbial profiles of CRC mice. Increased abundance of Akkermansia and Lactobacillus and decreased abundance of Enterococcus were found to be associated with RVA monotherapy. The enrichment of Acinetobacter and Mucispirillum in CRC mice was suppressed by the combination treatment. A serum metabolomics study revealed that linoleic acid metabolism and β-oxidation of long-chain fatty acids were modulated by both RVA-containing regimens. Conclusions: These findings demonstrate that oral administration of RVA, either alone or combined with current clinical therapeutic methods, may represent a promising strategy for CRC treatment. Targeted modulation of gut microbiota and fatty acid metabolism may provide promising approaches to hinder CRC progression.