Bopu powder^® and Sangrovit^® were developed from Macleayacordata and are widely used in agriculture and animal husbandry, but their impurities have been rarely reported in the literature. Impurity analysis is of great importance to the quality and safety of veterinary drugs. In this study, high-performance liquid chromatography/quadrupole time-of-flight mass spectrometry (HPLC-Q-TOF-MS) combined with a screening method was used to screen and characterize the impurities in Bopu powder^® and Sangrovit^®. A total of 58 impurities were screened from Bopu powder^® and Sangrovit^® using the screening strategies, of which 39 were identified by their accurate m/z value, characteristic MS/MS data, and fragmentation pathways of references. This established method was used for impurity analysis for the first time and proved to be a useful and rapid tool to screen and identify the impurities of Bopu powder^® and Sangrovit^®, especially for those at trace levels in a complex sample. In addition, this study marks the first comprehensive research into impurities in these two products and has great significance for the systematic detection of impurities in other plant-derived drugs.

In traditional Chinese medicine, Angelica dahurica is a valuable herb with numerous therapeutic applications for a range of ailments. There have not yet been any articles on the methodical assessment and choice of the best reference genes for A. dahurica gene expression studies. Real-time quantitative PCR (RT-qPCR) is widely employed as the predominant method for investigating gene expression. In order to ensure the precise determination of target gene expression outcomes in RT-qPCR analysis, it is imperative to employ stable reference genes. In this study, a total of 11 candidate reference genes including SAND family protein (SAND), polypyrimidine tract-binding protein (PTBP), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), actin (ACT), TIP41-like protein (TIP41), cyclophilin 2 (CYP2), elongation factor 1 α (EF1α), ubiquitin-protein ligase 9 (UBC9), tubulin β-6 (TUB6), thioredoxin-like protein YLS8 (YLS8), and tubulin-α (TUBA) were selected from the transcriptome of A. dahurica. Subsequently, three statistical algorithms (geNorm, NormFinder, and BestKeeper) were employed to assess the stability of their expression patterns across seven distinct stimulus treatments. The outcomes obtained from these analyses were subsequently amalgamated into a comprehensive ranking using RefFinder. Additionally, one target gene, phenylalanine ammonia-lyase (PAL), was used to confirm the effectiveness of the selected reference genes. According to the findings of this study, the two most stable reference genes for normalizing the expression of genes in A. dahurica are TIP41 and UBC9. Overall, our research has determined the appropriate reference genes for RT-qPCR in A. dahurica and provides a crucial foundation for gene screening and identifying genes associated with the biosynthesis of active ingredients in A. dahurica.

Sanguinarine (SAN), as the main active component of a traditional Chinese veterinary medicine, has been widely used in the animal husbandry and breeding industry. However, the metabolites of SA are still uncertain. Therefore, this research aimed to investigate the metabolites of SA based on rats in vivo. The blood, feces, and urine of rats were collected after the oral administration of 40 mg/kg SAN. Ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS/MS) was employed to identify the metabolites of SAN. The elemental composition of sanguinarine metabolites was inferred by analyzing their exact molecular weight, and the structures of the metabolites were predicted based on their fragment ions and cleavage pathways. A total of 12 metabolites were identified, including three metabolites in the plasma, four in the urine, and nine in the feces. According to the possible metabolic pathways deduced in this study, SAN was mainly metabolized through reduction, oxidation, demethylation, hydroxylation, and glucuronidation. This present research has summarized the metabolism of SAN in rats, which is helpful for further studying the metabolic mechanism of SAN in vivo and in vitro.

Lilium lancifolium Thunb., as a predominant variety of medicinal and edible lilies, has long been renowned in traditional medicine for “moistening the lungs, relieving coughs, and calming the mind to soothe the heart.” The bitter taste formation in L. lancifolium is predominantly attributed to secondary metabolites such as alkaloids, this study explores an alternative mechanism underlying taste divergence among Lilium brownii var. viridulum, and Lilium pumilum DC, proposing a foundational scientific question: Are peptides one of the important sources of bitterness in Lilium lancifolium Thunb.? Peptidomic analysis identified 8479 peptide sequences, with 46.27% upregulated in L. lancifolium flesh. Through high-throughput molecular docking with the bitter taste receptor TAS2R14, 214 candidate bitter peptides were identified, showing the strongest average binding affinity (−119.73 kcal/mol). Molecular dynamics simulations further demonstrated that four of these peptides formed stable interactions with key residues in TAS2R14. Cellular assays confirmed TAS2R14 activation by these peptides, as indicated by enhanced EGFP reporter fluorescence, upregulation of downstream signaling molecules (GNAT1, PLCB2, TRPM5), decreased cAMP levels, and increased IP₃ accumulation. Transcriptomic analysis further indicated that bitter peptides mediate taste transduction primarily through neuroactive receptor interaction pathways. These findings represent the first identification of bitter peptides as a key source of bitterness in L. lancifolium and elucidates their transduction mechanism combining peptidomics, computational simulation, and cellular validation. Our study provides a methodological framework for exploring flavor substances in other plant-derived foods.