BACKGROUND AND PURPOSE:Euphorbia pekinensis (EP) is known to cause significant intestinal toxicity, primarily manifesting as severe diarrhoea, yet the precise molecular mechanisms and the active components responsible have remained elusive. This study aimed to identify the diarrheal constituents of EP and elucidate the molecular pathway through which they induce gut toxicity. EXPERIMENTAL APPROACH:The laxative effects of EP components were assessed in vivo using mouse models and diarrhoea-related indicators, with histological analysis of intestinal tissue. Ex vivo rabbit intestinal tract assays were employed to study smooth muscle contraction. The underlying mechanism was investigated using intestinal organoid fluorescence co-localization and analysis of tryptophan metabolites in mice to determine the role of enterochromaffin (EC) cells and serotonin (5-HT). KEY RESULTS:We identified specific glycosphingolipids (GSLs), including a novel hexosylceramide (HexCer), as the primary toxic agents in EP. These GSLs act as direct agonists of the TRPA1 ion channel on intestinal EC cells. This activation triggers a TRPA1-mediated influx of Ca2+ into EC cells, leading to excessive 5-HT release. The resulting localized overstimulation of 5-HT receptors causes aberrant intestinal smooth muscle contraction and epithelial hypersecretion, culminating in severe diarrhoea. CONCLUSION AND IMPLICATIONS:This research reveals that the gut toxicity of EP is driven by a previously unrecognized GSL-TRPA1-5-HT signalling pathway in the intestinal epithelium. These findings provide a clear mechanistic basis for EP-induced diarrhoea and highlight a potential new target for managing gut toxicity.

Oligosaccharides in Atractylodes macrocephala Koidz. (AMK) have a wide range of clinical effects, especially in terms of improving immunity. In this study, the oligosaccharide fingerprint of AMK was first established by high-performance liquid chromatography coupled with evaporative light-scattering detection. This method was confirmed to be accurate and reliable. Twenty-six batches of samples from four regions were detected, and the oligosaccharide fingerprint similarity of AMK was evaluated (> 0.900). Second, the oligosaccharide profile of AMK was analyzed using high-performance liquid chromatography coupled to high-resolution tandem mass spectrometry. Fourteen inulin-type oligosaccharides with a degree of polymerization of 2–15 were identified, including sucrose, 1-kestose, and nystose. Finally, two representative compounds, 1-kestose and nystose, were selected for quantification analysis. The established method had good regression equations, precision, repeatability, and stability. The average contents of 1-kestose and nystose were 0.688% and 0.827%, respectively. This study provides valuable information on the quality evaluation and discrimination of different varieties of AMK.

Gelatins are important and frequently-used food additives, but the quality control of gelatins is always an intractable problem in routine analysis. In this work, a new strategy for simultaneously rapid identification and quantification of gelatins from various species using ultrasound assisted digestion-UPLC-MS/MS was described. Fourteen peptide markers were used for authenticity of gelatins by the presence or absence of these species-specific peptides. In the meantime, adulteration ratios could be calculated based on the peak area of different species peptide markers. With 10 g/L trypsin and ultrasound application, digestion time could be decreased from >12 h (traditional method) to 5 min, and the determination of animal gelatins could be achieved in 20 min with a single analysis run. Twenty-five commercial gelatin samples were screened, among which two deer-hide gelatin samples were adulterated with 900 g/kg and 265 g/kg cattle-hide gelatin (CHG); two donkey-hide gelatin (ACC) samples were adulterated with 66 g/kg and 381 g/kg horse-hide gelatin; and one ACC sample was adulterated with 786 g/kg CHG. This strategy provides an efficient and sensitive authentication and traceability of gelatin-containing products, and could also be applied to processed meat or protein food.

The essential oil from Curcumae wenyujin (CWEO) has antibacterial activity, but the antibacterial compounds in the CWEO have not been identified. The spectrum-effect relationship between chemical compositions and pharmacological activity is regarded as a potentially useful tool to screen bioactive compounds. The binary chromatographic fingerprint of CWEO was established by gas chromatography/mass spectrometry (GC-MS) and high-performance liquid chromatography/diode-array (HPLC-DAD). A data-level information integration method was used to investigate the chemical characteristics encoded in two chromatographic fingerprints. Additionally, the antibacterial activity against Escherichia coli (E. coli）was determined by the liquid dilution method. The spectrum-effect relationship between the binary chromatographic fingerprint and antibacterial activity was investigated with principal component analysis (PCA) and orthogonal projection to latent structures (OPLS) analysis. Compared with a single data matrix, the combined data matrix allowed more accurate identification of the strong outliers in the PCA. Additionally, the results of OPLS were used to establish a spectrum-effect relationship with a closer correlation than that generated with the single data matrix. The results indicated that germacrone, curdione, and furanodiene were detected in HPLC and GC-MS fingerprints, and may be the main antibacterial components. Abbreviations: CWEO, essential oil from C. wenyujin; MIC, minimum inhibitory concentration; OPLS, orthogonal projection to latent structures.

Paclitaxel-betulinic acid hybrid nanosuspensions (PTX-BA-NP) with increased anti-breast cancer activity were developed. The resultant nanosuspensions (NP) had a mean particle size of 282.54 +/- 5.4 nm, a polydispersity index of approximately 0.242 +/- 0.02, a zeta potential of -19.7 +/- 0.19 mV and a redispersibility index of 103.3 +/- 0.01%. The cumulative dissolution percentage of PTX coarse powder and PTX-BA-NP dried powder at 60 min were 15.4% and 90.8%, respectively. MCF-7 cell-based testing showed that treatment with PTX-BA-NP led to more PTX-BA-NP accumulation in the cytoplasm of breast cancer cells, less cell cycle arrest in the G2-M phase, more cell cycle arrest in the GO-G1 phase, more apoptosis-induced cell death and stronger inhibition of cell migration than paclitaxel nanosuspensions (PTX-NP). Biodistribution studies showed that tumor accumulation levels at 12 h in the PTX-BA-NP group were approximately 2.67- and 2.33-fold higher than the levels in the Taxol (R) and PTX-NP groups, respectively. in vivo antitumor efficacy demonstrated that PTX-BA-NP exerted the strongest tumor inhibition among the four groups, with a tumor inhibition rate of 47.79 +/- 2.28%, followed by PTX-NP (35.05 +/- 5.55%), Taxol (R) (22.67 +/- 6.01%) and betulinic acid nanosuspension (BA-NP) (14.38 +/- 6.02%). These findings indicate that PTX-BA-NP holds great promise for breast cancer therapy.

Anhydrosafflor yellow B (AHSYB) is one of the major active water-soluble pigments from Carthamus tinctorius, which has been found to inhibit ADP-induced platelet aggregation and possess significant antioxidant activity. However, the metabolic fate of AHSYB in vivo remains unknown. In order to explore whether AHSYB is extensively metabolized, the metabolites of AHSYB in plasma, urine, bile, and feces samples after intravenous administration to the rats were investigated by ultra-fast liquid chromatography/quadrupole time-of-flight mass spectrometry (UFLC/Qq-TOF-MS/MS) combined with Metabolitepilot (TM) software. In total, AHSYB and 22 metabolites including both phase I and phase II metabolism processes were found and tentatively identified from the bio-samples. The metabolic pathways were involved in oxidation, reduction, hydroxylation, methylation, dimethylation, O-acetylation, hydrolyzation, sulfation, glucuronidaton, glutathionation and combination with glucose. The results showed that the renal and biliary routes play an important role in the clearance and excretion of AHSYB as well as hepatocyte metabolism. All of these results were reported for the first time and would contribute to a further understanding of the in vivo intermediated processes and metabolic mechanism of AHSYB and its analogs. (C) 2016 Elsevier B.V. All rights reserved.

The fingerprints of the volatile oil from Curcuma wenyujin Y.H. Chen & C. Ling were established by high performance liquid chromatography (HPLC) and the anti-tumor activities of its volatile oil were evaluated in sarcoma 180 (S180) tumor-bearing mice by intraperitoneal administration. The spectrum-effect relationships between HPLC fingerprints and anti-tumor activities were investigated using partial least squares (PLS). The results showed that a close correlation existed between the spectrum-effect relationships. Furandiene, germacrone and curdione in the HPLC fingerprints might be the main anti-tumor components. The anti-tumor activities of the volatile oil from C. wenyujin were related with the main active constituents. This work provides a general model of the combination of HPLC and anti-tumor effect in vivo to study the spectrum-effect relationships of the volatile oil from C. wenyujin, which would be helpful to demonstrate the sesquiterpenoids of C. wenyujin have in vivo anti-tumor activities.

Ultra-flow liquid chromatography/quadrupole-time-of-flight mass spectrometry (UFLC/Q-TOF MS) method combined with metabolitepilot(MT) software was used for analysis of the metabolites of liguzinediol in dogs. Urine, bile, feces and plasma samples were collected after intravenous administration of 8 mg/kg liguzinediol to healthy dogs. Besides liguzinediol, seven metabolites were detected and identified by UFLC/Q-TOF MS method. The results showed that liguzinediol had some main metabolic pathways in dogs including oxidation, sulfation, cysteine conjugation, N-acetylcysteine conjugation and glucuronidation.