V.Native mass spectrometry (native MS) has seen tremendous development and an increase in application over the past decade for the study of proteins and protein complexes. Although conventionally performed using a static nanospray emitter in an offline fashion, native MS has been increasingly applied in hyphenated methods, where a wide variety of separation techniques are directly coupled to online native MS detection. Those new developments have greatly expanded the utility of native MS in protein biopharmaceutical characterization. Analytical hydrophobic interaction chromatography (HIC) method, although frequently used for the characterization of monoclonal antibodies (mAbs) and antibody-drug-conjugates (ADCs), has rarely been explored for online coupling with native MS. This is largely due to the high salt concentrations used in HIC analysis that are not compatible with direct MS detection. In this study, we overcame this challenge via an innovative makeup and splitting flow design and successfully achieved d identity elucidation for the HIC-UV method used in quality control. Ulcerative colitis (UC), an immune system disease, is characterized by long duration and easy relapse. Sophora flavescens (S. flavescens), also named "Kushen", is a traditional Chinese medicine, widely used to treat UC in clinics. Alkaloids and flavonoids are the main constituents of S. flavescens. Previous studies indicated that the effects of S. flavescens against UC mainly attribute to its alkaloids. In view of the clinical applications of its flavonoids and our preliminary experiments on the effects of S. flavescens treatment, we speculated that flavonoids also could exert an anti-UC effect, but its efficacy and mechanism are still not yet to be revealed. Herein, we examined the pharmacodynamic effects of the ethyl acetate (EtOAc) extract of S.  https://www.selleckchem.com/products/U0126.html  flavescens EtOAc (SFE) against dextran sodium sulfate-induced UC rats for the first time. Pharmacodynamic effects indicated that SFE could significantly alleviate the loss in the body weight and shortening of the colon length, reduce colon bleeding and improve colon tissue damage of UC rats. A total of 28 prototypes and 41 metabolites were unambiguously or tentatively detected in rat's plasma and urine. Among them, 28 prototypes and 3 phase I metabolites shared 40 UC targets, the targets contributed to 51 metabolic pathways in 5 modules. Additionally, genistein, formononetin, isokurarinone, kurarinone, maackiain, kushenol N, trifolirnizin, kuraridin and norkurarinone were suggested to be potential active compounds in SFE for treating UC by comprehensively investigating the results of network pharmacology analysis, metabolic analysis in vivo, and previous researches. Finally, a combination of metabolic analysis in vivo with network pharmacology can elucidate the material basis and pharmacodynamic effect of traditional Chinese medicines, and lay the foundation for further clarify the anti-UC mechanism of SFE. In this article, we report a comprehensive characterization of volatile and polar constituents extracted from the aerial parts of Thymus munbyanus subsp. coloratus, a shrub that is used as culinary ingredient and as traditional medicine in Algeria, mainly to treat respiratory and gastrointestinal disorders and endocrine dysfunctions. Headspace solid phase microextraction (HS-SPME) coupled with gas chromatography-mass spectrometry (GC-MS) was used to assess volatile constituents, whereas the phytochemical composition of solid residues obtained from extraction with solvents at diffrent polarity was obtained by an integrated Nuclear Magnetic Resonance (NMR) and liquid chromatography coupled with tandem mass spectrometry (LC-MSn) approach. Fourty-five apolar ccompounds were identified, mainly oxygenated monoterpenes (65.8%), sesquiterpene hydrocarbons and nonoterpene hydrocarbons (18.6 and 14.5%, respectively). On the other hand, LC-MSn and NMR analyses revealed the presence of aglyconic and glycosilated flavonoids, phenylpropanoid derivatives and triterpenoid acids related to oleanolic acid, mainly in the methanol, dichloromethane and hexane extracts. Overall, these data indicate that Thymus munbyanus subsp. coloratus could be a potential source of antioxidants and bioactive compounds, and our results represent a starting point for further research on this plant species. Therapeutic monoclonal antibodies can potentially induce unwanted immune responses, resulting in the production of anti-drug antibodies (ADAs). The binding of ADAs to drugs and subsequent formation of immune complexes (ICs) can trigger various responses, dependent on the size, concentration, and subclass of ADAs. To better understand the impact of ADAs on pharmacokinetics, pharmacodynamics, and toxicological profiles, a bioanalytical method was developed for the detection of ICs between human monoclonal immunoglobulin G (IgG) and ADAs in biological samples. Regarding the experimental procedure, in brief, the human antibody-specific ICs and unbound human antibody in biological samples are separated through blue native polyacrylamide gel electrophoresis (BN-PAGE). The target fractions are then cut from the gel, followed by in-gel trypsin digestion and subsequent liquid chromatography tandem-mass spectrometry (LC-MS/MS) to monitor the human IgG-specific peptide. This method was able to detect various types of human antibodies with a lower limit of detection of 10 μg/mL in monkey serum. The assay performance for the detection of ICs was demonstrated using spiked samples, and pre-incubated ICs in monkey serum were clearly detected. Taken together, these findings indicate that our method enables a semi-quantitative analysis for estimating the ratio of human antibody included ICs in comparison to the total antibody. This method was successfully applied to an in vivo study using mice, and the data helped explain the unexpectedly rapid clearance of a humanized antibody due to the formation of large ICs. The combination of the separation of ICs by BN-PAGE and the detection of the human IgG-specific peptide by LC-MS/MS is a useful general bioanalytical approach for the detection of ICs in animals.