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idely used detection methods for activity analysis of type Ⅱ RIPs include depurination assay based on N-glycosidase activity and cytotoxicity assay. Both the methods provide simple, rapid, and sensitive analysis of type Ⅱ RIP toxicity, and complement other detection methods. Owing to the importance of type Ⅱ RIP toxins, the Organization for the Prohibition of Chemical Weapons (OPCW) has proposed clear technical requirements for the identification and analysis of relevant samples. We herein reviewed the structural characteristics, mechanism of action, and the development and application of type Ⅱ RIP detection methods; nearly 70 studies on type Ⅱ RIP toxins and their detection methods have been cited. In addition to the technical requirements of OPCW for the unambiguous identification of biotoxins, the trend of future development of type Ⅱ RIP-based detection technology has been explored.Ionic liquids are low-temperature molten salts with almost no vapor pressure, and they are composed of organic cations and inorganic anions. Ionic liquids are characterized by the properties of good chemical stability, high solubility, designable structure, high conductivity and so on. The physicochemical properties of an ionic liquid depend on the nature and size of the cation and anion, which confer unique characteristics; hence, these reagents are also termed "designed extractants." As a new class of green solvents, ionic liquids are potential replacements to traditional volatile organic solvents used for extraction; for this reason, ionic liquids have attracted the attention of scientists. Research on the methods of preparation and applications of ionic liquids is being diversified, and they are extensively used in catalytic chemistry, photoelectron chemistry, materials chemistry, analytical chemistry, etc. By functional guiding design, the structures of ionic liquids, especially the imidazole ring cation. Literature survey shows that the distribution of the binding active sites of ionic liquid-immobilized materials to the target molecules is more uniform, which increases the adsorption capacity of the materials. The adsorption efficiency of ionic liquid-immobilized materials is related to the type of ionic liquid, amount of adsorption material, concentration of the sample solution, adsorption temperature, solution pH, flow rate of the eluent, and type and amount of the eluting solvent. The existing disadvantages of ionic liquids, such as simple structures, insufficient basic theoretical research, and unsatisfactory extraction degree in complex matrixes would also be discussed. The corresponding solutions would be presented with the aim of providing guidance for the application of ionic liquid-immobilized materials in the separation and analysis of targets in complex matrices, thus paving the way for a new direction in the field of extraction and separation.Polymeric monolithic columns are fabricated by in situ polymerization of the corresponding monomer, crosslinkers, porogenic solvents and radical initiators within a mold. Compared with the conventional packed solid phase extraction adsorbents, polymeric monolithic columns with a continuous porous structure process distinctive advantages of rapid mass transfer and excellent permeability, which facilitates the extraction of trace amounts of the target from the matrix even at high flow velocities. Besides, these materials can be easily fabricated in situ within various cartridges, avoiding a further packing step associated with packed particulate adsorbents. Additionally, the abundant monomer availability, flexible porous structure, and wide applicable pH range make monoliths versatile for use in separation science. Thus, polymeric monolithic columns have been increasingly applied as efficient and promising extraction media for sample pretreatment food, pharmaceutical, biological and environmental analyses. Howeanomaterials, such as carbonaceous nanoparticles, metallic materials and metal oxides, metal-organic frameworks, covalent organic frameworks and inorganic nanoparticles have been extensively explored as hybrid adsorbents in the modes of solid phase extraction, solid phase microextraction, stir bar sorption extraction and on-line solid phase extraction. This review specifically summarizes the fabrication methods for nanomaterial incorporated polymeric monoliths and their application to the field of sample pretreatment. The existing challenges and future possible perspectives in the field are also discussed.Microextraction is a rapidly developing sample preparation technology in the field of analytical chemistry, which is seeing widespread application. Accurate sample preparation can not only save time but also improve the efficiency of analysis, determination, and data quality. At present, sample pretreatment methods must be rapid, allow for miniaturization, automation, and convenient online connection with analytical instruments. To meet the requirements of green analytical methods and improve the extraction efficiency, microextraction techniques have been introduced as suitable replacements to conventional sample preparation and extraction methods. Microextraction using a packed sorbent (MEPS) is a new type of sample preparation technology. this website The MEPS equipment was prepared using microsyringe with a volume of 50-500 μL, including MEPS syringes and MEPS adsorption beds (barrel insert and needle, BIN), which is essentially similar to a miniaturized solid phase extraction device. The BIN contains the adsorbent and widely used in the analysis of bio-matrix samples. MEPS devices could also play an important role in field pretreatment and analysis.In "shotgun" proteomics strategy, the proteome is explained by analyzing tryptic digested peptides using liquid chromatography-mass spectrometry. In this strategy, the retention time of peptides in liquid chromatography separation can be predicted based on the peptide sequence. This is a useful feature for peptide identification. Therefore, the prediction of the retention time has attracted much research attention. Traditional methods calculate the physical and chemical properties of the peptides based on their amino acid sequence to obtain the retention time under certain chromatography conditions; however, these methods cannot be directly adopted for other chromatography conditions, nor can they be used across laboratories or instrument platforms. To solve this problem, in recent years, deep learning was introduced to proteomics research for retention time prediction. Deep learning is an advanced machine-learning method that has extraordinary capability to learn complex relationships from large-scale data. ning are discussed.Protein glycosylation is a ubiquitous and important biological process involved in various molecular functions and biological pathways. It also yields important biomarkers for clinical diagnoses. However, glycopeptide analysis is challenging due to low abundance, low ionization efficiency, and glycan heterogeneity. In the present study, a method based on hydrophilic interaction liquid chromatography (HILIC) was developed for the selective enrichment of glycopeptides using a novel metal-organic framework (MOF) nanocomposite (AuGC/ZIF-8). Dual functionalization with glutathione and cysteine has resulted in an ultra-hydrophilic MOF, with synergistic effects and lower steric hindrance, providing more affinity sites for the glycopeptide enrichment. Horseradish peroxidase (HRP) was used as a model glycoprotein, and AuGC/ZIF-8 was used to enrich glycopeptides prior to analysis by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). AuGC/ZIF-8 displayed outstanding performance at enriching HRP glycopeptides, with high enrichment capacity (250 μg/mg), high selectivity in mixtures containing bovine serum albumin (BSA) (HRP-BSA (1∶200, mass ratio)), and high sensitivity at very low content (0.3 ng/μL). Thus this MOF holds promise for in-depth, comprehensive glycoproteomic and related analysis.Bioactive peptides play important roles in promoting human health, such as lowering blood pressure, blood sugar and blood lipid, anti-obesity, and anti-cancer. Thus, exploring functional bioactive peptides and developing efficient production technologies are of crucial importance. Herein, we review the development of function discovery and production technology for natural bioactive peptides. Presently, the top-down and bottom-up approaches are mainly used for the function discovery and production of natural active peptides. The top-down approach includes the direct extraction and identification for functional discovery, and the direct extraction, enzymatic hydrolysis and microbial fermentation for production. The bottom-up approach includes the polypeptide modification and database mining for functional discovery, and the chemical synthesis, enzyme synthesis, recombinant expression and cell-free synthesis for production. The top-down approach is usually associated with complicated process, lower efficiency, higher cost, harder quality control, and uncertain functionality, while the bottom-up approach is more suitable for the development of peptide drugs but difficult to be used for functional foods. With the technology development of sequencing and mass spectrometry, it is easier to obtain the proteomic information of various organisms at the molecular level. Based on the proteomic information, the top-down and bottom-up approaches can be combined to overcome the disadvantages of using these two approaches alone, thus providing a new strategy for the rapid development and production of natural active peptides.Angucyclines/angucyclinones are a large group of polycyclic aromatic polyketides and their producers are widely distributed in nature. This family of natural products attracts great attention because of their diverse biological activities and unique chemical structures. With the development of synthetic biology and the exploitation of the actinomycetes from previously unexplored environments, angucyclines/angucyclinones-like natural products with new skeletons were continuously discovered, thus enriching the structural diversity of this family. In this review we summarize the new angucyclines/angucyclinones analogues discovered in the last decade (2010-2020) by using different strategies, such as changing cultivation conditions, genetic modification, genome mining, bioactivity-guided compound isolation, and fermentation of actinomycetes from underexplored environments. We also discuss the role of synthetic biology in the discovery and development of new compounds of the angucycline/angucyclinone family.Streptomyces are major sources of bioactive natural products. Genome sequencing reveals that Streptomyces have great biosynthetic potential, with an average of 20-40 biosynthetic gene clusters each strain. However, most natural products from Streptomyces are produced in low yields under regular laboratory cultivation conditions, which hamper their further study and drug development. The production of natural products in Streptomyces is controlled by the intricate regulation mechanisms. Manipulation of the regulatory systems that govern secondary metabolite production will strongly facilitate the discovery and development of natural products of Streptomyces origin. In this review, we summarize progresses in pathway-specific regulators from Streptomyces in the last five years and highlight their role in improving the yields of corresponding natural products.