Identification and characterization of camptothecin tailoring enzymes in Nothapodytes tomentosa.

Camptothecin is a complex monoterpenoid indole alkaloid with remarkable antitumor activity. Given that two C-10 modified camptothecin derivatives, topotecan and irinotecan, have been approved as potent anticancer agents, there is a critical need for methods to access other aromatic ring-functionalized congeners (e.g., C-9, C-10, etc.). However, contemporary methods for chemical oxidation are generally harsh and low-yielding when applied to the camptothecin scaffold, thereby limiting the development of modified derivatives. Reported herein, we have identified four tailoring enzymes responsible for C-9 modifications of camptothecin from Nothapodytes tomentosa, via metabolomic and transcriptomic analysis. These consist of a cytochrome P450 (NtCPT9H) which catalyzes the regioselective oxidation of camptothecin to 9-hydroxycamptothecin, as well as two methyltransferases (NtOMT1/2, converting 9-hydroxycamptothecin to 9-methoxycamptothecin), and a uridine diphosphate-glycosyltransferase (NtUGT5, decorating 9-hydroxycamptothecin to 9-ß-D-glucosyloxycamptothecin). Importantly, the critical residues that contribute to the specific catalytic activity of NtCPT9H have been elucidated through molecular docking and mutagenesis experiments. This work provides a genetic basis for producing camptothecin derivatives through metabolic engineering. This will hasten the discovery of novel C-9 modified camptothecin derivatives, with profound implications for pharmaceutical manufacture.

Neonicotinoid insecticides in plant-derived Foodstuffs: A review of separation and determination methods based on liquid chromatography.

Neonicotinoids (NEOs) are the most widely used insecticides globally. They can contaminate or migrate into foodstuffs and exert severe neonic toxicity on humans. Therefore, lots of feasible analytical methods were developed to assure food safety. Nevertheless, there is a lack of evaluation that the impacts of food attributes on the accurate determination of NEOs. This review aims to provide a comprehensive overview of sample preparation methods regarding 6 categories of plant-derived foodstuffs. Currently, QuEChERS as the common strategy can effectively extract NEOs from plant-derived foodstuffs. Various enrichment technologies were developed for trace levels of NEOs in processed foodstuffs, and multifarious novel sorbents provided more possibility for removing complex matrices to lower matrix effects. Additionally, detection methods based on liquid chromatography were summarized and discussed in this review. Finally, some limitations were summarized and new directions were proposed for better advancement.

Purity Assessment of Dinotefuran Using Mass Balance and Quantitative Nuclear Magnetic Resonance.

Dinotefuran (DNT) belongs to the third-generation neonicotinoid pesticides, which are among the most common residuals in a variety of food commodities. To guarantee accurate quantification and traceability of results in food samples, certified reference materials (CRMs) are the indispensable benchmark. In this work, a DNT CRM was characterized and its purity was assessed by two independent methods, including mass balance (MB) and quantitative nuclear magnetic resonance spectroscopy (qNMR). The mass fraction of moisture was 0.33 mg/g, the inorganic impurity was 0.01 mg/g, and no detectable organic solvent was detected. Benzoic acid was chosen as the internal standard for qNMR. Its mass fraction was 997.9 mg/g and 992.9 mg/g by MB and qNMR, respectively. Eventually, the DNT CRM was assigned a mass fraction of 995 mg/g, with expanded uncertainty of 5 mg/g (k = 2). This CRM can be used to prepare calibrant solutions and is applicable to national routine monitoring of DNT residuals in agro-products and food.

Simultaneous Determination of Neonicotinoid and Carbamate Pesticides in Freeze-Dried Cabbage by Modified QuEChERS and Ultra-Performance Liquid Chromatography-Tandem Mass Spectrometry.

Dehydrated vegetables are popular in instant foods, but few reports have focused on their pesticide residues. This research developed and validated a modified QuEChERS method combined with ultra-performance liquid chromatography-tandem mass spectrometry to determine 19 kinds of neonicotinoid and carbamate pesticides in freeze-dried cabbage. Herein, acetonitrile/water (v/v = 2:1) was selected in the extraction step. Meanwhile, 4 g anhydrous magnesium sulfate and 1 g sodium chloride were applied to the partitioning step. Dispersive solid-phase extraction sorbents were selected, and liquid chromatography conditions were further optimized for dealing with the matrix effect. The limits of quantification ranged from 1.0 to 10.0 µg/kg. The validation results were acceptable, with average recoveries of 78.7-114.0% and relative standard deviations below 14.2%. The method recoveries were closely related to the volume proportion of water in the extractant. Finally, the developed method was applied to real freeze-dried cabbages and four pesticides (propamocarb, imidacloprid, acetamiprid, and thiacloprid) were detected in six samples.

Chlorantraniliprole in foods: Determination, dissipation and decontamination.

Chlorantraniliprole (CAP) is the first commercially available anthranilic diamide insecticide that targets ryanodine receptors. However, excessive use of CAP can lead to persistent contamination on treated foods and adverse effects on human wellness. The current review focuses on CAP residue analysis in foods by using chromatographic techniques. QuEChERS (quick, easy, cheap, effective, rugged and safe) is the most widely used sample preparation strategy and liquid chromatography tandem mass spectrometry is the predominant analytical method for various food matrices including vegetable, fruit, grain, fish and so on. Moreover, this review summarizes the dissipation pattern of CAP on foods and found it usually dissipates fast on plant in open-field environment. For decontamination, common culinary cleaning methods could effectively remove CAP from vegetables. Finally, some new directions are proposed for better advancement.

Impurity Profiling of Dinotefuran by High Resolution Mass Spectrometry and SIRIUS Tool.

Dinotefuran (DNT) is a neonicotinoid insecticide widely used in pest control. Identification of structurally related impurities is indispensable during material purification and pesticide registration and certified reference material development, and therefore needs to be carefully characterized. In this study, a combined strategy with liquid chromatography high-resolution mass spectrometry and SIRIUS has been developed to elucidate impurities from DNT material. MS and MS/MS spectra were used to score the impurity candidates by isotope score and fragment tree in the computer assisted tool, SIRIUS. DNT, the main component, worked as an anchor for formula identification and impurity structure elucidation. With this strategy, two by-product impurities and one stereoisomer were identified. Their fragmentation pathways were concluded, and the mechanism for impurity formation was also proposed. This result showed a successful application for combined human intelligence and machine learning, in the identification of pesticide impurities.

Impurity identification in thiamethoxam by high resolution mass spectrometry and computer assisted elucidation.

Thiamethoxam (TMX) is a widely used neonicotinoid insecticide in pest control. Identification of structurally related impurities is very important during certified reference material development and pesticide registration, thus it needs to be carefully characterized. In this study, a combined strategy with liquid chromatography-high resolution mass spectrometry and computer assisted elucidation (SIRIUS) has been developed for the impurity elucidation in TMX material. MS and MS/MS spectra were used to score the impurity candidates by isotope score and fragment tree in SIRIUS. TMX, the main component, worked as an anchor for formula identification and structure elucidation of impurity. With this strategy, four impurities were identified, including two byproducts (TMX-OCH3 and TMX-Cl) and two metabolites (clothianidin and TMX-urea). Their fragmentation pathways were concluded, and mechanism of impurity formation was also proposed. This result showed successful application of combining human intelligence with machine learning in impurity identification from chemicals.

The Rhododendron Plant Genome Database (RPGD): a comprehensive online omics database for Rhododendron.

BACKGROUND: The genus Rhododendron L. has been widely cultivated for hundreds of years around the world. Members of this genus are known for great ornamental and medicinal value. Owing to advances in sequencing technology, genomes and transcriptomes of members of the Rhododendron genus have been sequenced and published by various laboratories. With increasing amounts of omics data available, a centralized platform is necessary for effective storage, analysis, and integration of these large-scale datasets to ensure consistency, independence, and maintainability. RESULTS: Here, we report our development of the Rhododendron Plant Genome Database (RPGD; http://bioinfor.kib.ac.cn/RPGD/ ), which represents the first comprehensive database of Rhododendron genomics information. It includes large amounts of omics data, including genome sequence assemblies for R. delavayi, R. williamsianum, and R. simsii, gene expression profiles derived from public RNA-Seq data, functional annotations, gene families, transcription factor identification, gene homology, simple sequence repeats, and chloroplast genome. Additionally, many useful tools, including BLAST, JBrowse, Orthologous Groups, Genome Synteny Browser, Flanking Sequence Finder, Expression Heatmap, and Batch Download were integrated into the platform. CONCLUSIONS: RPGD is designed to be a comprehensive and helpful platform for all Rhododendron researchers. Believe that RPGD will be an indispensable hub for Rhododendron studies.