A multiplex approach of MS, 1D-, and 2D-NMR metabolomics in plant ontogeny: A case study on Clusia minor L. organs (leaf, flower, fruit, and seed).

INTRODUCTION: The genus Clusia L. is mostly recognised for the production of prenylated benzophenones and tocotrienol derivatives. OBJECTIVES: The objective of this study was to map metabolome variation within Clusia minor organs at different developmental stages. MATERIAL AND METHODS: In total 15 organs/stages (leaf, flower, fruit, and seed) were analysed by UPLC-MS and 1H- and heteronuclear multiple-bond correlation (HMBC)-NMR-based metabolomics. RESULTS: This work led to the assignment of 46 metabolites, belonging to organic acids(1), sugars(2) phenolic acids(1), flavonoids(3) prenylated xanthones(1) benzophenones(4) and tocotrienols(2). Multivariate data analyses explained the variability and classification of samples, highlighting chemical markers that discriminate each organ/stage. Leaves were found to be rich in 5-hydroxy-8-methyltocotrienol (8.5 µg/mg f.w.), while flowers were abundant in the polyprenylated benzophenone nemorosone with maximum level detected in the fully mature flower bud (43 µg/mg f.w.). Nemorosone and 5-hydroxy tocotrienoloic acid were isolated from FL6 for full structural characterisation. This is the first report of the NMR assignments of 5-hydroxy tocotrienoloic acid, and its maximum level was detected in the mature fruit at 50 µg/mg f.w. Seeds as typical storage organ were rich in sugars and omega-6 fatty acids. CONCLUSION: To the best of our knowledge, this is the first report on a comparative 1D-/2D-NMR approach to assess compositional differences in ontogeny studies compared with LC-MS exemplified by Clusia organs. Results derived from this study provide better understanding of the stages at which maximal production of natural compounds occur and elucidate in which developmental stages the enzymes responsible for the production of such metabolites are preferentially expressed.

Assessment of the Long-Range NMR C,H Coupling of a Series of Carbazolequinone Derivatives.

Synthesis, the complete 1H- and 13C-NMR assignments, and the long-range C,H coupling constants (nJC,H) of some hydrogen-deficient carbazolequinones, assessed by a J-HMBC experiment, are reported. In these molecules, the protons, used as entry points for assignments, are separated by several bonds with non-protonated atom carbons. Therefore, the use of long-range NMR experiments for the assignment of the spectra is mandatory; we used HSQC and HMBC. On the other hand, the measured heteronuclear (C,H) coupling constants 2J to 5J) allow us to choose the value of the long-range delay used in the HMBC experiment less arbitrarily in order to visualize a desired correlation in the spectrum. The chemical shifts and the coupling constant values can be used as input for assignments in related chemical structures.

Regioselective preparation and NMR spectroscopy study of 2-chloro-4-ethoxy-quinoline for the synthesis of 2-((3-aminopropyl)amino)quinolin-4(1H)-one.

Herein, we describe the C4-ethoxylation of 2,4-dichloroquinoline to prepare 2-chloro-4-ethoxy-quinoline (3), which is a prominent intermediate used for the synthesis of 2-substituted quinolones. To achieve this goal, we studied different conditions for the reaction between 2,4-dichloroquinoline and sodium ethoxide. We discovered that the use of 18-crown-6 ether as an additive and dimethylformamide as the reaction solvent allowed us to obtain the desired product 3 in very good yield and selectivity. In addition, a definitive distinction between the C2 and C4 ethoxylation products was achieved using 1 H─15 N heteronuclear multiple bond correlation. Compound 3 is an intermediate used for the synthesis of 2-((3-aminopropyl)amino)quinolin-4(1H)-one, which displays peculiar behavior during 1 H nuclear magnetic resonance analysis, such as the broadening of the H8 singlet and unexpected deuteration at the C8-position. Effort has been dedicated to understand these findings.

Isolation, characterisation and antibacterial activity studies of coumarins from Rhododendron lepidotum Wall. ex G. Don, Ericaceae / Estudos de isolamento, caracterização e atividade antibacteriana de cumarinas de Rhododendron lepidotum Wall. ex G. Don, Ericaceae

Six coumarins daphnin (1), daphnetin (2), daphnetin glucoside (3), rhodonetin (4), rhodonin (5) and umbelliferone (6) were isolated from the methanolic extract of Rhododendron lepidotum Wall. ex G. Don, Ericaceae (aerial part). The compounds and their acetyl derivatives were screened for antibacterial activity against Staphylococcus aureus ATCC-29213, methicillin resistant Staphylococcus aureus ATCC-15187, Escherichia coli ATCC-8739, Pseudomonas aeruginosa ATCC-9027 by microdilution method as compared to the reference ciprofloxacin. Compound 2 displayed the best antibacterial activity with MIC 125 μg/mL against S. aureus ATCC-29213 and MRSA ATCC-15187 followed by 4 which exhibited the MIC value of 250 μg/mL against all the four tested strains. All molecules showed better antibacterial activity than their acyl derivatives.

Seis cumarinas dafinina (1), dafinetina (2), dafinetina glicosídeo (3), rodonetina (4), rodonina (5) e umbeliferona (6) foram isoladas do extrato metanólico das partes aéreas de Rhododendron lepidotum Wall. ex G. Don, Ericaceae. Os compostos e seus derivados acetilados foram testados para verificar sua atividade antibacteriana contra Staphylococcus aureus ATCC-29213, Escherichia coli resistente à meticilina, Staphylococcus aureus ATCC-15187, ATCC-8739, Pseudomonas aeruginosa ATCC-9027, pelo método de microdiluição, usando ciprofloxacina como referência. A substância 2 apresentou a melhor atividade antibacteriana com o MIC 125 μg/mL contra S. aureus ATCC-29213 e MRSA ATCC-15187 seguido pela substância 4, que apresentou o valor de CIM de 250 μg/mL contra as quatro cepas testadas. Todas as moléculas apresentaram melhor atividade antibacteriana do que seus derivados acetilados.