A novel on-flow mass spectrometry-based dual enzyme assay

Seidl, Claudia; Vilela, Adriana Ferreira Lopes; Lima, Juliana M.; Leme, Gabriel Mazzi; Cardoso, Carmen L.

doi:10.1016/j.aca.2019.04.057

Cited by 13 publications

(17 citation statements)

References 32 publications

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“…The isolated metabolites 1 – 6 were tested for their ability to inhibit cholinesterase (ChEIs) (Table 3). The inhibition assays were carried out using an immobilized capillary enzyme reactor (ICER) based on acetylcholinesterase human recombinant and/or butyrylcholinesterase from human serum, respectively hu AChE-ICER and hu BChE-ICER (in accordance with published procedure) 25–27 . Compounds 2–6 were more active towards hu AChE than hu BChE, indicating a selective cholinesterase inhibition, and 4 was the most active compound, with 27.7% (100 μM) inhibition against hu AChE.…”

Section: Resultsmentioning

confidence: 99%

“…Test samples were submitted to a punctual cholinesterase inhibition screening assay. hu AChE and hu BChE were immobilized independently onto fused silica capillary (0.1 mm I.D × 0.375 mm × 30 cm) using glutaraldehyde as spacer ( hu AChE-ICER and hu BChE-ICER) 26,27 .…”

Section: Methodsmentioning

confidence: 99%

“…All analyses were performed at room temperature (21 °C). The enzymatic reaction was monitored by direct quantification of acetylcholine hydrolysis product, Ch ( m/z : [M + H] + 104.17) 26,27 .…”

Section: Methodsmentioning

confidence: 99%

“…2 U mL −1 of hu AChE and 0.5 U mL −1 of hu BChE were immobilized and the apparent kinetic constant (K Mapp = 70 μM) was obtained independently using acetylcholine (ACh) as substrate by varying the substrate concentration while measuring the Ch [M + H] + 104.17 m/z formation. GraphPad Prism 5 software was used to obtain Michaelis–Menten plots by nonlinear regression analysis 27 .…”

Section: Methodsmentioning

confidence: 99%

“…Fragmentation amplitude was set to 120%. Nitrogen was used as sheath gas and helium as collision gas 27 .…”

Section: Methodsmentioning

confidence: 99%

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Botryane terpenoids produced by Nemania bipapillata, an endophytic fungus isolated from red alga Asparagopsis taxiformis - Falkenbergia stage

Medina

Araújo

Batista

et al. 2019

Sci Rep

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A chemical study of the EtOAc extract of Nemania bipapillata (AT-05), an endophytic fungus isolated from the marine red alga Asparagopsis taxiformis - Falkenbergia stage, led to the isolation of five new botryane sesquiterpenes, including the diastereomeric pair (+)-(2 R ,4 S ,5 R ,8 S )-( 1 ) and (+)-(2 R ,4 R ,5 R ,8 S )-4-deacetyl-5-hydroxy-botryenalol ( 2 ), (+)-(2 R ,4 S ,5 R ,8 R )-4-deacetyl-botryenalol ( 3 ), one pair of diastereomeric botryane nor sesquiterpenes bearing an unprecedented degraded carbon skeleton, (+)-(2 R ,4 R ,8 R )-( 4 ) and (+)-(2 R ,4 S ,8 S )-( 5 ), which were named nemenonediol A and nemenonediol B, respectively, in addition to the known 4β-acetoxy-9β,10β,15α-trihydroxyprobotrydial ( 6 ). Their structures were elucidated using 1D and 2D NMR, HRESIMS and comparison with literature data of similar known compounds. The absolute configurations of 2 , 3 and 4 were deduced by comparison of experimental and calculated electronic circular dichroism (ECD) spectra, while those of 1 and 5 were assigned from vibrational circular dichroism (VCD) data. Compound 4 weakly inhibited acetylcholinesterase, whereas compound 1 inhibited both acetylcholinesterase and butyrylcholinesterase. Compounds 1 , 3 , 5 and 6 were tested against two carcinoma cell lines (MCF-7 and HCT-116), but showed no significant citotoxicity at tested concentrations (IC 50 > 50 µM).

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%