A high throughput method for detection of cyclooxygenase-2 enzyme inhibitors by effect-directed analysis applying high performance thin layer chromatography-bioassay-mass spectrometry.

A high throughput method was developed to detect bioactive molecules with inhibitory activity over cyclooxygenase (COX-2) enzyme applying effect-directed analysis and planar chromatography hyphenated with bioassay and mass spectrometry. The assay was based on the indirect measurement of arachidonic acid transformation into prostaglandin with the colorimetric co-substrate N,N,N',N'-tetramethyl-p-phenylenediamine. Inhibitory zones were observed as colorless bands over a blue background. Using a central composite design the critical factors like substrate concentration, enzyme: substrate ratio, reaction time, and co-substrate concentration were optimized. Optimal conditions were achieved with 0.03 mg/mL of arachidonic acid, 0.15 U/mL of COX-2, and 8.21 mg/mL of chromogenic reagent. Method usefulness was challenged analyzing fresh Chiloe's giant garlic (Allium ampeloprasum L) ethanol: water (8:2 v/v) extract, finding COX-2 inhibitors that were preliminarily identified as the isomers γ-glutamyl-S-allyl-l-cysteine and γ-glutamyl-S-(trans-1-propenyl)-L- cysteine.

Bioactive Compounds and Antioxidant Activity in the Fruit of Rosehip (Rosa canina L. and Rosa rubiginosa L.).

Rosehips (Rosa spp., Rosaceae) are wild rose bushes with more than 100 species. Its fruits vary in colour and size, depending on the species, and are recognised for their nutritional characteristics. Ten samples of Rosa canina L. and Rosa rubiginosa L. fruits were collected at different geographical points from Southern Chile. Nutrients such as crude protein and minerals and functional properties such as phenolic compounds, ascorbic acid, and also antioxidant activities were evaluated by HPLC-DAD-ESI-MS/MS. The results revealed a high content of bioactive compounds, primarily ascorbic acid (6.0 to 8.2 mg g-1 fresh weight (FW)), flavonols (427.9 ± 0.4 µg g-1 FW) and antioxidant activity. We established a relationship between the antioxidant activity using Trolox equivalent antioxidant capacity (TEAC), cupric reducing antioxidant capacity (CUPRAC) and 2,2-diphenyl radical (DPPH) methods and the concentration of uncoloured compounds, such as flavonols and catechin. This antioxidant activity was primarily associated with the samples from Gorbea, Lonquimay, Loncoche, and Villarrica localities, and all of them were of the species Rosa rubiginosa L. The results here obtained represent novel information of rosehip fruits. In this sense, the reported information about compounds and antioxidant activities in rosehip fruits allowed us to continue new lines of research in relation to the potential formulation of new functional foods and also in the treatment and/or prevention of some diseases.

Metabolic and antioxidant effects of inoculation with arbuscular mycorrhizal fungi in crops of flesh-coloured Solanum tuberosum treated with fungicides.

BACKGROUND: Solanum tuberosum tubers have higher content of phenolic compounds such as hydroxycinnamic acid derivatives (HCAD) and anthocyanins in coloured genotypes. The use of fungicides for crops is common, but there are few studies regarding the interaction of fungicides and arbuscular mycorrhizal fungi (AMF). Here, the AMF-plant interactions and the metabolic responses of three potato genotypes with different tuber colorations (VR808, CB2011-509 and CB2011-104) inoculated with Claroideoglomus claroideum (CC), Claroideoglomus lamellosum (HMC26) or Funneliformis mosseae (HMC7) were studied together with the use of the fungicides MONCUT (M) and ReflectXtra (R). Mycorrhizal traits, phenolic compound profiles and antioxidant activity (AA) were evaluated. RESULTS: Despite only two HCADs being identified, with 5-caffeolquinic acid the most abundant, four anthocyanins were detected only in purple potato genotypes. The anthocyanin and HCAD profiles, as well as AA, showed that the CB2011-104 genotype had better characteristics than the other genotypes, while VR808 and CB509 showed similar responses. The responses were dependent on the specific combinations of genotype, fungicide and the AMF strain, and generally showed better responses when colonized by AMFs. CONCLUSION: The three potato genotypes had differential responses depending on the inoculated AMFs and the fungicide applied before sowing, where the optimal combinations for antioxidant response, mycorrhization degree and performance were HMC26/R for VR808, HMC7/M for CB2011-509 and HMC26/M for CB2011-104. Our results suggest the existence of functional compatibility that can be registered as beneficial effects even at the genotypic level of the host regarding a specific AMF strain. © 2021 Society of Chemical Industry.

Effect of fertilization and arbuscular mycorrhizal fungal inoculation on antioxidant profiles and activities in Fragaria ananassa fruit.

BACKGROUND: The main methods used to increase the yield and quality of strawberry fruit produced in acidic soils with low P availability include root colonization by arbuscular mycorrhizal (AM) fungi and adjusting the type of P supplementation by means of traditional (TF) or organic (OF) fertilization, and adjusting nutrient doses. In this study, the antioxidant properties of strawberry fruit were evaluated under different treatments of fertilization (TF or OF) and different doses of P supplied at planting (0, 50 or 100% of the agronomic recommendation) and in the presence or absence of AM fungus as a bioinoculant. RESULTS: Fruits without fertilization treatments and with TF presented with higher anthocyanin concentrations. In general, higher values were obtained without AM colonization. However, spectrophotometric tests showed the highest activity and concentration in the AM-inoculated treatments. It is likely that phenolic compounds other than anthocyanins are present in the extracts. These other compounds could not be identified by the method used but could be detected by the Folin-Ciocalteu method. CONCLUSION: This study provides information that allows for improvements in strawberry fruit quality by agronomic management, with a potential beneficial effect on the health of consumers. © 2018 Society of Chemical Industry.