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Leaf Disc Assays for Rapid Measurement of Antioxidant Activity.

ABSTRACT: Antioxidant levels are key parameters for studies of food quality, stress responses, and plant health. Herein, we have demonstrated that excised leaf disc has both radical scavenging activity and reducing power, and used this concept to develop 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), and potassium permanganate reduction (PPR) leaf disc assays. Reaction time and reagent concentration for these assays were optimized using leaves from spinach, kale, collards, mustard, and watermelon. Further, these assays were validated for linearity and intra-assay precision. Ultra-high performance liquid chromatography coupled to an electrospray quadrupole time-of-flight mass spectrometer (UPLC/ESI-HR-QTOFMS) was used for phytochemical profiling and studying relative abundances of certain phenolic compounds in various leaf discs suspended and cell-free extracts. The mass spectral analysis showed that leaf disc suspended methanolic extracts had almost same phytochemical profiles to those of cell-free extracts. The DPPH leaf disc assay demonstrated better radical scavenging potential than the conventional cell-free extract method. By contrast, the observed antioxidant activity values in ABTS and PPR leaf disc assays were lower than those of conventional cell-free extract-based methods. In conclusion, the developed leaf disc assays are simple and rapid for the qualitative and comparative assessment of the antioxidant potential of leaf samples, as well as can be a good alternative to conventional cell-free extract based methods.

SUBMITTER: Kasote DM

PROVIDER: S-EPMC6374478 | biostudies-literature | 2019 Feb

REPOSITORIES: biostudies-literature

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Leaf Disc Assays for Rapid Measurement of Antioxidant Activity.

Kasote Deepak M DM Jayaprakasha Guddadarangavvanahally K GK Patil Bhimanagouda S BS

Scientific reports 20190213 1

Antioxidant levels are key parameters for studies of food quality, stress responses, and plant health. Herein, we have demonstrated that excised leaf disc has both radical scavenging activity and reducing power, and used this concept to develop 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), and potassium permanganate reduction (PPR) leaf disc assays. Reaction time and reagent concentration for these assays were optimized using leaves from spi ...[more]

PMID: 30760761

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Project description:BackgroundDiabetes mellitus type II (DMT-2) is a widely spread metabolic disorder both in developed and developing countries. The role of oxidative stress is well established in DMT-2 pathogenesis. The synthetic drugs for DMT-2 are associated with serious side complications. Antioxidant and α-glucosidase inhibitory actions of phytochemicals from various plant species are considered as an alternative to synthetic drugs for DMT-2 management. The present study aimed to evaluate the antioxidant activity, α-glucosidase inhibitory potential and phytochemical profiling of Hyophorbe lagenicaulis.MethodsThe total phenolic and flavonoid contents, in vitro antioxidant activity (α, α-diphenyl-β-picrylhydrazyl (DPPH) free radical scavenging and phosphomolybdenum method) and α-glucosidase inhibition of ultrasonicated hydroethanolic H. lagenicaulis leaf extracts were determined spectrophotometrically. The results of DPPH assay and α-glucosidase inhibition were reported in terms of IC50 value. The phytochemical profiling was accomplished by UHPLC-Q-TOF/MS/MS technique.Results and discussionFindings leaped 60% ethanolic extract as rich fraction regarding total phenolic and flavonoid contents. The 60% ethanolic fraction was a promising source of natural antioxidants and α-glucosidase inhibitory agents as indicated by anti-radical and enzyme inibitory activities. Kaempferol, rutin, hesperetin 5-O-glucoside, kaempferol-coumaroyl-glucoside, luteolin 3-glucoside, Isorhamnetin-3-O-rutinoside, trimethoxyflavone derivatives and citric acid were identified by UHPLC-Q-TOF-MS/MS. These compounds were believed to be responsible for the strong antioxidant and enzyme inhibitory activity of plant extracts. The extensive metabolite profiling of H. lagenicaulis was carried out the first time as never reported previously. The H. lagenicaulis might be an appropriate choice to manage diabetes mellitus in an alternate way. The findings may be further exploited extensively for toxicity evaluation to proceed with functional food development having antidiabetic attributes.

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Leaf Disc Assays for Rapid Measurement of Antioxidant Activity.

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Leaf Disc Assays for Rapid Measurement of Antioxidant Activity.

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