Analysis of Antioxidant Phytochemicals and Anti-Inflammatory Effect from Vitex rotundifolia L.f

Abstract

An extraction method using 80% EtOH was selected and applied to obtain the total extracts from leaves, flowers, fruits, twigs, and roots of Vitex rotundifolia L.f. based on the antioxidant activity-guided experiments. Subsequently, total extract from each part of V. rotundifolia was successfully partitioned into fractions, which were evaluated for their antioxidant and anti-inflammatory properties via DPPH, ABTS, and NO assays, respectively. Among them, EtOAc (E) and n-butanol (B) fractions showed the potent antioxidant activity and the methylene chloride (MC) fractions of roots, leaves, and fruits that exhibited strong scavenging activity on DPPH and ABTS radicals. In the anti-inflammatory assay, n-hexane (H) and MC fractions of leaves potently inhibited NO production in LPS-stimulated RAW264.7 cells, followed by E fractions derived from fruits, flowers, twigs, and roots, along with B fractions from flowers and twigs. Additionally, a comprehensive HPLC-decoupled MS profiling was established and validated using seven isolated marker compounds (1–7), which were identified by analysis of their UV, NMR, and MS data. The established method was also applied for quantification of these marker compounds in each organ collected from different locations, and to assess their antioxidant capacity by a screening DPPH-HPLC method. Principal component analysis suggested the botanical organs from this plant correlated with the marker compound contents in association with bioactivity. The study results are a prelude to further studies involving the active fractions and provide a comprehensive insight into the functional products of this plant against oxidative diseases.

Analysis of Antioxidant Constituents from Ilex rotunda and Evaluation of Their Blood–Brain Barrier Permeability

“…The HPLC-DPPH method can be used to rapidly assess pure antioxidant compounds in complex mixtures [ 17 ]. The above-established method was used to determine antioxidant compounds based on reduced peak areas between DPPH treated and untreated groups.…”

“…The control sample was prepared by mixing 100 μL methanol with 100 μL of the extract. Both mixtures were analyzed using the same established analytical methods [ 17 ]. The outflow was monitored at 254 and 326 nm wavelength.…”

“…Therefore, the present study reports approaches of searching for antioxidant constituents in I. rotunda by 1,1-diphenyl-β-picrylhydrazine (DPPH) and 3-ethylbenzothiazoline-6-sulfonic acid diammonium salt (ABTS) radical scavenging assays. These eight marker compounds isolated from an active fraction of I. rotunda extract were validated using the established method, and antioxidant constituents were rapidly identified through the high-performance liquid chromatography-1,1-diphenyl-β-picrylhydrazine (HPLC-DPPH) method [ 17 ]. Finally, a parallel artificial membrane permeability assay for blood–brain barrier (PAMPA-BBB) was applied to assess brain-penetrable antioxidants from I. rotunda extract [ 18 ].…”

Anti-Inflammatory Activity of Compounds Derived from Vitex rotundifolia

“…rotundifolia is a beach plant distributed along the coastal line ranging from Mediterranean regions to Central Asia [ 7 ]. Previous studies have reported that this plant contains a variety of chemical derivatives including caffeoylquinic acids, lignans, essential oils, flavonoids, terpenoids, and iridoids with its active constituents showing many biological activities such as antioxidative [ 8 ], anticancer [ 9 ], and anti-inflammatory [ 10 ] activities. In traditional medicine, this plant is used to treat inflammation, rheumatic pain, migraine, and infections [ 11 ].…”

“…In traditional medicine, this plant is used to treat inflammation, rheumatic pain, migraine, and infections [ 11 ]. Our previous study [ 8 ] revealed that total extracts and fractions of V . rotundifolia possess potent antioxidant and anti-inflammatory capacities.…”