ABSTRACT
Objective: To establish a quick method of ultra-performance liquid chromatography-quadrupole time-of-fight mass spectrometry (UHPLC-Triple-TOF-MS) for the identification of phenolic acids (diffractaic acid, usnic acid, orsellinic acid, etc.) in Usneae Filum. Methods: The separation was performed on the chromatographic column of Phenomenex Luna 3u C18 (150 mm × 2.0 mm, 3 μm), and the mobile phase was methanol (0.05% formic acid)-0.05% formic acid solution (4 mmol ammonium acetate), with a gradient elution at a flow rate of 0.3 mL/min. The column temperature was 30℃, and negative ion mode was used for TOF-MS. Results: Seventeen compounds were identified or tentatively characterized based on the retention time and MS spectra. They are depsides, two benzo furan, ant multi substituted single benzene as well. And the preliminary fragmentation rules are summarized. Conclusion: The results demonstrate that UHPLC-Triple-TOF-MS method is novel, quick, and efficient for the identification of the compounds in Usneae Filum.
ABSTRACT
Diffractaic acid (DA) is a naturally occurring depside derivative found in several lichen species. It has a wide range of important biological effects such as analgesic and antiviral properties, although its cytotoxic, cytogenetic and oxidative effects have not been investigated in human blood tissue yet. Therefore, increasing concentrations (1, 5, 10, 25, 50, 100 and 200 mgL-1) of DA was added into human whole blood cultures. 3-(4.5-dimethylthiazol-2-yl)-2.5-diphenyl tetrazolium bromide (MTT) assay was used to assess the cell viability and/or cytotoxicity and genotoxic damage potential of DA using chromosome aberration (CA) and micronucleus (MN) tests were performed. In addition, oxidative alterations were determined by the total antioxidant capacity (TAC) and total oxidant status (TOS) assays. The results revealed that DA reduced cell viability at higher concentrations than 50 mgL-1. The all tested concentrations of DA were non-genotoxic. In vitro treatments with DA led to increases of TAC levels in the cultured blood cells without changing the TOS levels as compared to the control group. Consequently, DA exhibited a significant non-mutagenic and antioxidant potential in vitro.