Polyphenol Composition of Extracts of the Fruits of Laserpitium Krapffii Crantz and Their Antioxidant and Cytotoxic Activity

Abstract

During inflammation, reactive oxygen species (ROS) are produced in large amounts, causing oxidative stress. Several studies confirm that plant extracts rich in phenolic compounds may inhibit ROS production. For that reason, the aim of this work is the qualitative and quantitative analysis of phenolic acids and flavonoids in the etheric (LAEN) and methanolic (LAM) extracts of the fruits of Laserpitium krapffii Crantz, as well as their antioxidative and cytotoxic properties. Liquid chromatography-electrospray tandem mass spectroscopy (LC-ESI-MS/MS) enabled the identification of 12 phenolic acids and nine flavonoids. Both tested extracts scavenged ROS in a concentration-dependent manner. Stronger activity was observed for the methanolic extract. The cytotoxic effect of both extracts in increasing concentrations on five types of cancer cell lines was also investigated. The cytotoxicity was estimated using trypan blue vital staining. It was found that the analyzed extracts induced the apoptosis of the cells of all the tested cell lines. In conclusion, our results present that the fruits of L. krapffii can be a source of valuable compounds with protective effects against oxidative damage.

Keywords: LC-MS/MS; Laserpitium; antioxidants; cytotoxic activity; phenolics.

Figure 1

The chromatograms in multiple reaction mode (MRM) of flavonoid aglycones occuring in the methanolic extract of L. krapffii fruits: 1-luteolin; 2-eriodictyol; 3-quercetin; 4-kaempferol.

Figure 2

The chromatogram in MRM of flavonoid glycosides occurring in the methanolic extract of L. krapffii fruits: 1-rutin; 2-hyperoside; 3-isoquercetin; 4-kaempferol-3-O-rutinoside; 6-astragalin.

Figure 3

The valuation of the HL-60 cell line viability exposed to 24 h of increasing concentrations of: (A) — LAEN extract and (B) — LAM extract.

Figure 4

The valuation of the HL-60/MX1 cell line viability exposed to 24 h of increasing concentrations of: (A)—LAEN extract and (B)—LAM extract.

Figure 4

The valuation of the HL-60/MX1 cell line viability exposed to 24 h of increasing concentrations of: (A)—LAEN extract and (B)—LAM extract.

Figure 5

The valuation of the HL-60/MX2 cell line viability exposed to 24 h of increasing concentrations of: (A)—LAEN extract and (B)—LAM extract.

Figure 6

The valuation of the CCRF-CEM cell line viability exposed to 24 h of increasing concentrations of: (A)—LAEN extract and (B)—LAM extract.

Figure 7

The valuation of the CEM/C1 cell line viability exposed to 24 h of increasing concentrations of: (A)—LAEN extract and (B)—LAM extract.

Figure 8

Antioxidant activity of the etheric (LAEN) and methanolic (LAM) extracts against: (A) 2,2-diphenyl-1-picrylhydrazyl radical (DPPH), and (B) 2,2′-azino-bis-(3-ethyl-benzthiazoline-6-sulfonic acid) (ABTS). Data are expressed as means ± SEM; obtained in three independent experiments, assayed in triplicate. Statistical significance of differences was calculated between both extracts; * p < 0.05.

Figure 9

Scavenging effect of the etheric (LAEN) and methanolic (LAM), in the concentration range of 2 to 50 μg/mL, on O₂^•− scavenging [%] in the xanthine/xanthine oxidase system (A). Effects of the LAM (B) and LAEN (C) on the xanthine/xanthine oxidase system [%]: nitrobluetetrazolium (NBT) reduction determines the scavenging of O₂^•−; uric acid production determines the oxidase inhibition. Statistical significance of differences was calculated between both extracts or between uric acid production and NBT reduction; * p < 0.05.

Figure 10

Scavenging effects of the etheric (LAEN) and methanolic (LAM) on hypochlorous acid (HOCl)-mediated oxidation of 5-thio-2-nitrobenzoic acid (TNB). Data are expressed as means ± SEM, obtained in at least three independent experiments, and assayed in triplicate. Statistical significance of differences was calculated between both extracts; * p < 0.05.