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. 2017 Jan 13;14(2):130-141.
doi: 10.21010/ajtcam.v14i2.14. eCollection 2017.

ANALYSIS OF PHENOLIC AND FLAVONOIDS OF WILD EPHEDRA ALATA PLANT EXTRACTS BY LC/PDA AND LC/MS AND THEIR ANTIOXIDANT ACTIVITY

Fuad Al-Rimawi  1 , Saleh Abu-Lafi  2 , Jehad Abbadi  3 , Ayman A A Alamarneh  1 , Raneen A Sawahreh  4 , Imad Odeh  1
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ANALYSIS OF PHENOLIC AND FLAVONOIDS OF WILD EPHEDRA ALATA PLANT EXTRACTS BY LC/PDA AND LC/MS AND THEIR ANTIOXIDANT ACTIVITY

Fuad Al-Rimawi et al. Afr J Tradit Complement Altern Med. .

Abstract

Background: Ephedra is among Palestinian medicinal plants that are traditionally used in folkloric medicine for treating many diseases. Ephedra is known to have antibacterial and antioxidant effects. The goal of this study is to evaluate the antioxidant activity of different extracts from the Ephedra alata plant growing wild in Palestine, and to analyze their phenolic and flavonoid constituents by HPLC/PDA and HPLC/MS.

Materials and methods: Samples of the Ephedra alata plant grown wild in Palestine were extracted with three different solvents namely, 100% water, 80% ethanol, and 100% ethanol. The extracts were analyzed for their total phenolic content (TPC), total flavonoid content (TFC), antioxidant activity (AA), as well as phenolic and flavonoids content by HPLC/PDA/MS.

Results: The results revealed that the polarity of the extraction solvent affects the TPC, TFC, and AA of extracts. It was found that both TPC and AA are highest for plant extracted with 80% ethanol, followed by 100% ethanol, and finally with 100% water. TFC however was highest in the following order: 100% ethanol > 80% ethanol > water. Pearson correlation indicated that there is a significant correlation between AA and TPC, but there is no correlation between AA and TFC. Simultaneous HPLC-PDA and UHPLC-MS analysis of the ethanolic plant extracts revealed the presence of Luteolin-7-O-glucuronide flavone, Myricetin 3-rhamnoside and some other major polyphenolic compounds that share myricetin skeleton.

Conclusion: Ephedra alata extract is rich in potent falvonoid glycosidic compounds as revealed by their similar overlaid UV-Vis spectra and UHPLC-MS results. On the basis of these findings, it is concluded that Ephedra alata constitutes a natural source of potent antioxidants that may prevent many diseases and could be potentially used in food, cosmetics, and pharmaceutical products.

Keywords: Ephedra Alata; HPLC; LC/MS; antioxidant activity; flavonoid content; phenolic content.

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Figures

Figure 1
Figure 1
Overlaid HPLC-PDA chromatograms of crude 100% ethanol Ephedra alata extract using infusion (red) and sonication (blue) methods at 350 nm.
Figure 2
Figure 2
Overlaid HPLC-PDA chromatograms of crude water (blue), 80% ethanol (green) and 100% ethanol (red) extracts of Ephedra alata at 350 nm. The overlaid UV-Vis spectra of the main peaks are depicted at the right corner.
Figure 3
Figure 3
Overlaid HPLC-PDA chromatograms of Ephedra alata crude 100% ethanol at 350 nm (black) and 472 nm (blue). The overlaid UV-Vis spectra of the main peaks eluted later between 64-68 minutes are depicted at the right corner.
Figure 4
Figure 4
Figure 4 Overlaid chromatograms of phenolic and flavonoids standards mixture and 100% ethanol extract of Ephera alata at 272 nm (A) and 350 nm (B) respectively. The overlaid UV-Vis spectra of the main peaks are depicted at the right corner of chromatogram (B).
Figure 5
Figure 5
UHPLC of the Ephedra alata ethanol extract (A) and the (+)-ESI mass spectrum of Luteolin-7-Oglucuronide flavonoid.
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