Phytochemical profile and a preliminary in ovo screening of the ethanolic extract of Coffea arabica green seeds

Abstract

Aims: Coffea arabica has garnered significant attention for its potential therapeutic applications due to its rich phytochemical profile, including chlorogenic acids, flavonoids, and alkaloids like caffeine, which are associated with antioxidant, anti-inflammatory, and antimicrobial effects. This study aimed to (i) evaluate the phytochemical composition of an ethanolic extract derived from green seeds of Coffea arabica and (ii) perform a preliminary in ovo screening to determine its mucosal tolerability, irritant potential, and pro-angiogenic activity using the chick embryo chorioallantoic membrane (CAM) model.

Methods: An ethanolic extract was prepared from C. arabica green seeds and subjected to LC-MS analysis for the identification and quantification of polyphenols. The antioxidant capacity was assessed through the DPPH radical-scavenging assay, comparing extract performance to ascorbic acid as a standard. In ovo testing was conducted using the HET-CAM assay to observe potential irritation on the CAM surface and to evaluate angiogenic activity. Chick embryos were monitored for vascular changes, irritant signs (hemorrhage, lysis, coagulation), and overall tolerability post-application over six days.

Results: Phytochemical analysis confirmed the presence of chlorogenic acid and 4-O-caffeoylquinic acid in the extract. The extract exhibited substantial antioxidant activity (66.38%), though slightly lower than that of ascorbic acid (97.36%). In the HET-CAM assay, no signs of irritation or toxicity were observed, and the extract was well tolerated for up to six days post-application. Additionally, the extract promoted angiogenesis, with increased vascularization observed, suggesting a stimulatory effect on neovascularization without inducing tissue damage.

Conclusions: The ethanolic extract of Coffea arabica green seeds demonstrates promising antioxidant and pro-angiogenic properties, alongside high mucosal biocompatibility. These findings support its potential applications in therapeutic and cosmetic formulations, particularly those targeting tissue regeneration and skin repair. Further studies are recommended to explore the underlying mechanisms and to confirm efficacy in more advanced biological models.

Keywords: Arabica coffee; FT-IR spectroscopy; HET-CAM assay; LC-MS investigation; angiogenesis.

Figure 1

The main steps involved in the obtainment of the tested extract.

Figure 2

HPLC chromatogram of ethanolic extract from C. arabica green seeds.

Figure 3

FT-IR spectra of C. arabica ethanolic extract.

Figure 4

The time-dependent AOC of ethanol extract of C. arabica green seed.

Figure 5

Stereomicroscopic images showing the effects induced by C. arabica green seed ethanolic extract on CAM using the HET-CAM anti-irritant assay. The images document the absence of hemorrhage, vascular lysis, and coagulability after the application of the tested extract. Scale bars represent 500 μm.

Figure 6

Stereomicroscopic images showing the effects produced by ethanolic extract of C. arabica ethanolic extract on CAM; images are shown initially at 0 h, 24 h, 48 h, and 72 h post-treatment, controlling for DMSO solvent and C. arabica ethanolic extract. Scale bars represent 500 μm.